The researchers prepared 80% ethanol extracts from seeds collected in Saudi Arabia’s Jazan region, then quantified selected phytochemical classes and individual phenolics. Total phenolics, flavonoids, alkaloids and terpenes were measured with colorimetric assays, while HPLC–DAD was used to analyze phenolic compounds. The biological testing included DPPH and ABTS radical-scavenging assays, FRAP, inhibition of α-amylase, α-glucosidase and butyrylcholinesterase, antimicrobial testing against four bacterial reference strains and one yeast strain, biofilm assays, and cytotoxicity measurements in Caco-2 colorectal adenocarcinoma cells and Wi38 normal lung fibroblasts.

Coffee seed extract contained 172.46 ± 7.17 mg gallic acid equivalents per gram of dry extract for total phenolics, 157.36 ± 3.34 mg quercetin equivalents per gram for total flavonoids, 10.16 ± 0.27 mg/g of alkaloids and 12.41 ± 0.26 mg limonene equivalents per gram for total terpenes. The corresponding loquat values were 40.00 ± 1.58 mg GAE/g, 3.24 ± 1.02 mg QuE/g, 4.25 ± 0.24 mg/g and 2.30 ± 0.04 mg/g. Chlorogenic acid was the dominant measured phenolic in both extracts, at 63,681.37 ± 1,245.6 µg/g in coffee and 5,848.71 ± 98.3 µg/g in loquat.

The coffee extract recorded lower half-maximal inhibitory concentrations in the principal antioxidant and enzyme assays. Its reported IC₅₀ values were 0.42 ± 0.02 µg/mL for ABTS radical scavenging and 0.66 ± 0.03 µg/mL for DPPH, compared with 1.85 ± 0.06 and 2.10 ± 0.08 µg/mL, respectively, for loquat. Coffee also showed lower IC₅₀ values for α-glucosidase inhibition, at 10.20 ± 0.15 µg/mL versus 48.50 ± 1.10 µg/mL, and butyrylcholinesterase inhibition, at 3.20 ± 0.08 µg/mL versus 6.90 ± 0.12 µg/mL. These measurements demonstrate activity in the specific in vitro assays but do not establish the inhibition mechanism or effects in people.

Both extracts inhibited the tested microorganisms, although the quantitative endpoints generally favored coffee. The reported minimum inhibitory concentration of coffee extract against Staphylococcus aureus was 7.8 µg/mL, compared with 31.25 µg/mL for loquat extract. Against C. albicans, the corresponding values were 15.62 and 62.5 µg/mL. The study used reference strains, including Enterococcus faecalis, S. aureus, Escherichia coli, Salmonella typhi and C. albicans, rather than clinical or drug-resistant isolates.

Established-biofilm testing produced the study’s most prominent loquat result. At 75% of the minimum fungicidal concentration, loquat extract reduced measured C. albicans biofilm biomass by 94.12 ± 0.8%. Coffee extract reduced the same measure by 96.68%. These crystal-violet measurements cover both living and dead microorganisms and extracellular material, so they cannot by themselves distinguish inhibition of biofilm formation from cell killing. Parallel viability assessments indicated that the lowest tested concentrations were sublethal, but additional imaging and viability methods would be needed to characterize the effect more precisely.

In the cell assays, coffee extract showed differential cytotoxicity between the two tested cell lines, with a reported Caco-2 IC₅₀ of 78.5 ± 0.2 µg/mL and a Wi38 IC₅₀ of 181.3 ± 2.2 µg/mL, producing a preliminary selectivity index of 2.31. Loquat extract also reduced Caco-2 cell viability, but was more toxic to the normal fibroblast line. The study assessed one cancer cell line and one normal cell line over 48 hours; it did not establish broad anticancer activity, apoptosis, therapeutic selectivity or safety in humans.

Analysis of 12 phenolics quantified in at least one extract found strong inverse associations between chlorogenic acid concentration and several IC₅₀ measurements. Those correlations are compatible with chlorogenic acid contributing to the observed activity, but they do not demonstrate causation. The chemical analysis was limited to 18 phenolic standards, and a major chromatographic peak remained unidentified. Caffeine and loquat triterpenoids were not quantified, leaving their proposed contributions unconfirmed.

The study was also constrained by its use of one seed source and crude extracts, plus the absence of pharmacokinetic, animal or clinical testing. The findings therefore identify measurable in vitro differences between the two seed extracts. Coffee seed extract showed the broader set of stronger assay results, while loquat seed extract was distinguished by its reduction of established C. albicans biofilm biomass. Broader microbiological, chemical and in vivo studies are needed to determine how those laboratory observations translate beyond the tested systems.

Subject of Research: Comparative in vitro phytochemical and biological assessment of coffee and loquat seed extracts

Article Title: Comparative phytochemical characterization and in vitro biological activities of coffee (Coffea arabica) and loquat (Eriobotrya japonica) seed extracts

Article References: Al-Rajhi, A. M. H., Alsalamah, S. A., Almotayri, A. M., Almimoni, N. M., Zeleei, R. I., Al Abboud, M. A., Shater, A.-R. M., Mashraqi, A., Sahagi, E. M., Masmali, I., & Abdelghany, T. M. (2026). Comparative phytochemical characterization and in vitro biological activities of coffee (Coffea arabica) and loquat (Eriobotrya japonica) seed extracts. Scientific Reports, 16(1), Article 32073. https://doi.org/10.1038/s41598-026-72286-4

Image Credits: AI Generated

DOI: 10.1038/s41598-026-72286-4

Keywords: coffee seed extract, loquat seed extract, chlorogenic acid, antioxidant activity, enzyme inhibition, antimicrobial activity, Candida albicans, biofilm reduction, cell cytotoxicity, in vitro study, Comparative, phytochemical

Polyphenols have long attracted interest for wound therapy because they combine several beneficial properties in a single molecule. Chlorogenic acid (CGA), found in coffee and many fruits, exhibits antioxidative, anti-inflammatory, and antibacterial activities. Quercetin (QCT), a flavonoid with broad pharmacological effects, modulates oxidative stress, inflammatory responses, and cellular signalling pathways. Both compounds act as antimicrobials with limited potential for resistance development, a crucial advantage in an era when conventional antibiotics are losing efficacy. Yet both face a fundamental delivery problem: poor water solubility, low stability, or limited dermal bioavailability restrict their clinical use. CGA is chemically unstable and prone to degradation, while QCT is so poorly soluble that it struggles to exert biological activity at all.

Liposomes offer a compelling solution. Composed of phosphatidylcholine from soybean, these vesicles resemble biological membranes, providing high biocompatibility, low cytotoxicity, and non-irritant properties suitable for application to damaged or inflamed skin. Hydrophilic compounds can be carried in the aqueous core, while lipophilic ones partition into the lipid bilayer. The Tromsø team, led by Lisa Myrseth Hemmingsen and Nataša Škalko-Basnet, used the thin-film hydration method to prepare both CGA-loaded liposomes (CGA-LP) and QCT-loaded liposomes (QCT-LP), then tailored vesicle size through membrane extrusion for CGA-LP and probe sonication for QCT-LP, depending on each compound’s physicochemical properties.

All formulations landed within the target range of 200–300 nanometres, a size considered favourable for dermal therapy because it enhances retention within wound tissue and supports penetration into bacterial biofilms. Entrapment efficiency varied dramatically between the two compounds, reflecting their opposing chemistries. CGA-LP showed a low entrapment efficiency of about 23 percent, commonly observed for small hydrophilic molecules held in the aqueous core. QCT-LP, in contrast, achieved an entrapment efficiency of roughly 98 percent, consistent with the lipophilic nature of QCT and its incorporation into the lipid bilayer. Notably, the loaded formulations remained more stable over four weeks of storage than their empty counterparts, possibly because polyphenols interact with phospholipid headgroups through hydrogen bonding and polar interactions, increasing membrane cohesion and limiting vesicle fusion or aggregation.

In vitro release studies using Franz diffusion cells maintained at 32 degrees Celsius, simulating skin surface temperature, revealed the sustained-release behaviour that makes liposomes attractive for chronic wounds. Free CGA permeated rapidly, with about 60 percent crossing the membrane within 24 hours, whereas CGA-LP released only about 30 percent over the same period. QCT-LP released about 50 percent over 24 hours, while free QCT in methanol showed lower permeation of roughly 13 percent. The slower release from liposomes is attributable to diffusion of the payload through the lipid bilayer, providing more consistent exposure, a sought property for chronic wound therapy where prolonged therapeutic concentrations are needed. Sustained release might also maintain concentrations at the infection site for extended periods, potentially enhancing antimicrobial activity while minimizing off-target effects and reducing the frequency of dressing changes.

