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This work addressed the challenge of reducing sucrose content while maintaining sweetness perception in food systems. Based on the idea that sucrose sweetness is mainly captured from the dissolved fraction at the crystal surface during oral processing, rather than from the entire crystal mass, a protein-assisted strategy was proposed to partially replace internal sucrose with protein while preserving perceived sweetness. Soy protein isolate (SPI) and whey protein isolate (WPI), with and without heat treatment, were first characterized and evaluated for their interactions with mucin and their effects on mucin microrheological behavior. The results showed that both proteins interacted with mucin and altered the interfacial and microrheological properties of the mucin system, while heat treatment further modified these effects through structural changes in the proteins. Based on this, protein-sucrose composite particles were prepared using a protein-assisted particle formation process. Structural characterization showed that the particles retained the characteristic crystalline phase of sucrose while containing detectable protein-associated regions. Ex vivo porcine tongue experiments demonstrated that the protein-sucrose composite particles reduced burst release and prolonged sucrose retention on the tongue surface. Application in chocolate further showed that these particles enhanced sweetness perception compared with pure sucrose particles at the same formulation level. Overall, this work provides a novel sucrose reduction strategy by using proteins as inert fillers and oral-interface modulators, thereby improving the sweetness perception efficiency of sucrose and offering new insight for the development of reduced-sugar foods.

Tibetan Plateau, which was once a severe endemic area of Kashin-Beck disease (KBD), was low with selenium in the soil geochemical background and with harsh environment, thus resulting in selenium deficiency in local foods while high environmental-derived oxidative stress and inflammatory responses in human body. Diet was the major selenium source for humans, while external environment conditions could influence selenium consumption and disturb the selenium metabolism. This study explored the influence on urinary selenium of Tibetan population from the pespevtive of selenium intake, represented by food patterns and selenium concentration, and from the perspective of selenium metabolism caused by environmental stress. From 2019 to 2022, 572 food consumption questionnaires of Tibetan residents, matched urine samples, staple food samples, and local agricultural soil and water samples were collected. Environmental stressors, including absolute oxygen concentration, temperature-humidity index and ultraviolet radiation intensity were considered. Statistical analysis and Random Forest (RF) were used to explore the association, contribution importance and interactive effect of the factors above on urinary selenium concentrations. Urinary selenium concentrations of Tibetan residents were still generally low, declined with age whereas no difference was found between genders. In terms of food consumption patterns, dietary diversity score (DDS) and non-local food intake, poultry consumption frequency in particular were positively associated with urinary selenium, exerted a pro-linear relationship. Highland barley and wheat flour selenium concentrations showed positive impact on the urinary selenium concentrations, and the magnitude of impact was more obvious when the highland barley selenium concentration exceeded 0.04 µg/g, and the wheat flour selenium concentration was lower than 0.051 µg/g. The soil and water selenium concentrations also positively influenced the urinary selenium concentrations. Environmental stressors exerted distinct influencing curves on urinary selenium concentrations, with higher ultraviolet radiation intensity (UVRI) and lower environmental comfort induced by cold and dry conditions positively regulating urinary selenium concentrations, while oxygen concentration exhibited a marginal effect only when it reached a certain threshold. Food diversity, non-local food intake, staple food selenium concentrations and local agricultural selenium concentrations were all important factors for elevating the selenium status among Tibetan residents. Environmental stressors regulated the urinary selenium concentrations differently due to the divergent action pathways on the selenium depletion and metabolism. Correspondingly, increasing the food diversity through introducing non-local food, increasing the selenium level of local food and elevating the human selenium reference standards were suggested.

