Introduction Excessive consumption of sugar-sweetened beverages is a major contributor to dental caries and a global public health concern due to its negative effects on oral health. Phenolic compounds found in edible plants have shown promising effects on caries prevention. Several studies indicate that these bioactive substances can inhibit caries formation through various mechanisms. However, studies which consider the complex interplay of the various factors are missing. In particular, human studies on the caries reducing effects of phenolic compounds in the co-presence of fermentable sugars are lacking. Objective This human intervention study investigated the effect of daily consumption of an apple juice-based drink enriched with phenolic compounds on caries development, compared to the exact same drink without phenolic compounds. Methods 30 healthy male participants were enrolled in a double-blind, randomized, cross-over design. Caries development was measured using lesion depth in bovine tooth enamel samples installed in a removable dental appliance. A mixed model was used to analyze the data, including treatment effects, various oral parameters and interaction terms. Results The analysis showed that consumption of the drink enriched with phenolic compounds resulted in a significant reduction in enamel lesion depth compared to the control, after adjustment for oral parameters and their interactions. Conclusion Enrichment of phenolic compounds could be a promising strategy to mitigate the cariogenic effects of fermentable sugars in beverages. Analyzing the complex interactions between the oral parameters and treatment we bridge the translational gap of in vitro studies and achieve physiological relevance.

Cloudy apple juices produced by spiral filter press technology have recently been reported to contain potentially health-promoting levels of flavan-3-ols. Due to the novelty of these juices, the impact of commonly applied juice clarification treatments on the concentration of flavan-3-ols and other phenolic compounds was comprehensively investigated using RP-UHPLC-DAD and HILIC-FLD. Besides cloudy apple juices with high levels of phenolic compounds, a phenolic compound extract was prepared to further enrich the juices. Subsequent clarification was performed using cross-flow ultrafiltration modules with and without prior addition of gelatin, silica sol, bentonite, and activated charcoal. Ultrafiltration alone lowered phenolic compound concentrations of the apple juice by 9

Due to their importance for sensory perception, volatiles of blackcurrant (Ribes nigrum L.) juices obtained using three pressing systems in two subsequent years were assessed by HS-SPME–GC–MS. The 35 volatiles assigned comprised 15 terpenes, 10 terpenoids, six esters, two aldehydes and each one alcohol and norterpenoid. The aroma impact compound 4-methoxy-2-methyl-2-butanethiol (4MMB) was additionally quantitated by GC–SIM-MS after derivatization and solid phase extraction. Spiral filter press processing under exclusion of oxygen resulted in higher total concentrations of volatiles in the juices (3022 and 4865 μg/100 mL in 2019 and 2020, respectively) compared to those obtained using the conventional systems horizontal filter press and decanter (860–2122 μg/100 mL). Multivariate statistics permitted the clustering of the samples according to both, the pressing systems and year of processing and to deduce tentative marker compounds mostly contributing to their differentiation. Spiral filter pressed juices stood out for elevated levels of especially oxidation-sensitive terpenes. Levels of 4MMB ranged between 3.6 and 4.8 ng/100 mL among all samples. However, descriptive sensory analyses (n = 15 trained panelists) found only insignificant differences among the descriptor intensities comparing blackcurrant juices and nectars produced thereof for sensory analyses, each produced with the aforementioned three technologies. By contrast, consumer acceptance and preference tests (n = 51 consumers) showed a significant preference of the decanter made juice and the nectar derived thereof, accompanied by low ratings for the spiral filter pressed juices.

