ITPRIPL1 binds CD3ε to impede T cell activation and enable tumor immune evasion.

Cancer immunotherapy has transformed treatment possibilities, but its effectiveness differs significantly among patients, indicating the presence of alternative pathways for immune evasion. Here, we show that ITPRIPL1 functions as an inhibitory ligand of CD3ε, and its expression inhibits T cells in the tumor microenvironment. The binding of ITPRIPL1 extracellular domain to CD3ε on T cells significantly decreased calcium influx and ZAP70 phosphorylation, impeding initial T cell activation. Treatment with a neutralizing antibody against ITPRIPL1 restrained tumor growth and promoted T cell infiltration in mouse models across various solid tumor types. The antibody targeting canine ITPRIPL1 exhibited notable therapeutic efficacy against naturally occurring tumors in pet clinics. These findings highlight the role of ITPRIPL1 (or CD3L1, CD3ε ligand 1) in impeding T cell activation during the critical "signal one" phase. This discovery positions ITPRIPL1 as a promising therapeutic target against multiple tumor types.

Recent advancements in the utilization of ultrasonic technology for the curing of processed meat products: A comprehensive review.

Curation meat products involves multiple stages, including pre-curing processing (thawing, cleaning, and cutting), curing itself, and post-curing processing (freezing, and packaging). Ultrasound are nonthermal processing technology widely used in food industry. This technology is preferred because it reduces the damages caused by traditional processing techniques on food, while simultaneously improving the nutritional properties and processing characteristics of food. The utilization of ultrasonic-assisted curing technology has attracted significant attention within the realm of meat product curing, encouraging extensive research efforts. In terms of curing meat products, ultrasonic-assisted curing technology has been widely studied due to its advantages of accelerating the curing speed, reducing nutrient loss, and improving the tenderness of cured meats. Therefore, this article aims to comprehensively review the application and mechanism of ultrasound technology in various stages of meat product curing. Furthermore, it also elaborates the effects of ultrasonic-assisted curing on the tenderness, water retention, and flavor substances of the meat products during the curing process. Besides, the implication of the ultrasound in the processing of meat curation plays a potent role together with other technologies or methods. The use of ultrasound technology in the process of meat curation was analyzed, which might be a theoretical insight for the industrialization prospects of the meat product.

Discovery of galectin-8 as an LILRB4 ligand driving M-MDSCs defines a class of antibodies to fight solid tumors.

LILRB4 is an immunosuppressive receptor, and its targeting drugs are undergoing multiple preclinical and clinical trials. Currently, the absence of a functional LILRB4 ligand in solid tumors not only limits the strategy of early antibody screening but also leads to the lack of companion diagnostic (CDx) criteria, which is critical to the objective response rate in early-stage clinical trials. Here, we show that galectin-8 (Gal-8) is a high-affinity functional ligand of LILRB4, and its ligation induces M-MDSC by activating STAT3 and inhibiting NF-κB. Significantly, Gal-8, but not APOE, can induce MDSC, and both ligands bind LILRB4 noncompetitively. Gal-8 expression promotes in vivo tumor growth in mice, and the knockout of LILRB4 attenuates tumor growth in this context. Antibodies capable of functionally blocking Gal-8 are able to suppress tumor growth in vivo. These results identify Gal-8 as an MDSC-driving ligand of LILRB4, and they redefine a class of antibodies for solid tumors.

The effects of different hydrocolloids on lotus root starch gelatinization and gels properties.

