Search Results (58)

Background/Objectives: Rare earth elements (REEs) are emerging environmental pollutants, with human exposure increasing due to recent industrial and technological activities. While most studies have focused on detecting REEs in human fluids, their presence in tissues remains understudied. Aortic valve degeneration is known to facilitate the adsorption of various chemical elements; however, the occurrence of REEs in human valves has not yet been investigated. This exploratory study aimed to determine the presence of REEs in the aortic valves of patients with aortic stenosis undergoing surgical valve replacement. It also analyzed potential correlations between REE levels in the valves, epicardial adipose tissue, serum, and selected disease markers. Methods: Samples of aortic valve, epicardial adipose tissue, and serum were collected from 20 adult patients undergoing elective aortic valve replacement. The concentrations of 14 REEs in these samples were measured using inductively coupled plasma mass spectrometry. Biochemical and clinical parameters of the patients were also considered to explore potential associations with the determined REE levels. Results: Total REEs, heavy REEs, and light REEs in aortic valves, epicardial fat, and serum were not intercorrelated. Moreover, for any sample type, they were not significantly related to the patient’s demographics (age and sex), clinical characteristics (body mass index, heart failure severity, and systolic pressure gradients), kidney function (estimated glomerular filtration rate), and biochemical markers (creatinine, lipoprotein(a), total cholesterol, HDL, LDL, and fibrinogen). Smoking was the only factor influencing REE burden in studied patients, with active smokers revealing 61% higher serum REE concentrations and past smokers exhibiting 133% higher REE valvular deposition. Conclusions: The findings suggest that REE accumulation in aortic valve tissues occurs independently of systemic and clinical parameters but may be promoted by smoking, highlighting the need to investigate the underlying mechanisms of REE deposition. Given the small sample size and the cross-sectional, hypothesis-generating design, these observations should be interpreted with caution and treated as preliminary. Larger, longitudinal studies are needed to validate these results and explore potential causal relationships. Further research should also include the tissue originating from individuals without aortic stenosis for comparison. A deeper understanding of the pathways and health risks associated with REEs in cardiovascular tissues may offer valuable insights into their broader implications for human health. Full article

The effects of cellulase–xylanase synergistic treatment combined with twin-screw extrusion on the physicochemical, functional, and in vitro fermentation characteristics of buckwheat bran dietary fiber (BBDF) were investigated. Compared to single enzymatic hydrolysis, the synergetic modification was more effective in promoting the soluble DF (SDF) ratio (increased from 10.68% to 32.67%), functional properties, and prebiotic activities of BBDF and decreasing the insoluble DF (IDF) content. Under 0.6% (w/w) cellulase and xylanase with mild extrusion conditions (40–80 °C), the modified BBDF exhibited the highest capacities for glucose and cholesterol adsorption. FTIR and XRD experiments indicated that the enzymatic extrusion destroyed the intermolecular interactions of BBDF. Furthermore, enzymatically extruded BBDFs showed 2.2-fold higher short-chain fatty acid (SCFA) yields during in vitro fecal fermentation (total SCFAs: 87.8 mM vs. 40.0 mM in control), with butyrate production reaching 2.5 mM (+76.3%), among which the mildly extruded BBDFs exhibited superior prebiotic effects. Full article

Novel bread formulations with natural improvers have become an essential part of improving the quality of bakery products. In the present study, novel bread improvers made using Gagome kelp cellulose (GC) were systemically evaluated, and cellulose-improved dough (GC-dough), gluten (GC-gluten), and gluten protein and starch (GC-starch) were all studied. The results indicated that the water and oil holding capacity, cholesterol-adsorptive capacity, and the unsaturated fat and saturated fat adsorptive capacities of GC had increased. GC also showed high glucose adsorptive capacity, antioxidant activity, α-amylase inhibition, and glucose diffusion inhibition activity. Furthermore, the color of the GC-dough was improved with the addition of the GC, which also affected the content of glutenin, the water holding capacity in GC-gluten, and the solubility of GC-starch. In addition, the cross-linked network formed by GC could be observed in the GC-bread, indicating an improvement in texture and sensory evaluation. Bread with 1% (m/m) added GC provided the highest sensory characteristics and the best cold storage stability, which suggests that it is the best strategy for further study. The results might show a potential application of by-products of marine origin in commercial bakery production. Full article

