Wine tannins and their aggregation/release with lipids and proteins: Review and perspectives for neurodegenerative diseases.

Tannins are amphiphilic molecules, often polymeric, which can be generally described as a core containing hydrophobic aromatic rings surrounded by hydroxyl groups. They have been known for millennia and are part of human culture. They are ubiquitous in nature and are best known in the context of wine and tea tasting and food cultures. However, they are also very useful for human health, as they are powerful antioxidants capable of combating the constant aggressions of everyday life. However, their mode of action is only just beginning to be understood. This review, using physicochemical concepts, attempts to summarize current knowledge and present an integrated view of the complex relationship between tannins, proteins and lipids, in the context of wine drinking while eating. There are many thermodynamic equilibria governing the interactions between tannins, saliva proteins, lipid droplets in food, membranes and the taste receptors embedded in them. Taste sensations can be explained using these multiple equilibria: for example, astringency (dry mouth) can be explained by the strong binding of tannin micelles to the proline-rich proteins of saliva, suppressing their lubricating action on the palate. In the presence of lipid droplets in food, the equilibrium is shifted towards tannin-lipid complexes, a situation that reduces the astringency perceived when consuming a tannic wine with fatty foods, the so-called "camembert effect". Tannins bind preferentially to taste receptors located in mouth membranes, but can also fluidify lipids in the non-keratinized mucous membranes of the mouth, which can impair the functioning of taste receptors there. Cholesterol, present in large quantities in keratinized mucous membranes, stiffens them and thus prevents tannins from disrupting the conduction of information through other taste receptors. As tannins assemble and disassemble depending on whether they are in contact with proteins, lipids or taste receptors, a perspective on their potential use in the context of neurodegenerative diseases where fibrillation is a key phenomenon will also be discussed.

Paper chromatography approach for the assessment of interaction between red wine and whole saliva.

In-mouth interaction of red wine compounds with salivary proteins is a primary event allegedly responsible for eliciting the mouth-feel sensation of astringency. Those interactions have been currently associated with precipitation of salivary protein/polyphenol complexes. However, such single physicochemical evidence for interaction does not account for the complexity of astringency. This study aimed to develop a paper chromatography method to assess interactions between red wine and the salivary protein fraction using stepwise series of red wine/saliva binary mixtures from 100% wine to 100% saliva ("Alpha and Omega series"). Aliquots of each one of the mixtures were spotted on a cellulose membrane to scrutinize independently the distribution areas of wine components (naturally pink-colored) and salivary protein (stained blue in Coomassie Brilliant R-250). This double target detection revealed interactions between saliva and red wine components along most of the quantitative Alpha and Omega series, a point of equivalence corresponding to maximum interactivity for both complex reactants and a non-diffusible sub-fraction of saliva displaying the highest interactivity. The results indicate a novel way to assess quantitatively physicochemical interactions between red wines and human saliva but also provide new lights to approach the identification of molecular salivary structures involved in triggering astringency.

Study of Serial Exposures of an Astringent Green Tea Flavonoid Extract with Oral Cell-Based Models.

It is well known that repeated exposure to phenolic compounds (PCs) raises astringency perception. However, the link between this increase and the oral cavity's interactions with salivary proteins (SPs) and other oral constituents is unknown. To delve deeper into this connection, a flavonoid-rich green tea extract was tested in a series of exposures to two oral cell-based models using a tongue cell line (HSC3) and a buccal mucosa cell line (TR146). Serial exposures show cumulative PC binding to all oral models at all concentrations of the green tea extract; however, the contribution for the first and second exposures varies. The tongue mucosal pellicle (HSC3-Mu-SP) may contribute more to first-stage astringency (retaining 0.15 ± 0.01 mg mL-1 PCs at the first exposure), whereas the buccal mucosal pellicle (TR146-Mu-SP) retained significantly less (0.08 ± 0.02 mg mL-1). Additionally, increased salivary volume (SV+), which simulates the stimulation of salivary flow brought by a food stimulus, significantly enhances PC binding, particularly for TR146 cells: TR46-Mu-SP_SV+ bound significantly higher total PC concentration (0.17 ± 0.02 mg mL-1) than the model without increased salivary volume TR146-Mu-SP_SV- (0.09 ± 0.03 mg mL-1). This could be associated with a higher contribution of these oral cells for astringency perception during repeated exposures. Furthermore, PCs adsorbed in the first exposure to cell monolayer models (+TR146 and +HSC3) change the profile of PCs bound to these models in the second exposure. Regarding the structure binding activity, PCs with a total higher number of hydroxyl groups were more bound by the models containing SP. Regarding the SP, basic proline-rich proteins (bPRPs) may be involved in the increased perception of astringency upon repeated exposures. The extent of bPRP precipitation by PCs in mucosal pellicle models for both cell lines (HSC3 and TR146) in the second exposure (76 ± 13 and 83 ± 6%, respectively) was significantly higher than in the first one (25 ± 14 and 5 ± 6%, respectively).