Antioxidant activity was assessed using two complementary radical scavenging assays, DPPH and ABTS·+, with vitamins C and E serving as reference antioxidants. CGA exhibited concentration-dependent scavenging between 5 and 25 micromolar, reaching a plateau at higher concentrations, and showed higher activity than vitamin C at 10 micromolar, confirming radical scavenging potential comparable to the reference vitamins. QCT displayed high DPPH activity across the tested range, likely favoured by the lipophilic nature of the DPPH radical, but only about 20 percent scavenging in the ABTS·+ assay regardless of concentration, a comparatively low response that may reflect the assay’s compatibility with hydrophilic antioxidants and possible interactions between QCT and formulation components.

Biocompatibility studies on murine macrophages, cells that play a central role in wound healing, revealed no cytotoxicity for either liposomal formulation. Almost all formulations maintained cell viability above 80 percent, the generally regarded non-toxic threshold, at lipid concentrations up to 50 micrograms per millilitre. In a striking contrast, free QCT at equivalent concentrations exhibited cytotoxic effects, underscoring how liposomal entrapment can transform a problematic compound into a tolerable one. The anionic and neutral liposome compositions, similar to cell membranes, generally exhibit low cytotoxicity, reinforcing the suitability of these systems for topical use.

Antimicrobial evaluation against S. aureus MSSA 476 produced nuanced results. In the conventional broth microdilution assay, only QCT-LP showed measurable inhibitory activity, with a minimum inhibitory concentration of 61.25 micrograms per millilitre. Free QCT could only be tested up to 10 micrograms per millilitre due to limited solubility, at which it achieved just 25.5 percent inhibition. CGA and CGA-LP showed no measurable MIC within the tested range. However, when the researchers turned to isothermal microcalorimetry, a technique that monitors bacterial metabolic activity in real time through heat flow measurements, a different picture emerged. CGA-LP induced a greater reduction in total metabolic rate than free CGA, with more pronounced suppression at lower concentrations of 30 to 120 micrograms per millilitre, suggesting that entrapment might enhance the interaction or uptake of CGA by bacterial cells at sub-inhibitory concentrations. Interestingly, the empty liposomes alone also decreased measurable metabolic activity, implying that components of the liposomal carrier exert subtle metabolic pressure on S. aureus independently of the active compound.

The discrepancy between the two antimicrobial assays likely reflects their different readouts: metabolic activity can persist even when bacterial growth is inhibited, and decreases in metabolic activity do not always correspond directly to reductions in viable bacteria. The microcalorimetry findings point toward the added value of real-time metabolic monitoring in revealing nuanced effects that endpoint MIC measurements cannot capture, potentially influencing bacterial physiology and response to treatment in ways that conventional testing overlooks.

The Tromsø team concludes that liposomal entrapment of CGA and QCT was successful, enabling sustained release and modulating the delivery and biological performance of both polyphenols. QCT-LP demonstrated measurable antibacterial activity in a conventional assay, while CGA-LP altered bacterial metabolism via microcalorimetry. Both formulations showed suitable biocompatibility with murine macrophages, supporting their topical potential. The researchers caution, however, that direct evaluation of wound-healing efficacy in biologically relevant wound models remains necessary to confirm therapeutic relevance. Additional studies in relevant in vitro and in vivo wound models will be required to establish effects on oxidative stress, inflammation, and wound healing processes. Still, the findings suggest that liposomal delivery could address two of the most stubborn challenges in chronic wound care, oxidative stress and bacterial infection, with natural compounds that bacteria are unlikely to develop resistance against.

Subject of Research: Liposomal delivery of polyphenols for chronic wound therapy

Article Title: Harnessing liposomal delivery to boost polyphenols’ potential in chronic wound therapy

Article References: Hemmingsen, L. M., Kristensen, M., Dang, C. T. L., Salamonsen, M., Fredheim, E. G. A., & Škalko-Basnet, N. (2026). Harnessing liposomal delivery to boost polyphenols’ potential in chronic wound therapy. Frontiers in Drug Delivery, 6, Article 1839747. https://doi.org/10.3389/fddev.2026.1839747

Image Credits: AI Generated

DOI: 10.3389/fddev.2026.1839747

Keywords: liposomes, polyphenols, chronic wounds, chlorogenic acid, quercetin, antimicrobial resistance, drug delivery, wound healing, Harnessing, liposomal, delivery, boost

The research team, led by Hyun-Jin Na and Mi Jeong Sung, with Min-Sun Kim as co-author, used the fruit fly Drosophila melanogaster to dissect how redox imbalance derails intestinal stem cell function during aging. The fly midgut has become a premier model for this kind of work because its stem cells can be tracked, genetically manipulated, and compared across the lifespan with remarkable precision. As flies age, their intestinal stem cells proliferate excessively, the epithelial barrier deteriorates, and reactive oxygen species accumulate, a triad of defects that mirrors what happens in mammalian intestines, including our own.

Chlorogenic acid, abbreviated CGA, is an ester of caffeic acid and quinic acid that ranks among the most abundant polyphenols in the human diet. Coffee drinkers consume hundreds of milligrams daily, and the compound has attracted attention for anti-inflammatory, antioxidant, and metabolic effects in contexts ranging from diabetic vascular damage to chronic kidney disease. What remained unclear was whether CGA could specifically protect the stem cell compartment of an aging intestine, and if so, through what molecular channel the protection flows.

To answer these questions, the researchers deployed three complementary fly models: naturally aging flies, flies subjected to hydrogen peroxide-induced oxidative stress, and flies fed a high-sucrose diet that accelerates intestinal decline. Across all three conditions, CGA treatment was associated with reduced accumulation of reactive oxygen species, diminished DNA damage response signaling, and suppression of the aberrant stem cell proliferation that characterizes stressed and aged guts. The compound also improved intestinal barrier integrity, a functional readout with direct relevance to age-related leaky gut, and reduced stress-associated autophagic responses in the epithelium.

The genetic centerpiece of the study is CncC, the Drosophila ortholog of the mammalian transcription factor NRF2. NRF2 sits at the apex of cellular antioxidant defense, binding to antioxidant response elements in DNA and switching on genes that neutralize oxidants, repair damage, and maintain proteostasis. Earlier work from the Jasper and Bohmann laboratories had established that Keap1-Nrf2 signaling controls intestinal stem cell proliferation in flies, making the pathway a natural candidate for mediating dietary protection. When the team analyzed flies lacking functional CncC, the protective effects of CGA on stem cell dynamics and oxidative stress markers were lost, demonstrating that the compound requires this pathway to exert its benefits.

Importantly, the authors are careful about what their data do and do not show. The study does not establish direct activation of CncC by the parent CGA molecule or by its metabolites. This distinction matters because polyphenols often act indirectly, for example by generating a mild oxidative signal that triggers the cell’s own defense systems, or through gut microbial conversion into more active species. The requirement for CncC-dependent capacity, rather than proof of direct binding, positions CGA as an upstream modulator whose precise molecular target remains an open question for future work.

Transcriptomic analysis using RNA sequencing added a genome-wide dimension to the findings. The researchers profiled gene expression changes associated with CGA treatment and found alterations in pathways related to antioxidant defense, genome maintenance, and cell cycle regulation. These transcriptional signatures align with the observed phenotypes: fewer reactive oxygen species, less DNA damage signaling, and normalized stem cell division. The data processing pipeline relied on established bioinformatics tools, including Fastp for read preprocessing, STAR for alignment, Salmon for transcript quantification, and DESeq2 for differential expression analysis, lending methodological rigor to the pathway-level conclusions.

The broader significance of the work lies in connecting dietary bioactive compounds to a specific stem cell aging mechanism. Intestinal epithelial homeostasis depends on a delicate balance between stem cell self-renewal and differentiation, and oxidative stress tips this balance toward hyperproliferation and barrier failure. By showing that CGA attenuates this redox-driven dysregulation through a CncC-dependent route, the study extends the redox-protective effects of dietary polyphenols from general cytoprotection into the realm of tissue-specific stem cell aging. Because NRF2 is conserved from flies to humans, the findings provide a mechanistic framework that can be tested in mammalian models and, eventually, in human nutrition studies.

Several caveats temper the enthusiasm. The work was conducted entirely in flies, and dose translation from fly food to human diets is far from straightforward. The authors note that their data do not prove direct CncC activation, leaving open the possibility that metabolites, microbiome interactions, or indirect stress-signaling pathways mediate the effect. Moreover, while barrier function improved and autophagic stress responses declined, the long-term consequences of CGA supplementation on lifespan and healthspan in aging animals remain to be fully characterized in this system.

Even so, the study offers a compelling proof of principle that a common dietary molecule can engage an evolutionarily conserved stress-response pathway to preserve stem cell function in an aging organ. As populations worldwide age, interventions that maintain intestinal integrity could have outsized effects on healthy longevity, given the gut’s role in immunity, metabolism, and systemic inflammation. The Korea Food Research Institute team’s findings, supported by the institute’s Main Research Program, suggest that the humble polyphenol in a morning cup of coffee may be doing more than providing antioxidants in the bloodstream; it may be helping to keep the gut’s regenerative engine running smoothly as the years accumulate.

