Variation and fractionation of δ2H, δ18O, and δ17O stable isotopes from irrigation water to soil, grapes, and wine for the traceability of geographical origins.

To investigate the variation and fractionation of stable isotopes from irrigation water to soil, grapes, and wine, δ2H, δ18O, and δ17O in different samples from 10 regions in China were determined using a water isotope analyser. The values were significantly different among regions according to the chemometric analysis. All isotopes were significantly and positively correlated with irrigation water-soil and grape-wine. A significant water isotopic fractionation effect was observed from the irrigation water to the soil, grapes, and wine. Stable isotope distribution characteristics correlated with longitude, latitude, altitude, temperature, precipitation, station pressure and wind speed. The linear discriminant analysis (LDA), random forest (RF), support vector machine (SVM), and feed-forward neural network (FNN) models 58.33-100 %, 80-100 %, 53.33-100 %, and 73.33-100 % accurate for distinguishing the geographical origins of all samples from training and test data, respectively. These findings provide a theoretical basis for authenticating the geographic origin of Chinese wines using stable isotope analysis.

Insight into the growth and metabolic characteristics of indigenous commercial S. cerevisiae NX11424 at high and low levels of yeast assimilable nitrogen based on metabolomic approach.

Yeast assimilable nitrogen (YAN) is one of the important factors affecting yeast growth and metabolism. However, the nitrogen requirement of indigenous commercial S. cerevisiae NX11424 is unclear. In this study, metabolomics was used to analyze the metabolite profiles of the yeast strain NX11424 under high (433 mg/L) and low (55 mg/L) YAN concentrations. It was found that yeast biomass exhibited different trends under different YAN conditions and was generally positively correlated with the initial YAN concentration, while changes of key biomarkers of yeast strain NX11424 at different stages of fermentation showed a similar trend under high and low YAN concentrations. The YAN concentration affected the metabolite levels of the yeast strain NX11424, which resulted in the significant difference in the levels of pyruvic acid, α-oxoglutarate, palmitoleic acid, proline, butane-2,3-diol, citrulline, ornithine, galactinol, citramalic acid, tryptophan, alanine, phosphate and phenylethanol, mainly involving pathways such as central carbon metabolism, amino acid metabolism, fatty acid metabolism, purine metabolism, and energy metabolism. Yeast strain NX11424 could utilize proline to produce protein under a low YAN level. The intracellular level of citrulline and ornithine under high YAN concentration was higher than that under low YAN level. Yeast strain NX11424 is more suitable for fermentation at lower YAN level. The results obtained here will help to rational utilize of YAN by S. cerevisiae NX11424, and is conducive to precise control of the alcohol fermentation and improve wine quality.

Influence of flor yeast starters on volatile and nitrogen compounds during a controlled biological aging.

Fino Sherry wine undergoes biological aging carried out by a velum of flor yeast within a traditional dynamic system known as "criaderas and solera". The complex microbiota of biofilm-forming Saccharomyces cerevisiae strains play a crucial role in shaping the distinctive organoleptic profile of these types of wines. For this reason, the aim of this study is to analyze the changes produced by different flor yeast strains in the volatilome and the aminogram of different wines from the criaderas and solera system during biological aging in the laboratory, simulating a flor yeast velum condition at different stages of the system. Results suggest that each strain metabolizes wine differently, finding that depending on the wine, some strains are better suited for the process than others. In addition, it is found that the content of biogenic amines in Fino Sherry wines, previously attributed to malolactic bacteria, varies according to the yeast strain metabolizing the wine, suggesting that flor yeast could be used to modify biogenic amines content during biological aging. Results indicate that the use of selected flor yeast starters in biological aging may be of interest to modulate some parameters during Fino Sherry wine aging.

Oxygen alters redox cofactor dynamics and induces metabolic shifts in Saccharomyces cerevisiae during alcoholic fermentation.

