Antarctic krill has the largest biomass of any wild animal species on Earth. Krill oil extracted from Antarctic krill is rich in phospholipids， and is a major natural source of marine phospholipids. In marine phospholipids， eicosapentaenoic acid （EPA） and docosahexaenoic acid （DHA） are esterified at the sn-2 position of glycerol. This unique structural configuration endows them with the dual physiological functions of both phospholipids and Ω-3 polyunsaturated fatty acids. Compared with terrestrial phospholipids， marine phospholipids exhibit superior bioactivity in several key aspects， including neurotrophic support， anti-atherosclerotic effects， and lipid metabolism regulation， thereby conferring significant application value in pharmaceuticals， health foods， and functional foods. However， pharmaceutical excipient applications require phospholipid purity above 90%， a standard also required for high-end pharmaceutical formulations and active pharmaceutical ingredients. Currently， commercial krill oil products vary significantly in quality. Their total phospholipid content ranges from only 30% to 60%. Therefore， purifying phospholipids from Antarctic krill is essential. This process holds practical value for improving the quality standards of marine phospholipids. Hydrophilic interaction liquid chromatography （HILIC） separates polar compounds through multiple mechanisms， relying on polar interactions between the hydrophilic headgroups of phospholipids and the stationary phase， as well as partitioning within the water-rich layer on the stationary phase surface. In this study， a HILIC-based method was developed for the purification of phospholipids from Antarctic krill oil. Industrial-scale preparative chromatography requires stable stationary phases. Three HILIC-compatible stationary phases were systematically evaluated. These were unmodified bare silica gel， Diol （3% carbon）， and Diol-H （7% carbon）. Diol showed favorable retention and resolution. Therefore， Diol was selected as the optimal stationary phase. The superior performance of the Diol phase compared to Diol-H suggests that a moderate carbon content is crucial. Excessive hydrophobicity in Diol-H likely interfered with the specific hydrogen bonding interactions required for separating polar lipid headgroups. Ethanol and water were chosen as the mobile phases， because these solvents have low toxicity. Ethanol-water proportions were investigated to develop a three-step elution program： washing with 100% ethanol， elution with 95% ethanol， and column re-equilibration with 80% ethanol. Based on chromatographic plate and rate theories， a Diol column （250 mm×4.6 mm） was used. Isocratic elution with 95% ethanol was applied. The relationship between flow rate， theoretical plate number， and peak width was evaluated. An optimal flow rate was selected. Peak broadening under high sample loading was examined. This determined the maximum loading capacity. It was optimized at 8% of the stationary phase mass. Separation remained efficient at this level. Resolution did not decrease significantly. The final purification protocol involved washing with 2 column volumes of 100% ethanol， eluting the target phospholipid fraction with 5.5 column volumes of 95% ethanol， and re-equilibrating the column with 2.5 column volumes of 80% ethanol. Finally， scale-up separation was performed using a DAC50 column. The crude oil contained 57.88% phospholipids. Four fractions were collected based on online chromatograms. Each fraction was concentrated， dried， and then analyzed. The recovery of the purified product was 49.15%. Purity was measured using the molybdenum blue colorimetric method （GB/T 5537-2008）. Purity reached 95.42%. The overall phospholipid recovery was 81%. EPA and DHA were quantified via external standard method （GB 5009.168-2016）. EPA and DHA contents were 18.5% and 7.18%， respectively. This method efficiently prepares pharmaceutical-grade phospholipids. The process is environmentally friendly and easily scalable for industrial manufacturing. This approach supports the purification of marine phospholipids from Antarctic krill oil and the high-purity concentrate serves as a reference material. It supports clinical research on krill-derived lipids and promotes applications in drug delivery and nutraceuticals. This work improves quality standards for marine lipid products and contributes to the sustainable use of Antarctic krill resources. Preparative liquid chromatography is a mature technology widely used for separating complex samples like drugs and natural products. This study investigated HILIC for purifying phospholipids from Antarctic krill oil. The research provides a technical basis for selecting preparation methods.