In this study, xanthan gum (XG), sodium alginate (SA), guar gum (GG), and gum Arabic (GA), were used to modify Lotus root starch (LRS). The incorporation XG, SA, and GG significantly (p < 0.05) influence the swelling power (SP) of LRS, among which the 1.5 % of XG exhibited the highest value of 25.84 g/g at 90 °C. Gelatinization analysis revealed that XG raised the final viscosity (FV) and lowered the breakdown (BD), while SA significantly increased peak viscosity (PV) and BD. Furthermore, GG and GA exhibited a substantial reduction in setback (SB). The incorporation of XG, SA, and GG enhanced the rheological and structural properties (e.g., gel strength and elasticity) of LRS. Particularly, XG demonstrated a more prominent effect, while GA exhibited an opposite trend. Moreover, the structural analyses revealed that hydrophilic colloids have no impact on the functional group and crystal structure of the LRS. However, complex system exhibited the more stable hydrogen bonding. The addition of 1.5 % XG exhibited the most stable hydrogen bonding and highest water binding affinity. Overall, the results demonstrated the effect of different hydrophilic colloids on LRS, offering a theoretical basis for LRS applications and novel insights for the use of starches and hydrocolloids.

A comprehensive review of ultrasonic assisted extraction (UAE) for bioactive components: Principles, advantages, equipment, and combined technologies.

The increasing focus on health and well-being has sparked a rising interest in bioactive components in the food, pharmaceutical, and nutraceutical industries. These components are gaining popularity due to their potential benefits for overall health. The growing interest has resulted in a continuous rise in demand for bioactive components, leading to the exploration of both edible and non-edible sources to obtain these valuable substances. Traditional extraction methods like solvent extraction, distillation, and pressing have certain drawbacks, including lower extraction efficiency, reduced yield, and the use of significant amounts of solvents or resources. Furthermore, certain extraction methods necessitate high temperatures, which can adversely affect certain bioactive components. Consequently, researchers are exploring non-thermal technologies to develop environmentally friendly and efficient extraction methods. Ultrasonic-assisted extraction (UAE) is recognized as an environmentally friendly and highly efficient extraction technology. The UAE has the potential to minimize or eliminate the need for organic solvents, thereby reducing its impact on the environment. Additionally, UAE has been found to significantly enhance the production of target bioactive components, making it an attractive method in the industry. The emergence of ultrasonic assisted extraction equipment (UAEE) has presented novel opportunities for research in chemistry, biology, pharmaceuticals, food, and other related fields. However, there is still a need for further investigation into the main components and working modes of UAEE, as current understanding in this area remains limited. Therefore, additional research and exploration are necessary to enhance our knowledge and optimize the application of UAEE. The core aim of this review is to gain a comprehensive understanding of the principles, benefits and impact on bioactive components of UAE, explore the different types of equipment used in this technique, examine the various working modes and control parameters employed in UAE, and provide a detailed overview of the blending of UAE with other emerging extraction technologies. In conclusion, the future development of UAEE is envisioned to focus on achieving increased efficiency, reduced costs, enhanced safety, and improved reliability. These key areas of advancement aim to optimize the performance and practicality of UAEE, making it a more efficient, cost-effective, and reliable extraction technology.

miR-100-5p activation of the autophagy response through inhibiting the mTOR pathway and suppression of cerebral infarction progression in mice.

In recent years, the association between microRNAs (miRNAs) and autophagy in cerebral infarction (CI) has attracted increasingly more attention. The mammalian target of the rapamycin (mTOR) pathway is a key protein regulating the autophagy response. miR-100-5p can bind to the mTOR protein, but its role in CI remains unclear yet. This experiment aims to clarify the role of miR-100-5p in CI. Bioinformatics analysis was performed to screen differentiated expressed functional genes between CI tissue and normal tissue specimens. In vivo experiments: the mouse model of CI was established by middle cerebral artery occlusion (MCAO) methods, After being treated with miR-100-5p-overexpressing lentivirus, the amount of terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL)-positive fluorescence and the fluorescent expression level of mTOR protein were significantly inhibited in the CI region. Western blotting analysis showed that miR-100-5p inhibited the protein expression level of phosphorylated mTOR and total mTOR and enhanced the expression of autophagy-related proteins Beclin, microtubule-associated protein light chain 3II (LC-3II), and autophagy-related gene 7 (ATG-7). For in vitro experiment, after the BV-2 cells were successfully infected with the control lentivirus and miR-100-5p-overexpression lentivirus, they were stimulated with 1% hypoxia and low-glucose medium in a tri-gas incubator for 24 h. It was found that miR-100-5p could significantly lower the protein expression level of phosphorylated mTOR and total mTOR, and increase the expression of the Beclin, LC-3II, ATG-7 autophagy related proteins. miR-100-5p promotes the autophagy response through binding to mTOR protein, thereby inhibiting apoptosis and delaying the progression of CI.