Environmental conditions significantly impact the metabolism of Saccharomyces cerevisiae, a Crabtree-positive yeast that maintains a fermentative metabolism in high-sugar environments even in the presence of oxygen. Although the introduction of oxygen has been reported to induce alterations in yeast metabolism, knowledge of the mechanisms behind these metabolic adaptations in relation to redox cofactor metabolism and their implications in the context of wine fermentation remains limited. This study aimed to compare the intracellular redox cofactor levels, the cofactor ratios, and primary metabolite production in S. cerevisiae under aerobic and anaerobic conditions in synthetic grape juice. The molecular mechanisms underlying these metabolic differences were explored using a transcriptomic approach. Aerobic conditions resulted in an enhanced fermentation rate and biomass yield. Total NADP(H) levels were threefold higher during aerobiosis, while a decline in the total levels of NAD(H) was observed. However, there were stark differences in the ratio of NAD+/NADH between the treatments. Despite few changes in the differential expression of genes involved in redox cofactor metabolism, anaerobiosis resulted in an increased expression of genes involved in lipid biosynthesis pathways, while the presence of oxygen increased the expression of genes associated with thiamine, methionine, and sulfur metabolism. The production of fermentation by-products was linked with differences in the redox metabolism in each treatment. This study provides valuable insights that may help steer the production of metabolites of industrial interest during alcoholic fermentation (including winemaking) by using oxygen as a lever of redox metabolism.

Effect of commercial Saccharomyces cerevisiae and non-Saccharomyces yeasts on the chemical composition and bioaccessibility of pineapple wine.

Alcoholic fermentation is one of man's most efficient food preservation processes, and innovations in this area are a trend in food science and nutrition. In addition to the classic Saccharomyces yeasts, various other species may have desirable characteristics for obtaining fruit wines. This study investigated the profile of non-Saccharomyces commercial yeasts compared with S. cerevisiae regarding pineapple wine's chemical composition and bioaccessibility. The fermentation profile of the yeasts Lachancea thermotolerans, Brettanomyces bruxellensis, Brettanomyces lambicus, and S. cerevisiae was evaluated for sugar and alcohol content, and the pineapple wines obtained were analyzed for amino acids, phenolics, and organic acids by HPLC and volatile profile by GC/MS. All yeast strains were able to produce ethanol and glycerol at acceptable levels. L. thermotolerans produced higher levels of lactic acid (0.95 g/L) and higher consumption of free amino acids. B. bruxellensis produced higher levels of individual phenolics and ethanol 109 g/L. The alcoholic fermentation process improved the bioaccessibility of phenolics such as catechin (237 %), epigallocatechin gallate (81 %), procyanidin B1 (61 %) and procyanidin B2 (61 %). The yeasts differed in their volatile profiles, with Brettanomyces and Lachancea producing higher levels of compounds associated with pineapple aroma, such as ester ethyl butyrate (260-270 µg/L). These results demonstrate the importance of choosing the yeast strain for the conduction of alcoholic fermentation and that the yeasts Brettanomyces and Lachancea showed technological potential in obtaining pineapple wines. This study contributes to developing processes for obtaining fruit wines by highlighting two non-Saccharomyces yeast species with technological potential for alcoholic fermentations.

Mannoproteins modulate olfactrory perception and copigmentation of organoleptic-active-components in wines: Effects and potential molecular mechanisms.

In this research, accelerated aroma release experiments and malvidin-3-O-glucoside copigmentation experiments in model red wine solutions were designed to investigate the abilities and molecular mechanisms of mannoproteins in modulating olfactory/chromatic properties of red wines. Results indicate that under orthonasal condition, mannoprotein MP2 was promising aroma modulator due to its predictable behaviors in expelling and retaining the aroma compounds during different periods. Low field nuclear magnetic resonance and molecular dynamic simulation proved that the modulation ability of MP2 should be explained by its transitionary interacting preferences with water/aroma compound molecules. Retronasal results show that the release of aroma compounds and olfactory perceptions were irregular and difficult to predict, probably due to the complexity of the retronasal condition. All mannoproteins protected malvidin-3-O-glucoside and quercetin via the formation of binary/ternary complexes, and quercetin was found prior to be protected than malvidin-3-O-glucoside. Principal mannoprotein A0A6C1DV26 might be the critical malvidin-3-O-glucoside protector. With the presence of quercetin, principal mannoproteins B3LQU1/B5VL26 in mannoprotein MP1 might exhibit intramolecular and/or intermolecular mechanisms that strengthened the hyperchromic effect, thus enhanced the copigmentation.

Synergetic effect of Accentuated Cut Edges (ACE) and macerating enzymes on the phenolic composition of Marquette red wines.