Supported lipid bilayers (SLBs) that model neuronal membranes are needed to explore the role of membrane lipids in the misfolding and aggregation of amyloid proteins associated with neurodegenerative diseases, including Parkinson’s and Alzheimer’s disease. The neuronal membranes include not only phospholipids, but also significant amounts of cholesterol, sphingomyelin, and gangliosides, which are critical to its biological function. In this study, we explored the conditions for the formation of an SLB, for the five-component lipid mixture composed of zwitterionic 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), anionic 1,2-dioleoyl- sn-glycero-3-phospho-L-serine (DOPS), nonionic cholesterol (Chol), zwitterionic sphingomyelin (SM), and anionic ganglioside (GM), using the quartz crystal microbalance with dissipation monitoring (QCM-D) technique, by varying experimental parameters such as pH, buffer type, temperature, vesicle size, and osmotic stress. SLB formation from this multicomponent lipid system was found challenging because the vesicles adsorbed intact on the quartz crystal and failed to rupture. For most of the variables tested, other than osmotic stress, we found no or only partial vesicle rupture leading to either a supported layer of vesicles or a partial SLB that included unruptured vesicles. When osmotic stress was applied to the vesicles already adsorbed on the surface, by having a different salt concentration in the rinse buffer that follows vesicle flow compared to that of the dilution buffer during vesicle flow and adsorption, vesicle rupture increased, but it remained incomplete. In contrast, when osmotic stress was applied during vesicle flow and adsorption on the surface, by having different salt concentrations in the dilution buffer in which vesicles flowed compared to the hydration buffer in which vesicles were prepared, complete vesicle rupture and successful formation of a rigid SLB was demonstrated. The robustness of this approach to form SLBs by applying osmotic stress during vesicle adsorption was found to be independent of the number of lipid components, as shown by SLB formation from the 1-, 2-, 3-, 4-, and 5-component lipid systems. Full article

Objectives: The industrial production of influenza vaccines is facing significant challenges, particularly in improving virus production efficiency. Despite advances in cell culture technologies, our understanding of the production characteristics of high-yield suspension cell lines remains limited, thereby impeding the development of efficient vaccine production platforms. This study aims to investigate the key features of STAT1 knockout suspension-adapted MDCK cells (susMDCK-STAT1-KO) in enhancing influenza A virus (IAV) production. Methods: Suspension-adapted susMDCK-STAT1-KO cells were compared to suspension-adapted wild-type MDCK cells (susMDCK) for IAV production. Virus quantification, gene expression analysis, and cholesterol deprivation assays were performed. Metabolite profiles, viral RNA quantification, and lipid and dry weight measurements were also conducted to assess the viral replication and release efficiency. Results: The susMDCK-STAT1-KO cells exhibited significantly improved virus adsorption (64%) and entry efficiency (75%) for the H1N1 virus, as well as accelerated viral transcription and replication for both the H1N1 and H9N2 viruses. Virus release was identified as a limiting factor, with a 100-fold higher intracellular-to-extracellular viral RNA ratio. However, the STAT1-KO cells showed a 2.39-fold higher release rate (750 virions/cell/h) and 3.26-fold greater RNA release for the H1N1 virus compared to wild-type cells. A gene expression analysis revealed enhanced lipid metabolism, particularly cholesterol synthesis, as a key factor in viral replication and release. Cholesterol deprivation resulted in reduced viral titers, confirming the critical role of intracellular cholesterol in IAV production. Conclusions: This study demonstrates the enhanced influenza virus production capacity of susMDCK-STAT1-KO cells, with significant improvements in viral yield, replication, and release efficiency. The findings highlight the importance of STAT1-mediated immune modulation and cholesterol metabolism in optimizing virus production. These insights provide a foundation for the development of more efficient vaccine production platforms, with implications for large-scale industrial applications. Full article

As a small-molecule gelator used as a stabilizer in gel emulsions, it has numerous advantages, such as low dosage, independence from phase ratios, and ease of control. In this study, a cholesterol derivative (CSA) was designed and synthesized to be used as a stabilizer for gel emulsions. Gelation experiments demonstrated that this small molecule could gelate various organic solvents, including linear alkanes, toluene, isoamyl alcohol, and acetone. Based on these gelation experiments, a series of gel emulsions were prepared with water as the dispersed phase and an organic solvent immiscible with water as the continuous phase. Finally, the gelation behavior of the gelator/water/toluene and gelator/water/cyclohexane systems was investigated, exploring the effects of different systems and varying water content within the same system on the structure and stability of the gel emulsions. Studies have shown that the gel emulsion prepared from the gelator/water/toluene system exhibits superior stability, likely due to the molecular self-assembly behavior of this cholesterol derivative exhibited in the water/toluene biphasic system. The research results provide a basis for using gel emulsions as templates to prepare porous materials and adjust their internal structure, ultimately laying a solid foundation for applying these porous materials in fields such as adsorption and catalysis. Full article