Subject of Research: Chlorogenic acid protection of aging intestinal stem cells via the CncC/NRF2 oxidative stress pathway in Drosophila

Article Title: Oxidative stress-associated intestinal stem cell dysfunction during aging is attenuated by chlorogenic acid in a CncC-dependent manner

Article References: Na, H.-J., Kim, M.-S., & Sung, M. J. (2026). Oxidative stress-associated intestinal stem cell dysfunction during aging is attenuated by chlorogenic acid in a CncC-dependent manner. GeroScience. https://doi.org/10.1007/s11357-026-02586-0

Image Credits: AI Generated

DOI: 10.1007/s11357-026-02586-0

Keywords: chlorogenic acid, intestinal stem cells, oxidative stress, NRF2, CncC, Drosophila, aging, gut barrier, DNA damage response, autophagy, polyphenols, GeroScience

The research team, led by scientists at the University of Primorska in Slovenia together with collaborators at the Institute for Medicinal Plants Research Dr Josif Pančić in Serbia, set out with a simple but commercially significant question: does the plant material that survives essential oil extraction still contain anything worth extracting? Immortelle is cultivated across Italy, Spain, Portugal, Corsica, and the coastal stretches of Slovenia and Croatia primarily for its essential oil, which accumulates in glandular trichomes on the flowering shoots. The oil yield is remarkably low, typically between 0.5 and 0.8 percent of the processed biomass, which means that for every kilogram of precious oil, producers generate well over a hundred kilograms of dry leftover plant matter.

Chemically, this outcome makes sense. Steam distillation is designed to pull out volatile, hydrophobic molecules such as alpha-pinene, geranyl acetate, and the ester 2-phenylethyl tiglate, which give immortelle oil its characteristic scent and its documented skin-related bioactivities. Those compounds dissolve poorly in water, so they should largely remain locked in the plant matrix or escape with the oil itself. But the non-volatile phenolic fraction — hydroxybenzoic acids, hydroxycinnamic acids, and flavonoid derivatives — follows different solubility rules, and because the plant material is not finely ground before distillation, a substantial share of these intracellular compounds can survive the steam treatment intact.

To test that hypothesis, the researchers prepared hot-water infusions from immortelle biomass both before and after distillation, steeping one gram of dried material in 200 milliliters of boiling water for ten minutes, exactly the way a consumer would brew a tea bag. They then quantified fifteen phenolic compounds by high-performance liquid chromatography with diode-array detection and measured total phenolic content with the Folin-Ciocalteu assay. The results were striking: infusions from distilled biomass contained the major phenolic constituents at concentrations statistically indistinguishable from those extracted from undistilled material. Total phenolic content actually came out slightly higher in the distilled sample, at 28.89 milligrams of gallic acid equivalents per gram versus 26.99 for the undistilled herb, and the radical-scavenging capacity measured by the DPPH assay followed the same pattern.

The dominant compounds in both infusions were a rosmarinic acid derivative at roughly 50 micrograms per gram, vanillic acid at about 27 micrograms per gram, and chlorogenic acid at around 15 micrograms per gram. These are molecules with meaningful nutritional pedigrees. Rosmarinic acid has shown a favorable safety profile in human trials at oral doses of 500 milligrams, with reported benefits for cognitive performance and sleep quality. Chlorogenic acid has been implicated in fat oxidation and body-composition regulation in cell models and clinical studies of enriched coffee, while caffeic and p-coumaric acids have been linked to improved vascular endothelial function in adlay tea research.

Knowing that a residue is chemically rich is one thing; proving that people will happily drink it is another. Phenolic compounds are notorious for imparting bitterness, and earlier work by the same group found that plain immortelle infusions scored only 3.14 out of 6 for taste acceptability. To address this, the team blended the distilled biomass with Vitalplant, a commercially available botanical mixture containing buckthorn bark, peppermint leaves, caraway fruit, and parsley fruit, a formulation chosen both for its aromatic masking potential and for prior preclinical evidence of metabolic benefits. The blends were analyzed alongside the individual components, and the mixtures containing immortelle roughly doubled the phenolic content and antioxidant activity compared with Vitalplant alone, confirming the residue as the dominant contributor of bioactivity.

Interestingly, distillation left a subtle fingerprint on the detailed phenolic profile of the blends. The mixture with undistilled immortelle contained more chlorogenic acid, caffeic acid, and one luteolin derivative, while the mixture with distilled biomass delivered more rosmarinic acid, its derivative, and a second luteolin derivative. The authors caution that these shifts may reflect changes in solvent accessibility within the plant matrix after oil removal, or interactions among the blended herbs, rather than simple loss during distillation. Either way, the total phenolic payload was essentially preserved, which is the number that matters most for a functional ingredient.

The team then took the concept into a human trial. Thirty-two adults aged 40 to 60, all carrying at least two traits of metabolic syndrome, drank 200 milliliters of one of three beverages every morning for one week in a randomized, partially crossover design: Vitalplant plus distilled immortelle, Vitalplant plus undistilled immortelle, or Vitalplant alone. Sensory scoring revealed a genuine advantage for the distilled material. The blend with distilled immortelle earned an overall impression score of 4.15 and a taste-liking score of 3.70, both better than the undistilled blend’s 3.27 and 2.95, and its overall impression was not significantly different from Vitalplant alone, which scored around 5. The researchers speculate that removing volatile constituents during distillation may strip out compounds that amplify bitterness perception, though they note this explanation remains tentative because the volatiles were not directly measured.

Physiologically, the one-week intervention was deliberately short, and most measures stayed flat. No significant between-beverage differences emerged for body composition, blood lipids, glucose, or inflammatory markers, and none of the within-group changes in gastrointestinal symptoms or appetite survived correction for multiple comparisons. One notable exception was blood pressure: the group drinking the distilled-immortelle blend showed a significant reduction in both systolic pressure, down 4.05 millimeters of mercury, and diastolic pressure, down 3.38 millimeters of mercury, changes that remained significant after adjustment. Because there was no significant difference between the beverages on this endpoint, the authors urge caution, noting that regression to the mean and measurement variability cannot be excluded. Crucially, no adverse effects appeared in any group, supporting the short-term safety of beverages made from post-distillation biomass.

The broader implications reach well beyond immortelle. From an ecological standpoint, a preliminary study has suggested that dumping distillation residues on soil can depress plant-available nitrogen, possibly because residual phenolics inhibit nitrification and alter microbial communities, so extracting those compounds before land application could ease that agronomic risk while capturing commercial value. The authors also argue that the same valorization logic should apply to other dual-purpose aromatic crops such as sage, fennel, rosemary, and lavender, plants that are distilled for oil yet also have traditions as foods and herbal remedies. Longer intervention periods, comprehensive chemical profiling including mass-spectrometric identification of the unidentified peaks, and formal sensory optimization of bitterness and astringency remain the next steps. But the core proof of concept now stands: the fragrant waste pile outside the distillery is not waste at all, and with a simple pot of boiling water it can be converted into a phenolic-rich infusion that people can actually drink.

Subject of Research: Valorization of post-distillation Helichrysum italicum biomass as a phenolic-rich functional beverage ingredient

Article Title: Valorization of post-distillation Helichrysum italicum biomass for functional beverage infusions

Article References: Kenig, S., Arbeiter, A. B., Stupar, A., Mišan, A., Zugic, A., Nešić, I., Tadić, V., Bandelj, D., Pražnikar, Z. J., & Petelin, A. (2026). Valorization of post-distillation Helichrysum italicum biomass for functional beverage infusions. Food Chemistry: X, Article 104604. https://doi.org/10.1016/j.fochx.2026.104604

Image Credits: AI Generated

DOI: 10.1016/j.fochx.2026.104604

Keywords: Helichrysum italicum, immortelle, essential oil distillation residue, phenolic compounds, functional beverage, food waste valorization, circular bioeconomy, antioxidant activity, rosmarinic acid, chlorogenic acid, sensory evaluation, clinical trial

The research team, led by Ayşe Gül Filik and corresponding author Gökhan Filik of the university’s Agricultural Biotechnology Department, together with Olgay Kaan Tekin, set out to answer a question that has become increasingly urgent in modern animal science: can native plant resources rich in bioactive compounds serve as effective, natural alternatives to synthetic feed additives? Consumer pressure for so-called clean label animal products, combined with rising feed costs and concerns over chemical residues, has pushed researchers worldwide to screen local flora for phytogenic additives — plant-derived compounds that might improve animal health and product quality without pharmaceutical inputs. Viburnum opulus, which grows wild across Europe, North Africa, northern Asia, and particularly in the Turkish provinces of Kayseri, Sivas, Tokat, Kırşehir, and Ankara, presented an attractive candidate because of its unusually dense phytochemical profile.