One of the challenges of cold-hardy grape cultivars is their typical low content of tannins, alongside the presence of anthocyanin diglucoside and high acidity, which can lead to unbalanced red wines. This study hypothesized that the combination of Accentuated Cut Edges (ACE) and macerating enzymes would improve phenolics extraction from grape skins after disruption. The effects of those two winemaking techniques, either used separately or together, on red wine quality characteristics were investigated at crushing, bottling, and after six or nine months of aging. Overall, the combination of treatments improved the concentration of monomeric phenolics (20 %) and tannins (21 %) after nine months of aging. ACE or enzyme treatment separately applied had little impact on phenolics extraction in finished wines. This study exhibited a potential strategy to modify phenolics profile through the synergistic effect of ACE and macerating enzymes by causing cellular breakdown in a cold-hardy red grape cultivar.

Identification of Two New Taste-Active Compounds in Oak Wood: Structural Elucidation of Potential ß-Methyl-γ-octalactone Precursors and Quantification in Spirits.

Barrel aging is a crucial stage that influences the taste of wines and spirits, particularly increasing their sweetness and bitterness. This increase is caused by nonvolatile compounds released from oak wood. To search for such molecules, we performed a taste-guided inductive fractionation protocol using several analytical techniques. By using HRMS and NMR, two new galloylated derivatives were elucidated. Their enzymatic hydrolysis revealed the formation of ß-methyl-γ-octalactone, indicating that they are potential precursors. The taste properties of these isomers revealed a sweet and bitter taste for P-WL-1 and P-WL-2, respectively. An LC-HRMS quantification method was performed to evaluate the influence of aging parameters such as botanical origin and toasting process on their concentrations. Several spirits were also analyzed to confirm their presence in this matrix. These results improve the understanding of the molecular markers responsible for the taste of beverages.

Effects of different beverages on the color stability and fluorescence of resin composites: in situ study.

OBJECTIVE: To evaluate the color change (ΔE) and fluorescence (FL) of resin composites after the consumption of beverages. MATERIALS AND METHODS: One hundred and sixty disc-shaped specimens (6 × 2 mm) of Filtek Z350 XT (FZX) and Forma (FOR) resin composites were randomized for use in devices by five volunteers who ingested 100 ml daily of red wine (RW), beer (BE), energy drink (ED) and water (WT) over 15 days. The data were collected from digital photographs and analyzed using the CIE-Lab and RGB scales to obtain ΔE and FL and statistics via two-way ANOVA (for ΔE) and ANOVA for repeated measures (for FL), α = 0.05. RESULTS: The highest ΔE values were obtained for FZX and FOR in the RW (ΔE = 10.5 for FZX and ΔE = 9.90 for FOR) and BE (ΔE = 6.3 for FZX and ΔE = 6.1 for FOR) drinks. For FL, there were different levels of change between the composites, with a much more significant reduction in FL intensity with RW. CONCLUSION: Beverages have the potential to stain the composites evaluated, especially alcoholic beverages. CLINICAL SIGNIFICANCE: Before carrying out an esthetic treatment with resin composites, it is ideal to investigate the types of drinks consumed by patients, for greater predictability of treatment.

Protective effects of Chinese bayberry pomace wine against oxidative stress on Drosophila melanogaster.

The Chinese bayberry pomace wine (CPW) was prepared with the assisted fermentation of lactic acid bacteria and acetic acid bacteria, and its antioxidant effect on Drosophila melanogaster was researched. After mixed fermentation, CPW had a better color, which means there was more retention of anthocyanins, and the functional activity of anthocyanins could enhance the antioxidant capacity of flies. We found that the lifespan of flies exposed to CPW was prolonged, and the reproductive capacity of these flies was decreased. The food intake of flies was also influenced by CPW with gender differences. Furthermore, CPW alleviated the excessive proliferation of the intestinal precursor cells of H2O2-induced flies and activated the transcription level of antibacterial peptide genes. CPW had a protective effect on H2O2-induced acute injury flies, with an increased survival rate, enhanced SOD and CAT activities, and decreased malondialdehyde (MDA) content in flies. The expression of oxidative stress-related genes including CuZn-SOD, Mn-SOD, and CAT was also significantly upregulated by CPW, but the downregulation effect of CPW on age-related gene expression such as methuselah (MTH), the target of rapamycin (TOR) and ribosomaiprotein S6 kinase (S6K) was sex-specific. These results suggested that CPW played an important role in anti-oxidative stress injury, which was beneficial to promoting the reuse of by-products from Chinese bayberry processing.

