The RNA m5C methyltransferase NSUN1 modulates human malaria gene expression during intraerythrocytic development.

Introduction: Plasmodium falciparum is the most damaging malaria pathogen and brings a heavy burden to global health. Host switching and morphological changes in P. falciparum are dependent on an effective gene expression regulatory system. C5 methylation of cytosines is a common RNA modification in eukaryotes, and the NSUN family are essential m5C modification executors. Currently, little is known about this family in Plasmodium spp. In this study, we focus on exploring the function of PfNSUN1 protein. Methods: An efficient CRISPR/Cas9 gene editing technique was applied to construct the PfNSUN1 knockdown strain. The knockdown efficiency was confirmed by growth curves and western blot experiments. The knockdown transcriptome data was acquired to find differentially expressed genes, and target genes of PfNSUN1 protein were identified by RNA immunoprecipitation and high-throughput sequencing experiments. Results: The efficiency of PfNSUN1 protein down-regulated was about 34%. RNA-seq data revealed that differentially expressed genes were mainly down-regulated. And there were 224, 278, 556 genes that were down-regulated with more than 2-fold changes and p-adj<0.05 at ring, trophozoite and schizont stages, respectively. PfNSUN1 protein was significantly enriched on 154 target genes, including 28S ribosomal RNA and pfap2-g5 transcription factor. Discussion: PfNSUN1 is a crucial RNA post-transcriptional modification protein in P. falciparum. It plays a pivotal role in regulating gene expression and parasite growth by targeting 28S ribosomal RNA and pfap2-g5 transcription factor.

Recent Advances in the Inhibition of Membrane Lipid Peroxidation by Food-Borne Plant Polyphenols via the Nrf2/GPx4 Pathway.

Lipid peroxidation (LP) leads to changes in the fluidity and permeability of cell membranes, affecting normal cellular function and potentially triggering apoptosis or necrosis. This process is closely correlated with the onset of many diseases. Evidence suggests that the phenolic hydroxyl groups in food-borne plant polyphenols (FPPs) make them effective antioxidants capable of preventing diseases triggered by cell membrane LP. Proper dietary intake of FPPs can attenuate cellular oxidative stress, especially damage to cell membrane phospholipids, by activating the Nrf2/GPx4 pathway. Nuclear factor E2-related factor 2 (Nrf2) is an oxidative stress antagonist. The signaling pathway regulated by Nrf2 is a defense transduction pathway of the organism against external stimuli such as reactive oxygen species and exogenous chemicals. Glutathione peroxidase 4 (GPx4), under the regulation of Nrf2, is the only enzyme that reduces cell membrane lipid peroxides with specificity, thus playing a pivotal role in regulating cellular ferroptosis and counteracting oxidative stress. This study explored the Nrf2/GPx4 pathway mechanism, antioxidant activity of FPPs, and mechanism of LP. It also highlighted the bioprotective properties of FPPs against LP and its associated mechanisms, including (i) activation of the Nrf2/GPx4 pathway, with GPx4 potentially serving as a central target protein, (ii) regulation of antioxidant enzyme activities, leading to a reduction in the production of ROS and other peroxides, and (iii) antioxidant effects on LP and downstream phospholipid structure. In conclusion, FPPs play a crucial role as natural antioxidants in preventing LP. However, further in-depth analysis of FPPs coregulation of multiple signaling pathways is required, and the combined effects of these mechanisms need further evaluation in experimental models. Human trials could provide valuable insights into new directions for research and application.

Saccharomyces cerevisiae oral immunization in mice using multi-antigen of the African swine fever virus elicits a robust immune response.