Taste modulation induced by interactions among specific compounds remains poorly understood. L-histidine and creatinine are both bitter compounds abundant in dried bonito dashi. This study investigated whether ten bitter compounds could shift the taste quality of L-histidine from bitterness toward sweetness, as assessed by sensory evaluation using a fixed-sum method. When presented alone, 4% L-histidine elicited bitterness accompanied by weak sweetness, whereas iso-bitter creatinine evoked predominantly bitterness. Remarkably, pre-stimulation with creatinine shifted the taste quality of L-histidine toward sweetness. Similar effects were observed after pre-stimulation with isoleucine, tryptophan, phenylalanine, and creatine, but not with histidine, valine, leucine, caffeine, or carnosine. This effect was transient, as increasing the interval after creatinine pre-stimulation restored the bitterness of L-histidine. In addition, 0.1% NaCl altered the bitterness-to-sweetness balance of L-histidine toward sweetness after pre-stimulation with creatinine. The taste quality of histidine alone was highly sensitive to pH: bitterness peaked at pH 7.0, whereas sweetness increased under alkaline conditions. In contrast, varying the pH of the pre-stimulation solution between 5.0 and 8.0 did not affect the modulation of L-histidine taste. These findings demonstrate a previously unrecognized histidine-specific form of taste modulation, suggesting interactions among naturally coexisting bitter compounds may dynamically alter taste qualities in traditional broths and other complex foods.

This study presents the first comprehensive, multisectoral assessment of India's nitrogen (N) budget, quantifying N inputs, flows, circularity, and losses across ten economic sectors. We present a scalable Environmentally Extended Input-Output (EEIO) framework, trace 198 N flows to estimate the conversion potential of input N to reactive nitrogen (Nr) losses, and estimate sectoral N use efficiency (NUE). The total N-input in India is estimated at ∼50 Tg-N yr-1 in 2020, dominated by Haber-Bosch fixation (38%), imports (33%), biological N fixation (27%), and fossil fuel combustion (18%). Cropland and industry together receive ∼90% of total N inputs; however, industry exhibits higher NUE (62%) than cropland (56%), given tighter regulations, whereas agriculture operates as a semiclosed loop with substantial losses. Shifts in dietary patterns toward animal protein and processed foods exacerbate feed-food competition and amplify Nr emissions. Overall, 52% of N inputs are converted to inert N2, 37% are emitted as Nr, and 10% of Nr accumulate within sectors. Atmospheric Nr losses (∼9 Tg-N) are primarily as NH3 (63%), NOx (32%), and N2O (5%), while hydrosphere losses (∼9 Tg-N) occur via runoff (67%) and leaching (33%). Key Nr processes include biomass burning (13 Tg-N), runoff (11 Tg-N), and combustion (8 Tg-N). These findings highlight critical intervention points for reducing N inefficiencies and mitigating environmental Nr load in India's evolving N economy.

This study revealed an integrated investigation of the cytotoxic potential targeting leukaemia of an active fraction, enriched in (9Z)-5,8,11-trihydroxyoctadec-9-enoic acid (OA), obtained from Abelmoschus sagittifolius roots, a promising Vietnamese traditional plant. OA-enriched fraction exhibited moderate cytotoxic activity against Molt4 cancer cell line (IC50 = 42 µg/mL), showing greater sensitivity compared to K562, U937, THP1, and Jurkat cancer cell lines. Network pharmacology results suggested that OA might exhibit its anti-leukaemia activity primarily via altering the PI3K/Akt signalling pathway. Molecular docking further demonstrated strong binding affinities of OA towards ESR1, PPARG, and PIK3CA, mediated by hydrogen bonding and hydrophobic interactions. Besides, simulated digestion studies indicated the limited bioavailability and bioaccessibility of OA. The ADMET study also indicated that OA is suited as a lead or bioactive component in standardised functional foods or phytopharmaceuticals than a systemic small-molecule drug. Further research should be done to confirm its therapeutic uses for developing functional and nutraceutical supplements.