Response surface methodology (RSM) was used to optimize the extraction process of naringin. The central component design included three parameters of extraction, namely temperature (X₁), solid–liquid ratio (X₂), and extraction time (X₃). The optimum extraction temperature was 67 °C; the ratio of feed to solvent was 54:1 mL/g, and the extraction time was 2.8 h. According to the best extraction conditions, naringin was processed to verify the accuracy of the model. Five parallel experiments were set up, and a yield of 3.248% naringin was obtained, which was equivalent to the predicted yield of 3.256%. Naringin was purified to obtain naringin-refined products using DM101 macroporous adsorption resin. Naringin dihydrochalcone was synthesized following catalytic hydrogenation of purified naringin. The structures of naringin and naringin dihydrochalcone were determined via Fourier infrared spectrometer and nuclear magnetic resonance spectrometry. In vitro determination of the lipid-lowering activity of naringin dihydrochalcone was also conducted. Further focusing on HepG2 cells, a high cholesterol-induced high-fat HepG2 cell model was established. We measured the effects of different concentrations of naringin dihydrochalcone on intracellular lipids in denatured HepG2 cells and further validated the lipid-lowering effect of naringin at the cellular level. The results showed that naringin dihydrochalcone has a potential application in functional foods for lowering blood lipids. Full article

Amphotericin B (AmB) causes toxicity to the erythrocyte membrane, leading to hemolysis, which limits the clinically effective dose for AmB intravenous therapy in invasive fungal infections. The molecular mechanism by which AmB adheres to the membrane of erythrocytes is the key factor in causing AmB to be toxic to the membrane of erythrocytes, but it is not yet fully understood; the mechanism by which AmB adheres to the liquid microdomains with higher fluidity formed by cholesterol and unsaturated phospholipids remains especially unclear. This study examined the adsorption of AmB at different concentrations, 5, 45, 85, and 125 $\mathsf{\mu }\mathrm{g}/\mathrm{m}\mathrm{L}$, on unsaturated phospholipid membranes containing 50 mol% cholesterol. The thermodynamic properties and structure of DOPC monolayers and DOPC/cholesterol mixed monolayers at different concentrations of AmB have been investigated using the Langmuir monolayer model and the BAM method. The impact of varying concentrations of AmB on the hydrophilic and hydrophobic domains of the DOPC bilayers and the DOPC/cholesterol mixed bilayers have also been discussed using large unilamellar vesicle liposomes and fluorescence techniques. It is shown that for AmB concentrations greater than 5 $\mathsf{\mu }\mathrm{g}/\mathrm{m}\mathrm{L}$, with an increase in AmB’s concentration, the reorganization time for the DOPC/cholesterol monolayer increases, and the elastic modulus of the DOPC/cholesterol mixed monolayer decreases. In particular, when AmB’s concentration is higher than 85$\mathsf{\mu }\mathrm{g}/\mathrm{m}\mathrm{L}$, the liquid-condensed phase domains on the DOPC/cholesterol monolayer reduce significantly and the liquid-expanded phase domain enlarges from the BAM images. When the AmB concentration reaches 5 $\mathsf{\mu }\mathrm{g}/\mathrm{m}\mathrm{L}$, the disorder of the hydrophobic and hydrophilic domains of the DOPC/cholesterol bilayer increases as the AmB concentration increases. The way in which AmB interacts with the DOPC/cholesterol mixed membrane is related to the concentration of AmB. The higher the concentration of AmB, the more likely it is to remove cholesterol from the unsaturated phospholipid membrane. The results are helpful to understand the mechanism of AmB’s toxicity to the erythrocyte’s membrane, which has a guiding value for seeking ways to reduce the AmB’s toxicity. Full article

The incorporation of okara, a by-product of soybean milk production, into gluten-free products such as crispy waffles poses challenges due to the absence of gluten’s viscoelastic properties and the high fiber content of okara. This study aimed to evaluate the effects of okara flour on the rheological properties, physical attributes, and sensory qualities of gluten-free waffles. Waffle batters with varying levels of okara flour (10%, 20%, 30%, and 40%) were prepared, and their rheological properties were analyzed using oscillatory shear and creep-recovery tests. Physical properties, proximate composition, cholesterol and glucose adsorption capacities, storage stability, and sensory attributes were also assessed. The results demonstrated that increasing okara flour content improved batter elasticity and viscosity (with complex viscosity reaching up to 10,923 Pa·s for 40% okara flour) but decreased spread ratio by up to 45% and increased moisture content by approximately 2.7%. Higher okara content also led to a 16% decrease in brightness (L*) and increased hardness, reaching 325.26 g/s at 40% substitution. Sensory evaluation revealed that waffles with 30% okara flour were preferred for their texture and overall liking, with a score of 7.43 compared to higher substitution levels. Cholesterol and glucose adsorption capacities were high in okara flour, contributing to potential health benefits. Storage stability tests showed acceptable moisture content, water activity, and microbiological safety over 60 days, though hardness decreased by about 42%. In conclusion, okara flour enhances the nutritional profile of gluten-free waffles, but its impact on texture and flavor requires careful formulation adjustments to optimize consumer acceptance. Full article