Ethanol as a switch to induce soybean lipophilic protein self-assembly and resveratrol delivery.

Protein-based nanoparticles or nanocarriers of emulsion systems have piqued the interest of nutrition and health care goods. As a result, this work examines the characterisation of ethanol-induced soybean lipophilic protein (LP) self-assembly for resveratrol (Res) encapsulation, particularly the influence on emulsification. By varying the ethanol content ([E]) in the range of 0-70% (v/v), the structure, size, and morphology of LP nanoparticles may be adjusted. Similarly, the self-assembled LPs have a strong [E] dependency on the encapsulation efficiency of Res. For [E] = 40% (v/v), Res had the highest encapsulation efficiency (EE) and load capacity (LC) of 97.1% and 141.0 µg/mg nanoparticles, respectively. Most of the Res was encapsulated by the hydrophobic core of LP. Moreover, for [E] = 40% (v/v), LP-Res showed significantly improved emulsifying properties, independent of low-oil or high-oil emulsion systems. Furthermore, the ethanol-induced production of appropriate aggregates increased emulsion system stability, hence increasing Res retention during storage.

Galectin-8 alters immune microenvironment and promotes tumor progression.

Galectin-8 (Gal-8), encoded by LGALS8 gene, is a unique member of the Galectin family with diverse biological functions, including tumor-modulating capabilities. Recently, evidence has accumulated supporting an essential role for Gal-8 in regulating innate and adaptive immunity, with high expression in tumors and other immune dysregulation diseases. This study reveals the role of Gal-8-induced tumor immunosuppression by analyzing animal models and clinical data of tumor-infiltrating cells. In Gal-8 expressing tumor, we found that suppressive immune cells, including Tregs and MDSCs, expanded while CD8+ cells decreased, providing direct evidence that Gal-8 regulates the tumor immune microenvironment. In addition, we not only analyzed the expression of Gal-8 in clinical samples of breast and colorectal cancer but also classified the tissue expression patterns. Further analysis revealed that Gal-8 correlates with lymph node metastasis and immunophenotyping. Consistent with animal experiments, our analysis of LGALS8 gene expression showed its negative association with infiltrated active CD8+ T cells and immune stimulatory modulators in cancers. Our study identified the potential prognostic and therapeutic value of Gal-8, and further research on developing corresponding targeted therapeutic strategies is awaited.

The novel amylase function of the carboxyl terminal domain of Amy63.

α-amylase plays a crucial role in regulating metabolism and health by hydrolyzing of starch and glycogen. Despite comprehensive studies of this classic enzyme spanning over a century, the function of its carboxyl terminal domain (CTD) with a conserved eight ß-strands is still not fully understood. Amy63, identified from a marine bacterium, was reported as a novel multifunctional enzyme with amylase, agarase and carrageenase activities. In this study, the crystal structure of Amy63 was determined at 1.8 Å resolution, revealing high conservation with some other amylases. Interestingly, the independent amylase activity of the carboxyl terminal domain of Amy63 (Amy63_CTD) was newly discovered by the plate-based assay and mass spectrometry. To date, the Amy63_CTD alone could be regarded as the smallest amylase subunit. Moreover, the significant amylase activity of Amy63_CTD was measured over a wide range of temperature and pH, with optimal activity at 60 °C and pH 7.5. The Small-angle X-ray scattering (SAXS) data showed that the high-order oligomeric assembly gradually formed with increasing concentration of Amy63_CTD, implying the novel catalytic mechanism as revealed by the assembly structure. Therefore, the discovery of the novel independent amylase activity of Amy63_CTD suggests a possible missing step or a new perspective in the complex catalytic process of Amy63 and other related α-amylases. This work may shed light on the design of nanozymes to process marine polysaccharides efficiently.