Perovskite quantum dots (PQDs) have emerged as highly promising nanomaterials for fluorescence-based sensing owing to their exceptional optical characteristics, including near-unity photoluminescence quantum yields, narrow emission linewidths, broad absorption cross-sections, and compositional tunability. Despite these advantages, three core challenges continue to limit their practical applications: poor stability in the presence of moisture, oxygen, or heat, and the toxicity of lead-based compositions. Particularly in the context of analytical sensing applications, PQDs show an inherent lack of molecular selectivity toward specific analytes. Although reviews providing a general overview of PQD synthesis or optical properties have been published, a consolidated discussion of sensor designs and mechanisms in PQD-based systems is lacking. Importantly, reviews focusing on design strategies for stability and selectivity, and the emerging role of molecularly imprinted polymers (MIPs) as a route to analyte-specific recognition is lacking. This review addresses that gap by systematically covering the foundations of PQD photophysics, including radiative recombination mechanisms and exciton dynamics, followed by the major sensing mechanisms in PQD-based systems: photoinduced electron transfer (PET), Förster resonance energy transfer (FRET), intramolecular charge transfer (ICT), and excited-state intramolecular proton transfer (ESIPT). Besides, we critically evaluate important design strategies such as surface functionalization, encapsulation, compositional doping, and hybrid material integration. A key focus of this review is a discussion on MIP-PQD hybrid systems, with in-depth discussion of the fluorescence modulation mechanisms that arise from analyte rebinding to imprinted cavities. Finally, representative applications in environmental monitoring, biomedical diagnostics, and food safety are reviewed, and future directions toward lead-free, portable, and AI-integrated sensing platforms are outlined.

Benzalkonium chloride (BAC) is a widely used disinfectant, but its improper application can induce Salmonella Typhimurium (S. Typhimurium) to develop tolerance to subsequent stressors. In this study, we demonstrated that adaptation of S. Typhimurium to BAC conferred both direct protection and cross-protection against cold stress, resulting in significantly enhanced survival at -20°C compared with non-adapted cells. BAC-adapted S. Typhimurium also exhibited a survival advantage on chicken meat during frozen storage at -20 °C. Membrane fatty acid analysis indicated that adaptation of S. Typhimurium to BAC led to a 2.2-fold increase in the levels of cyclopropane fatty acids. Moreover, BAC-adapted S. Typhimurium showed a significant 8.0% increase in cell surface hydrophobicity. RT-qPCR analysis demonstrated upregulation of efflux pump genes tolC, acrB, and mdfA (3.6-, 3.4-, and 3.0-fold, respectively) and cold shock-related genes cspA, cspE, and rpoS (2.4-, 4.0-, and 2.6-fold, respectively). These findings suggest that upregulation of cold shock genes in BAC-adapted cells was associated with the observed cross-protection. Taken together, adaptation of S. Typhimurium to BAC promoted the development of direct and cross-protective phenotypes, which may increase the risk of cross-contamination and salmonellosis in the food industry, posing a serious threat to food safety and public health.

Uncontrolled use of tobramycin (TOB) raises serious health concerns, including nephrotoxicity and ototoxicity, making it imperative to develop simple and sensitive detection methods. This study introduces a novel enzyme-free and label-free fluorescent biosensor based on a cascade signal amplification strategy, integrating an entropy-driven DNA circuit (EDC) with catalytic hairpin assembly (CHA). The mechanism is initiated by the specific binding of TOB to its aptamer, which induces a conformational change and exposes a priming sequence. This sequence triggers a strand displacement reaction to release an initiator strand, subsequently activating the CHA process to generate fuel strands. These fuel strands drive the EDC, resulting in the liberation of numerous G-quadruplex sequences from the stem of hairpin probes. The released G-quadruplexes bind to thioflavin T (ThT), yielding a significantly enhanced fluorescence signal proportional to the TOB concentration. The proposed biosensor exhibits outstanding analytical performance with a detection limit of 92 fM and a wide linear range from 3 pM to 30 nM. Furthermore, the method effectively distinguishes TOB from other interfering antibiotics, highlighting its high specificity. It also demonstrates practical applicability by achieving satisfactory recovery rates in milk sample assays. With its high sensitivity, cost-effectiveness, and operational simplicity without the need for complex enzymes or labels, the proposed strategy offers a promising platform for food safety supervision and clinical diagnostics.