That profile is what makes the fruit scientifically compelling. Analyses of Viburnum opulus berries have revealed high concentrations of polyphenols, flavonoids, tannins, carotenoids, and vitamin C. The dominant phenolic compound is chlorogenic acid, which can account for between 54 and 96 percent of the fruit’s total phenolic content and is regarded as a powerful antioxidant and antimicrobial agent. The fruit is also rich in organic acids, particularly malic and quinic acids, and contains notable concentrations of free amino acids such as aspartic and glutamic acid. Previous laboratory and pharmacological studies have linked Viburnum opulus extracts to lipid metabolism regulation, reductions in free fatty acids, cholesterol homeostasis effects, and even anticancer and antidiabetic activity, but almost none of this work had examined the plant as a direct dietary supplement in poultry. The Turkish team’s experiment was designed to fill that gap with a dose-response approach.

The trial involved 176 laying Japanese quails aged 30 weeks, a species favored in poultry nutrition research for its short generation interval and high egg production capacity. The birds — 132 females and 44 males — were randomly allocated into four dietary treatment groups, each with 11 replicate cages containing three females and one male. The cage, not the individual bird, served as the experimental unit for all performance and production measurements, a design choice that guards against pseudo-replication in statistical analysis. The control group received a standard basal diet formulated to meet quail nutritional requirements with 177 grams per kilogram of crude protein and 2800 kilocalories per kilogram of metabolizable energy. The three treatment groups received the same basal diet supplemented with Viburnum opulus fruit powder, abbreviated VOFP, at inclusion levels of 5, 10, and 15 grams per kilogram of feed. The fruit itself was harvested at full maturity from the university’s research fields, air-dried for a week, finished in a ventilated oven at 65 degrees Celsius for 48 hours, and ground through a one-millimeter sieve in an ultra-centrifuge mill to ensure homogeneous mixing. The feeding experiment ran for eight weeks under a 16:8 light-dark photoperiod with feed and water available ad libitum.

Over the 56-day trial, the researchers tracked body weight, body weight gain, feed intake, and feed conversion ratio weekly, collected eggs twice daily, and subjected eggs gathered on the third day of each week to a battery of quality analyses. External parameters included egg weight, dimensions measured with digital calipers, shape index, shell thickness at three points along the shell, and breaking resistance determined with a texture analyzer. Internal quality was assessed through albumen and yolk heights, albumen and yolk indices, and the Haugh unit, the internationally recognized standard for egg interior quality. Yolk color was quantified spectrophotometrically using CIELAB parameters. Critically, the team also monitored cholesterol metabolism by analyzing yolk cholesterol concentrations at the start of the trial and during weeks two, four, six, and eight, using a spectrophotometric extraction method adapted from earlier phytogenic additive research. Data were analyzed with general linear models, and orthogonal polynomial contrasts were applied to detect dose-response relationships across the supplementation levels.

The headline finding on performance was reassuring: dietary VOFP supplementation produced no statistically significant adverse effects on body weight, body weight gain, or feed conversion ratio, indicating that the birds tolerated even the highest inclusion level well. There were, however, subtle shifts in feeding behavior. Total feed intake was highest in the 5 gram per kilogram group at 7888.0 grams, while the 15 gram per kilogram group consumed the least at 7115.6 grams, a statistically significant difference. The authors suggest this reduction at the highest dose may reflect the fruit’s acidic profile — its abundant malic and quinic acids may slightly inhibit palatability — though the free amino acids in the fruit could simultaneously support protein metabolism. Egg production itself was statistically unchanged across groups, but the 5 gram per kilogram group achieved the highest total egg weight at 1828.60 grams and produced notably more afternoon-laid eggs than the control and highest-dose groups.

The most striking results emerged in egg internal quality. Supplementation at 5 grams per kilogram significantly increased egg yolk height, which reached 13.55 millimeters, and elevated the yolk index compared with controls, both indicators of a richer, more standing yolk. Albumen weight and albumen index were also highest at this dose. Among yolk color parameters, the b* value, which reflects yellowness, peaked at 19.74 in the 10 gram per kilogram group — an attribute with real market value, since consumers associate deeper yolk color with superior eggs. Shell characteristics told a more nuanced story: shell weight decreased with supplementation, reaching its lowest value of 1.22 grams in the 5 gram per kilogram group, yet shell thickness and breaking resistance were unaffected, suggesting that mineral metabolism and structural integrity were preserved even as shell mass shifted. The Haugh unit remained statistically unchanged across all treatments.

The cholesterol results are the most tantalizing and the most cautiously framed. Across the trial, average yolk cholesterol concentrations declined numerically in all supplemented groups, but statistical significance appeared only once: in the sixth week, when cholesterol in the 15 gram per kilogram group fell to 41.04 milligrams per gram of yolk, down from 52.31 milligrams per gram in the control group. The authors interpret this as a potential time-dependent effect of the fruit powder on cholesterol metabolism, plausibly connected to the phenolic compounds that earlier cell studies have linked to lipid accumulation and fatty acid metabolism pathways. Yet they are careful to note that the specific phytochemical composition of the powder used in this trial was not characterized, so the mechanism remains hypothetical rather than confirmed. The transient nature of the effect — significant in week six but not sustained across the full eight weeks — underscores how much remains unknown about how plant bioactives interact with avian physiology over time.

What the study ultimately delivers is preliminary but valuable evidence that Viburnum opulus fruit powder is a viable, well-tolerated phytogenic feed additive for laying quails, with the low dose improving selected internal egg quality traits and the high dose showing a potential, if inconsistent, cholesterol-modulating effect. The researchers call for follow-up work incorporating phytochemical characterization, hepatic cholesterol metabolism markers, antioxidant status measurements, and gut microbiota analyses to pin down the mechanisms at play. They also point to food science findings showing that encapsulation can stabilize the fruit’s bioactive compounds, hinting at strategies to enhance and prolong its effects in feed. For a fruit long celebrated in Anatolian folk tradition and now attracting attention in pharmacology laboratories worldwide, the journey from village juice press to scientific feed supplement is still at its beginning — but these quail eggs offer an early, measurable sign that the berry’s bioactive promise may extend from human health into the food chain itself.

Subject of Research: Effects of dietary Viburnum opulus fruit powder supplementation on performance, egg quality, and yolk cholesterol in laying Japanese quails

Article Title: The effects of Viburnum opulus L. fruit powder on the performance, egg traits, and egg cholesterol of laying Japanese quails

Article References: Filik, A. G., Filik, G., & Tekin, O. K. (2026). The effects of Viburnum opulus L. fruit powder on the performance, egg traits, and egg cholesterol of laying Japanese quails. Archives Animal Breeding, 69(3), 507-515. https://doi.org/10.5194/aab-69-507-2026

Image Credits: AI Generated

DOI: 10.5194/aab-69-507-2026

Keywords: Viburnum opulus, Japanese quail, phytogenic feed additive, egg quality, egg yolk cholesterol, poultry nutrition, chlorogenic acid, clean label, feed conversion ratio, Haugh unit, Anatolia, animal science

The central problem the team set out to solve is a familiar one in crop genetics. Conventional reference genomes are linear, meaning they represent a single consensus sequence against which all other varieties are compared. Such references capture only a limited slice of the diversity that exists within a species, and variants that fall outside the reference sequence can be missed entirely. Graph-based pangenomes address this limitation by encoding the genomes of many individuals simultaneously, allowing single-nucleotide polymorphisms, insertions and deletions, and large structural variants to be represented side by side. But the very comprehensiveness that makes large graphs powerful also makes them computationally demanding, and in some cases they emphasize structural changes at the expense of smaller variants that nonetheless matter for breeding.

MiniTG1.0 took a deliberately economical approach. Rather than aggregating dozens or hundreds of genomes, the researchers built the graph from just two carefully chosen parents: Solanum lycopersicum TS-RS1, a cultivated tomato, and Solanum pimpinellifolium TS-RS2, its wild relative. The two lineages differ substantially in fruit weight, sugar and acid composition, and disease-related chemistry, which makes the contrasts between them informative. The graph integrated 5.76 million variants spanning SNPs, INDELs, and structural variants across the population the team would go on to analyze, providing a dense but tractable map of the differences separating the cultivated and wild genomes.

To connect that variation to traits, the researchers generated 230 seventh-generation recombinant inbred lines, a population in which chromosomes from the two parents have been shuffled and fixed through repeated selfing. Each line carries a unique mosaic of the parental genomes, so any trait difference among the lines can be traced back to the chromosomal segments inherited from each parent. By combining genome-wide association studies across this population with transcriptomic data, metabolite profiling, and targeted functional experiments, the team created a pipeline that moves from statistical association to experimentally verified gene function within a single framework.

