Food Chemistry

Cloudberry (Rubus chamaemorus L.)-derived nanovesicles (NVs) represent a promising but still poorly characterized class of plant-derived vesicles with potential relevance for skin-related applications. Here, we isolated cloudberry fruit-derived NVs and investigated their physicochemical and molecular properties, cellular uptake, cytocompatibility, and functional effects in human dermal fibroblasts (HDF) and HaCaT keratinocytes. Nanoparticle tracking analysis and transmission electron microscopy confirmed a nanosized vesicle preparation with characteristic round morphology, while protein quantification supported reproducible isolation of NV-associated material. In vitro, cloudberry NVs showed concentration-dependent effects on cell viability and proliferation, with lower doses being better tolerated. Labelled NVs were internalized by both HDF and HaCaT cells in a time-dependent manner. Under oxidative stress conditions, cloudberry NVs reduced H2O2-induced senescence-associated {beta}-galactosidase staining in HDFs and exerted cytoprotective effects in both cell lines, alongside measurable cell-free antioxidant activity in the DPPH assay. In scratch wound-healing assays, cloudberry NVs modulated wound closure in a dose-dependent manner, with the lowest tested concentration showing the most favorable response. UHPLC-MS/MS-based proteomics and metabolomics further indicated the presence of diverse secondary metabolites and stress-related protein cargo. Together, these results support the view that cloudberry-derived NVs are biologically active plant nanovesicles with potential utility in skin-related regenerative applications. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=126 SRC="FIGDIR/small/741293v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@c9094dorg.highwire.dtl.DTLVardef@81a1b5org.highwire.dtl.DTLVardef@9fafe2org.highwire.dtl.DTLVardef@1d421d3_HPS_FORMAT_FIGEXP M_FIG C_FIG

Cassava and cocoyam are major dietary staples in sub-Saharan Africa, commonly processed by natural fermentation before consumption. Although fermentation reduces antinutritional compounds and improves food quality, its molecular effects remain poorly characterized. We used untargeted mass spectrometry-based metabolomics with a computational annotation pipeline to compare fermentation-induced molecular remodeling in the two tubers, which showed distinct responses. In cassava, 718 of 773 significant features (92.9%) were depleted, indicating a predominantly catabolic process. In cocoyam, the response was more balanced, with 385 of 1,013 features (38.0%) enriched, including di- and tripeptides consistent with proteolytic processing. Class analysis, molecular networking, and pathway enrichment revealed tuber-specific signatures: cassava was dominated by purine metabolism, whereas cocoyam showed stronger enrichment of amino acid pathways. Cyanogenic glycoside-related features were depleted, consistent with detoxification. Biotransformation prediction also suggested putative fermentation products absent from current databases, highlighting the under-characterized chemistry of these tubers.

LC/MS-based food lipidomics provides detailed information on intact lipid species, but the resulting datasets are often difficult to translate into concepts directly useful for food quality, processing, nutritional profiling and authenticity assessment. Here, we present Lipid Food Profile (LFP), a module of the LipidOne platform designed to convert annotated LC/MS lipidomics data into interpretable food-relevant lipid indices. LFP applies an in silico hydrolysis strategy to reconstruct acyl, alkyl and alkenyl chains from intact lipid species while preserving their lipid-class origin. The reconstructed information is then summarized into index categories related to food lipid quality, compositional balance, omega balance, oxidative stability, chain remodelling and ether-linked chain contribution. The interpretative value of LFP was evaluated using three published food lipidomics datasets addressing different analytical questions: X-ray-induced lipid remodelling in Chlorella vulgaris, spatial lipid heterogeneity in Mugil cephalus bottarga, and geographical-origin assessment of camel milk. Across these case studies, LFP recovered the main conclusions of the original lipidomics investigations, including treatment-associated lipid remodelling, inner-outer layer differences in bottarga and regional variation in camel milk. Importantly, LFP reorganized these findings into a smaller number of food-oriented indices, providing additional information on saturation balance, oxidative susceptibility, chain architecture and classification potential. Overall, LFP provides an interpretative layer for LC/MS food lipidomics that complement conventional fatty-acid analysis and molecular-species-based interpretation. By translating complex lipidomic tables into structured lipid index profiles, the module may support more accessible and chemically meaningful analysis of food composition, processing effects, lipid quality and exploratory traceability applications. LFP is freely accessible through the LipidOne web platform (LipidOne.eu). HighlightsO_LILipid Food Profile translates LC/MS food lipidomics into interpretable lipid indices. C_LIO_LIThe workflow preserves chain and lipid-class information without chemical hydrolysis. C_LIO_LIPublished case studies show that LFP recovers and extends previous interpretations. C_LIO_LILFP supports food quality, processing and exploratory origin/authenticity assessment. C_LIO_LIThe module complements conventional fatty-acid analysis and molecular lipidomics. C_LI

Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), the most abundant protein on Earth, is an attractive and sustainable food ingredient owing to its favourable nutritional and techno-functional properties. Leafy vegetables are particularly rich sources of RuBisCO; however, large-scale vegetable production generates substantial quantities of residual biomass throughout agri-food supply chains. Drying is widely used to stabilise this biomass and facilitate storage, transport, and handling, yet most reported RuBisCO extraction methods have been developed for fresh material and are poorly suited to dried feedstocks. Here, we present a simple, scalable, and cost-effective process for the recovery of food-grade RuBisCO from dried leafy biomass. Using spinach, rocket, and kale as model systems, efficient protein extraction was achieved from both freshly dried leaves and commercially available leaf powders without the need for resource-intensive processing. Application of the method to spinach yielded approximately 75 mg of high-purity RuBisCO per 100 g fresh-leaf equivalent, corresponding to an extraction efficiency of [~]70%, which increased to [~]90% following supplementation with 20 mM CaCl2. The recovered protein fraction also exhibited favourable foaming capacity and foam stability, demonstrating its potential as a functional food ingredient. This work provides a practical route for the valorisation of dried vegetable residues and supports the development of circular, waste-to-value supply chains for sustainable plant protein production.

Marine macroalgae are valuable sources of bioactive compounds. In this study, we thus investigated the chemical composition and biological activity of an extract from the green seaweed Ulva lacinulata, composed of small bioactive compounds. Comprehensive compositional analyses and high-resolution mass spectrometry revealed its diverse molecular profile composed in particular of peptides/amino acid derivatives, saccharides, low-chain fatty diacids, oxylipins and minerals. Its anti-inflammatory activity was assessed after 6 h pre-treatment in LPS-stimulated cultured RAW 264.7 macrophages, showing that it significantly and dose-dependently reduced the expression and/or secretion of pro-inflammatory cytokines such as TNF-alpha; and IL-6, and targeted the NF-kB signaling cascade. It modulated the SIRT1-AMPK signaling axis and increased the LC3-II/LC3-I ratio, supporting the activation of a controlled autophagic response. This work highlighted the potential of this marine-derived extract as a safe and effective functional ingredient for the development of functional food and/or dietary supplements targeting chronic low-grade inflammation.

Kakadu plum (Terminalia ferdinandiana), an Australian native fruit, is among the richest known dietary sources of vitamin C and hydrolysable tannins, yet its capacity to protect the intestinal epithelium against oxidative stress remains largely unexplored. This study optimised the extraction of bioactive compounds from freeze-dried Kakadu plum powder and evaluated their antioxidant activity using both chemical and cellular antioxidant in vitro assay. Phenolic compounds were extracted using three solvents (water, 80% ethanol, and 80% methanol) combined with shaking, ultrasound, or microwave assistance. Solvent, rather than processing technique, was the dominant determinant of antioxidant capacity: ethanol and methanol maximised total phenolic content, total flavonoid content, and DPPH radical-scavenging activity, whereas water extracts showed the highest ferric-reducing antioxidant power. Twenty-four phenolic compounds identified by HPLC-ESI-QTOF-MS/MS were mapped by network pharmacology to nine core oxidative-stress targets, and cross-species molecular docking predicted conserved binding of key phenolics to canine orthologs of PTGS2 and MMP2. In an H2O2-induced oxidative-stress in vitro cell model using canine small intestinal epithelial cells, both water (less than 25 ug/mL) and ethanol (less than 250 ug/mL) extracts significantly suppressed intracellular reactive oxygen species (ROS) in a dose-dependent manner, with the ethanol extract effective across a wider concentration range. This work demonstrated that Kakadu plum extract could be a promising natural, multi-target antioxidant ingredient for canine intestinal health, and provided a reference for future in vivo research.