Investigating the role of tartaric acid in wine astringency.

Previous studies acknowledged that tartaric acid-imparted low-pH contributed to the enhancement of astringency, but in-depth studies are lacking and the underlying mechanisms are not clearly understood. This work introduced new insight into the effect of tartaric acid on astringency perception from the perspectives of complex formation, protein secondary structure, chemical bond type and salivary layer fluidity by establishing models using proteins (α-amylase, salivary proteins) and tannic acid. Results demonstrated that tartaric acid affects wine astringency by two mechanisms: a) Tartaric acid compound directly affects the wine astringency by forming ternary complexes and causing the protein structure to stretch by changing the hydrogen bond and hydrophobic bond between protein-polyphenol complexes. b) pH affected astringency by increasing the fluidity of the salivary layer rather than increasing the consumption of the salivary layer. The findings provide valuable information to the wine industry to regulate wine astringency by the management of tartaric acid.

Histomorphometric study of ethanolic extract of Graptophyllum pictum (L.) Griff. leaves on croton oil-induced hemorrhoid mice: A Javanese traditional anti-hemorrhoid herb.

ETHNOPHARMACOLOGY RELEVANCE: Graptophyllum pictum (L.) Griff., known as "handeuleum" in West Java and "Daun Ungu" in Indonesia, is traditionally used to cure hemorrhoids. AIM OF THE STUDY: The purpose of this study is to prove its effectiveness scientifically using anorectal histological parameters in Croton oil-induced hemorrhoid mice. MATERIALS AND METHODS: In vivo tests were performed by observing histomorphologic changes in mice anorectal tissue induced by croton oil. In addition, in vitro assay was performed for evaluating antioxidant activity, astringency property, and hemostasis-associated activity. The antioxidant activity was measured using a DPPH radical scavenging assay. The total flavonoid and phenolic contents were also determined spectrophotometrically. RESULTS: The in vivo assay showed that the oral-topical combination use of the ethanolic extract of G. pictum leaves demonstrated significant improvement on the croton oil-induced anorectal damage better than the single application by oral or topical application. CONCLUSION: These results showed that G. pictum has potent anti hemorrhoid activity, especially for the combinational use of oral and topical administration.

Effect of Different Enological Tannins on Oxygen Consumption, Phenolic Compounds, Color and Astringency Evolution of Aglianico Wine.