The performance gains over a linear reference were substantial. Joint analysis of the three variant classes explained 18.18 percent more average trait heritability than the linear reference genome, and it improved the mapping of established loci controlling trichomes and growth habit. The graph also placed the H and SELF-PRUNING genes closer to their expected trait signals than the linear reference did, indicating that the compact graph preserved, and in some respects sharpened, the resolution needed to locate genes underlying complex phenotypes. For a breeding-oriented tool, that combination of efficiency and precision is the key selling point.

Fruit weight provided the most striking demonstration. The analysis recovered the well-known fw2.2 region, a classic locus in tomato genetics, and additionally uncovered a previously unknown fruit-weight region the team designated fw6.4. Within this new locus, the gene SlILL6 emerged as a strong candidate. When the researchers overexpressed SlILL6, the resulting plants produced fruits weighing roughly 62 to 63 grams, compared with approximately 101 grams in wild-type plants, a reduction of nearly forty percent. The evidence supports a role for SlILL6 in limiting fruit growth through altered cell expansion, adding a new entry to the short list of genes known to govern tomato fruit size.

The study also extended its reach into fruit chemistry. Metabolite profiling detected 1,258 compounds across the population and identified 128 that accumulated at different levels in the two parental backgrounds. A metabolome-based genome-wide association study then connected variation in chlorogenic acid, an antioxidant phenolic compound, to the gene SlCGT. This connection proved functionally meaningful in the context of disease. Using CRISPR-Cas9 gene editing, the team created knockout lines that accumulated more chlorogenic acid and, when challenged with Botrytis cinerea, the fungus responsible for gray mold, developed smaller lesions. Overexpression lines showed the opposite pattern, accumulating less chlorogenic acid and suffering larger lesions. The researchers further showed that chlorogenic acid directly suppresses fungal growth, closing the loop between gene, metabolite, and phenotype.

The authors emphasized that the study demonstrates how a smaller, strategically designed genome graph can deliver both computational efficiency and biological precision. By pairing a wild parent with a cultivated parent and a segregating population, the framework allowed the team to follow inherited variation directly into fruit traits rather than searching through broad diversity panels alone. The discovery of SlILL6 and SlCGT, they noted, shows how the same pipeline can move from statistical association to experimentally tested function, creating a clearer route from pangenomic data to concrete breeding decisions involving fruit size, quality-related metabolites, and resistance to postharvest disease.

The practical implications for molecular design breeding are considerable. SlILL6 could be evaluated as a target for adjusting fruit size, giving breeders a lever on yield that complements the classical fw2.2 locus. SlCGT offers a route to raising chlorogenic acid levels and strengthening resistance to gray mold, and notably the researchers found this could be done without reducing soluble solids in the tested lines, addressing a common trade-off between defense chemistry and fruit quality. MiniTG1.0 itself supplies a compact analytical resource for tracking small and large variants together, which may help breeders recover useful alleles from wild germplasm more efficiently than current approaches allow.

The researchers are careful to note the boundaries of the work. The findings were generated from two parental genomes, greenhouse-grown recombinant lines, and targeted functional tests, so field trials across diverse environments and genetic backgrounds will be needed to confirm agronomic performance, disease protection, and any trade-offs before these targets enter commercial breeding programs. Even so, the study sketches a template that other crop teams may follow: instead of building ever-larger pangenomes, choose informative parents, build a compact graph, and let a purpose-designed population carry the variation into traits that breeders can actually use. If that template proves portable, the quiet efficiency of MiniTG1.0 may matter as much as the individual genes it revealed.

Subject of Research: Graph-based pangenomics of tomato fruit size, metabolite composition, and fungal disease resistance

Article Title: Tomato pangenome maps genes for fruit size and defense

Article References: Tomato pangenome maps genes for fruit size and defense. (n.d.). Original publication

Image Credits: AI Generated

DOI: Not provided

Keywords: tomato, pangenome, graph genome, fruit weight, SlILL6, SlCGT, chlorogenic acid, gray mold, Botrytis cinerea, recombinant inbred lines, CRISPR-Cas9, molecular breeding

The appeal of the approach lies in its elegant simplicity. Polyphenol oxidase, the copper-containing enzyme responsible for the browning that occurs when an apple is sliced, catalyzes the two-electron oxidation of o-diphenols such as chlorogenic acid into their corresponding o-quinones. These quinones are highly electrophilic and readily react with nucleophilic amino acid residues, including lysine, cysteine, and tryptophan, on proteins and peptides, forming covalent adducts. In other words, the very reaction that food processors spend fortunes trying to prevent becomes, under controlled conditions, a precise and environmentally friendly tool for building novel peptide-phenolic conjugates.

Coffee itself provides an exceptionally rich reservoir of phenolic substrates. Chlorogenic acids, esters of caffeic and quinic acids, can account for up to twelve percent of the dry weight of a coffee bean and are the principal contributors to the beverage’s antioxidant capacity. Using HPLC coupled with UV/Vis detection and mass spectrometry, the team profiled the phenolic composition of green Uganda coffee beans and identified eight major chlorogenic acid derivatives. Three caffeoylquinic acid isomers, 3-CQA, 4-CQA, and 5-CQA, were detected with a common molecular ion at m/z 353, while two feruloylquinic acid isomers appeared at m/z 367 and three dicaffeoylquinic acids at m/z 515. Among these, 5-caffeoylquinic acid emerged as the dominant phenolic constituent, confirming its central role in coffee chemistry and its suitability as the primary substrate for conjugation reactions.

The choice of enzyme source proved equally consequential. Five fresh apple varieties purchased from a German market, Jonagold, Boskoop, Golden Delicious, Cox Orange, and Braeburn, were screened for both polyphenol oxidase activity and chlorogenic acid content. The results revealed striking cultivar-dependent differences. Braeburn juice exhibited by far the highest enzyme activity, measured at 6262.4 units per liter using a 4-methylcatechol-proline adduct assay based on the Lambert-Beer law, with excellent linearity over the reaction period. Jonagold and Golden Delicious showed intermediate activities of roughly 3600 and 3400 units per liter, while Boskoop and Cox Orange lagged dramatically behind, with reductions of approximately 94 and 91 percent relative to Braeburn. Chlorogenic acid content also varied widely, with Boskoop containing the most at 325.3 micrograms per milliliter of juice and Cox Orange the least at 63.6 micrograms per milliliter.

Because the primary goal was to identify an efficient natural source of the enzyme, Braeburn was selected as the biocatalyst for the modification experiments, a choice the authors justified by noting that its chlorogenic acid content remained sufficiently high to support enzymatic oxidation while its catalytic potential far exceeded that of the other cultivars. The researchers also explored how storage conditions affect enzyme performance, and the findings carry practical significance for any future food application. Apples frozen at minus 20 degrees Celsius before juicing yielded juices with remarkably high polyphenol oxidase activity, reaching 15208.6 units per liter, compared with only 1780.7 units per liter for apples held at room temperature. The authors attribute this effect to ice crystal formation during freezing, which disrupts cellular membranes and releases membrane-bound enzyme into the juice matrix. Subsequent storage of the juices at minus 20 degrees Celsius for up to two weeks produced only minor losses in activity, indicating that frozen apple juice could serve as a stable, practical reservoir of the biocatalyst.

With the phenolic donor and the enzymatic catalyst characterized, the team turned to the conjugation reactions themselves. Ground green coffee was infused in boiling water to prepare an extract, which was then combined with the milk peptides BioZate 1 and glycomacropeptide under two different conditions: an alkaline pathway at pH 9, a common reference method for driving phenolic-protein conjugation, and an enzymatic pathway in which freshly pressed Braeburn apple juice supplied approximately 18.8 units of polyphenol oxidase per reaction at a natural pH of 4.7. Both reactions ran for 24 hours at room temperature with continuous stirring and exposure to air, after which the products were dialyzed, lyophilized, and stored for analysis.

Quantification of free chlorogenic acid derivatives by HPLC revealed substantial depletion of the phenolic pool over the reaction period. At the start, 5-CQA predominated at roughly 47 to 52 milligrams per gram of protein across all treatments, with the other isomers present at considerably lower levels. After 24 hours, total CQA content fell significantly in every sample. The alkaline-modified BioZate 1 conjugate showed the greatest decline, dropping from 67.8 to 26.1 milligrams per gram of protein, while the corresponding glycomacropeptide sample fell from 69.0 to 40.8. Dicaffeoylquinic acids were nearly eliminated in the alkaline BioZate 1 sample. The authors caution, however, that this reduction reflects the overall conversion of phenolic substrates, encompassing oxidation, isomerization, self-polymerization, and condensation, rather than a direct measure of covalent binding efficiency alone. Notably, alkaline conditions promoted isomerization of 5-CQA into its 3- and 4-CQA forms, a well-documented side reaction that the enzymatic route largely avoids.