The structural and vibrational responses of glycerol-water clusters are closely associated with their local hydrogen-bonding environments. In this work, glycerol-water clusters with 1:1, 1:2, and 1:3 cluster compositions were investigated using density functional theory at the B3LYP/6-311 + G(d,p) level combined with the integral equation formalism polarizable continuum model (IEFPCM). Four continuum dielectric environments with ε = 24.852, 32.613, 35.688, and 46.826 were considered while keeping the explicit glycerol-water compositions unchanged. Optimized structural parameters, simulated Raman spectra, potential-energy-distribution-assisted vibrational assignments, and interaction region indicator (IRI) visualization were jointly analyzed to characterize dielectric-environment-dependent variations in hydrogen bonding. The calculations indicate that changes in the continuum dielectric environment are accompanied by variations in H···O distances, O-H···O angular directionality, and OH stretching frequencies. With increasing hydration number, the vibrational response becomes increasingly influenced by multiple glycerol-water and water-water contacts rather than by a single geometric descriptor. These results provide molecular-level computational insight into the relationship among local hydrogen-bond geometry, electronic characteristics, and Raman response in the investigated glycerol-water cluster models.

Azomycin (2-nitroimidazole), a compound synthesized by Streptomyces eurocidicus, is medically significant as a build block for the drug applied in treatment for drug-resistant tuberculosis and Chagas disease, as well as tumor hypoxia imaging. Despite its clinical value, the optimized bioprocess parameters for its production remain largely unexplored. This study evaluated the influence of various carbon and nitrogen sources (specifically glucose, starch, soybean oil, and casamino acids) on azomycin yields during submerged cultivation. Batch fermentations revealed that azomycin biosynthesis is phase-shifted, initiating only after the primary carbon source is near depletion. Notably, although soybean oil served as an excellent carbon source for biomass accumulation, it yielded no detectable azomycin production. By contrast, supplementing the media with free amino acids (casamino acids) substantially enhanced product synthesis. Using an optimized semi-defined medium of glucose and casamino acids, a titer of 46.34 mg/L was achieved in 250 mL DASbox system. Scaling this configuration to an instrumented 3 L bioreactor resulted in a final concentration of 69.18 mg/L over 7 days, marking an 89% increase in the product-to-biomass yield (Yp/x) compared to the smaller scale tested. Liquid chromatography-mass spectrometry (LC-MS) verified the identity of the extracted 2-nitroimidazole. These findings demonstrate that the precision balancing of glucose and organic nitrogen is essential to unlock and maximize the biosynthetic potential of wild-type S. eurocidicus.

Soybean meal (SBM) and soybean oil (SO) are important aquafeed ingredients, but both are prone to oxidation. However, few studies have compared the effects of oxidized SBM (OSM) and oxidized SO (OSO) in fish. In this study, OSM and OSO were generated by SBM and SO under the same conditions and then used to evaluate their effects on liver antioxidant capacity, inflammation, and apoptosis in Megalobrama amblycephala. A total of 144 fish (initial body weight: 14.37 ± 0.13 g) were randomly divided into three groups and reared for 8 weeks. Both OSM and OSO increased plasma low-density lipoprotein cholesterol (LDL-C) and decreased high-density lipoprotein cholesterol (HDL-C). Oil Red O staining confirmed hepatic lipid accumulation in both groups. The OSO significantly increased plasma MDA levels, while hepatic protein carbonyl (PC) levels were significantly elevated only in the OSM group. Meanwhile, both treatments significantly increased interleukin-1β (IL-1β) levels and decreased transforming growth factor-β (TGF-β) levels. TUNEL assays confirmed increased hepatocyte apoptosis, associated with the downregulation of pi3k, akt, and nf-κb and changes in caspase family gene expression. Notably, OSO upregulated caspase-8 expression, whereas OSM significantly upregulated caspase-9 and caspase-3 expression, suggesting that OSM may be associated with the regulation of mitochondrial apoptosis-related genes. In conclusion, OSM and OSO induce hepatic lipid accumulation, oxidative stress, and inflammation, and the downregulation of pi3k, akt, and nf-κb expression may be associated with hepatocyte apoptosis. Overall, OSM and OSO exhibited different hepatic response patterns.

Plant leaves are not only vital for material exchange with the environment, but also act as primary sites for the deposition, permeation, and action of exogenous functional molecules in agricultural applications such as pest control, nutrient supplementation, and post-harvest processing. Visualizing the spatial distribution of exogenous molecules in plant leaves is of great importance. Traditional methods like chromatography and mass spectrometry may require destroying samples and make it difficult to obtain in situ distribution information. Typical Raman spectroscopy suffers from fluorescence interference and low sensitivity. This study uses tobacco leaves as a model and applies stimulated Raman Scattering (SRS) microscopy combined with Lasso spectral unmixing to achieve label-free, high-resolution, rapid visualization, and semi-quantitative analysis of propylene glycol and glycerol. The results show that SRS can clearly distinguish the characteristic spectra of tobacco, propylene glycol, and glycerol, visually presenting the distribution differences of polyols on the surface and inside the leaf. Besides, the co-localization of the two components and the analysis of their relatively mass ratios are also realized. This method provides a universal technical solution for detecting the distribution of exogenous small molecules in plant leaves.