Grapevine downy mildew caused by Plasmopara viticola is a deleterious vine disease currently controlled using synthetic and copper-based fungicides. Environmental concerns surrounding such fungicides necessitate sustainable alternatives like grape cane extracts. However, studies into their efficacy and effects on the product remain limited. This study aimed to assess the open-field efficacy of a novel grape cane extract formulation, alone and combined with a copper agent, against downy mildew, and to examine its impact on berry and must composition. The results were compared to standard fungicides frequently used in integrated and organic viticulture. In 2022, the grape cane extract formulation reduced downy mildew severity by 78 % on berry clusters, comparable to the maximum acceptable dose of copper for organic farming in Germany (3 kg/ha/a). Combining the grape cane extract with the copper agent was more effective than the copper agent alone, but not more effective than the grape cane extract alone. In 2023, disease development was marginal, despite artificial inoculation. Compositional differences detectable by FTIR were insignificant between musts derived from all treatments, increased copper levels were observed whenever copper had been applied. The phenolic content in freeze-dried, de-seeded berries of 2022 was significantly lower after copper (14.6 mg/g dry weight DW) and grape cane extract treatments (6.6 mg/g DW) when compared to the control (36.1 mg/g DW), with discrepancies in stilbenoid and flavan-3-ol levels in particular, as analysed by HPLC-DAD. However, these differences were not confirmed in 2023, with results showing no correlations between different treatments and phenolic contents. These findings suggest that applications of the phenolic grape cane extract do not lead to artificially altered levels or relevant residues of the active constituents; i.e., the phenolic compounds in the product. In brief, grape cane extracts might represent a promising natural alternative to controlling grapevine downy mildew, particularly in organic viticulture, which, to date, heavily relies on (eco-)toxic copper-pesticides. Further multi-year studies including wine analyses are warranted to follow the entire flow towards the final product.

Protein removal with aspergillopepsin in combination with short-time heating has recently emerged as an opportunity for winemakers to prevent protein haze in wine. In this study, we first treated a protein-rich must and wine with two different aspergillopepsin preparations along an identical control treatment without enzyme addition on a pilot plant scale. While treating wines was rather ineffective, treating musts was effective in reducing protein concentrations by up to 86%, leading to haze-stable wines according to the common heat test. Varying thermal loads in further experiments, a rather mild heat treatment of 65 degrees C for 30 s compared to those in earlier studies was found sufficient for enzymatic protein degradation. Treating must from a fungus-resistant variety (cv. Fidelio) was as successful as treating must from a blend of conventional varieties (cv. Gewurztraminer/Muller-Thurgau). Triangle sensory tests found no clear evidence of an effect on the quality by heat treatments with and without enzyme dosage.

Potentially health-promoting concentrations of flavan-3-ols were previously shown to be retained in apple juices produced with the emerging spiral filter press. Due to the novelty of this technology, the factors governing the stability of flavan-3-ol-rich apple juices have only scarcely been studied. Therefore, we produced flavan-3-ol-rich apple juices and concentrates (16, 40, 70 degrees Brix) supplemented with ascorbic acid (0.0, 0.2, 1.0 g/L) according to common practice. Flavan-3-ols (DP1-7) and twelve flavan-3-ol reaction products were comprehensively characterized and monitored during storage for 16 weeks at 20 and 37 degrees C, employing RP-UHPLC- and HILIC-DAD-ESI (-)-QTOF-HR-MS/MS. Flavan-3-ol degradation followed a second-order reaction kinetic, being up to 3.5-times faster in concentrates (70 degrees Brix) than in single strength juices (16 degrees Brix). Furthermore, they diminished substantially faster compared to other phenolic compounds. For instance, after 16-weeks at 20 degrees C, the maximum loss of flavan-3-ols (-70 %) was greater than those of hydroxycinnamic acids (-18 %) and dihydrochalcones (-12 %). We observed that flavan-3-ols formed adducts with sugars and other carbonyls, such as 5-(hydroxymethyl) furfural and the ascorbic acid-derived L-xylosone. Increased degradation rates correlated particularly with increased furan aldehyde levels as found in concentrates stored at elevated temperatures. These insights could be used for optimizing production, distribution, and storage of flavan-3-ol-rich apple juices and other foods and beverages.