BACKGROUND: In the wine industry, in addition to condensed tannins of grape origin, other commercial tannins are commonly used. However, the influence of oxygen uptake related to different tannin additions during the post fermentative phase in wine has not been completely investigated. In this study, we evaluated the influence of four different commercial tannins (namely, condensed tannins, gallotannins, ellagitannins and tea tannins) during four saturation cycles. METHOD: Wine samples were added with four different tannin classes (30 g/hL) as to have 5 different experimental samples: control, gallotannins (GT), condensed tannins (CT), ellagitannins (ET), and tea tannins (TT). The chemical composition of the four commercially available tannin mixtures was defined by means of NMR and high-resolution mass spectrometry. After the addition of tannins, each wine sample was oxidized by air over four cycles of saturation. During the experiment oxygen consumption rate (OCR), sulfur dioxide consumption, acetaldehyde production, phenolic compounds, chromatic characteristics, astringency measured by the reactivity towards saliva proteins and astringency subqualities were evaluated. RESULTS: The experiment lasted 52 days. The addition of tannins influenced the oxygen consumption on the 1st day of the saturation cycles and, in the case of TT, a higher total consumption of oxygen was also detected. Acetaldehyde increased during the experiment while the native anthocyanins decreased throughout the oxidation process. CONCLUSION: Wines added with tannins featured improved color intensities with respect to the control; the addition of TT, GT and ET slightly promoted the formation of short polymeric pigments; the astringency, determined before and at the end of the experiment, decreased in all the samples, including the control wine, and mostly in the ET and GT samples.

Characterization of Bitter and Astringent Off-Taste Compounds in Potato Fibers.

Applying the sensomics approach, a combination of activity-guided fractionation and taste dilution analysis (TDA) followed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), ultrahigh-performance liquid chromatography time-of-flight mass spectrometry (UHPLC-TOF-MS), and one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy (1D/2D NMR) allowed the elucidation of key off-taste compounds in potato dietary fiber isolates. Previously already having been described as off-taste compounds in potato tubers, saponins α-chaconine and α-solanine were shown to be also major contributors to overall off-taste in potato fiber isolates. Moreover, fatty acids as well as fatty acid oxidation products, namely, E-9,10,13-trihydroxy-octadec-11-enoic acid as well as newly identified compounds hexadecyl(E/Z)-ferulate and octadecyl(E/Z)-ferulate, were shown to be key inducers to off-taste in the isolates, exhibiting taste recognition thresholds between 18 and 981 µmol/L. This paper demonstrates the isolation, structure determination, quantitation as well as sensory attributes of these key off-taste compounds.

Rheological study of tannin and protein interactions based on model systems.

The interaction between wine tannins and saliva proteins is responsible for wine astringency perception, producing a depletion of salivary proteins and changes on oral friction. In sensorial terms, astringency is described as a dryness and puckering sensation in the mouth, which is related to the "structure" or "body" of red wines. However, these last descriptors, as structure or body, are perceived during wine tasting and commonly related to wine viscosity. To address these differences on sensory response, we hypothesize that tannin-protein interactions could be a key factor involved in the viscosity of red wines/saliva mixtures, just as they are for astringency. We used a rheological method to study the impact of tannin-protein interaction on the viscosity of model wine-saliva systems. Mixtures of model saliva based on mucin and typical astringent compounds, as commercial tannins and gallic acid, were evaluated for their rheological behavior. The viscometric flow of the fluid mixtures was determined, and subsequently, the viscosity was evaluated at a shear rate of 60 s-1 . It was observed that red wines/saliva mixtures exhibit non-Newtonian flow and ascending tannin doses led to an increase in the apparent viscosity. Nephelometric analysis demonstrate that tannin-mucin aggregates were formed, which suggests that these complexes were potentially responsible for the viscosity increases, modifying the rheological behavior of these mixtures. Results from this work propose that tannin-protein interactions are also involved in the underlying mechanism of thickness perception of red wines and rheology could be a complementary instrumental technique for wine mouthfeel characterization.

Interaction between Ellagitannins and Salivary Proline-Rich Proteins.

The first contact of tannins with the human body occurs in the mouth, where some of these tannins are known to interact with salivary proteins, in particular with proline-rich proteins (PRPs). These interactions are important at a sensory level, especially for astringency development, but could also affect the biological activities of the tannins. This study gathers information on the relative affinity of the interaction, complex stoichiometry, and tannin molecular epitopes of binding for the interactions between the families of PRPs (bPRPs, gPRPs, and aPRPs) and three representative ellagitannins (castalagin, vescalagin, and punicalagin). These interactions were studied by saturation-tranfer difference NMR and microcalorimetry. The effect of the PRP-ellagitannin interaction on their antioxidant ability was also assessed by ferric reduction antioxidant power (FRAP) assays. The results support a significant interaction between the studied tannins and PRPs with binding affinities in the micromolar range. Punicalagin was always the ellagitannin with higher affinity. aPRPs were the salivary PRPs with higher affinity. Moreover, it was observed that when ellagitannins are present in low concentrations (5-50 µM), as occurs in food, the antioxidant ability of these tannins when complexed with salivary PRPs could be significantly impaired.