Hydrogels are promising candidates for flexible and wearable sensors, while the inherent feature of high water content makes them prone to be dehydrated and frozen, leading to the function failure. The existing approaches to improve the lifespan of hydrogel sensors are mainly hindered by complex preparation, limited efficiency, and especially weakened comprehensive performances. In this study, inspired by the separation-synergy mode of the skin, a kind of hydrogel-in-hydrogel sensor was fabricated through a facile in-situ photopolymerization of methacrylated hyaluronic acid. The internal sodium alginate/polyvinyl alcohol hydrogel loaded with Ag nanowires served as the sensor while the external glycerol-water packaging hydrogel played crucial roles in freezing resistance and water retention. Benefiting from the moderate sensing sensitivity and conformal adhesion on human skin, the hydrogel-in-hydrogel sensor was effective in monitoring of human motions and transmission of Morse code. More importantly, this sensor was demonstrated to be reliable in low-temperature environments (-20 °C, > 6 h) and at room temperature (up to 150 days), significantly enhancing its service life. This work provides a new paradigm for developing environment-tolerant and durable hydrogel sensors.

Commercial food packaging materials are mainly synthesized from non-renewable petroleum-based polymers. Starch-based bioplastics have been developed as an alternative material since they are renewable and environmentally friendly. In this research, composite films derived from sago starch as matrix and microcrystalline cellulose (MCC) as filler with the addition of glycerol as plasticizers have been prepared. Ginger extract was also added as an antioxidant agent. The effect of ultrasonication at different time durations of MCC on the physicochemical properties of the composite film was investigated. Ultrasonication broke down the agglomerated MCC into smaller particle sizes and improved the dispersion in the starch matrix, as shown in the FESEM photograph. The mechanical properties were enhanced after ultrasonication. In addition, the ultrasonicated composite films showed better barrier properties toward water vapor compared to the untreated film. In addition, the developed antioxidant film containing ginger extract exhibited a great inhibition toward browning and shrinking on sliced apples during storage at ± 4 °C in a cooling room for 10 days. Overall, the composite films were biodegraded in two weeks. These biodegradable composite films can be a promising alternative for food-grade and eco-friendly packaging materials.

Vaping, often advertised as a safe alternative to traditional tobacco smoking, has recently been subjected to active scientific research due to its potential adverse effects on human health. This narrative review synthesises selected recent literature to provide a comprehensive view of the systemic risks associated with the use of electronic cigarettes. It lists key chemical components (propylene glycol, vegetable glycerine, nicotine, and flavourings) and thermal decomposition products of vape aerosol (formaldehyde, acrolein, propylene oxide, glycidol) and evaluates their multi-systemic toxic effects and clinical manifestations, as well as specific effects on the respiratory, cardiovascular, immune, and nervous systems. Current data suggest that vaping can lead to a wide range of acute and chronic diseases, including the e-cigarette or vaping-associated lung injury (EVALI) and obliterative bronchiolitis and increase the risk of developing oncological, cardiovascular, and neurological diseases. Particularly sensitive is the developing adolescent brain. A growing body of evidence indicates that vaping is not "less harmful" but merely "differently" harmful and calls for further long-term research and strengthened public health regulation.

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are n-3 polyunsaturated fatty acids with EPA receiving increasing attention due to its documented beneficial effects on cardiovascular and cerebrovascular health. In this study, an effective enzymatic transesterification employing multi-step addition of water was developed to produce EPA-enriched glycerides from glycerol and fatty acid ethyl esters. Lipozyme 435 showed the best performance in EPA-enriched glyceride synthesis. Furthermore, multi-step water addition significantly enhanced glyceride synthesis efficiency, and the underlying mechanism was investigated using molecular dynamics simulations. Under the optimized conditions (65 °C, 6 h, 2.0 mbar, an ethyl ester to glycerol molar ratio of 3.5:1, and a lipase dosage of 6%), water was added at 2 h and 4 h, with 5% (w/w) relative to the mass of ethyl esters. The triacylglycerol (TAG) content in the product reached 82.31%, while the EPA and DHA contents in the glyceride fraction were 60.50% and 20.57%, respectively. A novel finding of this study is that the catalytic performance of the lipase in the transesterification is significantly influenced by the multi-step addition of water under vacuum.