Inflammasomes are multi-protein complexes, which are formed in response to tissue injury, infections, and metabolic stress. However, aberrant inflammasome activation has been linked to several inflammatory diseases. Anthocyanins have been reported to attenuate NLR family pyrin domain-containing 3 (NLRP3) inflammasome activation, but the influence of grape/blueberry anthocyanins and especially their gut-derived metabolites on NLRP3 inflammasome activation in human monocytes remains unclear. Therefore, human leukemic monocytes (THP-1 cells, Tohoku Hospital Pediatrics-1 cells) were preincubated with different concentrations of grape/blueberry anthocyanins, homovanillyl alcohol, or 2,4,6-trihydroxybenzaldehyde (THBA) before the NLRP3 inflammasome was activated by lipopolysaccharide and/or nigericin. Apoptosis-associated speck-like protein containing a CARD (ASC) speck formation, as well as ASC and NLRP3 protein expression, were determined using flow cytometry. Caspase-1 activity was measured in cultured cells, and pro-inflammatory cytokine secretion was determined using enzyme-linked immunosorbent assays. Anthocyanins and their metabolites had no effect on ASC or NLRP3 protein expression. However, THBA significantly inhibited ASC speck formation in primed and unprimed THP-1 monocytes, while caspase-1 activity was significantly declined by grape/blueberry anthocyanins. Furthermore, reduced inflammasome activation resulted in lower pro-inflammatory cytokine secretion. In conclusion, our results show for the first time that grape/blueberry anthocyanins and their gut-derived metabolites exert anti-inflammatory effects by attenuating NLRP3 inflammasome activation in THP-1 monocytes.

The retention of flavan-3-ols and other phenolic compounds during apple juice production at pilot plant scale (200 kg, cv. Boskoop) was investigated. An oxygen-excluding spiral filter press and a conventional decanter were used with and without pectinase mash treatment. Phenolic compounds were comprehensively identified and quantitated by RP-UHPLC and HILIC, both coupled to DAD-FLD and DAD-ESI(-)-QTOF-HR-MS/MS. These techniques combined with using a NIST cocoa flavan-3-ol standard allowed for the first time an individual quantification of flavan-3-ol fractions (DP 1-7) in apple juices. Spiral filter-pressed juices were exposed to less oxidation and exhibited four times higher total phenolic compound levels than decanter-made juices (1016 vs. 262 mg/L). Apple juices derived from pectinase-treated mashes had lower total phenolic compound levels than their non-treated counterparts. However, those made by spiral filter press still retained significantly higher concentrations (780 vs. 104 mg/L). Flavan-3-ols were especially well retained by spiral filter press processing, reaching unprecedentedly high concentrations of up to 713 mg/L. A 280 mL serving of non-treated spiral filter-pressed juice would therefore suffice to provide the daily intake of 200 mg flavan-3-ols, equaling the dose of cocoa flavan-3-ols associated with an authorized European health claim for healthy blood flow.

Chemotherapy with irinotecan (CPT-11), the pro-drug of the highly cytotoxic SN-38, is among the standard-of-care treatments for colorectal cancer. To counteract undesired toxic side effects on healthy tissue such as the intestinal epithelium, the use of preparations rich in polyphenols with anti-oxidative and anti-inflammatory properties such as anthocyanins has been proposed. In the present study, the question of whether non-tumorigenic human epithelium cells (HCEC-1CT) can be protected against the cytotoxic impact of SN-38 by anthocyanin-rich polyphenol extracts without compromising the desired therapeutic effect against tumor cells (HCT-116) was addressed. Hence, single and combinatory effects of anthocyanin-rich polyphenol extracts of elderberry (EB), bilberry (Bil), blackberry (BB) and black currant (BC) with the chemotherapeutic drug SN-38 were investigated. Out of the extracts, BB showed the most potent concentration-dependent cytotoxicity alone and in combination with SN-38, with even stronger effects in non-tumorigenic HCEC-1CT cells. In cytotoxic concentrations, BB decreased the level of DNA/topoisomerase I covalent complexes in HCEC-1CT cells below base level but without concomitant reduction in SN-38-induced DNA strand breaks. The herein reported data argue towards an interference of anthocyanins with successful treatment of cancer cells and a lack of protective properties in healthy cells.