African swine fever virus (ASFV) is one of the most complex viruses. ASFV is a serious threat to the global swine industry because no commercial vaccines against this virus are currently available except in Vietnam. Moreover, ASFV is highly stable in the environment and can survive in water, feed, and aerosols for a long time. ASFV is transmitted through the digestive and respiratory tract. Mucosal immunity is the first line of defense against ASFV. Saccharomyces cerevisiae (SC), which has been certified by the U.S. Food and Drug Administration and has a generally recognized as safe status in the food industry, was used for oral immunization in this study. ASFV antigens were effectively expressed in recombinant SC strains with high DNA copy numbers and stable growth though surface display technology and chromosome engineering (δ-integration). The recombinant SC strains containing eight ASFV antigens-KP177R, E183L, E199L, CP204L, E248R, EP402R, B602L, and B646L- induced strong humoral and mucosal immune responses in mice. There was no antigenic competition, and these antigens induced Th1 and Th2 cellular immune responses. Therefore, the oral immunization strategy using recombinant SC strains containing multiple ASFV antigens demonstrate potential for future testing in swine, including challenge studies to evaluate its efficacy as a vaccine against ASFV.

Synthesis and performance of a nanosensing platform for homocysteine detection: A series of iridium(III) complexes containing aldehyde group as probe and MOF as supporting substrate.

The low cost and simple detection method for Hcy (homocysteine) is highly desired in analytical and biological fields since Hcy has been regarded as a bio-marker for multiple diseases. In this work, five Ir(C^N)2(N^N)+ compounds having -CHO group in their C^N or N^N ligand were synthesized and tried for Hcy sensing. Electron-donating groups such as -NH2 and -CH3 were incorporated into the C^N or N^N ligand. Their geometric structure, electronic structure, and optical parameters (with or without Hcy) were analyzed and compared carefully to explore their Hcy sensing potential. The sensing mechanism was revealed by NMR titration and theoretical simulation as a cyclization reaction between the -CHO group and Hcy. The optimal compounds, which showed increased emission quantum yield (2.5-fold) and emission blue-shift (by âˆ¼ 100 nm) upon Hcy, were then covalently grafted into a porous host bio-MOF-1. Linear working plots were fitted, with good selectivity, LOD of 0.15 µM, and response time of 33 s. The novelty of this work was the eye-sensitive emission color change of this nanosensing platform from red (without Hcy) to green (with Hcy).

Dietary Sources, Stabilization, Health Benefits, and Industrial Application of Anthocyanins-A Review.

Natural phytochemicals are well known to protect against numerous metabolic disorders. Anthocyanins are vacuolar pigments belonging to the parent class of flavonoids. They are well known for their potent antioxidant and gut microbiome-modulating properties, primarily responsible for minimizing the risk of cardiovascular diseases, diabetes, obesity, neurodegenerative diseases, cancer, and several other diseases associated with metabolic syndromes. Berries are the primary source of anthocyanin in the diet. The color and stability of anthocyanins are substantially influenced by external environmental conditions, constraining their applications in foods. Furthermore, the significantly low bioavailability of anthocyanins greatly diminishes the extent of the actual health benefits linked to these bioactive compounds. Multiple strategies have been successfully developed and utilized to enhance the stability and bioavailability of anthocyanins. This review provides a comprehensive view of the recent advancements in chemistry, biosynthesis, dietary sources, stabilization, bioavailability, industrial applications, and health benefits of anthocyanins. Finally, we summarize the prospects and challenges of applications of anthocyanin in foods.

Mechanism of ultrasound-induced soybean/egg white composite gelation: Gel properties, morphological structure and co-aggregation kinetics.

This study aims to develop ultrasound-assisted acid-induced egg white protein (EWP)-soy protein isolate (SPI) composite gels and to investigate the mechanistic relationship between the co-aggregation behavior of composite proteins and gel properties through aggregation kinetics monitored continuously by turbidity. The results showed that the inclusion of EWP caused the attenuation of gel properties and maximum aggregation (Amax) because EWP could aggregate with SPI at a higher rate (Kapp), which impeded the interaction between SPI and the formation of a continuous gelling network. In the EWP-dominated system, SPI with higher molecular weights also increased the fractal dimension of gels. Ultrasound improved properties of composite gels, especially the SPI-dominated system. After ultrasound treatment, the small, uniform size of co-aggregates and the decrease in potential led to an increase in the aggregation rate and formation of dense particles, consistent with an increase in gelation rate and texture properties. Excessively fast aggregation generated coarse chains and more pores. Still, the exposure of free sulfhydryl groups assisted the gel structure units to form a compact network through disulfide bonding. On the whole, the study could provide theoretical support for a deeper understanding on the interaction mechanism and gelation of composite proteins.