Structural evidence came from MALDI-TOF mass spectrometry. The unmodified BioZate 1 peptide displayed characteristic peaks at m/z 7089.2, 7581.3, 12463.5, and 14173.7, while both modified forms exhibited new signals at approximately m/z 9300 and 18600, consistent with the incorporation of coffee-derived phenolics and, in the alkaline case, possible cross-linking or partially degraded species. Glycomacropeptide, which shows a sharp native peak at m/z 6802.6, gained new peaks at roughly m/z 9300 and 9500 after treatment under either condition, indicating the formation of higher-molecular-mass peptide-polyphenol complexes. The authors are careful to note that mass spectrometry alone cannot distinguish true covalent attachment from cross-linking or aggregation, and that definitive identification of binding sites will require high-resolution LC-MS/MS and peptide mapping in follow-up studies.

To estimate the extent of stable phenolic association, the researchers employed reverse-phase HPLC with 8 molar urea, which disrupts non-covalent interactions such as hydrogen bonding and hydrophobic association, leaving behind a fraction considered consistent with covalent modification. The alkaline-treated BioZate 1 conjugate retained the highest bound CQA level at 14.95 micrograms per milligram of protein, significantly exceeding its enzymatic counterpart, and the same pattern held for glycomacropeptide, where the alkaline sample reached 10.14 micrograms per milligram. The authors attribute the superior alkaline performance to the greater nucleophilicity of lysine and cysteine residues at high pH, which facilitates stronger interactions with oxidized phenolic intermediates.

Yet the enzymatic route retains compelling advantages despite its somewhat lower conjugation yield. Alkaline treatment promotes non-specific oxidation, chlorogenic acid isomerization, and potential damage to susceptible amino acid residues, whereas polyphenol oxidase-mediated oxidation proceeds under mild, food-relevant conditions and generates reactive quinones through a naturally occurring enzymatic mechanism. The use of apple-derived enzyme also aligns with clean-label processing strategies that consumers increasingly demand. The authors acknowledge a limitation: because a heat-inactivated apple juice control was not included, the specific contribution of the enzyme cannot be fully separated from other heat-sensitive components of crude juice, and future experiments with purified or inactivated enzyme preparations will be needed. Even so, the study establishes a credible foundation for a new class of peptide-phenolic conjugates built from coffee and milk, two of the world’s most familiar food matrices. Before these hybrid molecules reach functional food formulations, their antioxidant activity, digestibility, bioaccessibility, stability, and safety must be rigorously evaluated, work the researchers indicate is already underway in dedicated follow-up investigations.

Subject of Research: Enzymatic conjugation of coffee chlorogenic acids with milk bioactive peptides using apple-derived polyphenol oxidase

Article Title: Conjugation of Coffee Phenolics With Milk Bioactive Peptides: Characterization of Coffee and Apple Phenolics and Evaluation of Polyphenol Oxidase as a Natural Biocatalyst

Article References: Hamadneh, B. N., Mustafa, E. D., Alkafafy, M. E., Khojah, E., Samra, B. N., Wapet, D. E. M., Mahmoud, M. M., Badawy, W. Z., Khalil, M., Salama, M. A., Abdin, M., & Ali, M. (2026). Conjugation of Coffee Phenolics With Milk Bioactive Peptides: Characterization of Coffee and Apple Phenolics and Evaluation of Polyphenol Oxidase as a Natural Biocatalyst. Food Science & Nutrition, 14(10), Article e72435. https://doi.org/10.1002/fsn3.72435

Image Credits: AI Generated

DOI: 10.1002/fsn3.72435

Keywords: chlorogenic acid, polyphenol oxidase, milk peptides, coffee phenolics, BioZate 1, glycomacropeptide, apple juice, biocatalyst, MALDI-TOF mass spectrometry, HPLC-MS, functional foods, protein-phenolic conjugation

The clinical problem the researchers targeted is widespread. Non-steroidal anti-inflammatory drugs such as indomethacin relieve pain, fever, and inflammation, but they do so partly by inhibiting cyclooxygenase-1, an enzyme that helps produce the prostaglandins that keep the gastric mucosa healthy. Stripping away that protection leaves the stomach wall exposed to acid and pepsin, and indomethacin additionally ignites inflammatory signaling cascades that kill epithelial cells. Current countermeasures, chiefly proton pump inhibitors like omeprazole, suppress acid secretion by blocking the gastric H+/K+-ATPase, but long-term use of these drugs carries its own concerns, motivating the search for gentler, food-based alternatives.

Petasites japonicus offered an intriguing candidate. The plant has traditionally been used in East Asia for tension headaches and gastrointestinal spasms, and in Europe and North America for migraines, bronchial asthma, and seasonal allergic rhinitis. Its leaves and stems are rich in phenolic acids, including chlorogenic acid and fukinolic acid, along with several dicaffeoylquinic acid derivatives. To capture these compounds efficiently, the team reflux-extracted one kilogram of dried, ground plant material with either water or 50 percent aqueous ethanol at 50 degrees Celsius for five hours. High-performance liquid chromatography then quantified the phenolic content of each preparation.

The solvent choice mattered considerably. The 50 percent ethanol extract contained 42.48 milligrams of fukinolic acid per gram, compared with 25.35 milligrams in the water extract, while chlorogenic acid levels were nearly identical at roughly 13.5 milligrams per gram. Adding in the dicaffeoylquinic derivatives, the ethanol extract delivered a total polyphenol content of 80.21 milligrams per gram against 53.31 for water. On that chemical basis, the researchers selected the ethanol extract for all biological testing, reasoning that its richer phenolic payload would better represent the plant’s active potential.

In the laboratory, the extract proved strikingly protective. When human gastric adenocarcinoma AGS cells and Hs746T gastric carcinoma cells were exposed to 800 micromolar indomethacin for three hours, viability collapsed to 55.4 percent and 69.2 percent of control levels respectively. Pretreatment with the extract for one hour beforehand significantly blunted that damage, lifting viability to 71.9 percent in AGS cells at 50 micrograms per milliliter and 82.4 percent in Hs746T cells at 100 micrograms per milliliter. Crucially, the extract showed no cytotoxicity on its own across the entire 1 to 100 micrograms per milliliter range tested, and the researchers noted that dietary polyphenols naturally reach comparable concentrations in the gastric lumen before absorption.

Molecular analysis revealed three converging mechanisms. First, indomethacin sharply reduced MUC5AC, the predominant mucin glycoprotein that forms the stomach’s protective mucus barrier, dropping it to 0.65-fold in AGS cells and 0.48-fold in Hs746T cells; extract pretreatment restored those levels substantially. Second, the extract suppressed indomethacin-driven upregulation of acid secretion machinery, including the histamine H2 receptor gene HRH2 and subunits of the proton pump, ATP4A and ATP4B, which assemble into the H+/K+-ATPase that pumps acid into the stomach. Third, it dampened inflammation: indomethacin had boosted expression of inducible nitric oxide synthase and cyclooxygenase-2 and triggered phosphorylation of the transcription factor NF-κB and its inhibitor IκB-α, but the extract pulled all of these markers back down in a dose-dependent manner. Upstream, the extract also quieted the MAPK kinases ERK, JNK, and p38, whose indomethacin-induced phosphorylation reached as high as 8.24-fold in AGS cells before treatment brought it down.

The animal experiments translated these cellular findings into whole-body protection. Fifty male Sprague-Dawley rats were divided into control, indomethacin, omeprazole, and two extract-dose groups, with the test substances given orally once daily for seven days before a single 75 milligrams per kilogram dose of indomethacin. After quality-control exclusions, seven rats per group were analyzed. Macroscopic examination showed extensive hemorrhagic lesions in the indomethacin group, but the ulcer index fell to 0.58-fold at 50 milligrams per kilogram of extract and 0.40-fold at 100 milligrams per kilogram. Histological scoring told the same story: severe mucosal erosion, epithelial disruption, submucosal edema, and leukocyte infiltration in the indomethacin group scored 4.0, while extract treatment reduced the score to 1.3 and 0.7 at the two doses, with the lower dose matching omeprazole’s 1.3.

Tissue analyses confirmed the same molecular signature seen in culture. Indomethacin cut gastric MUC5AC protein to 0.58-fold of control, but extract restored it to 0.85-fold and 0.99-fold at the two doses. The drug raised Hrh2 mRNA to 3.28-fold and Atp4b to 1.16-fold; the higher extract dose brought these down to 1.88-fold and 0.85-fold respectively, the latter essentially matching control levels. Inflammatory chemistry followed suit: gastric nitric oxide rose from 3.52 to 5.86 micromolar under indomethacin and TNF-α from 1.39 to 1.98 picograms per milliliter, while extract treatment reduced both, and phosphorylated NF-κB, IκB-α, ERK, JNK, and p38 all declined dose-dependently in tissue lysates.

