The physiological benefits and post-prandial retention mechanisms of meat aroma compounds remain poorly understood. In this study, simulated roasted duck-derived biomimetic myosin-triglyceride-maltose micro/nanoparticles loaded with furfuryl mercaptan (FurMeSH; final concentration, 1 mg/L) were fractionated into homogeneous particles (HPs), microfiltered particles (MPs), and ultrafiltered particles (UPs). In Caenorhabditis elegans, bioactivity showed clear size dependence. MPs-FurMeSH showed the strongest protective trend under heat and oxidative stress, primarily by alleviating the carrier-induced stress observed in the unloaded MPs group and by reducing reactive oxygen species (ROS) relative to the stressed baseline. Integrated metabolomic and microbial profiling associated this protection with improved neuromuscular behavioral performance and shifts in the nematode-associated microbial community. During in vitro digestion, MPs structurally evolved into micelle-like colloidal structures and showed favorable FurMeSH retention/release behavior. Caco-2 assays further showed that digestion products of MPs-FurMeSH were associated with enhanced FurMeSH-related transport via a potential carrier-mediated pathway, while attenuating extensive Phase II glucuronidation. These findings demonstrate that food-derived microstructures can function as natural carriers, offering a mechanistic link between flavor retention and nutrient-like bioactivity and providing a matrix-design strategy for future aroma-based functional foods.

Obesity is a chronic metabolic disorder closely associated with dyslipidemia, insulin resistance, low-grade inflammation, and gut microbiota dysbiosis. Mulberry leaves are rich in bioactive proteins, but whether mulberry leaf albumin-type protein can improve diet-induced obesity remains unclear. In this study, ultrafiltered mulberry leaf albumin-type protein (UMP) was prepared and its anti-obesity effects were evaluated in high-fat diet (HFD)-fed C57BL/6J mice. UMP contained 87.12 ± 0.52 g/100 g protein, 2.52 ± 0.00 g/100 g polyphenols, and 8.21 ± 1.49 g/100 g polysaccharides, with two major albumin-type protein bands of approximately 14 and 52 kDa. Structural analysis showed that UMP was mainly composed of β-turns and α-helices. In HFD-fed mice, daily administration of UMP for 16 weeks reduced body weight gain by 3.85 g and 5.63 g in the low- and high-dose groups, respectively, without affecting food intake. Biochemical assays, glucose and insulin tolerance tests, and histological analysis showed that UMP improved insulin responsiveness, alleviated serum dyslipidemia, reduced hepatic lipid accumulation, and decreased circulating alanine aminotransferase, aspartate aminotransferase, and lipopolysaccharide levels. Histological analysis and nuclear magnetic resonance-based short-chain fatty acid quantification further showed that UMP protected colonic morphology and increased colonic short-chain fatty acid levels. Gut microbiota analysis showed that UMP restored microbial diversity, reduced the Firmicutes/Bacteroidota ratio, and enriched potentially beneficial genera, including Ileibacterium and norank_f_Muribaculaceae. Fecal biochemical assays suggested that UMP promoted fecal free fatty acid excretion and partially improved bile acid-related metabolic alterations. Untargeted serum metabolomics revealed that UMP reshaped metabolic pathways related to lipid turnover, bile acid signaling, glucose utilization, and glucuronidation. Correlation analysis linked UMP-enriched bacterial taxa with key metabolites involved in fatty acid and energy metabolism. Together, these findings indicate that UMP attenuates HFD-induced obesity through coordinated regulation of gut microbiota, intestinal metabolites, and systemic metabolic homeostasis. UMP may therefore represent a promising functional dietary protein for the prevention of obesity-related metabolic disorders.