[Impact of the Mediterranean diet and the consumption of fermented beverages on subjective well-being in different contexts in Spain]. / Impacto de la dieta mediterránea y del consumo de bebidas fermentadas en el bienestar subjetivo en diferentes contextos en España.

Introduction: The relationship between the Mediterranean diet, the consumption of fermented beverages (beer and wine), and subjective well-being in Spain was investigated. Using a cross-sectional study with 2,752 participants, dietary patterns and emotional well-being were evaluated in contexts such as work, the gym, and bars/restaurants. The results showed that context significantly impacts dietary habits and well-being. Participants at the gym adhered more to the Mediterranean diet and reported fewer negative emotions compared to other contexts. Correlation maps indicated that beer consumption was associated with a higher intake of red meat, while wine was related to a higher consumption of fish and seafood. Additionally, the study found that participants following the Mediterranean diet experienced higher levels of emotional well-being, with less frequent negative emotions such as anxiety and anger, and higher levels of life satisfaction, especially in contexts like the gym. These findings suggest that public health interventions should consider contextual factors to promote healthy eating habits and improve emotional well-being, highlighting the importance of the Mediterranean diet in health and the prevention of psychological problems.

Introducción: Se investigó la relación entre la dieta mediterránea, el consumo de bebidas fermentadas (cerveza y vino) y el bienestar subjetivo en España. Utilizando un estudio transversal con 2752 participantes, se evaluaron los patrones dietéticos y el bienestar emocional en contextos como el trabajo, el gimnasio y bares o restaurantes. Los resultados mostraron que el contexto tiene un impacto significativo en los hábitos dietéticos y el bienestar. Los participantes del gimnasio siguieron más la dieta mediterránea y reportaron menos emociones negativas en comparación a los de otros contextos. Los mapas de correlación indicaron que el consumo de cerveza se asociaba con una mayor ingesta de carne roja, mientras que el vino se relacionaba con un mayor consumo de pescado y mariscos. Además, el estudio encontró que los participantes que seguían la dieta mediterránea experimentaban mayores niveles de bienestar emocional, con menor frecuencia de emociones negativas, como la ansiedad y la ira, y mayores niveles de satisfacción con la vida, especialmente en contextos como el gimnasio. Estos hallazgos sugieren que las intervenciones de salud pública deberían considerar los factores contextuales para fomentar hábitos alimenticios saludables y mejorar el bienestar emocional, resaltando la importancia de la dieta mediterránea en la salud y en la prevención de problemas psicológicos.

The Role of Indigenous Yeasts in Shaping the Chemical and Sensory Profiles of Wine: Effects of Different Strains and Varieties.

The microbial terroir is an indispensable part of the terroir panorama, and can improve wine quality with special characteristics. In this study, eight autochthonous yeasts (Saccharomyces cerevisiae), selected in Huailai country, China, were trailed in small-scale and pilot fermentations for both white (Riesling and Sémillon) and red (Cabernet Sauvignon and Syrah) wines and evaluated by GC-MS analysis and the rate-all-that-apply (RATA) method. Compared to commercial yeast strains, the indigenous yeasts were able to produce higher concentrations of ethyl esters and fatty acid ethyl esters, and higher alcohol, resulting in higher odor activity values of fruity, floral attributes. Marked varietal effects were observed in the pilot fermentation, but yeast strains exerted a noticeable impact in modulating wine aroma and sensory profile. Overall, indigenous yeast could produce more preferred aroma compounds and sensory characteristics for both white and red wines, demonstrating the potential for improving wine quality and regional characteristics.

Alcohol Intake and Prevalent Kidney Stone: The National Health and Nutrition Examination Survey 2007-2018.