Bicontinuous bigels are promising solid fat replacers, but their inherent thermodynamic metastability easily causes phase separation and structural collapse. Existing strategies mostly rely on passive adjustment of phase volume ratios or single gelator concentrations. To address this limitation, this study proposed an active regulation strategy targeting the aqueous phase microenvironment to achieve controllable and stable bicontinuous phases. With Zein-gelatin (GA) hydrogels and beeswax-glycerol monostearate (GMS)-medium chain triglyceride (MCT) oleogels as materials, the regulatory effects of Zein-to-GA mass ratios (0:10 to 8:2) coupled with dynamic oil-to-water ratios (4:6 to 6:4) on bigel structure and properties were systematically investigated. Results demonstrated that the introduction of Zein endowed bigels with a uniform interpenetrating bicontinuous microstructure. At a 4:6 Zein-to-GA mass ratio and a 5:5 oil-to-water ratio, the system exhibited excellent mechanical and rheological properties via the synergy of hydrogen bonds, hydrophobic interactions and covalent cross-linking. The hardness (1006.93 g), cohesiveness (0.87), gumminess (874.70), water-holding capacity (WHC, 79.63%), and oil-holding capacity (OHC, 82.63%) all peaked. After application in reduced-fat chocolate sauce, the total fat content (TFC) decreased to 22.46% with a fat reduction rate (FRR) of 49.81%, and the product exhibited excellent spreadability. After 30 days of storage, the peroxide value (PV) was 0.62 meq/kg and the thiobarbituric acid reactive substances (TBARS) value was 1.89 mg/kg, indicating good lipid oxidation stability. This study confirms that aqueous phase microenvironment regulation is an effective strategy to stabilize bicontinuous bigels, providing theoretical support for their reduced-fat food applications.

Emerging technologies have been widely applied to modify complex protein systems; however, their effects are generally evaluated at the whole-protein level, limiting the understanding of the mechanisms governing individual protein fractions. Soy protein functionality is primarily determined by the β-conglycinin (7S) and glycinin (11S) fractions, yet their distinct responses to ohmic heating and consequent interfacial performance remain poorly understood. In this study, isolated 7S and 11S fractions were subjected to ohmic heating under different electric field strengths (control, 35, 40, 45, and 50 V∙cm[-1]) and evaluated for electrothermal properties, secondary structure, free sulfhydryl groups, surface hydrophobicity, ζ-potential, particle size, solubility, interfacial adsorption, and emulsion stability using 1-octanol, medium-chain triglycerides (MCT), and n-octane as oil phases. Ohmic heating induced fraction-dependent structural changes by modulating protein unfolding and aggregation. The 7S fraction showed greater conformational sensitivity, with increased free sulfhydryl content, α-helix to β-sheet conversion, and particle growth from approximately 0.4 to 2.7 μm, whereas 11S maintained comparatively stable soluble protein levels across most ohmic-heating conditions. These structural changes translated into distinct interfacial behaviors depending on the oil phase. MCT provided the most favorable balance between adsorption kinetics, emulsion stability, and rheological structuring, particularly for the 11S fraction treated at intermediate electric field strengths, whereas n-octane promoted stronger hydrophobic interactions and flocculation, and 1-octanol produced less structured emulsions. These findings demonstrate that ohmic heating can selectively tailor the structural and interfacial functionality of soybean protein fractions, providing new insights for designing protein ingredients with targeted emulsifying properties.