Proanthocyanidin content as an astringency estimation tool and maturation index in red and white winemaking technology.

Selecting the appropriate type of barrel for wine maturation but also deciding on the optimum maturation length, is a challenge for winemakers. As different types of barrel woods emerge, it is of great importance for a guideline to be established, which could facilitate winemaking decisions. Since the sensory perception of the finished wine, and particularly the intensity of astringency, is a decisive factor for the quality of a barrel-aged wine, in this experiment, the structural characteristics of wine proanthocyanidins were determined and their correlation with astringency was established. According to the results obtained, the proanthocyanidin content and the type of subunit that is dominant in tannin chains could be used to construct an astringency estimation model. The findings could provide winemakers with a useful tool when deciding how long to mature a specific type of wine in a specific wood container without making it appear coarse and astringent.

Wine astringency reduces flavor intensity of Brussels sprouts.

The bitterness of vegetables is a leading reason why they are avoided by children and some adults. Bitterness is perceived via TAS2R receptors located on the tongue. In contrast, astringency is a mouthfeel rather than a taste, and is perceived as a dry, puckering sensation. To date few reports have suggested any interactions between the two processes even though they often occur simultaneously in many real foods. In this study, we have used Brussels sprouts as an exemplar bitter vegetable and examined the influence of a number of different interventions on perceived intensity. Subjects rated the intensity of Brussels sprouts before and after three interventions: gravy, red wine, and water. Only red wine caused a significant (p < .0001) decrease in VAS scale, from 5.5 to 3.5 on a 10-point labeled magnitude scale. The results suggest the astringency of the red wine affected the perception of bitter in the Brussels sprout. Some possible mechanisms are discussed. PRACTICAL APPLICATIONS: This report reveals a possible insight into how bitterness is perceived in humans. By using astringency to affect salivary proteins, we suggest they may play a role in the detection of bitterness. This may be by helping to transport bitterness compounds to the taste bud receptors or a separate mechanism. Potentially this also opens up new ways to block bitterness.

An integrative salivary approach regarding palate cleansers in wine tasting.

Wine sensory sessions normally involve the tasting of several samples, to remove food residues from the mouth the use of palate cleansers (PC) is needed. Until now, there is no agreement on the best PC to use during wine tasting sessions. The aim of this work is to study the relationship between the components retained in saliva after wine tasting and the remnant sensory feeling (astringency, alcohol, and acidity). For that, different common PC (water, carbonated water, and milk) were tested and saliva samples (expectorated and scraped) from nine trained panelists were collected after wine with and without PC trials. Results showed that after palate cleansing and not cleansing, astringency, alcoholic and acidity perception were influenced by time, PC and panelist. Astringency perception showed the greatest intensity in comparison to alcoholic and acidity. Milk was the only PC which reduced quantifiable polyphenols in expectorated saliva, as well as reducing astringency feelings. Although compositions of expectorated and scraped saliva correlated between them, polyphenols accumulated in the expectorated saliva significantly more. Retained polyphenols were correlated with astringency perception, but no correlation was found with salivary proteins. These findings assessed the astringency build-up effect during wine tasting due to polyphenols accumulation in saliva, remarking the importance of an adequate PC selection. All things considered, the present work confirmed the relationship between after-swallow mouthfeel perception and mouth residues instrumentally quantified. Also, milk has proven to be the most effective of the three PC. PRACTICAL APPLICATIONS: During tasting the accumulation of residues from previous wine samples tasted, could mislead the judgment of wine sensory qualities by oenologists. Therefore, between tasting samples it is highly important to choose the right PC. However, until now the selection of PC remains empirical, therefore in this work, we proposed to study the residues in saliva by using different PC and quantifying instrumentally, the wine residues. The methodology selected to quantify the wines residues in saliva was quick and easy to use. Furthermore, instrumental results were related with the sensory feeling of mouth cleanliness without considering individual panel member's preferences of PC. In this study, to remove astringency feeling, milk was shown to be the best cleanser in comparison with water, carbonated water or nothing, but oenologist/winemakers could use this instrumental methodology in saliva to select which one is the best among their current PC used.