BACKGROUND Organic viticulture seeks sustainable alternatives for eco-toxic copper fungicides to control downy mildew caused by Plasmopara viticola. (Poly)phenol-rich extracts of agricultural byproducts are known to possess antifungal activity, but high production costs often limit their actual implementation. RESULTS Herein, we developed and produced novel ligninsulfonate-based grape cane extract formulations and an apple extract on pilot plant scale, including a detailed (poly)phenol characterization by HPLC-PDA-MS. Our grape cane extract formulations alone reduced downy mildew disease severity in greenhouse trials by 29-69% in a dose-dependent manner, while a standard application of the copper-based agent alone reached ca. 56%. When applied together, disease severity was diminished by 78-92%, revealing a synergistic effect that depended on the mixture ratio. Combining grape cane formulations with the apple extract, additive effects were found (80% disease severity reduction). CONCLUSION The studied plant extracts are proposed to both substitute for and synergistically reinforce copper fungicides in grapevine downy mildew control. This article is protected by copyright. All rights reserved.

Volatiles, descriptive sensory profiles as well as consumer acceptance and preference of juices from red-fleshed ‘Weirouge’ apples produced in 2019 and 2020 with three different dejuicing systems were assessed. HS–SPME–GC–MS analyses revealed differences in the profiles of volatiles in juices processed in an oxygen-reduced atmosphere with an innovative spiral filter press as compared to those obtained using conventional systems, i.e., horizontal filter press and decanter. A total of 49 volatiles was tentatively assigned and permitted a clustering of the samples according to vintage and processing technology by multivariate statistics. Tentative markers to differentiate the individual samples were deduced from the multivariate models. In both years, each three 1,3-dioxanes and C 6 alcohols were revealed as discriminative markers of horizontal filter pressed juices. Descriptive sensory analysis by trained panelists revealed higher intensity scores of ‘oxidized’ and ‘apple-like’ orthonasal odors in juices produced by horizontal filter press and decanter as compared to those obtained by spiral filter press. The visual appearance of the spiral filter pressed juices was significantly higher rated compared to those obtained by conventional pressing systems as revealed by an untrained consumer panel ( n = 65). In contrast, both odor and taste were lower rated, ultimately resulting in a clear-cut higher acceptance and preference of the decanter-made juices, followed by those obtained by horizontal and spiral filter press.

The comprehensive identification of the proteome content from a white wine (cv. Silvaner) is described here for the first time. The wine protein composition isolated from a representative wine sample (250 L) was identified via mass spectrometry (MS)-based proteomics following in-solution and in-gel digestion methods after being submitted to size exclusion chromatographic (SEC) fractionation to gain a comprehensive insight into proteins that survive the vinification processes. In total, we identified 154 characterized (with described functional information) or so far uncharacterized proteins, mainly from Vitis vinifera L. and Saccharomyces cerevisiae. With the complementarity of the two-step purification, the digestion techniques and the high-resolution (HR)-MS analyses provided a high-score identification of proteins from low to high abundance. These proteins can be valuable for future authentication of wines by tracing proteins derived from a specific cultivar or winemaking process. The proteomics approach presented herein may also be generally helpful to understand which proteins are important for the organoleptic properties and stability of wines.

Blackcurrant juices were produced with an oxygen-excluding spiral filter press and two conventional pressing systems exerting common oxygen exposure at pilot plant scale (200 kg) in technological duplicate in two subsequent years ( n = 4). In contrast to findings on previously studied fruits, contents of total and individual anthocyanins were not significantly higher after spiral filter (1636–1872 mg/L) than after conventional pressing (1529–2083 mg/L). During storage for 52 weeks at 4, 20 and 37 °C, anthocyanin levels depended strongly on storage temperature, but not on the used pressing system (conventional vs. oxygen-reduced). Other juice quality parameters like ascorbic acid behaved likewise. Furthermore, a D-optimal design of experiments was used to study the influence and interactions of the factors oxygen, ascorbic acid, sugar and pH value on anthocyanin levels during a heat treatment in a model solution made with preparatively isolated blackcurrant anthocyanins. In agreement with our production trials at pilot plant scale, oxygen levels were only marginally important for the rate of anthocyanin degradation, but the influence of sugar content, ascorbic acid content and pH value was highly relevant as discussed in this report.