Ultrasound-assisted alkali removal of proteins from wastewater generated during oil bodies extraction.

In this study, an ultrasonic-assisted alkaline method was used to remove proteins from wastewater generated during oil-body extraction, and the effects of different ultrasonic power settings (0, 150, 300, and 450 W) on protein recovery were investigated. The recoveries of the ultrasonically treated samples were higher than those of the samples without ultrasonic treatment, and the protein recoveries increased with increasing power, with a protein recovery of 50.10 % ± 0.19 % when the ultrasonic power was 450 W. Amino acid analysis showed that the amino acids comprising the recovered samples were consistent, regardless of the ultrasonic power used, but significant differences in the contents of amino acids were observed. No significant changes were observed in the protein electrophoretic profile using dodecyl polyacrylamide gel, indicating that sonication did not change the primary structures of the recovered samples. Fourier transform infrared and fluorescence spectroscopy revealed that the molecular structures of the samples changed after sonication, and the fluorescence intensity increased gradually with increasing sonication power. The contents of α-helices and random coils obtained at an ultrasonic power of 450 W decreased to 13.44 % and 14.31 %, respectively, whereas the ß-sheet content generally increased. The denaturation temperatures of the proteins were determined using differential scanning calorimetry, and ultrasound treatment reduced the denaturation temperatures of the samples, which was associated with the structural and conformational changes caused by their chemical bonding. The solubility of the recovered protein increased with increasing ultrasound power, and a high solubility was essential in good emulsification. The emulsification of the samples was improved well. In conclusion, ultrasound treatment changed the structure and thus improved the functional properties of the protein.

Expression patterns of ABCE model genes during flower development of melon (Cucumis melo L.).

In production, most cultivars of melon are andromonoecious and characterized by carrying both male and bisexual flowers on the same plant. In this study, four A-class genes (CmAP1a, CmAP1b, CmAP2a and CmAP2b), two B-class genes (CmAP3 and CmPI), two C-class genes (CmAGa and CmAGb) and four E-class genes (CmSEP1,2,3,4) were identified in melon. However, no D-class gene of melon was identified. The conserved domains of ABCE function proteins showed relatively high similarity between Arabidopsis and melon. The expression patterns of ABCE homeotic genes in different flower buds of melon suggested that transcripts of CmAP1a, CmPI and CmSEP1 in bisexual buds were significantly lower than that in male flower buds, while the expression levels of CmAGa, CmAGb and CmSEP4 in bisexual flower buds were significantly higher than that in male flower buds. There was no significant difference in expression levels of other ABCE model genes between male buds and bisexual buds. Subsequently, qRT-PCR was performed in different floral organs of bisexual flowers in melon. For A class genes, CmAP1a and CmAP1b showed the highest accumulation in sepals than petals, stamens and pistil, while CmAP2a and CmAP2b revealed the highest expression in pistil than other three floral organs. For B class genes, CmAP3 and CmPI were highly accumulated in petals and stamens though CmAP3 also showed abundant accumulation in pistil. For C class genes, the expression levels of CmAGa and CmAGb were higher in stamens and pistil than that in sepals and petals. For E class genes, CmSEP1 showed higher expression level in sepals and petals than stamens and pistil. CmSEP2, CmSEP3 and CmSEP4 showed the highest accumulation in pistil than other floral organs. These results provided a theoretical basis for studying the function of ABCE homeotic genes in floral organs development of melon.

A Dual-Function Wearable Electrochemical Sensor for Uric Acid and Glucose Sensing in Sweat.