This study demonstrates that inoculation with the Enterococcus wangshanyuanii strain F4 in a germ-free black soldier fly larval (BSFL) system enhances sulfamethoxazole (SMX) degradation, larval growth, and substrate conversion. Following inoculation with strain F4, the net SMX degradation rate reached 37.08%, and the net substrate consumption rate reached 46.12%, both representing significant improvements compared to the control group. Metagenomic analysis revealed that strain F4 modulated the BSFL gut microbial community structure and enriched functional genes associated with organic pollutant degradation. Accordingly, the activities of key degradation enzymes in the larval gut, including catechol-1,2-dioxygenase (C12O), catechol-2,3-dioxygenase (C23O), and peroxidase (POD), were significantly elevated following inoculation. Taken together, these findings suggest a synergistic effect between Enterococcus wangshanyuanii F4 and the host during the degradation process, which significantly enhances the removal of SMX by black soldier fly larvae. This provides a theoretical basis for the use of symbiotic microbial augmentation strategies in antibiotic bioremediation.

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS), driven by genetic and environmental determinants. The gut microbiome of people with MS (pwMS) is distinct and influences disease through immunomodulatory metabolite production. Circulating metabolites are altered in pwMS, but identifying microbial-metabolic drivers remains challenging. We previously showed that colonization by the gut commensal Limosilactobacillus reuteri exacerbates disease in the experimental autoimmune encephalomyelitis (EAE) model of MS in a tryptophan-dependent manner. Here, we integrated microbiomic and metabolomic data sets from a longitudinal EAE study utilizing high- and low-tryptophan diets in mice colonized with or without L. reuteri. Gut microbiome dynamics, under short- and long-term alterations in tryptophan bioavailability, were affected by diet, microbiome context, or disease. During short-term dietary intervention, L. reuteri colonization exerted a greater impact on microbiome composition than tryptophan bioavailability. With longer dietary exposure and EAE progression, high dietary tryptophan and L. reuteri colonization synergized to elicit profound microbiota changes, including alterations in Lachnospiraceae, Blautia, and Akkermansia. Integration of metabolomic and microbiomic data sets using joint Robust Aitchison PCA revealed clusters of associated metabolites and microbiota enriched for functional pathways, including bile acid and tryptophan metabolism. Metabolites outperformed microbiota in predicting EAE severity, identifying p-cresols and indoles as top disease-associated metabolites. Treatment with p-cresol or 3-indoleglyoxylic acid exacerbated EAE, enhanced proinflammatory T-cell responses, and increased cerebellar pathology. These data demonstrate that dietary responses are shaped by gut microbiome composition and that integrated microbiomic-metabolomic analyses can identify the drivers of disease worsening in MS.IMPORTANCEMultiple sclerosis (MS) is a multifactorial disease influenced not only by genetics but also by environmental factors, potentially including diet and the composition of the gut microbiome. We show that interactions between diet and commensal gut microbiota profoundly impact the levels of immunomodulatory systemic metabolites, including several that are associated with disease in people with MS (pwMS). Importantly, we demonstrate that individual gut microbiota-produced metabolites are sufficient to worsen disease in a mouse model of MS. Integration of gut microbiome and blood metabolite data sets, combined with subsequent predictive modeling, may bolster biomarker identification and the capacity to predict disease severity in pwMS, as compared to the performance of individual data sets alone. These findings highlight metabolites as key mediators linking diet and the gut microbiota to neuroinflammation. Importantly, this work suggests that targeting microbial metabolites or modifying diet-microbiome interactions may represent new strategies to reduce disease activity in MS and related autoimmune disorders.

Fish have the capacity to bioaccumulate metals such as mercury through the food web, posing potential health risks to aquatic ecosystems and humans. This study aimed to assess total mercury (THg) concentrations in different tissues of freshwater fish and evaluate the potential risk for consumers. Perch (Perca fluviatilis) and rudd (Scardinius erythrophthalmus), representing different trophic levels, were collected from Koronowo Reservoir and Lake Wierzchucińskie Duże in summer and autumn. THg accumulation followed the order: muscle > liver > gills, with significant differences observed between tissues. In perch, THg concentrations were 0.061 μg/g (muscle), 0.037 μg/g (liver), and 0.028 μg/g (gills). In rudd, THg levels were 0.050 μg/g (muscles), 0.034 μg/g (liver), and 0.022 μg/g (gills). The target hazard quotient (THQ) values were below 1, indicating that their consumption does not pose a significant health risk to humans. Estimated safe weekly intake was 1,839 g for perch and 2,249 g for rudd for dorsal muscle.