The association of alcohol intake with kidney stone disease (KSD) is not clear based on current clinical evidence. We examined the National Health and Nutrition Examination Survey (NHANES) 2007-2018 and used logistic regression analyses to determine the independent association between alcohol intake and prevalent KSD. In total, 29,684 participants were eligible for the final analysis, including 2840 prevalent stone formers (SFs). The mean alcohol intake was 37.0 ± 2.4 g/day among SFs compared to 42.7 ± 0.9 among non-SFs (p = 0.04). Beer [odds ratio (OR) = 0.76, 95% CI: 0.61-0.94, p = 0.01] and wine (OR = 0.75, 95% CI: 0.59-0.96, p = 0.03) intakes were strongly associated with lower odds of prevalent KSD, while liquor intake had no association. Furthermore, the effects of beer and wine intakes on stone formation were dose-dependent. The OR for comparing participants drinking 1-14 g/day of beer to non-drinkers was 1.41 (95%CI: 0.97-2.05, p = 0.07), that of >14-≤28 g/day of beer to non-drinkers was 0.65 (95% CI: 0.42-1.00, p = 0.05), that of >28-≤56 g/day of beer to non-drinkers was 0.60 (95% CI: 0.39-0.93, p = 0.02), and that of >56 g/day of beer to non-drinkers was 0.34 (95% CI: 0.20-0.57, p < 0.001). Interestingly, the effect of wine intake was only significant among participants drinking moderate amounts (>14-28 g/day), with an OR of 0.54 (95% CI: 0.36-0.81, p = 0.003) compared to non-drinkers, but this effect was lost when comparing low-level (1-14 g/day) and heavy (>28 g/day) wine drinkers to non-drinkers. These effects were consistent in spline models. This study suggests that both moderate to heavy beer intake and moderate wine intake are associated with a reduced risk of KSD. Future prospective studies are needed to clarify the causal relationship.

Consumption of Sylimarin, Pyrroloquinoline Quinone Sodium Salt and Myricetin: Effects on Alcohol Levels and Markers of Oxidative Stress-A Pilot Study.

BACKGROUND: Alcohol abuse is one of the most common causes of mortality worldwide. This study aimed to investigate the efficacy of a treatment in reducing circulating ethanol and oxidative stress biomarkers. METHODS: Twenty wine-drinking subjects were investigated in a randomized controlled, single-blind trial (ClinicalTrials.gov. Identifier: NCT06548503; Ethical Committee of the University of Padova (HEC-DSB/12-2023) to evaluate the effect of the intake of a product containing silymarin, pyrroloquinoline quinone sodium salt, and myricetin (referred to as Si.Pi.Mi. for this project) on blood alcohol, ethyl glucuronide (EtG: marker for alcohol consumption) and markers of oxidative stress levels (Reactive Oxygen Species-ROS, Total Antioxidant Capacity-TAC, CoQ10, thiols redox status, 8-isoprostane, NO metabolites, neopterin, and uric acid). The effects of the treatment versus placebo were evaluated acutely and after 1 week of supplementation in blood and/or saliva and urine samples. RESULTS: Si.Pi.Mi intake reduced circulating ethanol after 120 min (-33%). Changes in oxidative stress biomarkers, particularly a TAC (range +9-12%) increase and an 8-isoprostane (marker of lipidic peroxidation) decrease (range -22-27%), were observed too. CONCLUSION: After the administration of Si.Pi.Mi, the data seemed to suggest a better alcohol metabolism and oxidative balance in response to wine intake. Further verification is requested.

Precursors consumption preferences and thiol release capacity of the wine yeasts Saccharomyces cerevisiae, Torulaspora delbrueckii, and Lachancea thermotolerans.

The aromatic profile of wine determines its overall final quality, and among the volatile molecules that define it, varietal thiols are responsible for shaping the distinctive character of certain wine varieties. In grape must, these thiols are conjugated to amino acids or small peptides in a non-volatile form. During wine fermentation, yeasts play a principal role in expressing these aromatic compounds as they internalise and cleavage these precursors, releasing the corresponding free and aroma-impacting fraction. Here, we investigate the impact of three wine yeasts (Saccharomyces cerevisiae, Torulaspora delbrueckii and Lachancea thermotolerans) on thiol releasing in synthetic grape must fermentations supplemented with different cysteinylated (Cys-4MSP and Cys-3SH) and glutathionylated (GSH-4MSP and GSH-3SH) precursors. We demonstrate higher consumption levels of cysteinylated precursors, and consequently, higher amounts of thiols are released from them compared to glutathionylated ones. We also report a significant impact of yeast inoculated on the final thiols released. Meanwhile T. delkbrueckii exhibits a great 3SHA releasing capacity, L. thermotolerans stands out because of its high 3SH release. We also highlight the synergic effect of the co-inoculation strategy, especially relevant in the case of S. cerevisiae and L. thermotolerans mixed fermentation, that has an outstanding release of 4MSP thiol. Although our results stem from a specific experimental approach that differs from real winemaking situations, these findings reveal the potential of unravelling the specific role of different yeast species, thiol precursors and their interaction, to improve wine production processes in the context of wine aroma enhancement.