Flavourzyme-assisted two-stage hydrolysis is a recognized debittering strategy for protein hydrolysates, yet peptidomic-level evidence for the underlying mechanism-particularly the role of pH control mode-remains limited. This study compared pH-stat and free-fall pH Flavourzyme treatments (3 and 4 h) of an Alcalase-derived rice protein hydrolysate (A6; degree of hydrolysis by the o-phthalaldehyde method [DH-OPA] 24.69%) using nanoLC-ESI-Q-TOF de novo sequencing to characterize peptide-profile changes associated with bitterness reduction. Formal sensory evaluation (15 trained panelists, paired comparison with Bonferroni correction for 10 comparisons) of the ultrafiltered (<2500 Da) fractions placed A6-s3 (pH-stat, 3 h) at the least-bitter end of the ranking, although it did not differ significantly from A6-s4 after multiplicity correction. The apparent ranking differed from that observed in the preliminary evaluation of unfiltered hydrolysates, which used a smaller panel and a different sensory design. De novo sequencing identified 963-1226 peptides per hydrolysate. A6-s3 exhibited the lowest mean Q value (1430 cal mol[-1]; Kruskal-Wallis, p < 10-22) and the lowest predicted bitter peptide proportion (49.6%). A6 and A6-s3 shared only one exact PEAKS-assigned sequence among >1900 combined detections, indicating extensive differences in the detectable peptide profiles. Database-assisted annotation against the Oryza sativa proteome showed a markedly lower assignment rate for A6-s3 (0.2%) than for A6 (52.8%, predominantly glutelins); this was treated as a descriptive analytical observation because I/L ambiguity, de novo sequencing uncertainty, stochastic data-dependent acquisition (DDA) sampling, and database coverage may contribute. A6-s3 also showed the lowest C-terminal strongly hydrophobic residue frequency (62.3%; I, L, F, W, Y, P) and the highest N-terminal amino acid diversity (Shannon entropy, 3.91 bits). These findings indicate that maintaining pH during Flavourzyme treatment was associated with a more favorable bitterness-related peptide profile. The results are compatible with established terminal processing accompanied by broader changes in the detectable peptide population, but they do not establish or quantify a distinct contribution from internal cleavage.

Emerging micropollutants in drinking water sources represents a growing challenge for water treatment systems. Bisphenol-A (BPA) and 17β-estradiol (E2) are endocrine disruptors widely detected in aquatic matrices that are not efficiently removed by conventional treatment. This study evaluated, at the bench scale, operational performance and rejection of BPA and E2 by three nanofiltration membranes-NFM1, NFM2 and NFM3-operating at 8 bar and using ultrafiltered water from the Lago Norte Water Treatment Plant (WTP), Brasília/DF, Brazil, spiked with both compounds at 150-250 µg/L, as the feed matrix. Hydraulic parameters, such as permeate flux and water permeability, were assessed alongside rejection. NFM1 exhibited the highest permeate fluxes (136.1 and 171.7 L/h·m[2]); however, it showed the lowest rejection (E2: 57-73%; BPA: 28-60%). The NFM2 membrane showed intermediate rejection behavior (E2: 86-89%; BPA: 67-91%) but presented the lowest permeate flux (51.2 to 63.3 L/h·m[2]). The NFM3 membrane presented the highest rejection and greatest operational stability (E2: 90-95%; BPA: 95-97%), with a permeate flux of 59.1 to 67.0 L/h·m[2]. Size exclusion was the predominant removal mechanism, though adsorption also contributed during the initial hours of operation. The results confirm a trade-off between permeate production and contaminant rejection, with no single membrane outperforming all others across all criteria.

Treated water systems are critical infrastructures for public health, but can be vulnerable to contamination, whether intentional or accidental. The ability to detect water-transmitted pathogens and biological agents in potable water is critical to effectively respond to and remediate a contamination event or waterborne outbreak. The Protocol for the Recovery and Concentration of Biological Agents in Water samples, or water processing protocol (WPP), was developed for use during such events. The WPP can be used on either 100-L of water concentrated by ultrafilters in the field or unconcentrated 1-L grab samples. The WPP is pathogen agnostic, as is comprised of two workflows that use size selection to capture and concentrate biological agents from parasites down to toxins. A single-laboratory evaluation was conducted by CDC, followed by a multi-laboratory validation (MLV) study with five state public health laboratories. Prior to the MLV, state laboratory participants received in-person training which was followed by training exercises performed in their laboratories. WPP performance was assessed by percent recovery using REXEED-25S ultrafiltered tap water samples seeded with B. anthracis and MS2 coliphage. A bridging study also evaluated different ultrafilter manufacturers for a robust supply chain during an emergency response. The WPP demonstrated ∼50% recovery efficiency for both targets across all filters. During the bridging study, ELISIO-25H performed slightly better for B. anthracis recovery and participants reported a better user experience with ELISIO-25H compared to Xevonta Hi 23 filters. The WPP serves as an essential method for public health readiness to adequately respond to and remediate water system contamination events and waterborne outbreaks.