Anti-oxidant, -inflammatory, and -carcinogenic activities of bioactive plant constituents, such as anthocyanins, have been widely discussed in literature. However, the potential interaction of anthocyanin-rich extracts with routinely used chemotherapeutics is still not fully elucidated. In the present study, anthocyanin-rich polyphenol extracts of blackberry (BB), bilberry (Bil), black currant (BC), elderberry (EB), and their respective main anthocyanins (cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, cyanidin-3-O-rutinoside, and cyanidin-3-O-sambubioside) were investigated concerning their cytotoxic and DNA-damaging properties in murine CT26 cells either alone or in combination with the chemotherapeutic agent SN-38. BB exerted potent cytotoxic effects, while Bil, BC, and EB only had marginal effects on cell viability. Single anthocyanins comprised of the extracts could not induce comparable effects. Even though the BB extract further pronounced SN-38-induced cytotoxicity and inhibited cell adhesion at 100–200 µg/mL, no effect on DNA damage was observed. In conclusion, anti-carcinogenic properties of the extracts on CT26 cells could be ranked BB >> BC ≥ Bil ≈ EB. Mechanisms underlying the potent cytotoxic effects are still to be elucidated since the induction of DNA damage does not play a role.

Stilbenoids in grape cane (Vitis vinifera L.) are bioactive compounds relevant for plant defense and the potential valorization of this byproduct. Our screening of grape cane from 102 varieties showed constitutive stilbenoid levels in a wide range (557-7748 mg/kg of dry weight). Analyses of genetically distinct clones of selected cultivars unraveled that intravarietal variability (e.g., cv. Riesling, 3236-6541 mg/kg) was higher than that across samples from a single clone but different vineyard positions (3017-3710 mg/kg). Furthermore, stilbenoid levels in samples obtained in October, December, and February (3 years, 2017-2019) showed pronounced quantitative and qualitative variability and the highest yields upon December pruning. For instance, vitisin B and ε-viniferin in cv. Pinot Noir and Accent were predominant in 2017 and 2019 (both >90% of total stilbenoids) but not in 2018 (both <55%) when temperatures were high and precipitation low. In brief, we report the variability of stilbenoid levels in grape cane depending upon genetic and environmental factors.

In this work, two vintages (2019 and 2020) of red-fleshed ‘Weirouge’ apples were processed with the innovative spiral filter press technology to investigate juice production in an oxygen-reduced atmosphere. After pressing, a more brilliant red color and appreciably higher amounts of oxidation-sensitive constituents (ascorbic acid, anthocyanins, and colorless (poly)phenols) were seen in spiral filter pressed juices compared to those produced with conventional systems (horizontal filter press and decanter). In a subsequent stability study (24 weeks storage at 4, 20, and 37 °C), the color and phenolic compounds were monitored and differences in the juices produced with the different pressing-systems were widely maintained during the storage period. The analyses of the anthocyanins and colorless (poly)phenols were conducted by UHPLC-DAD-ESI-QTOF-HR-MS/MS and UHPLC-DAD. The spiral filter press emerged as a promising technology for the production of juices with a more attractive color and a better retention of oxidation-sensitive constituents during processing and storage compared to conventional juices.

Organic and biodynamic management practices in viticulture have been sharply increasing worldwide for almost two decades and are seen as one possibility for environmentally friendly production. Consumers often presume that the quality of organically grown crops is different or higher compared to conventionally grown crops.In the current study, wine grape quality under integrated, organic and biodynamic viticulture was assessed in a long-term field trial in Geisenheim, Germany, starting seven years after conversion. Treatments substantially differed in their amino acid concentration in juice, flavonol content and their composition of hydroxycinnamic acids. Organic and biodynamic treatments showed significantly higher concentrations of all amino acids in juice and a significantly higher concentration of flavonols in berry skins. Major drivers of these changes are reductions of yield and pruning weight under organic and biodynamic management on the one hand and increased light exposure of bunches in the respective treatments on the other hand.Linking amino acid composition in juice, grape skin flavonoid and hydroxycinnamate concentration to canopy structure, vigour and yield enables growers to actively control and manipulate wine grape quality in the different management systems.