A coupled physics-based model is developed to describe secondary breakup of levitating droplets in a high-intensity ultrasonic standing wave field. The model uniquely integrates acoustic radiation pressure-induced droplet deformation, parametric excitation of capillary waves, radial oscillations and collective interaction of cavitation bubbles, and energy accumulation in the system. The methodology involves simultaneous numerical solution of the Mathieu-Hill equation, the Rayleigh-Plesset equation with inter-bubble pressure coupling, and energy conservation principles. A critical gas volume fraction α∼cr∼≈ 0.35-0.38 is identified as the bifurcation threshold between two distinct breakup regimes. Below this value, capillary-wave fragmentation dominates, yielding a fragmentation time of ∼250-300 ms and a minimum droplet size limited by the capillary wavelength (∼24.7 μm for water at 22 kHz). Above the threshold, collective bubble collapse prevails, reducing fragmentation time to ∼10-15 ms and producing droplets an order of magnitude smaller (∼1 μm), as predicted by a Weber-criterion formulation incorporating bubble wall collapse velocity. Analytical expressions for threshold acoustic pressures required to achieve minimum droplet size are derived and validated numerically. For thin disk-shaped droplets (kh<< 1, relevant for droplets < 0.5 mm), the critical excitation pressure is frequency-independent and can reach 84 kPa, whereas for larger droplets it ranges between 15-19 kPa at 22-44 kHz. Predicted final droplet radii for water (28 μm), glycerol (22 μm), and ethanol (18 μm) at 35 kPa and 22 kHz agree quantitatively with experimental data. The proposed model offers a predictive framework for controlling aerosol dispersity by tailoring gas content, initial droplet size, and acoustic field parameters, thereby enabling energy-efficient, contactless atomization technologies with tunable droplet characteristics.

Diacylglycerol (DAG) are functional lipids with benefits including weight reduction and improved blood lipid profiles. However, enzymatic synthesis optimization is constrained by the rigid design of conventional response surface methodology (RSM). This study experimentally implemented and evaluated an Active-Learning Sequential Surrogate-Based Optimization workflow for sunflower oil DAG synthesis. AL-SSBO sequentially incorporated replicate measurements under recommended conditions and achieved a DAG content of 53.77% after five predefined optimization rounds. The AL-SSBO yielded a prediction error of only 0.10 percentage points, compared with 1.43 for RSM. Bootstrap-based inference supports the observed performance advantage of AL-SSBO over RSM. Optimal conditions were 11.93 h, 68.56 °C, 8.93 wt% lipase loading, and an oil-to-glycerol weight ratio of 11.71. Analysis of exploration-exploitation behavior and spatial sampling showed that the recommended conditions became increasingly concentrated near a high-DAG region, consistent with retrospective stabilization within the investigated domain.

Edwardsiella piscicida is a devastating bacterial pathogen in aquaculture, and in-depth elucidation of its pathogenic mechanisms is essential for establishing effective control strategies; the identification of novel regulatory factors can deepen our understanding of bacterial pathogenesis. Our previous work identified the functionally uncharacterized protein ETAE_2516 as a potential target of the response regulator CpxR of the CpxRA two-component system, a key virulence regulator in this pathogen. Bioinformatic analysis and functional prediction indicated that ETAE_2516 is an AraC-family transcription factor that is potentially involved in glycerol metabolism; it was therefore designated GlvR (Glycerol-linked Virulence Regulator). In this study, an in-frame glvR deletion mutant (ΔglvR) and its complemented strain (ΔglvRC) were constructed by gene knockout, and their phenotypes in metabolism, stress resistance, biofilm formation, and pathogenicity were systematically characterized. Deletion of glvR did not affect basal bacterial growth but significantly increased sensitivity to oxidative stress and led to intracellular accumulation of reactive oxygen species. The ΔglvR mutant exhibited markedly impaired growth in glycerol-supplemented medium, and the expression of glycerol metabolism-associated genes was significantly downregulated, confirming that GlvR positively regulates glycerol metabolism. Loss of GlvR also repressed the expression of flhC, the master regulator of the flagellar regulon, thereby impairing bacterial motility. Infection assays using tilapia epithelial (TSE-04) cells, RAW 264.7 macrophages, and a tilapia (Oreochromis niloticus) infection model demonstrated that GlvR is essential for bacterial adhesion to host epithelial cells, resistance to host serum killing, intracellular survival within macrophages, and tissue colonization, and all of these defects were restored to wild-type levels in the complemented strain, verifying that they are specifically attributable to the loss of glvR. Analysis of the upstream regulatory mechanism further revealed that CpxR binds directly to the glvR promoter region and positively activates its transcription. Collectively, these findings demonstrate that the CpxR-GlvR regulatory axis functions as a central hub integrating carbon metabolism, oxidative stress adaptation, and virulence expression, thereby providing a novel perspective for understanding the host adaptation mechanisms of E. piscicida and a potential target for anti-virulence strategies against edwardsiellosis.