Simultaneous detection of uric acid and glucose using a non-invasive approach can be a promising strategy for related diseases, e.g., diabetes, gout, kidney disease, and cardiovascular disease. In this study, we have proposed a dual-function wearable electrochemical sensor for uric acid and glucose detection in sweat. The sensor with a four-electrode system was prepared by printing the ink on a common rubber glove. CV and chronoamperometry were used to characterize the prepared sensor's electrochemical sensing performance. The sensors exhibited the linear range from 0 to 1.6 mM and 0 to 3.7 mM towards uric acid and glucose electrochemical sensing in phosphate-buffered solution, with the corresponding limit of detection of 3.58 µM and 9.10 µM obtained, respectively. Moreover, the sensors had shown their feasibility of real sample sensing in sweat. The linear detection range for uric acid (0 to 40 µM) and glucose (0 to 1.6 mM) in the sweat can well cover their concentration range in physiological conditions. The prepared dual-function wearable electrochemical sensor features easy preparation, fast detection, high sensitivity, high selectivity, and the practical application potential in uric acid and glucose sensing.

Highly biologically active and pH-sensitive collagen hydrolysate-chitosan film loaded with red cabbage extracts realizing dynamic visualization and preservation of shrimp freshness.

Accurate and efficient detection of food freshness is of great significance to guarantee food safety. Herein, pH sensitive colorimetric films with considerable biological activities have been prepared by combining red cabbage anthocyanin extracts (RCE) with collagen hydrolysate-chitosan (CH-CS) matrix film. The formation mechanism of CH-CS-RCE films was discussed by SEM, FT-IR and XRD, which showed that RCE was successfully fixed in CH-CS film through hydrogen bonding and electrostatic interaction. The CH-CS-RCE films exhibited good mechanical properties, high barrier ability, excellent thermal stability, significant antioxidant and antimicrobial activity, and especially sensitive response to pH and ammonia. Fickian diffusion was the main mechanism for the release of RCE from CH-CS-RCE films and such release mechanism facilitated the maintenance of functional features of films. During the storage of shrimps at 4 °C, CH-CS-RCE2% showed a remarkable preservation effect on shrimps, and their shelf life was prolonged from 2 d to 5 d. Furthermore, CH-CS-RCE2% provided a dynamic visual color switching to detect the freshness of shrimp, realizing real-time monitoring of freshness. Color information (RGB) extracted via smartphone APP was used to enhance the accuracy and universality of freshness indication. Thus, this multifunctional film has great potential in food preservation and freshness monitoring.

Dithiothreitol-induced reassembly of soybean lipophilic protein as a carrier for resveratrol: Preparation, structural characterization, and functional properties.

We employed dithiothreitol (DTT) to reassemble soy lipophilic protein (LP) and increased its solubility for encapsulating resveratrol (Res); we subsequently added hydroxypropyl methylcellulose (HPMC) to further stabilize Res. Physicochemical characterization, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and spectral analysis revealed that DTT triggered the breakage and reassembly of the disulfide bond. Consequently, the solubility of LP increased from 38.64 % to 71.49 %, and the number of free sulfhydryl groups increased to 7.84 mol·g-1. Furthermore, the encapsulation efficiency and structure of reassembled LP nanoparticles loaded with Res were found to be closely related to the DTT concentration used for induction. When HPMC was added, the LP-Res complex demonstrated spontaneous self-assembly, and the pH and temperature stability of the Res in the nanoparticles improved. An in vitro digestion simulation revealed that the reassembled LP was an efficient carrier for Res delivery. Particularly, HPMC improved the bioavailability and sustained release of Res.

Development of a monoclonal antibody to ITPRIPL1 for immunohistochemical diagnosis of non-small cell lung cancers: accuracy and correlation with CD8+ T cell infiltration.