Nisin is a promising antimicrobial peptide widely used in food preservation due to its efficacy against Gram-positive spoilage and pathogenic bacteria. Although Nisin is increasingly applied in the food sector, the biopeptide suffers from instability within food matrixes and can rapidly lose its antimicrobial potential following interaction with food biomolecules. Thus, it is necessary to investigate approaches that can be employed to extend the stability and activity of Nisin. Hence, the aim of this study was to develop and characterise a chitosan-alginate polyelectrolyte microencapsulation system capable of enhancing Nisin stability while retaining antimicrobial activity. The microencapsulation of Nisin was achieved by pre-gelation of alginate using calcium chloride and subsequent direct electrostatic interaction between cationic Nisin and chitosan with pre-gelled anionic alginate at pH 5.0. Following microcapsule formation, physicochemical and structural characterisation was performed using Zeta potential determination and measurement of the polydispersity index (PDI) via dynamic light scattering. SEM micrographs were used to confirm morphology, while Fourier-transform infrared (FTIR) spectroscopy and high-performance liquid chromatography (HPLC) were utilised to assess chemical integrity and functional group preservation of encapsulated Nisin. Following this, stable microcapsules with diameters ranging from 150-200 nm and smooth surface morphology were obtained. Microcapsule formation was strongly influenced by formulation parameters, particularly pH, calcium ion concentration, and chitosan content, with deviations from optimal acidic conditions (< pH 5.0) resulting in aggregation, increased polydispersity, and reduced encapsulation efficiency. The microcapsules were monodispersed (PDI {approx} 0.30) and electrostatically stable, exhibiting a Zeta potential of approximately +36 mV. These microcapsules remained stable over a prolonged storage period of 21 days under refrigerated conditions while retaining antimicrobial activity against Bacillus cereus. Encapsulation efficiency reached approximately 65%, confirming effective retention of Nisin within the polymer matrix. Overall, the findings demonstrate that chitosan-alginate ionic gelation is a non-denaturing and effective encapsulation strategy for extending the functional stability of Nisin. These microcapsules show strong potential as natural antimicrobial delivery systems for food and beverage applications, particularly in acidic food matrices, with implications for improved food safety and shelf-life extension.