Proteinuria is a well-established risk factor for chronic kidney disease progression, and its reduction is associated with improved kidney outcomes. During proteinuric chronic kidney diseases, the abnormal passage of plasma-derived proteins through the damaged glomerular filtration barrier can lead to the activation of various pathophysiological processes in the tubular compartment. These mainly comprise complement activation and excessive proximal tubule reuptake of filtered proteins, such as albumin and associated lipids, which trigger cytotoxic effects through the dysregulation of multiple signaling pathways. Collectively, these signaling events can induce proximal tubular cell apoptosis or the acquisition of injury phenotypes, characterized by the paracrine release of bioactive substances that promote infiltration of immune cells and fibroblast-to-myofibroblast differentiation, eventually contributing to the development of tubulointerstitial fibrosis and progressive loss of kidney function. In this review, we provided an up-to-date overview of the complex mechanisms behind the toxic effects of ultrafiltered proteins on the tubulointerstitium under proteinuric conditions. The mechanistic insights that have been gained recently could be valuable for the future development of new classes of drugs to be used in combination with, or as an alternative to, the currently approved anti-proteinuric treatments in patients who are poorly responsive or unable to tolerate them, respectively.

Fucoidan shows promise for food and therapeutic applications; however, inadequate purification leads to inconsistent composition and over-claimed bioactivity. This study rigorously fractionated eight brown algae species from Hokkaido, Japan, using anion-exchange chromatography to generate three primary fractions (FN1, FN2, and FN3). While fractionation enriched sulfate content, it resulted in a >90% reduction in measured antioxidant activity. Multivariate analysis identified the interaction between sulfate and saccharide contents as the principal determinant of this residual activity. Makombu-derived FN3 was then enzymatically digested, treated with activated charcoal, and ultrafiltered to yield FN3 UF, which contained 48.9% sulfate and negligible glucuronic acid, as determined by high-performance anion-exchange chromatography with pulsed amperometric detection. Overall results suggest that the high activity typically reported is largely extrinsic, driven by co-extracted impurities, whereas the true intrinsic activity is trace (~1 μg/mg). Synchrotron small-angle X-ray scattering revealed that this refinement triggers an expansion of the Bragg distance from 12.6 nm (alginate-rich FN1) to 21.7 nm (sulfated FN3), providing the first direct physical evidence that sulfate-driven electrostatic repulsion governs fucoidan's nanostructural organization in solution. These findings establish a previously undocumented physical baseline, proving that rigorous chemical refinement is a mandatory prerequisite for reliable structure-function evaluations of fucoidan.

Hydrophobic bioactive compounds, such as curcuminoids, β-carotene and long-chain lipids, as well as amphiphilic structural lipids (milk fat globule membrane (MFGM)-associated phospholipids), often exhibit low bioaccessibility due to poor aqueous solubility and/or susceptibility to degradation, which limits their effective use in functional foods. Buttermilk, a dairy byproduct enriched with proteins, lipids and MFGM components, provides a structurally complex, amphiphilic matrix that can enhance micellar solubilization, protect hydrophobic and amphiphilic compounds during digestion and thereby modulate their potential bioavailability. This systematic review and meta-analysis, conducted and reported in accordance with the PRISMA 2020 guidelines, synthesizes quantitative data from in vitro gastrointestinal digestion studies to evaluate the impact of buttermilk and related matrices (e.g., buttermilk yogurt, ultrafiltered buttermilk, and composite nanosystems) on the bioaccessibility of hydrophobic compounds and MFGM phospholipids compared with aqueous or non-buttermilk controls. We identified a limited but growing body of in vitro evidence indicating that buttermilk-based matrices generally increase the intestinal bioaccessibility of curcuminoids, β-carotene, omega-3 fatty acids, vitamin and MFGM phospholipids relative to non-buttermilk systems, with particularly pronounced effects in structured emulsions, yogurts, ultrafiltered buttermilk and MFGM-enriched nanosystems. Rather than a single effect size, the data point to a compound- and matrix-dependent spectrum of improvements, influenced by both the chemical nature of the bioactive and the supramolecular organization of the dairy matrix. Mechanistically, the available findings support a plausible hypothesis that buttermilk enhances bioaccessibility via MFGM-mediated micellar solubilization, interfacial protection against pH- and enzyme-driven degradation and favorable lipid partitioning, although these pathways remain to be confirmed in dedicated mechanistic and in vivo studies. Methodological heterogeneity and the exclusive reliance on in vitro models are important limitations, but overall, the synthesis supports buttermilk and MFGM-rich ingredients as sustainable, food-grade carriers for lipophilic nutraceuticals and highlights the importance of dairy matrix structure in the design of functional delivery systems.

An efficient endolysosomal pathway is crucial to mediate the reabsorption and processing of ultrafiltered solutes including low-molecular-weight (LMW) proteins by epithelial cells lining the proximal tubule (PT) of the kidney. The zebrafish pronephros is used as a model system for congenital or acquired disorders that impair endolysosomal processing in PT cells, causing inappropriate loss of solutes and LMW proteins in urine. Here, we describe a new reporter ½vdbp-NanoLuc zebrafish line, in which vitamin D-binding protein is coupled to NanoLuc luciferase for detection of PT dysfunction and LMW proteinuria. We demonstrate the reliability and value of the ½vdbp-NanoLuc biosensor in fish models of monogenic endolysosomal diseases, gentamicin and cisplatin-induced nephrotoxicity, and metal contamination. This novel reporter system yields mechanistic insights into cadmium- and copper-induced PT dysfunction and provides a platform for drug screening.