Improving wine fermentation efficiency of Torulaspora delbrueckii by increasing the ploidy of yeast inocula.

The life cycle of most non-conventional yeasts, such as Torulaspora delbrueckii (Td), is not as well-understood as that of Saccharomyces cerevisiae (Sc). Td is generally assumed to be haploid, which detracts from some biotechnological properties compared to diploid Sc strains. We analyzed the life cycle of several Td wine strains and found that they were mainly diploid during exponential growth in rich medium. However, most cells became haploid in stationary phase, as observed for Sc haploid heterothallic strains. When transferred and incubated in nutrient-deficient media, these haploid cells became polymorphic, enlarged, and transitioned to diploid or polyploid states. The increased ploidy, that mainly results from supernumerary mitosis without cytokinesis, was followed by sporulation. A similar response was observed in yeasts that remained alive during the second fermentation of base wine for sparkling wine making, or during growth in ethanol-supplemented medium. This response was not observed in the Sc yeast populations under any of the experimental conditions assayed, which suggests that it is a specific adaptation of Td to the stressful fermentation conditions. This response allows Td yeasts to remain alive and metabolically active longer during wine fermentation. Consequently, we designed procedures to increase the cell size and ploidy of haploid Td strains. Td inocula with increased ploidy showed enhanced fermentation efficiency compared to haploid inocula of the same strains.

Bioaccessible Phenolic Alkyl Esters of Wine Lees Decrease COX-2-Catalyzed Lipid Mediators of Oxidative Stress and Inflammation in a Time-Dependent Manner.

Lipophenols, phenolic compounds esterified with fatty alcohols or fatty acids, provide greater health benefits upon dietary ingestion of plant-based foods than unesterified (poly)phenols. Based on this premise, the present study aimed to demonstrate the role of gastrointestinal enzymes (pepsin, pancreatin, and pancreatic lipase) in releasing alkyl gallates and trans-caffeates from wine lees, providing bioactive compounds with enhanced capacities against oxidative stress (OS) and para-inflammation. The UHPLC-ESI-QqQ-MS/MS-based analysis revealed ethyl gallate and ethyl trans-caffeate as the most prominent compounds (1.675 and 0.872 µg/g dw, respectively), while the bioaccessibility of the derivatives of gallic and caffeic acids was dependent on the alkyl chain properties. The de novo formation of alkyl gallates during gastric and intestinal digestion resulted from intestinal enzyme activity. Moreover, the in vitro capacity of bioaccessible alkyl esters of gallic and trans-caffeic acids to reduce cyclooxygenase-2 concentration and modulate oxilipins related to OS (8-iso-PGF2α) and inflammation (PGF2α and PGE2) was demonstrated in a time-dependent manner. In conclusion, the presence of alkyl esters of gallic and trans-caffeic acids in wine lees and their subsequent formation during digestion of this byproduct emphasize their value as a source of antioxidant and anti-inflammatory compounds, encouraging the consideration of wine lees as a valuable ingredient for health-promoting coproducts.

Curse or blessing: Growth- and laccase-modulating properties of polyphenols and their oxidized derivatives on Botrytis cinerea.

Infection of grapevines with the grey mold pathogen Botrytis cinerea results in severe problems for winemakers worldwide. Browning of wine is caused by the laccase-mediated oxidation of polyphenols. In the last decades, Botrytis management has become increasingly difficult due to the rising number of resistances and the genetic variety of Botrytis strains. During the search for sustainable fungicides, polyphenols showed great potential to inhibit fungal growth. The present study revealed two important aspects regarding the effects of grape-specific polyphenols and their polymerized oxidation products on Botrytis wild strains. On the one hand, laccase-mediated oxidized polyphenols, which resemble the products found in infected grapes, showed the same potential for inhibition of growth and laccase activity, but differed from their native forms. On the other hand, the impact of phenolic compounds on mycelial growth is not correlated to the effect on laccase activity. Instead, mycelial growth and relative specific laccase activity appear to be modulated independently. All phenolic compounds showed not only inhibitory but also inductive effects on fungal growth and/or laccase activity, an observation which is reported for the first time. The simultaneous inhibition of growth and laccase activity demonstrated may serve as a basis for the development of a natural botryticide. Yet, the results showed considerable differences between genetically distinguishable strains, impeding the use of a specific phenolic compound against the genetic variety of wild strains. The present findings might have important implications for future understanding of Botrytis cinerea infections and sustainable Botrytis management including the role of polyphenols.