Effect of the addition of mannoproteins on the interaction between wine flavonols and salivary proteins.

It has been suggested that the addition of flavonols (i.e. white grape skins) improves and stabilizes the color of red wines. However, it has been shown that flavonol glycosides produce a mouth-drying and mouth-coating sensation at very low threshold concentrations. Moreover, the addition of polysaccharides to wines is a practice addressed to improve the smoothness and roundness and correct excessive astringency, so we have studied the effect of the addition of yeast mannoproteins (MP) on the interaction between quercetin 3-glucoside and human salivary peptides. Sensory analysis showed the first evidence of the mannoprotein smoothing effect when the flavonol is added to wine. Additionally, MP/SP/polyphenol interactions were studied using fluorescence spectroscopy, dynamic light scattering and isothermal titration calorimetry. Results obtained indicate not only the existence of interactions between mannoproteins and flavonols but also between mannoproteins and salivary proteins (SP), suggesting a possible formation of protein/polyphenol/polysaccharide ternary complex.

Mechanisms of astringency: Structural alteration of the oral mucosal pellicle by dietary tannins and protective effect of bPRPs.

The interaction of tannins with salivary proteins is involved in astringency. This paper focussed on saliva lining oral mucosae, the mucosal pellicle. Using a cell-based model, the impact of two dietary tannins (EgC and EgCG) on the mucosal pellicle structure and properties was investigated by microscopic techniques. The role of basic Proline-Rich-Proteins (bPRPs) in protecting the mucosal pellicle was also evaluated. At low (0.05 mM) tannin concentration, below the sensory detection threshold, the distribution of salivary mucins MUC5B on cells remained unaffected. At 0.5 and 1 mM, MUC5B-tannin aggregates were observed and their size increased with tannin concentration and with galloylation. In addition, 3 mM EgCG resulted in higher friction forces measured by AFM. In presence of bPRPs, the size distribution of aggregates was greatly modified and tended to resemble that of the "no tannin" condition, highlighting that bPRPs have a protective effect against the structural alteration induced by dietary tannins.

Interactions between wine phenolic compounds and human saliva in astringency perception.

Astringency is a complex perceptual phenomenon involving several sensations that are perceived simultaneously. The mechanism leading to these sensations has been thoroughly and controversially discussed in the literature and it is still not well understood since there are many contributing factors. Although we are still far from elucidating the mechanisms whereby astringency develops, the interaction between phenolic compounds and proteins (from saliva, oral mucosa or cells) seems to be most important. This review summarizes the recent trends in the protein-phenol interaction, focusing on the effect of the structure of the phenolic compound on the interaction with salivary proteins and on methodologies based on these interactions to determine astringency.

Effect of a commercial tannin on the sensorial temporality of astringency.

Astringency is a tactile sensation that is generated by a reduction in lubrication in the oral cavity and is generally attributed to the interaction of procyanidins or condensed tannins with salivary proteins. Several factors influence tannin-protein interactions, such as pH, alcohol, sweetness, oxygen and polyphenol content. A scarcely studied factor is the effect of the tannin content on the perception of astringency. The objective of this study was to evaluate the effect of different concentrations of commercial oenological tannin (COT) on the timing of the perception of astringency. For this model, a vinous solution enriched with three concentrations of COT was used. The samples were subjected to a storage period of three months. Additionally, a panel was trained in the perception of astringency in red wines using a method of temporal dominance of sensations (TDS). Astringency descriptors were selected, and the TDS method was used to characterize the astringency. The samples were evaluated using traditional descriptors in TDS and astringency descriptors in TDS. In traditional TDS curves, treatments with higher concentrations of COT showed a higher and more persistent dominance index in the descriptor astringency. Moreover, the stimulus duration variable increased as the COT level increased. Likewise, temporary astringency was dominant over alcohol. For astringency TDS, at low concentrations of COT, the soft and adhesive descriptors were dominant, whereas at high COT concentrations, aggressive and drying were perceived as the dominant descriptors. An increasing concentration of tannin in the vinous solution generated an increased duration and dominance of astringency and reduced the duration of the sensation of alcohol. Finally, the type of perceived astringency was closely related to the tannin concentration.