The binding of the HIV-1 Gag polyprotein to the plasma membrane is a critical step in viral replication. The association with membranes depends on the lipid composition, but its mechanisms remain unclear. Here, we report the binding of non-myristoylated Gag to lipid membranes of different lipid compositions to dissect the influence of each component. We tested the contribution of phosphatidylserine, PI(4,5)P2, and cholesterol to membrane charge density and Gag affinity to membranes. Taking into account the influence of the membrane surface potential, we quantitatively characterized the adsorption of the protein onto model lipid membranes. The obtained Gag binding constants appeared to be the same regardless of the membrane charge. Furthermore, Gag adsorbed on uncharged membranes, suggesting a contribution of hydrophobic forces to the protein–lipid interaction. Charge–charge interactions resulted in an increase in protein concentration near the membrane surface. Lipid-specific interactions were observed in the presence of cholesterol, resulting in a two-fold increase in binding constants. The combination of cholesterol with PI(4,5)P2 showed cooperative effects on protein adsorption. Thus, we suggest that the affinity of Gag to lipid membranes results from a combination of electrostatic attraction to acidic lipids, providing different protein concentrations near the membrane surface, and specific hydrophobic interactions. Full article

The convenient and sensitive detection of metabolites is of great significance for understanding human health status and drug development. Solid-phase electrochemiluminescence (ECL) enzyme electrodes show great potential in metabolite detection based on the enzyme-catalyzed reaction product hydrogen peroxide (H₂O₂). Herein, a solid-phase ECL enzyme sensor was fabricated based on a confined emitter and an immobilized enzyme using electrostatic nanocage array, constructing a platform for the sensitive detection of cholesterol. The electrostatic cage nanochannel consists of a bipolar and bilayer vertically aligned mesoporous silica film (bp-VMSF). The upper layer of bp-VMSF is an amino-modified, positively charged VMSF (p-VMSF), and the lower layer is a negatively charged VMSF (n-VMSF). The most commonly used ECL probe tris(bipyridine)ruthenium(II) (Ru(bpy)₃²⁺) is fixed in n-VMSF by electrostatic adsorption from n-VMSF and electrostatic repulsion from the upper p-VMSF, generating significantly enhanced and stable ECL signals. The successful preparation of the electrostatic cage was characterized by scanning electron microscopy (SEM) and electrochemical methods. After amino groups on the outer surface of bp-VMSF were derivatized with aldehyde, cholesterol oxidase (ChOx) molecules were covalently immobilized. The successful construction of the enzyme electrode was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). When the corresponding enzyme substrate, cholesterol, was present in the solution, the ECL signal of Ru(bpy)₃²⁺ was quenched by the enzyme-catalyzed reaction product H₂O₂, enabling the high-sensitivity detection of cholesterol. The linear range for detecting cholesterol was from 0.05 mM to 5.0 mM, with a limit of detection (LOD) of 1.5 μM. Full article

In this study, hot water treatment (WT), ultrasonic treatment (UT), ultrasonic-sodium hydroxide treatment (UST), ultrasonic-enzyme treatment (UET), and ultrasonic-microwave treatment (UMT) were used to treat sweet potatoes. The structural, physicochemical, and functional properties of the extracted soluble dietary fibres (SDFs) were named WT-SDF, UT-SDF, UST-SDF, UET-SDF, and UMT-SDF, respectively. Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), thermal properties, and Brunauer–Emmett–Teller (BET) analysis were employed. The structural results indicated that the UST-SDF exhibited the best thermal stability, highest crystallinity, and maximum specific surface area. Moreover, compared to hot water extraction, ultrasonic extraction, or ultrasonic extraction in combination with other methods, enhanced the physicochemical and functional properties of the SDF, including extraction yield, water-holding capacity (WHC), oil-holding capacity (OHC), glucose adsorption capacity (GAC), glucose dialysis retardation index (GDRI), sodium cholate adsorption capacity (SCAC), cholesterol adsorption capacity (CAC), nitrite ion adsorption capacity (NIAC), and antioxidant properties. Specifically, the UST-SDF and UMT-SDF showed better extraction yield, WHC, OHC, GAC, CAC, SCAC, and NIAC values than the other samples. In summary, these results indicate that UST and UMT could be applied as ideal extraction methods for sweet potato SDF and that UST-SDF and UMT-SDF show enormous potential for use in the functional food industry. Full article