Activation of the yeast Retrograde Response pathway by adaptive laboratory evolution with S-(2-aminoethyl)-L-cysteine reduces ethanol and increases glycerol during winemaking.

BACKGROUND: Global warming causes an increase in the levels of sugars in grapes and hence in ethanol after wine fermentation. Therefore, alcohol reduction is a major target in modern oenology. Deletion of the MKS1 gene, a negative regulator of the Retrograde Response pathway, in Saccharomyces cerevisiae was reported to increase glycerol and reduce ethanol and acetic acid in wine. This study aimed to obtain mutants with a phenotype similar to that of the MKS1 deletion strain by subjecting commercial S. cerevisiae wine strains to an adaptive laboratory evolution (ALE) experiment with the lysine toxic analogue S-(2-aminoethyl)-L-cysteine (AEC). RESULTS: In laboratory-scale wine fermentation, isolated AEC-resistant mutants overproduced glycerol and reduced acetic acid. In some cases, ethanol was also reduced. Whole-genome sequencing revealed point mutations in the Retrograde Response activator Rtg2 and in the homocitrate synthases Lys20 and Lys21. However, only mutations in Rtg2 were responsible for the overactivation of the Retrograde Response pathway and ethanol reduction during vinification. Finally, wine fermentation was scaled up in an experimental cellar for one evolved mutant to confirm laboratory-scale results, and any potential negative sensory impact was ruled out. CONCLUSIONS: Overall, we have shown that hyperactivation of the Retrograde Response pathway by ALE with AEC is a valid approach for generating ready-to-use mutants with a desirable phenotype in winemaking.

Staining- and aging-dependent changes in color, translucency, and gloss of cloud-shade and one-shade resin composites.

PURPOSE: To compare staining and artificial accelerating aging-dependent changes in color, translucency, and gloss of cloud-shade and one-shade resin composite. METHODS: The "cloud-shade" composites were Filtek Easy Match Universal Restorative (FU) and SimpliShade (SS): three shades each (light, medium and dark), while the "one shade" composites were Admira Fusion x-tra (AD) and Venus Diamond One (VE). Polymerized composite specimens (10 mm in diameter, 2 mm-thick) were polished using PoGo disks for 40 seconds. Color, translucency, and gloss measurements were performed before and after exposure to 3.8-day staining in coffee or wine (equivalent to a year of service with a 15-minute exposure per day), or exposure to 150 kJ/m² of accelerated aging (n= 5 per condition x three conditions, for a total of 15 specimens per shade). Color and translucency were measured using a benchtop spectrophotometer, and CIEDE2000 color differences (ΔE00) and translucency parameter (TP00) were calculated. Gloss (gloss units, GU) was measured using a small-area gloss meter. Gloss retention percentages after staining and aging were calculated. A two-way ANOVA was used to compare the effect of material and procedure, while a Tukey's post hoc multiple comparison test was used to assess differences among levels within each variable (α= 0.05). RESULTS: The AD exhibited the best color stability upon staining, while VE was the most stable when exposed to artificial aging. Statistically significant differences were recorded among materials and procedures (P< 0.001), and their interactions (P< 0.01). AD also exhibited the highest TP stability upon staining, while the TP of FU was the most stable when exposed to artificial aging. Statistically significant differences were recorded among materials and procedures (P< 0.001), and their interactions (P< 0.01). Gloss retention upon staining and aging was greater than 90% for all tested materials. Statistically significant differences were recorded among materials (P< 0.001), procedures (P< 0.05), and their interactions (P< 0.01). CLINICAL SIGNIFICANCE: Staining and artificial accelerated aging changes in color, translucency parameter, and gloss retention of cloud-shade and one-shade resin composites were material- and procedure-dependent.