Traditional Mongolian fermented foods have been extensively utilized for dietary regulation and the promotion of gastrointestinal health. However, spontaneous fermentation remains inherently unpredictable, leading to significant variations in microbial community dynamics, metabolite accumulation, and the consistency and quality of the final product. Drawing on the traditional preparation of Mongolian acidic foods, this study established a controlled production strategy for whole-wheat probiotic fermented soup (WWPFS) by combining enzymatic pretreatment with probiotic-directed fermentation. Physicochemical characterization, 16S rRNA gene-based microbial community profiling, LC-MS/MS-based untargeted metabolomics, safety evaluation, and an Escherichia coli-induced gut microbiota dysbiosis model were employed to optimize and comprehensively characterize the fermentation process of WWPFS. The optimized process established a reproducible fermentation system consistently dominated by Lactobacillus and Bacillus across independent fermentation batches. Compared with traditional spontaneous fermentation, probiotic-directed fermentation remodeled the physicochemical properties of the whole-wheat matrix, including carbon, nitrogen, phosphorus, sulfur, and mineral composition, and facilitated the accumulation of putatively annotated LC-MS/MS features, including DL-lactate, 1,4-D-xylobiose, diacetyl, and phenyllactic-acid-related features derivatives. Acute oral and 28-day repeated-dose toxicity evaluations showed no treatment-related adverse effects within the tested dose range and study duration. In the Escherichia coli-induced gut microbiota dysbiosis mouse model, microbial richness, diversity, and community structure differed among the experimental groups, and both low- and high-dose WWPFS groups showed significant shifts in overall gut microbial community composition relative to the model group after multiple-testing correction, together with directional recovery of selected model-responsive bacterial genera. Cross-system integration identified coordinated response patterns between fermentation-derived metabolite features and model-responsive gut bacterial taxa, supporting a potential metabolite-microbiota link in WWPFS-mediated gut microbiota modulation. In summary, probiotic-directed fermentation improved the controllability of the traditional Mongolian fermented food production process, reshaped its metabolic profile, and enhanced its potential to modulate the gut microbiota. These findings provide experimental evidence supporting the modernization of traditional Mongolian fermented foods and the development of probiotic-based functional foods.

This study aims to investigate changes in protein secondary structures (-helix, -sheet, random coils, and -turn) and macronutrient distribution in different cool-season oat varieties and steam-pressure toasting durations using synchrotron-based mid-infrared (Mid-IR) spectroscopy and Fourier Transform Infrared spectroscopy (FTIR) imaging. All oat samples, provided by the Crop Development Center at the University of Saskatchewan, were harvested over three consecutive years (2018, 2019, and 2020). The first experiment compared four oat varieties (CDC Arborg, CDC Nasser, CDC Haymaker, and Summit), while the second examined CDC Nasser oats subjected to steam-pressure toasting (SPT) at 121 for 0, 30, 60, 90, and 120 minutes. FTIR chemical imaging revealed that carbohydrates, proteins and lipids in the four oat varieties were mainly concentrated in the endosperm, aleurone layer and embryo, crease region, and remained unchanged after SPT. Peak-fitting deconvolution of the Amide I band (1700-1600 cm-1) and subsequent quantitative analysis revealed that the four oat varieties exhibited broadly similar protein secondary structure profiles, with statistically significant but subtle variety effects detected for -helix (P = 0.026), -turn (P = 0.047), and the -helix to -sheet ratio (P = 0.048), however, -sheet and random coil proportions did not differ significantly among varieties. In contrast, SPT induced pronounced structural rearrangements, with significant increase in -sheet proportion (P = 0.003) and significant decreases in random coil content (P = 0.026). Notably, 30 minutes of toasting was sufficient to significantly increase -sheet and decrease the random coil contents. These changes are consistent with heat-induced protein denaturation and intermolecular -sheet aggregation, where thermal energy breaks the hydrogen bonds that stabilize the disordered random coil conformation, causing the unfolded polypeptide chains to reassemble into highly ordered -sheet aggregates. After SPT, the peak centers of Amide I and II bands shifted to lower wavenumbers and both bands broadened while their intensities were maintained, reflecting the reorganization of the remaining protein into -sheet aggregates rather than any loss of amide-active protein. These findings suggest that, although genotype has a relatively minor effect on the protein secondary structure of oats, hydrothermal treatments fundamentally reorganize the protein matrix from a disordered to an ordered conformation, which may have implications for protein digestibility, solubility, and nutritional function.