Moderate acute malnutrition (MAM), affecting over 30 million children globally, substantially increases the risk of morbidity and mortality. In 2023, the World Health Organization recommended lipid-based nutrient supplements (LNS) for children requiring supplementation for MAM, either ready-to-use supplementary food (RUSF) or ready-to-use therapeutic food (RUTF). However, evidence on the optimal product (RUTF or RUSF) and dosing strategy for children with MAM remains limited. The Modified Dosages for Acute Malnutrition MAM (MODAM-MAM) trial evaluated whether one- or two-sachet daily RUTF dosing were non-inferior to the de facto standard of one RUSF sachet daily for MAM supplementation. We conducted a three-arm, parallel, individually randomized controlled trial in Ethiopia. Children aged 6-59 months with MAM (mid-upper arm circumference (MUAC) 11.5-12.4 cm or WHZ between -3 and -2, without edema) were randomized to: one 540-kcal sachet of RUSF/day ("1-RUSF"), one 500-kcal sachet of RUTF/day ("1-RUTF"), or two sachets of RUTF/day ("2-RUTF"). Both intervention arms (1-RUTF and 2-RUTF) were evaluated for non-inferiority to the standard (1-RUSF) using a prespecified 6-percentage-point margin. The primary outcome was anthropometric recovery, defined as both MUAC ≥ 12.5 cm and WHZ ≥ -2 for two consecutive visits within 16 weeks. In total, 2,414 children were analyzed in the intention-to-treat analysis. Recovery proportions were 84.8% (679/801) for 1-RUSF, 85.0% (674/793) for 1-RUTF, and 88.1% (721/818) for 2-RUTF. Risk differences for recovery relative to the standard of care (1-RUSF) were +0.2 percentage points (95% CI [-3.3, 3.7]; p = 0.944) for 1-RUTF and +3.4 percentage points (95% CI [0.0, 6.7]; p = 0.050) for 2-RUTF, with both intervention arms meeting the non-inferiority criteria. Anthropometric gains and median length of stay (56 days) were similar across arms. Ration use was comparable for 1-RUSF and 1-RUTF, while 2-RUTF required 1.6-fold more product. Study limitations include evaluation of a single RUSF formulation and earlier-than-typical case-finding due to intensive screening. Both experimental arms, one RUTF sachet daily and two RUTF sachets daily, are non-inferior to the standard dosing of one RUSF sachet daily for MAM supplementation. Although two sachets of RUTF daily yielded slightly higher recovery, the marginal benefits do not justify substantially greater resource demands. Because one RUTF and one RUSF sachet daily yielded similar treatment outcomes, either product can be considered an effective option for supplementation. Product choice may therefore be guided by supply, cost, and operational considerations. clinicaltrials.gov NCT06056089.

To test whether dietary antioxidants moderated effects of PM2.5 from coal mine fire smoke on respiratory symptoms. We conducted cross-sectional analyses of diet and respiratory symptom data from 448 members of the Hazelwood Health Study Adult cohort. Individual-level exposure to PM2.5 from the 2014 coal mine fire was determined by combining modelled fire-related PM2.5 estimates and time-location diaries from the time of the fire. Data were evaluated using logistic regressions to evaluate associations between PM2.5 and respiratory symptoms and whether this was moderated by meeting recommended daily antioxidant intakes. Vitamins A and E, magnesium, and zinc attenuated the association between PM2.5 and chronic cough and (excluding magnesium) chronic phlegm. Omega-3 fatty acids attenuated the association between PM2.5 and COPD. Higher-quality diets may reduce harms due to smoke exposure from landscape fires.