To probe how the plant’s signature compounds might act directly on drug targets, the team docked chlorogenic acid and fukinolic acid against the H+/K+-ATPase, the histamine H2 receptor, cyclooxygenase-2, and the TNF-α trimer, using omeprazole, cimetidine, and aspirin as reference ligands. The phenolic acids bound the proton pump through the residue Cys813 and the H2 receptor through Asp98, and their strongest predicted affinities came at cyclooxygenase-2, where chlorogenic acid scored −9.3 and fukinolic acid −9.0 kilocalories per mole, comfortably outperforming aspirin’s −6.7. Both compounds also showed moderate binding at the TNF-α trimer interface. The authors caution that docking is predictive and that the whole extract, not isolated compounds, produced the observed effects, with future work needed to isolate individual contributions.

The dose translation is perhaps the most striking practical detail. Applying US Food and Drug Administration body-surface-area conversion, the highest rat dose of 100 milligrams per kilogram corresponds to a human equivalent dose of 16.22 milligrams per kilogram, roughly 973 milligrams daily for a 60-kilogram adult. Based on the extraction yield, that amount equates to eating approximately 6.5 grams of fresh Petasites japonicus leaves and stems, an entirely ordinary dietary quantity. The authors acknowledge limitations, including the cancer-derived nature of the cell lines and the absence of direct measurements of gastric juice volume, pH, and acidity, and they call for follow-up studies using primary cells, gastric organoids, or pylorus-ligated models. Even so, the convergence of preserved mucus barriers, dampened acid-pump genes, and suppressed NF-κB and MAPK signaling across cells, rats, and simulations makes a compelling case that this everyday vegetable deserves attention as a functional food ingredient for protecting the stomach against painkiller injury.

Subject of Research: Gastroprotective effects of Petasites japonicus ethanol extract against NSAID-induced gastric ulcer

Article Title: Gastroprotective effects of a dietary Petasites japonicus ethanol extract against indomethacin-induced gastric injury via regulation of mucosal defense, acid secretion-related targets, and inflammation

Article References: Hwang, B., Hwang, S.-K., Kim, J.-Y., Song, J.-H., Son, Y.-J., Kim, H., Jung, W., Lee, J. J., Cho, S.-C., Chung, H., & Moon, S.-K. (2026). Gastroprotective effects of a dietary Petasites japonicus ethanol extract against indomethacin-induced gastric injury via regulation of mucosal defense, acid secretion-related targets, and inflammation. Journal of Agriculture and Food Research, 31, Article 103325. https://doi.org/10.1016/j.jafr.2026.103325

Image Credits: AI Generated

DOI: 10.1016/j.jafr.2026.103325

Keywords: Petasites japonicus, gastric ulcer, indomethacin, NSAIDs, MUC5AC, NF-κB, MAPK, proton pump, chlorogenic acid, fukinolic acid, functional food, gastroprotection

The research team, led by Charlène Sirvins, Pascale Goupy and Claire Dufour of INRAE in France, focused on four abundant plant phenolic compounds: chlorogenic acid, caffeic acid, ferulic acid and hydroxytyrosol, the principal phenolic aglycone of olives. These molecules are ubiquitous in the human diet, found in coffee, fruits, cereals, olive oil and red wine, and previous work had shown that plant extracts rich in such compounds can reduce volatile N-nitrosamines in dry-cured bacon and sausages. What remained unclear was precisely how each phenolic structure intercepts the nitrosation reaction, and whether the standard laboratory test used to measure this protective capacity was telling the whole truth.

To find out, the team built a simplified cured meat model built around N-acetyltryptophan, a tryptophan derivative chosen because it carries a useful light-absorbing chromophore and retains the amide bond found in covalently linked amino acids within real proteins. The model contained nitrite at 120 parts per million, matching the newly reduced industrial maximum, plus 30 micromolar of non-heme iron, a pro-oxidant naturally present in meat. The researchers then tracked the formation of N-acetyl-N-nitrosotryptophan, a model secondary N-nitrosamine, over time at two pH values: pH 5, simulating meat processing and the early stage of gastric digestion, and pH 2.5, representing the harshly acidic final stage of digestion in the stomach. Using ultra-high-performance liquid chromatography coupled to diode-array detection and ion-trap mass spectrometry, they followed not only the target nitrosamine but dozens of newly formed reaction products.

The baseline behavior of the model system was itself revealing. At pH 2.5, nitrosation of the tryptophan derivative was rapid, reaching a maximum conversion of 17.2 percent within 45 minutes, but the resulting nitrosamine proved unstable and reverted quantitatively back to its parent amine within hours through acid-catalyzed denitrosation. At pH 5, the reaction was slower and the product more persistent, with a maximum conversion of 3.6 percent. This matters because nitrosated amino acids and peptides formed in the acidic stomach could pass into the near-neutral environment of the duodenum, around pH 6.5, where they would be stabilized before intestinal absorption. In other words, the stomach is not merely a hostile environment for nitrosamines; it is a potential factory for them, and whatever survives the journey onward is the more dangerous cargo.

When the phenolic compounds entered the picture, the differences between them were striking. Caffeic acid and ferulic acid, the two unesterified hydroxycinnamic acids, were the champions, suppressing nitrosation by 85 percent within 15 minutes at pH 2.5 and achieving complete inhibition after 105 minutes. At pH 5, caffeic acid still held inhibition between 66 and 77 percent over the reaction period. Chlorogenic acid, which is simply caffeic acid esterified with quinic acid, performed markedly worse, managing only 24 percent inhibition at pH 2.5 and 16 to 28 percent at pH 5. Hydroxytyrosol fell in between. The lesson was clear and chemically elegant: the same reactive core can behave very differently depending on whether a bulky ester group blocks access to it.

The mass spectrometric detective work revealed two fundamentally different protection strategies at play. For chlorogenic acid and hydroxytyrosol, both bearing a catechol group, the story begins with nitrite-induced oxidation. Nitrite strips electrons from the catechol ring, generating a reactive quinone intermediate. That quinone can then dimerize with another phenol molecule, undergo nitration to form nitrochlorogenic acid or nitrohydroxytyrosol, or be attacked by nucleophiles. Each of these pathways consumes multiple nitrite ions; nitration of a single catechol unit alone requires three. Hydroxytyrosol proved especially voracious, scavenging at least three nitrite ions per molecule at both pH values, with nitrohydroxytyrosol emerging as its dominant product and a rich menagerie of dimers, trimers and covalent adducts with the tryptophan derivative accumulating alongside.

Caffeic and ferulic acid took an entirely different route. Instead of being oxidized, their propenoic acid side chains were directly attacked by the nitrosonium ion, the very electrophile that would otherwise nitrosate the amine. This creates a resonance-stabilized nitroso cation on the phenolic molecule, and here comes the surprise: the tryptophan derivative, present in threefold excess, turned out to be a stronger nucleophile than water or nitrite, and it covalently latched onto that cation. The result was a family of acetaldehyde oxime adducts, some still bearing the nitrosated amine, others stripped of it. In effect, caffeic and ferulic acid were acting as decoys, diverting the nitrosating machinery and then capturing the amine target itself before it could be converted into a free, mobile nitrosamine.

That capture, however, exposed a hidden flaw in the standard assay. Because the covalently bound tryptophan derivative escaped quantification, the apparent disappearance of the free amine inflated the measured antinitrosating capacity of caffeic and ferulic acid. When the researchers recalculated protection based on the number of nitrite ions actually scavenged by the five major end products of each reaction, the ranking shifted. Hydroxytyrosol emerged as superior to chlorogenic acid at pH 5, while the two catechol compounds matched each other at pH 2.5, and the unesterified hydroxycinnamic acids, despite their dazzling performance in the simple test, actually quench less nitrite per molecule. The team showed that the same bias applies to flavonoids such as rutin, quercetin and epicatechin studied previously, meaning the corrected framework offers a more honest yardstick for screening plant extracts intended for cured meat reformulation.

The practical implications reach beyond the laboratory beaker. Phenolic-rich extracts from olive, grape and rosemary have already been shown to lower volatile N-nitrosamines in sausages and bacon, and olive leaf extract rich in oleuropein, an acylated hydroxytyrosol derivative, has been tested in ripened industrial sausages as a partial nitrite replacement. The new mechanistic map explains why such extracts work and which structural features matter most: catechol groups that surrender electrons to nitrite, and free propenoic chains that intercept the nitrosonium ion directly. It also raises a nutritional angle worth savoring. Eating plant foods rich in these phenols alongside cured meats, or indeed alongside the dietary nitrate from leafy vegetables that fuels endogenous nitrite production in the stomach, could dampen the nitrosation of amino acids during digestion itself. There remain open questions, notably the toxicological significance of the structurally modified peptides released when phenols bind covalently to food proteins, but the study firmly establishes that the chemistry of protection is as intricate as the chemistry of harm, and that understanding both is the surest path to a safer plate.