Raffinose family oligosaccharides (RFOs) are abundant in side streams from food and feed production from legumes, and the transition to plant-based diets increases the volume of such side streams. RFOs in the diet tend to have negative impacts on the consumers gut (e.g., nausea, bloating, diarrhoea), and in many ways, RFOs are comparable to lactose as a side stream from the dairy industry and symptoms associated with lactose intolerance. On the contrary, galactooligosaccharides (GOS) are recognized as prebiotics, and in this study we used a {beta}-galactosidase from Niallia circulans to produce potential prebiotics from RFOs (acceptors) and lactose (donor), which we hypothesized to have a lower fermentability than unmodified RFOs. The transglycosylation reactions resulted in RFO-based -{beta}-GOS, with NMR characterization showing ({beta}1-4) galactosylations on the non-reducing galactose end of RFOs as the major product. In reactions with RFOs, the characteristics were comparable to reactions with lactose alone and the new -{beta}-GOS products made up the largest fraction (by weight). A screening of 11 relevant gut and food microbe strains revealed that the gut commensal Bacteroides ovatus metabolised these modified oligosaccharides for growth whereas other strains grew only after adaption and others did not use them at all. This implies that mixed-linkage -{beta}-GOS are less fermentable by some microbes compared to raffinose, while other (beneficial) bacteria can still ferment them. The enzymatic synthesis established here is an interesting approach to upgrade abundant food side streams towards new prebiotics in a world where functional foods and food waste reduction receive increasing attention. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/731070v1_ufig1.gif" ALT="Figure 1000"> View larger version (22K): org.highwire.dtl.DTLVardef@18e0e62org.highwire.dtl.DTLVardef@1525b4borg.highwire.dtl.DTLVardef@1e7be88org.highwire.dtl.DTLVardef@18df278_HPS_FORMAT_FIGEXP M_FIG C_FIG

Background: Natural compounds from avocado fruit (avocadene, avocadyne, and acetate derivatives) exhibit notable biological activity, although their molecular mechanisms remain unclear. The avocado-derived lipids exert potent nematocidal activity against several parasitic nematodes. In Caenorhabditis elegans (C. elegans), those compounds caused concentration-dependent toxicity, impairing first stage larval growth, egg hatching, and adult survival. Treated worms exhibited impaired mitochondrial respiration, reduced oxygen consumption, and elevated reactive oxygen species. These effects suggest that avocado lipids disrupt mitochondrial function and lipid metabolism, in part by inhibiting acetyl-CoA carboxylase, the rate-limiting enzyme of fatty acid biosynthesis. Methods: We investigated the effects of these compounds on the lipid profile of C. elegans and their association with endogenous lipid pools using NMR spectroscopy, click-chemistry-based fluorescence labeling, thin-layer chromatography (TLC), and microscopy. Results: Lipidomic analysis of stage 4 larvae (L4) and embryos treated with avocadene acetate revealed increased lipid NMR signals. Fluorescence-assisted TLC and NMR further suggested that avocadyne preferentially associates with triglyceride-linked fatty acids, particularly monounsaturated and flexible polyunsaturated chains, without detectable interactions with conformationally-constrained polyunsaturated species. Fluorescent avocadyne derivatives were efficiently internalized with distinct localization patterns in L4 larvae and embryonic cells. Conclusions: Overall, the lipid homeostasis remodeling of L4 larvae in response to lipotoxic shock was associated with phospholipid increase and remarkable lipid droplets onset, whereas embryos showed accumulation of lipids in enlarged droplets and developmental arrest.

Type 2 diabetes (T2D) is a prevalent metabolic disorder affecting millions worldwide, characterized by insulin resistance and impaired glucose homeostasis. While mammalian models are widely used, Drosophila melanogaster provides a powerful alternative due to its conserved insulin signaling pathways, genetic tractability, and suitability for high throughput studies. In addition to glucose dysregulation, lipid metabolism plays a crucial role in T2D pathophysiology, as alterations in lipid composition contribute to insulin resistance and metabolic dysfunction. Lipidomic studies have emerged as an essential approach to identify metabolic signatures and potential biomarkers for disease progression and therapeutic targeting. In this study, T2D like model was established by inducing insulin resistance through knockdown of the insulin receptor in brain insulin-producing cells using the dilp2-Gal4>UAS-InRRNAi system. This genetic manipulation resulted in significant metabolic dysregulation, including elevated glucose, trehalose, and triacylglyceride levels, along with increased oxidative stress indicators. Additionally, mRNA expression analysis of key insulin signaling components, including insulin receptor substrate 1, dilp2, dilp3, dilp5, and phosphorylated Akt, further validated the model. To further investigate metabolic alterations, Lipid profiling was performed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) in non targeted LC-MS-based metabolomics approach to identify lipid biomarkers associated with T2D. Multivariate statistical analyses, including PCA and PLS-DA, revealed distinct lipid signatures between wild-type and T2D flies. Notably, specific phosphatidylglycerol species PG 34:0, PG 34:4, PA 38:3, PIP 38:1, PIP2 38:6, and LPS 24:0 demonstrated an area under the curve (AUC) of 1, indicating their strong reliability as lipid biomarkers for T2D diagnosis.