Glycosylphosphatidylinositol-specific phospholipase D1 (Gpld1) is a membrane-associated enzyme that modulates diverse cellular processes through the cleavage of glycosylphosphatidylinositol (GPI)-anchored proteins. Although recent studies have linked circulating Gpld1 to exercise-induced rejuvenation, its cell-autonomous role in coordinating redox homeostasis, melanogenesis, and cellular aging has not been fully elucidated. To address this, a stable Gpld1 knockout (KO) was generated in B16F1 melanoma cells using CRISPR/Cas9-mediated genome editing to investigate whether loss of Gpld1 induces aging-associated phenotypes and redox imbalance. Gpld1 KO cells exhibited senescence-like features, as evidenced by increased senescence-associated β-galactosidase (SA-β-gal) activity, accompanied by disrupted redox homeostasis and markedly enhanced melanin synthesis. In the context of skin aging, hyperpigmentation often accompanies senescence-associated changes and thus represents a relevant phenotypic readout in this model. Gene and protein expression analyses revealed coordinated remodeling of multiple aging-associated signaling pathways. Specifically, SIRT1 and the redox repair enzyme MsrA were downregulated, whereas SIRT7, forkhead box O1 (FoxO1), phosphorylated FoxO1 (p-FoxO1), and Caspase-1 were markedly upregulated. Reduced steady-state reactive oxygen species (ROS) levels detected by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) likely reflect compensatory activation of antioxidant signaling pathways rather than the absence of oxidative stress. Collectively, these findings demonstrate that Gpld1 KO induces Senescence-like features, alters redox homeostasis, and, in melanocytic cells, manifests as enhanced melanogenesis. These results identify Gpld1 as a previously unrecognized regulator of oxidative stress-driven aging and pigmentation, suggesting its potential relevance to aging-related hyperpigmentation and redox imbalance.

The first COVID-19 case in Massachusetts was identified in January 2020, marking the start of an unprecedented pandemic that resulted in more than 20,000 deaths statewide. In response, Massachusetts rapidly developed and launched the Community Tracing Collaborative (CTC), the nation's first large-scale statewide contact tracing support system. Beginning in April 2020, the CTC, an entirely virtual operation, recruited and trained more than 4000 lay staff to conduct outreach to COVID-19 cases and contacts. CTC services included case investigation and contact tracing; referral for social supports such as food and other resources; outbreak investigation and response; and referrals for testing and treatment. We conducted a cohort analysis of COVID-19 cases and close contacts referred to the CTC, as well as contacts identified by CTC staff during tracing activities, between May 1, 2020, and October 31, 2021. We calculated the proportions of cases and contacts with successful initial interviews and, among those, successful completion of isolation or quarantine. During the study period, the CTC supported 551,797 cases and 304,928 contacts. Among individuals served, 8.6% identified as Black, 22.0% as Hispanic, 50.8% as female, and 11.6% as non-English speaking. Overall, 79.3% had a successful initial interview outcome, and among these, 72.6% successfully completed isolation or quarantine. The CTC demonstrated that a large-scale, integrated case investigation and contact tracing program with embedded supportive services can be rapidly deployed and sustained, even in an unprecedented public health emergency.

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This study presents a novel single-integrated cartridge-based multiplex chemiluminescence immunosensor (SIC-MCLIS) platform designed for rapid, simultaneous quantification of five key coagulation and fibrinolysis biomarkers-thrombomodulin (TM), thrombin-antithrombin complex (TAT), plasmin-α2-plasmin inhibitor complex (PIC), tissue plasminogen activator-inhibitor complex (t-PAIC), and D-dimer. This fully automated "sample-in to result-out" system integrates all reagents and consumables within a compact reagent cartridge and analyzer, enabling sensitive detection from only 200 μL of plasma within 25 min. The platform employs a streptavidin-biotin amplification strategy and features precise pipetting, uniform temperature control, efficient washing, and low background signal, ensuring high accuracy and reproducibility. The platform exhibits enhanced analytical performance, with limits of detection (LoD) of 0.5 ng/mL for TAT, 1.0 TU/mL for TM, 0.06 μg/mL for PIC, 0.75 ng/mL for t-PAIC, and 5 ng/mL for D-dimer. The repeatability coefficient of variation (CV) is below 8%, the within-laboratory CV is under 10%, bias remains within 8%, and linearity is demonstrated by an R2 value exceeding 0.99. A method comparison study conducted on 228 clinical samples against the reference system HISCL-5000 showed a strong correlation (R2 > 0.99) and agreement exceeding 96% within the Bland-Altman limits. Overall, this innovative multiplex immunoassay system offers a rapid, cost-effective, and accessible solution for comprehensive coagulation-fibrinolysis assessment, with significant potential to improve real-time clinical management of thrombotic and coagulopathic disorders, especially in emergency and point-of-care settings.