Subject of Research: Inhibition of N-nitrosamine formation by plant phenolic compounds in cured meat and gastric digestion models

Article Title: The antinitrosating capacity of hydroxycinnamic acids and hydroxytyrosol in a cured meat model unraveled through structural and kinetic investigations

Article References: Sirvins, C., Goupy, P., & Dufour, C. (2026). The antinitrosating capacity of hydroxycinnamic acids and hydroxytyrosol in a cured meat model unraveled through structural and kinetic investigations. Current Research in Food Science, 13, Article 101581. https://doi.org/10.1016/j.crfs.2026.101581

Image Credits: AI Generated

DOI: 10.1016/j.crfs.2026.101581

Keywords: N-nitrosamines, nitrite, cured meat, polyphenols, hydroxytyrosol, caffeic acid, ferulic acid, chlorogenic acid, food chemistry, nitrosation, gastric digestion, food safety

The challenge is real and surprisingly difficult to detect. While green coffee beans can often be told apart by eye, roasting and grinding erase the morphological features that distinguish the two species. Once beans are roasted, authentication requires chemistry. Traditional approaches rely on trained cuppers whose sensory evaluations are subjective and generally fail to detect Robusta adulteration until it exceeds roughly 35 percent of the blend. Laboratory methods such as high-performance liquid chromatography offer objectivity but demand expensive instruments, skilled operators, and long analysis times, with per-sample costs around 100 US dollars. Nuclear magnetic resonance spectroscopy can quantify marker compounds like 16-O-methylcafestol, a Robusta-specific molecule, but remains too slow and costly for routine screening.

The Taiwanese team, led by Bing-Chen Gu and Chia-Che Wu, took a different route. Their platform uses disposable screen-printed sensor chips with a three-electrode configuration: carbon electrodes to measure caffeine, chlorogenic acids, and trigonelline; a gold electrode for sugar-related measurements; and a silver/silver chloride reference electrode. After a small aliquot of brewed coffee is mixed with species-specific electrolytes, the device applies voltammetric techniques that exploit well-characterized reaction pathways. Caffeine and chlorogenic acids are oxidized at the electrode surface in the low-potential region around 200 to 300 millivolts. Trigonelline is first converted under alkaline conditions to electroactive nicotinic acid, which then produces a prominent oxidation peak near 1500 millivolts. Reducing sugars are detected through copper redox chemistry, in which the sugars reduce copper ions to cuprous oxide under alkaline conditions, generating a characteristic signal in the negative potential region near minus 250 millivolts.

The entire electrochemical measurement takes less than three minutes per sample once the coffee extract is prepared, using only microliter-scale volumes. The team validated the platform rigorously, establishing linear calibration over working ranges of 20 to 500 milligrams per deciliter for caffeine and chlorogenic acids, 10 to 100 for trigonelline, and 65 to 1250 for reducing sugars. Limits of detection ranged from 1.26 milligrams per deciliter for trigonelline to 19.27 for reducing sugars. Spike-recovery experiments in real coffee matrices returned recoveries between 97.2 and 110.3 percent for trigonelline and 92.9 to 113.9 percent for sugars, comfortably within the accepted 80 to 120 percent window. Testing across 15 independently manufactured sensor chips from three production batches yielded relative standard deviations between 6.23 and 9.17 percent, demonstrating that the disposable chips perform consistently despite batch-to-batch variation. Matrix effects, the distortion of signals by other compounds in the coffee, were minimal, with slope ratios remaining close to 100 percent.

Crucially, the electrochemical readings captured the expected chemistry of coffee roasting. Across representative samples, chlorogenic acids, trigonelline, and reducing sugars all declined progressively as roast level deepened, reflecting thermal degradation, Maillard reactions, and caramelization. Caffeine, by contrast, showed only weak dependence on roast level, and Robusta consistently maintained higher caffeine concentrations than Arabica, matching decades of chromatographic evidence. These roast-dependent trends confirmed that the portable sensor was generating chemically meaningful data rather than artifacts, providing a credible foundation for the machine learning stage.

For the classification task, the researchers assembled a dataset of 138 samples: 66 Arabica coffees spanning 11 varieties from Ethiopia, Kenya, and Colombia; 18 Robusta samples from Vietnam, India, and Brazil; and 54 experimentally prepared blends at ratios ranging from 95:5 to 30:70 Arabica to Robusta. All beans were roasted to controlled light, medium, and dark levels verified by Agtron measurements, and blends were formulated from beans roasted to matching levels to prevent roast differences from confounding the analysis. Five variables, caffeine, chlorogenic acids, trigonelline, reducing sugars, and roast level, were fed into two machine learning classifiers. The random forest model achieved 97.37 percent test accuracy, while XGBoost classified every test sample correctly, achieving 100 percent accuracy with perfect precision, recall, and F1-scores for all three classes. Feature importance analysis revealed caffeine as the single most influential variable, followed by chlorogenic acids and roast level, though caffeine alone proved insufficient, misclassifying blends as pure Arabica when used without the other descriptors.

The team also stress-tested their models with stricter validation designs. In a leave-one-origin-out analysis, where all samples from one coffee origin were withheld from training, the binary Arabica-versus-Robusta discrimination retained perfect accuracy, balanced accuracy, and macro-F1 of 1.000, suggesting the models generalize to origins never seen during training. External validation using compositional data extracted from previously published studies, comprising 17 Arabica and 3 Robusta samples, also produced 100 percent correct classification by the trained XGBoost model.

Predicting blend ratios quantitatively proved harder than simple classification, and the researchers were candid about the pitfalls. A neural network regression model trained on the same five features achieved a coefficient of determination of 0.980 on a randomly split test set, with root mean square error of 0.050 and mean absolute error of 0.032. But because random splitting can scatter samples from the same Arabica-Robusta pairing across both training and test sets, the team implemented a far more demanding leave-one-blend-pair-out validation, in which all 18 samples from one complete blend pairing were withheld from every stage of model development. Under this nested group-aware framework, which selected an ensemble of four neural network architectures from a pool of 48 candidates per fold, pooled performance remained strong: R-squared of 0.9121, root mean square error of 0.0676, and 47 of 54 held-out predictions, or 87.04 percent, falling within plus or minus 0.10 of the true blend fraction. Pair-specific R-squared values ranged from 0.8863 to 0.9353, with the largest errors concentrated at low blend ratios for one particular pairing. The drop from 0.980 to 0.9121 confirmed that random splitting had provided an optimistic estimate, while the retained performance demonstrated genuine transferability to unseen blend combinations.

The interpretability analysis added further confidence. SHAP analysis indicated that caffeine and trigonelline were the most influential variables driving the neural network’s blend-ratio predictions, with chlorogenic acids, sugars, and roast level contributing smaller but systematic effects. The model, in other words, relied on the full multivariate chemical fingerprint rather than gaming any single feature. Benchmark comparisons underscored the value of the neural network approach: multiple linear regression managed only an R-squared of 0.146, showing that simple linear combinations cannot capture the nonlinear relationships among composition, roasting, and blend ratio, while random forest regression reached 0.951 but remained limited by its piecewise, tree-based structure.

The implications extend well beyond coffee fraud. The platform requires only a small extract aliquot, completes measurement within three minutes, and operates on portable, potentially low-cost hardware, making it suitable for real-time quality control at roasteries, import checkpoints, and retail settings. The authors acknowledge limitations: the blended samples used components with similar roast levels, so the current model is validated primarily for matched-roast blends, and broader sampling across origins, processing methods, and harvest seasons will be needed before wider deployment. Independent external validation of blend-ratio regression also remains outstanding because no published dataset contained all five required variables. Still, the study demonstrates a compelling template for intelligent food authentication, one that could readily extend to other beverages and foodstuffs by expanding the target analytes and sample diversity. As adulteration schemes grow more sophisticated, the marriage of cheap disposable sensors and machine learning may prove one of the most practical defenses the food industry has.

Subject of Research: Portable electrochemical sensing with machine learning for coffee species authentication and blend ratio prediction

Article Title: Portable electrochemical sensing coupled with machine learning for rapid Arabica and Robusta coffee authentication and blend ratio prediction

Article References: Gu, B.-C., Chung, K.-J., Hong, Z.-W., Yang, N.-H., & Wu, C.-C. (2026). Portable electrochemical sensing coupled with machine learning for rapid Arabica and Robusta coffee authentication and blend ratio prediction. Food Chemistry: X, 39, Article 104523. https://doi.org/10.1016/j.fochx.2026.104523

Image Credits: AI Generated

DOI: 10.1016/j.fochx.2026.104523

Keywords: coffee authentication, Arabica, Robusta, electrochemical sensing, machine learning, food adulteration, XGBoost, artificial neural network, caffeine, chlorogenic acid, blend ratio prediction, portable biosensor