During household storage, expressed human milk can develop odor and flavor changes that trigger infant refusal and lead caregivers to discard their saved milk supply. We show that typical refrigeration and freezing conditions disrupt the milk fat globule membrane (MFGM), exposing milk lipids to lipases that catalyze hydrolysis and oxidation. A pectin-based formulation (PBF) maintains MFGM integrity during storage and following lipase challenge, suppressing production of glycerol, free fatty acid, and oxylipin byproducts without broadly affecting milk macronutrients, the proteome, and culturable microbial burden. Across an independent cohort of lactating individuals, lipase activity varied but tracked with maternal milk lipase gene expression, implicating endogenous lipolysis in stored-milk deterioration. In a blinded olfactory panel, PBF-treated, lipase-challenged milk smelled more like fresh milk than untreated controls. Together, these findings show that stabilizing the MFGM can protect stored human milk from lipase-driven deterioration, preserve sensory quality, and support use for infant feeding.

Background and AimsThe genus Solanum including the wild species S. dulcamara, is rich in specialized metabolites such as steroidal glycoalkaloids (SGAs). Yet, much of this chemical diversity remains poorly characterized. This study aims to provide a comprehensive assessment of intra-specific chemodiversity in S. dulcamara. Using a dataset generated from 42 globally distributed accessions, we tested whether metabolic profiles differ among plant organs. We postulated that metabolic richness and abundance vary across accessions. Additionally, we hypothesized that differences in geographic origin or altitude affect SGA chemodiversity. MethodsAn untargeted metabolomic approach was applied to leaf, flower and root samples of 42 S. dulcamara accessions. Plants were grown in the greenhouse, and the extracted metabolites were analyzed using UHPLC-HRMS/MS in positive and negative ionization modes. Data processing and metabolite annotation were performed with a tailored bioinformatics workflow. Multivariate analyses were performed to evaluate chemical variation across organs and accessions. Key ResultsOur analyses revealed both organ and accession-specific metabolic diversity. Principal component analysis and clustering analyses revealed metabolic differentiation between leaves, flowers and roots. Leaves showed the highest metabolite richness and abundance, while roots showed the lowest. Alkaloids, especially SGAs, dominated positive mode profiles in roots, whereas shikimates and phenylpropanoids were prominent in negative mode profiles. Based on the leaf and flower SGAs profiles, four chemotypes were identified. Analyses of flavonoid and cinnamic acid derivatives, however, did not reveal chemotypes. Feature-based molecular network analyses confirmed that metabolite clusters are associated with plant organs, but not with altitude or geographic origin of the accessions. ConclusionsThe intraspecific chemodiversity within S. dulcamara is mainly driven by organ and accession-specific metabolic differences. We identified four SGA leaf and flower chemotypes, suggesting possible functional and ecological roles of this aboveground chemodiversity. These insights may contribute to applied research in plant resistance breeding and crop production.