Introduction: Cancer biomarkers are substances or processes highly associated with the presence and progression of cancer, which are applicable for cancer screening, progression surveillance, and prognosis prediction in clinical practice. In our previous studies, we discovered that cancer cells upregulate inositol 1,4,5-triphosphate receptor-interacting protein-like 1 (ITPRIPL1), a natural CD3 ligand, to evade immune surveillance and promote tumor growth. We also developed a monoclonal ITPRIPL1 antibody with high sensitivity and specificity. Here, we explored the application of anti-ITPRIPL1 antibody for auxiliary diagnosis of non-small cell lung cancer (NSCLC). Methods: NSCLC patient tissue samples (n = 75) were collected and stained by anti-ITPRIPL1 or anti-CD8 antibodies. After excluding the flaked samples (n = 15), we evaluated the expression by intensity (0-3) and extent (0-100%) of staining to generate an h-score for each sample. The expression status was classified into negative (h-score < 20), low-positive (20-99), and high-positive (≥ 100). We compared the h-scores between the solid cancer tissue and stroma and analyzed the correlation between the h-scores of the ITPRIPL1 and CD8 expression in situ in adjacent tissue slices. Results: The data suggested ITPRIPL1 is widely overexpressed in NSCLC and positively correlates with tumor stages. We also found that ITPRIPL1 expression is negatively correlated with CD8 staining, which demonstrates that ITPRIPL1 overexpression is indicative of poorer immune infiltration and clinical prognosis. Therefore, we set 50 as the cutoff point of ITPRIPL1 expression H scores to differentiate normal and lung cancer tissues, which is of an excellent sensitivity and specificity score (100% within our sample collection). Discussion: These results highlight the potential of ITPRIPL1 as a proteomic immunohistochemical NSCLC biomarker with possible advantages over the existing NSCLC biomarkers, and the ITPRIPL1 antibody can be applied for accurate diagnosis and prognosis prediction.

Structure, rheology, and functionality of emulsion-filled gels: Effect of various oil body concentrations and interfacial compositions.

The purpose of this study is to investigate the impact of varied oil body (OB) concentrations and interfacial compositions on the network topology and rheological and functional aspects of composite whey protein isolate (WPI) gels. Particle size and ζ-potential analyzes of the mixed gel solutions containing the OBs extracted at pH 6.8 (6.8-OB) and 11.0 (11.0-OB) revealed a greater aggregation in the 6.8-OB-containing mixed gel solution. 6.8-OB and 11.0-OB generated particle aggregates and oil-drop-embedded network architectures in the WPI gel, respectively. FT-IR analyses showed that OBs stabilized the protein gels' polymeric matrix by hydrogen bonding, steric hindrance, and hydrophobic interactions. Rheology and texture showed that OBs hardened gels. Low-field nuclear magnetic resonance showed that excessive inclusion of OBs (30% of 6.8-OB and 35% of 11.0-OB) compromised gel integrity and freeze-thaw stability. This study found that OBs can be active fillers in protein gels for functional meals.

Fabrication of Antioxidant Pickering Emulsion Based on Resveratrol-Grafted Zein Conjugates: Enhancing the Physical and Oxidative Stability.

Lipid oxidation is still a major problem complicating the development of food emulsions. In this study, an antioxidant Pickering emulsion stabilized by resveratrol-grafted zein (Z-R) conjugates and pectin (P) complex particles was prepared. The hydrophilic pectin successfully adjusted the wettability of Z-R; when the mass ratio of Z-R to P was 2:1 (Z-R/P2:1), the three-phase contact angle was 90.68°, and the wettability of the particles was close to neutral. Rheological analysis showed that the emulsion formed an elastic gel structure. FTIR spectra indicated that there was a hydrogen bond and electrostatic interaction between Z-R and P. The disappearance of characteristic infrared peaks of corn oil was due to a dense protective film formed on the surface of oil drops by Z-R/P2:1 particles, which was confirmed by confocal laser scanning microscopy. The emulsion stabilized by Z-R/P2:1 had excellent physical stability at a wide range of pH values (4-9), salt ion concentrations (0.04-0.15 mol·L-1) and storage times (0-30 days). The anti-lipid oxidation ability of the emulsion was outstanding; after storage for 14 days at room temperature, the MDA content in the emulsion was only 123.85 µmol/kg oil. In conclusion, the Z-R/P2:1 particles prepared in this study can effectively stabilize a Pickering emulsion and expand the usability of the method for constructing antioxidant Pickering emulsions.

The Feed Additive Potassium Diformate Prevents Salmonella enterica Serovar Pullorum Infection and Affects Intestinal Flora in Chickens.