Enterococcus faecalis remains a primary cause of persistent endodontic infections, persisting in root canals through robust biofilm formation and antimicrobial resistance. This study characterized clinical E. faecalis oral isolates and evaluated the comparative efficacy of conventional endodontic medicaments against the probiotic and antibiofilm potential of food derived Lactic Acid Bacteria (LAB). Clinical isolates (JUDS 01, JUDS 02, JUDS 03, JUSA 05) exhibited a high prevalence of ace, agg, cyl, and gelE virulence genes, demonstrating significant multidrug resistance. Among conventional treatments, TAP exhibited the notable antibacterial zones of inhibition (23&#x2009;&#xb1;&#x2009;1-24&#x2009;&#xb1;&#x2009;0.5&#xa0;mm) and uniform, highly significant biomass reduction (p&#x2009;<&#x2009;0.0001), whereas Ca(OH)2, NaOCl, and DAP completely failed to reduce mature biofilms (p&#x2009;>&#x2009;0.05). Among the probiotic strains, Pediococcus acidilactici JUFB demonstrated great physiological stress tolerance, maintaining robust survival under lethal acid conditions (OD600&#x2009;=&#x2009;0.32 at pH 2.0) and high growth under bile stress (OD600&#x2009;=&#x2009;1.15 in 0.3% Oxgall). Remarkably, organic extracts of P. acidilactici JUFB achieved a maximum of up to 70% relative reduction in mature E. faecalis biofilm biomass (p&#x2009;<&#x2009;0.0001) at 0.12&#xa0;mg/ml optimal concentration, overcoming P. acidilactici JUFF and Streptococcus thermophilus JUBM. GC-MS analysis of this active extract identified unique peaks, prioritizing core functional micro metabolites including carbamimidoyl sulfanyl acetic acid and 2-ethyl-2-methyl-1,3-propanediol as putative antibiofilm agents. These findings indicate that while TAP remains a potent conventional therapy, P. acidilactici JUFB possesses higher probiotic attributes and powerful antibiofilm activity, supporting its use as a resistance mitigating therapeutic alternative.

Blueberry (Vaccinium spp.) has gained worldwide popularity as a fruit crop of exceptional flavor and high nutritional value. Fruit acidity, a critical determinant of consumer preference in rabbiteye blueberry, is primarily governed by vacuolar malic acid accumulation; however, the underlying regulatory mechanism remains unclear. In the current study, VaALMT9, encoding an aluminum-activated malate transporter, was identified from rabbiteye blueberry transcriptome data. The full-length coding sequence encodes a 574 amino acid protein harboring the characteristic WEP fingerprint motif of the ALMT family. Subcellular localization analysis suggested that VaALMT9 localizes to the endomembrane system, most likely the tonoplast (vacuolar membrane). Expression analysis revealed that VaALMT9 transcript levels paralleled malate accumulation during fruit development, with both peaking at the purple stage and declining at the late ripening stage. Virus-induced gene silencing of VaALMT9 in blueberry fruits significantly reduced its transcript levels (by 96.74%) and malate contents of the two independent silenced lines were remarkably decreased by 0.62&#xa0;mg/g and 0.50&#xa0;mg/g, which were reduced by 19.37% and 15.78% in comparison with those of the control. Conversely, VaALMT9-overexpressing tomato markedly increased malate accumulation by 13.79-44.37%. Moreover, yeast one-hybrid and dual-luciferase reporter assays revealed that VaMYB1R1 and VaWRKY21 bind to the VaALMT9 promoter and repress its transcriptional activity. In conclusion, VaALMT9 was identified as a key gene promoting malate accumulation, and is transcriptionally inhibited by VaMYB1R1 and VaWRKY21. The findings provide new insights into the molecular regulation underlying fruit acid accumulation and offer valuable gene targets for genetic improvement of fruit quality in this fruit crop.