Lavandula x intermedia (lavandin) is an economically important hybrid valued for its high essential oil yield; however, studies linking solvent polarity to its bioactivity and phenolic composition remain limited. This study evaluated the effects of nine extraction solvents on the antibacterial and antioxidant activities of L. x intermedia, determined total phenolic content (TPC), and examined the relationship between chemical composition and biological activity. Aerial parts were extracted by maceration. Antibacterial activity was assessed using the broth microdilution method against four bacterial strains, while antioxidant capacity was measured by the DPPH assay. TPC was determined using the Folin-Ciocalteu method, and phenolic compounds were analyzed via LC-MS/MS. Results showed that solvent polarity significantly influenced bioactivity. Diethyl ether extracts exhibited the highest TPC (267.65 mg GAE/g), strongest antioxidant activity (IC50: 72.26 {micro}g/mL), and notable antibacterial effects (MIC: 125 {micro}g/mL), especially against Gram-positive bacteria. A strong positive correlation (r = 0.887) was observed between phenolic content and antimicrobial activity. LC-MS/MS identified key compounds, including fumaric acid, resveratrol, and hydroxycinnamic acid. Overall, moderate-polarity solvents such as diethyl ether and methanol were most effective, highlighting L. x intermedia as a promising natural source for pharmaceutical and nutraceutical applications.

This study investigates electro-fermentation candida Tropicalis SY005 to address fermentation kinetics limitation during grape must fermentation. In comparison with non-stimulated control sample, EF substantially enhanced sugar depletion, TSS drop by day 3 and generated a strongly reduced state (ORP -100 to -143mV). The oxidation-reduction shift enhanced cellular NAD+ regeneration, reducing total fermentation duration from 264 h to 72 h. GC-MS analysis showed pronounced major characteristic volatile compound confirming substantial metabolic pathway shifts in flavor of glycolytic flux. Moreover moderate electric field promoted cellular membrane electropermeabilization substantially promoting bioactive extraction.

In many tropical countries, fermentation of palm sap into palm wine is an important fermented beverage contributing to local economies, tradition, and culture. Traditionally made in villages and families, palm sap is not inoculated with starter cultures and fermentation commences spontaneously. It is therefore possible that fermentation is influenced by multiple ecological factors, which affect microbial dynamics and thus flavour outcomes. Here, we studied microbial communities during fermentation of palm sap from three different palm tree species (palmyra, coconut, and sugar palm) on the island of Bali, Indonesia in both the wet and dry seasons. Our results suggest that season of collection has a strong influence on microbial dynamics and succession, and these changes positively correlate to metabolite concentration. The change of the season from the dry to wet season led to the loss of microbial diversity with lower richness in the dry season. The dominance of Saccharomyces cerevisiae was not affected by season and fermentation time and was dominant in all samples. Potential spoilage species, such as Candida tropicalis were negatively correlated to ester production and more abundant in the dry season. As microbial species varied in incidence and thus biochemical activity, the chemical groups of esters from their metabolism related to the change of season and fermentation time, while volatile compounds and small molecules were highly discriminated by season in the resultant wines. Ethyl octanoate was consistently different across all variables through comparison by three-way ANOVA and is proposed as a biomarker of seasonal variation in palm sap fermentation. These findings improve our understanding of microbial dynamics in palm sap fermentation, revealing flavour differentiation within season and suggests that strategies for microbial management, product development and quality assurance will elevate this traditional product into the future.

Branched-chain amino acids (BCAAs) are essential amino acids involved in protein synthesis and energy metabolism. Because animals cannot synthesize BCAA de novo, plant-derived BCAAs are important to human nutrition. Although mungbean sprouts are widely consumed as functional plant-based food materials, changes in individual BCAA accumulation and their transcriptional regulation during mungbean germination remain poorly understood. In this study, amino acid contents and transcriptomic profiles were analyzed at three germination stages, 8H, 24H, and 72H. Total BCAA content increased during germination, whereas individual BCAAs exhibited distinct temporal accumulation patterns. Isoleucine and valine increased until 72H, while leucine increased during early germination and decreased after 24H. Transcriptome analysis revealed time-dependent expression changes in BCAA biosynthesis and degradation genes associated with the leucine decrease after 24H. These findings suggest that 24H represents an important transition point for BCAA accumulation and compositional change during mungbean germination. This study provides molecular evidence for the regulation of BCAA metabolism during mungbean germination and supports the potential use of germinated mungbean as a plant-based amino acid resource.