Exopolysaccharides (EPS) produced by lactic acid bacteria are increasingly recognized for their structural diversity and functional properties, contributing to food texture and offering potential applications in health, pharmaceuticals, and biotechnology. In this study, we report the multi-gram scale production and characterization of a novel EPS from Lactiplantibacillus pentosus KW1. EPS was produced in a 100 L fed-batch bioreactor using an ultrafiltered semi-defined medium. Proteomic analysis confirmed consistent expression of proteins within the proposed EPS biosynthetic cluster throughout fermentation. The culture underwent heat treatment, two-phase separation, depth filtration, and ultrafiltration for EPS recovery. Size-exclusion chromatography and NMR analysis identified a high-molecular-weight EPS (EPS A, >800 kDa) and two wall teichoic acids with distinct structures. EPS A was obtained at 92 mg/L and characterized as a repeating tetramer composed of pyruvylated galactose (Pyr-Gal), GlcNAc, Gal, and GalNAc. Monosaccharide composition was determined by high-performance anion-exchange chromatography, and structural elucidation was achieved via two-dimensional NMR spectroscopy. All monosaccharides were confirmed as ᴅ-sugars by reversed-phase HPLC. This work provides a foundation for understanding EPS from L. pentosus and its potential role in improving texture and stability in fermented foods.

Antimicrobial resistance can be an indicator of anthropogenic contamination in surface waters and is a potential public health threat. Methodological standardization for characterization of antimicrobial resistance in the environment is lacking. Quantitative PCR (qPCR) is used for rapid assessment of antibiotic resistance genes (ARGs) from environmental sources, including surface water. Here we describe the validation and application of a qPCR assay for 47 bacterial gene targets intended for surface water samples. The qPCR assay displayed excellent sensitivity (97.66%) and specificity (98.71%) for detecting ARGs when compared to whole genome sequencing of bacterial isolates. The qPCR assay was able to detect up to 6/8 (75.0%) of ARGs spiked into sterile water at varying concentrations and four sample ultrafiltration volumes. Nineteen different ARGs were detected across six samples sites at three national parks in Alaska using ultrafiltered surface water samples. The number of unique ARGs detected was higher at sites within parks with greater visitation. The relative abundance of ARGs/16S from Exit Creek in Kenai Fjords National Park, downstream from a visitor center was greater than all other sampled sites. We have demonstrated a robust qPCR assay for monitoring ARGs in surface waters, including those that are minimally human impacted.

Type 2 diabetes mellitus (T2DM) remains a global health challenge, necessitating novel therapeutic and dietary strategies. This study optimized hydroalcoholic extraction parameters to maximize α-amylase inhibitory activity from five African medicinal plants: Combretum glutinosum (CG), Ziziphus mauritiana (ZM), Gymnosporia senegalensis (GS), Boscia senegalensis (BS), and Citrullus colocynthis (CC). A central composite design (CCD) modeled the effects of the liquid-to-solid (L/S) ratio (5-15 mL/g) and ethanol concentration (0-100%, v/v), identifying optimal conditions at low L/S ratios (5 mL/g) and moderate-to-high ethanol concentrations (40-100%) for GS, ZM, and CG, where inhibition levels exceeded 80-98% of α-amylase activity. Extracts from CG, ZM, and GS showed the strongest inhibition (IC50 values of 3.67, 9.8, and 2.25 mg/mL, respectively). Antioxidant capacities, evaluated by DPPH and FRAP assays, correlated strongly with total phenolic content (TPC), with ZM exhibiting superior DPPH (IC50 = 1.94 ± 0.16 mg/mL) and FRAP (IC50 = 4.34 ± 0.52 mg/mL) activities. Incorporation of optimized plant powders (3%, w/v) into bread dough significantly influenced textural and colorimetric properties. Mixture design analysis revealed that CG-rich formulations (>2%) yielding hardness exceed 6 N, while ZM-GS blends maintain 3 N, offering targeted firmness control. The addition of medicinal plants significantly increased the total phenolics content by 60% of doughs and thus caused a significant improvement in antioxidant activities. These functional enrichments suggest potential for developing hypoglycemic bakery products with improved sensory attributes. This integrative approach combining extraction optimization and food formulation offers promising avenues for natural antidiabetic agents and functional food development.