Cross-linking net aggregates of thermolabile thaumatin-like proteins (TLPs) and chitinases (CHIs) are the primary source of haze in white wines. Although bentonite fining is still routinely used in winemaking, alternative methods to selectively remove haze proteins without affecting wine organoleptic properties are needed. The availability of pure TLPs and CHIs would facilitate the research for the identification of such technological advances. Therefore, we proposed the usage of recombinant TLP (rTLP) and CHI (rCHI), expressed by Komagataella phaffii, as haze-protein models, since they showed similar characteristics (aggregation potential, melting point, functionality, glycosylation levels and bentonite adsorption) to the native-haze proteins from Vitis vinifera. Hence, rTLP and rCHI can be applied to study haze formation mechanisms on a molecular level and to explore alternative fining methods by screening proteolytic enzymes and ideal adsorptive resins.

Cancer mortality is mainly due to metastasis. Therefore, searching for new therapeutic agents suppressing cancer cell migration is crucial. Data from human studies regarding effects of anthocyanins on cancer progression, however, are scarce and it is unclear whether physiological concentrations of anthocyanins and their metabolites reduce cancer cell migration in vivo. In addition, interactions with chemotherapeutics like 5-fluorouracil (5-FU) are largely unknown. Thus, we combined a placebo-controlled, double-blinded, cross-over study with in vitro migration studies of colon cancer cell lines to examine the anti-migratory effects of plasma-isolated anthocyanins and their metabolites (PAM). Healthy volunteers (n = 35) daily consumed 0.33 L of an anthocyanin-rich grape/bilberry juice and an anthocyanin-depleted placebo juice for 28 days. PAM were isolated before and after intervention by solid-phase extraction. HT-29 and Caco-2 cells were incubated with PAM in a Boyden chamber. Migration of HT-29 cells was significantly inhibited by PAM from juice but not from placebo. In contrast, Caco-2 migration was not affected. Co-incubation with 5-FU and pooled PAM from volunteers (n = 10), which most effectively inhibited HT-29 migration, further reduced HT-29 migration in comparison to 5-FU alone. Therefore, PAM at physiological concentrations impairs colon cancer cell migration and may support the effectiveness of chemotherapeutics.

The effect of cultivar and environmental variations and their interaction on anthocyanin components of strawberry were assessed for six cultivars grown in five locations from North to South of Europe in two different years. To evaluate the impact of latitude- and altitude-related factors, daily mean (Tmean), maximum (Tmax) and minimum (Tmin) temperature and global radiation accumulated for 3, 5, 10 and 15 days before fruit sampling, was analyzed. In general, fruits grown in the south were more enriched in total anthocyanin and pelargonidin-3-glucoside (pel-3-glc), the most abundant anthocyanin in strawberry. Principal component analysis (PCA) provided a separation of the growing locations within a cultivar due to latitudinal climatic differences, temporary weather changes before fruit collection and cultivation technique. PCA also depicted different patterns for anthocyanin distribution indicating a cultivar specific reaction on the environmental factors. The linear regression analysis showed that pel-3-glc was relatively less affected by these factors, while the minor anthocyanins cyanidin-3-glucoside, cyanidin-3-(6-O-malonyl)-glucoside, pelargonidin-3-rutinoside and pelargonidin-3-(6-O-malonoyl)-glucoside were sensitive to Tmax. The global radiation strongly increased cya-3-mal-glc in ‘Frida’ and pel-3-rut in ‘Frida’ and ‘Florence’. ‘Candonga’ accumulated less pel-3-glc and total anthocyanin with increased global radiation. The anthocyanin profiles of ‘Gariguette’ and ‘Clery’ were unaffected by environmental conditions.