A saliva molecular imprinted localized surface plasmon resonance biosensor for wine astringency estimation.

Wine astringency was evaluated based on the interaction of two complex matrices (red wine and saliva) by combining localized surface plasmon resonance (LSPR) and molecular imprinted polymers (MIP) at gold nanodisks as an alternative to sensorial analysis. The main objective of the work was to simulate wine astringency inside the mouth by mimicking this biological system. The LSPR/MIP sensor provided a linear response for astringency expressed in pentagalloyl glucose (PGG) units in concentrations ranging from 1 to 140µmol/L. The sensor was also applied to wine samples correlating well with sensorial analysis obtained by a trained panel. The correlation of astringency and wine composition was also evaluated showing that anthocyanins may have an important role, not only for pigmentation but also in astringency.

Effect of Astringent Stimuli on Salivary Protein Interactions Elucidated by Complementary Proteomics Approaches.

The interaction of astringent substances with salivary proteins, which results in protein precipitation, is considered a key event in the molecular mechanism underlying the oral sensation of puckering astringency. As the chemical nature of orally active astringents is diverse and the knowledge of their interactions with salivary proteins rather fragmentary, human whole saliva samples were incubated with suprathreshold and isointensity solutions of the astringent polyphenol (-)-epigallocatechin gallate, the multivalent metal salt iron(III) sulfate, the amino-functionalized polysaccharide chitosan, and the basic protein lysozyme. After separation of the precipitated proteins, the proteins affected by the astringents were identified and relatively quantified for the first time by complementary bottom-up and top-down mass spectrometry-based proteomics approaches. Major salivary target proteins, which may be involved in astringency perception, are reported here for each astringent stimulus.

Effect of flavonols on wine astringency and their interaction with human saliva.

The addition of external phenolic compounds to wines in order to improve their sensory quality is an established winemaking practice. This study was aimed at evaluating the effect of the addition of quercetin 3-O-glucoside on the astringency and bitterness of wines. Sensory results showed that the addition of this flavonol to wines results in an increase in astringency and bitterness. Additionally, flavonol-human salivary protein interactions were studied using fluorescence spectroscopy, dynamic light scattering and molecular dynamic simulations (MD). The apparent Stern-Volmer (KsvApp) and the apparent bimolecular quenching constants (kqApp) were calculated from fluorescence spectra. The KsvApp was 12620±390M(-1), and the apparent biomolecular constant was 3.94×10(12)M(-1)s(-1), which suggests that a complex was formed between the human salivary proteins and quercetin 3-O-glucoside. MD simulations showed that the quercetin 3-O-glucoside molecules have the ability to bind to the IB937 model peptide.

Astringent Mouthfeel as a Consequence of Lubrication Failure.

Herein, we systematically investigate the origin of astringent mouthfeel when we eat unripe fruits, drink coffee or tea, from the perspective of lubrication by simulating the dynamic weak interaction on the tongue with model protein (mucoprotein, MP) and polyphenolic compounds (tannic acid, TA). Astringency was due to the protein-mediated lubrication failure when encountering polyphenolic molecules that normally exist, for example in unripe fruits, coffee, tea. The underlying molecular mechanism of oral tribology is widely present in nature and enables us to engineer a tongue-like polyacrylamide composite hydrogel that exhibits high TA sensitivity and to develop a scientific strategy for catching slippery fish using TA-containing gloves. These results provide novel and useful insights into the failure of biological boundary lubrication on soft tissue surface with the adsorbed proteins.