Agrocybe cylindracea dietary fiber (ADF) contains 95% water-insoluble dietary fiber, resulting in poor application performance. To address this issue, ADF was modified by four methods (cellulase, sodium hydroxide, high-temperature, and Lactobacillus fermentation) in this paper. By comparing the physicochemical properties, microstructures, monosaccharide compositions, and functional characteristics (antioxidant and α-glucosidase inhibitory activities in vitro) of all modified ADF samples, the optimal modification method was selected. Results showed that sodium hydroxide treatment was deemed the most effective modification method for ADF, as alkali-treated ADF (ADF-A) revealed a higher oil-holding capacity (2.02 g/g), swelling capacity (8.38 mL/g), cholesterol adsorption (6.79 mg/g), and α-glucosidase inhibitory activity (more than 70% at 0.4–0.6 mg/mL) than the other modified samples. The looser microstructure in ADF-A might be attributed to molecular rearrangement and spatial structure disruption, which resulted in smaller molecular sizes and decreased viscosity, hence improving ADF’s physicochemical and functional qualities. All these findings indicate the greater application potential of modified ADF products in food and weight-loss industries, providing a comprehensive reference for the industrial application of ADF. Full article

Muscadine grapes are characterized by their large and abundant seeds and hard and thick skins that contain significant amounts of dietary fiber (DF). The current study investigated the chemical constituents, molecular architecture, and physicochemical attributes of DF derived from Muscadine grapes (Granny Val and Alachua) and compared them with those derived from Shine Muscat and Kyoho. Using a combined enzymatic method, the total dietary fiber (TDF) was extracted and divided into two parts: soluble dietary fiber (SDF) and insoluble dietary fiber (IDF). TDF (mainly IDF, with a small fraction of SDF) was dominated by cellulose, followed by pectin and hemicellulose. In addition, Granny Val and Alachua had a significantly higher abundance of TDF and IDF compared with Shine Muscat and Kyoho. Moreover, Shine Muscat had significantly the highest abundance of SDF among the four grape varieties. Of note, IDF from Granny Val and Alachua exhibited a complex and dense texture on its surface, and notably outperformed Shine Muscat and Kyoho in terms of cholesterol, fatty acid, heavy metal adsorption, and antioxidant activity. Collectively, Muscadine grapes, i.e., Granny Val and Alachua in the current study, possessed elevated DF levels (predominantly IDF), and their enhanced bioactivity underscored their potential as a potential food ingredient for further use. Full article

Bacterial lipopolysaccharides (LPSs) are important indicators of a bacteria presence in any samples. They can therefore be used for the detection of microbiological contamination in food and dairy products. We performed a comparative analysis of different bacterial models by the application of liposomes containing LPS from Salmonella enterica serotype typhimurium on the surface of an 11-mercaptoundecanoic acid (MUA) monolayer chemisorbed on the gold surface of quartz crystal. Using quartz crystal microbalance with dissipation monitoring (QCM-D), we were able to monitor the formation of the lectin, concanavalin A (ConA), layer on the MUA surface. We determined the optimal concentration of the ConA for the layer formation. ConA of 0.3 mg/mL was selected as the most suitable adsorption of liposomes containing LPS. Using the Sauerbrey equation, we calculated that approximately 1.13 × 10¹² ConA molecules per cm² was adsorbed on the MUA surface, which closely corresponds to the 1.19 × 10¹² molecules per cm² by theoretical models. Later, mixed LPS liposomes containing dipalmitoyl phosphatidyl choline (DPPC), dipalmitoyl phosphatidyl ethanolamine (DPPE) and cholesterol successfully interacted with the ConA layer, which resulted in a decrease in the resonant frequency and an increase in dissipation. We compared the adsorption of liposomes with different fractions of LPS and containing LPS from different bacteria. Lack of any LPS in liposomes caused weaker adsorption on the ConA layer. Liposomes containing 50% LPS caused the most prominent adsorption and were suitable for interaction with DNA aptamers specific to certain LPS. The addition of the aptamers to the surface of ConA covered by LPS-containing liposomes resulted in a decrease in resonant frequency and an increase in the dissipation. Using the Kelvin–Voigt viscoelastic model and multiharmonic response of acoustic sensors, we also determined changes in viscoelastic values of the molecular films during interaction with liposomes and the ConA layer. Full article