Extensive studies have shown that potassium diformate (KDF), an antibiotic substitute used as a feed additive, improves animal growth performance, although there is less direct evidence of its preventive effect on bacterial infections and its influence on the intestinal flora of animals. In this study, the inhibition effect of KDF on Salmonella enterica serovar Pullorum, an important enteric pathogen causing pullorum disease, was investigated in vitro and on a chicken infection model. The effect of KDF on the diversities and structures of chicken duodenal and cecum flora were also investigated using 16S rRNA gene sequencing. The results showed that addition of 0.5% KDF in feed or 0.1% KDF in drinking water significantly reduced the bacterial loads and the degree of pathological changes in the cecum, improved digestion and reduced the pH of the gastrointestinal tract of chickens infected with S. pullorum. KDF also significantly modified the diversity and abundance of intestinal microflorae in chickens. In particular, it promoted the colonization of several probiotics, such as Bacteroides, Blautia, Ruminococcus_torques_group and Faecalibacteriumm, which are involved in maintenance of the intestinal barrier, modulation of inflammation, energy supply for intestinal cells and pathogen resistance. These results enrich the theoretical basis for the clinical application of KDF in chickens.

The Intracellular Interaction of Porcine ß-Defensin 2 with VASH1 Alleviates Inflammation via Akt Signaling Pathway.

Defensins are a major class of antimicrobial peptides that facilitate the immune system to resist pathogen infection. To date, only ß-defensins have been identified in pigs. In our previous studies, porcine ß-defensin 2 (PBD-2) was shown to have both bactericidal activity and modulatory roles on inflammation. PBD-2 can interact with the cell surface TLR4 and interfere with the NF-κB signaling pathway to suppress the inflammatory response. In this study, the intracellular functions of PBD-2 were investigated. The fluorescently labeled PBD-2 could actively enter mouse macrophage cells. Proteomic analysis indicated that 37 proteins potentially interacted with PBD-2, among which vasohibin-1 (VASH1) was further tested. LPS, an inflammation inducer, suppressed the expression of VASH1, whereas PBD-2 inhibited this effect. PBD-2 inhibited LPS-induced activation of Akt, expression and release of the inflammatory mediators vascular endothelial growth factor and NO, and cell damage. A follow-up VASH1 knockdown assay validated the specificity of the above observations. In addition, PBD-2 inhibited LPS-induced NF-κB activation via Akt. The inhibition effects of PBD-2 on LPS triggered suppression of VASH1 and activation of Akt, and NF-κB and inflammatory cytokines were also confirmed using pig alveolar macrophage 3D4/21 cells. Therefore, the data indicate that PBD-2 interacts with intracellular VASH1, which inhibits the LPS-induced Akt/NF-κB signaling pathway, resulting in suppression of inflammatory responses. Together with our previous findings, we conclude that PBD-2 interacts with both the cell surface receptor (TLR4) and also with the intracellular receptor (VASH1) to control inflammation, thereby providing insights into the immunomodulatory roles of defensins.

Soy lipophilic protein self-assembled by pH-shift combined with heat treatment: Structure, hydrophobic resveratrol encapsulation, emulsification, and digestion.

This study evaluates the effect of pH (pH 3 and 11) and heat treatment (60 °C) in modifying the soybean lipophilic protein (LP) for the development of an encapsulation system to co-deliver resveratrol (Res) and vitamin D3. The structural and functional properties of LP after the modification will change to varying degrees. Meanwhile, Res was loaded into the hydrophobic core of LP, and the resulting Res-loaded structures have a uniform particle size distribution and a high encapsulation efficiency (78%). When the amount of Res encapsulation increases, the emulsification and oxidation resistance of the Pickering emulsion increased; the interfacial tension and interfacial protein adsorption increased to 11.21 mN/m and 97.34%, respectively. During simulated gastrointestinal digestion, the Pickering emulsion prepared with LP-Res nanoparticles at pH 11, 60 °C (pH 11, 60 °C-LP-Res) effectively protected Res and vitamin D3 from degradation or precipitation, indicating a significant increase in bioavailability.