Apple scar skin disease (ASSD), caused by Apple scar skin viroid (ASSVd), is characterized by peel scarring, cracking, dappling, and fruit deformation, resulting in reduced fruit quality and marketability. Despite its economic importance, the physicochemical and metabolic alterations underlying disease progression remain poorly understood. To address this knowledge gap, apple fruits representing four stages of ASSD (healthy, lightly infected, moderately infected, and highly infected) were comprehensively characterized. ASSVd infection was confirmed by RT-PCR, amplicon sequencing, and phylogenetic analysis. Fruit morphology and quality attributes, including firmness, total soluble solids (TSS), pH, titratable acidity (TA), and total phenolic content (TPC), were evaluated. ASSVd infection significantly reduced fruit weight and firmness and altered TSS and TA, indicating progressive deterioration of fruit quality. To investigate the underlying metabolic changes, peel and pulp tissues were analysed separately using gas chromatography-mass spectrometry (GC-MS), while major soluble sugars were quantified by 1H nuclear magnetic resonance (1H NMR) spectroscopy. Integrated metabolomic analyses revealed distinct tissue-specific metabolic reprogramming during disease progression. Major soluble sugars declined significantly during early infection, followed by tissue-dependent recovery at later stages, whereas organic acids, amino acids, phenolics, lipids, polyols, and pentacyclic triterpenoids exhibited dynamic stage-dependent changes. Notably, lupeol accumulated progressively, whereas ursolic acid and oleanolic acid declined, indicating disease-associated alterations in host triterpenoid metabolism. Multivariate analyses demonstrated clear metabolic separation among disease stages. Lupeol, ursolic acid, and chlorogenic acid were identified as candidate discriminatory metabolites in the peel, whereas myo-inositol, chlorogenic acid, and aspartic acid were identified in the pulp. Collectively, these findings demonstrate that ASSD induces coordinated, tissue-specific physicochemical and metabolic reprogramming that compromises postharvest fruit quality and reshapes defence-associated metabolism. This study provides the first integrated metabolomic characterization of ASSD progression and identifies potential metabolic biomarkers for disease diagnosis and severity assessment. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/741181v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@92e453org.highwire.dtl.DTLVardef@1cc2436org.highwire.dtl.DTLVardef@15d7e27org.highwire.dtl.DTLVardef@1056568_HPS_FORMAT_FIGEXP M_FIG C_FIG

High dietary fructose consumption is a major contributor to the development of obesity and related cardiometabolic diseases, highlighting the need for accurate assessment of fructose metabolism in humans. Stable isotope tracer approaches, such as 13C6-fructose, require highly sensitive and specific analytical methods to quantify both concentrations and isotopic enrichments. In this study, we developed and validated a robust gas chromatography-triple quadrupole mass spectrometry (GC-QQQ)-based method for the simultaneous measurement of unlabeled and 13C6-fructose in human plasma. The method employs oximation and per-acetate derivatization, and demonstrates high specificity and accuracy. Intra- and inter-assay precision were below 10%, with no detectable carry-over, and a lower limit of quantification (LLOQ) of 0.1 nmol/mL for concentration and 0.02 molar percent excess (MPE%) for enrichment and no interference from glucose. We further compared data acquisition using multiple reaction monitoring (MRM) and selected ion monitoring (SIM). MRM showed superior performance at the low concentrations and enrichment levels characteristic of clinical plasma samples, resulting in improved sensitivity and lower LLOQs compared to SIM. Overall, this validated method provides a sensitive and reliable approach for fructose tracer studies in humans. Its application will facilitate robust investigations into fructose metabolism, and its role in metabolic dysregulation and obesity-related disease.