Egg White Peptides Accelerating the Wound Healing Process Through Modulating the PI3K-AKT Pathway: A Joint Analysis of Transcriptomic and Proteomic.

Wound healing is a multiphase process with a complex repair mechanism; trauma-repairing products with safety and high efficiency have a great market demand. Egg white peptides (EWP) have various physiological regulatory functions and have been proven efficient in ameliorating skin damage. However, their underlying regulation mechanism has not been revealed. This study further evaluated the EWP ameliorating mechanism by conducting a full-thickness skin wound model. Results demonstrated that EWP administration significantly inhibited the expression of pro-inflammatory and shortened the inflammatory phase. Besides, EWP can accelerate the secretion of growth factors (PDGF, VEGF, and TGF-ß1) in skin tissue, significantly increasing the regeneration of granulation tissue and endothelium in the proliferation phase, thereby promoting wound healing. After 400 mg/kg EWP interventions for 13 days postoperation, the wound healing rate reached 90%. The combination of transcriptomic and proteomic analyses demonstrated the ameliorating efficiency effects of EWP on wound healing. EWP mainly participates in the functional network with the PI3K-AKT signaling pathway as the core to accelerate wound healing. These findings suggest a promising EWP-based strategy for accelerating wound healing.

Dextran sulfate acting as a chaperone-like component on inhibition of amorphous aggregation and enhancing thermal stability of ovotransferrin.

The tendency of ovotransferrin (OVT) to unfold and aggregate under 60 °C severely restricted sterilization temperature during egg processing. Searching for efficient strategies to improve OVT thermal stability is essential for improving egg product quality and processing suitability. Here, we investigated the effect of sulfate polysaccharide (dextran sulfate, DS) on heat-induced aggregation of OVT. We found that DS can effectively suppress amorphous aggregation of OVT at pH 7.0 after heating. Strikingly, the addition of 5 µM DS fully suppressed insoluble aggregates formation of 0.5 mg/mL OVT. Structure analysis confirmed that DS preserves nearly the entire secondary and tertiary structure of OVT during heating. The steric hindrance effect arising from strong electrostatic interactions between OVT and DS, coupled with reduced OVT hydrophobicity, is the underlying mechanism in suppressing protein-protein interactions, thus enhancing thermal stability. These findings suggest DS could act as protein stabilizers and chaperones, enhancing the thermostability of heat-sensitive proteins.

Egg White Protein-Proanthocyanin Complexes Stabilized Emulsions: Investigation of Physical Stability, Digestion Kinetics, and Free Fatty Acid Release Dynamics.

Egg white proteins pose notable limitations in emulsion applications due to their inadequate wettability and interfacial instability. Polyphenol-driven alterations in proteins serve as an effective strategy for optimizing their properties. Herein, covalent and non-covalent complexes of egg white proteins-proanthocyanins were synthesized. The analysis of structural alterations, amino acid side chains and wettability was performed. The superior wettability (80.00° ± 2.23°) and rigid structure (2.95 GPa) of covalent complexes established favorable conditions for their utilization in emulsions. Furthermore, stability evaluation, digestion kinetics, free fatty acid (FFA) release kinetics, and correlation analysis were explored to unravel the impact of covalent and non-covalent modification on emulsion stability, dynamic digestion process, and interlinkages. Emulsion stabilized by covalent complex exhibited exceptional stabilization properties, and FFA release kinetics followed both first-order and Korsmeyer-Peppas models. This study offers valuable insights into the application of complexes of proteins-polyphenols in emulsion systems and introduces an innovative approach for analyzing the dynamics of the emulsion digestion process.

The level of sulfate substitution of polysaccharide regulates thermal-induced egg white protein gel properties: The characterization of gel structure and intermolecular forces.

Sulfated polysaccharides exhibit great potential for regulating protein-protein interactions. In the present study, three sulfated microcrystalline cellulose (MCS) with different degrees of sulfate substitution (DSS: 0.33, 0.51, 0.61) were synthesized and the effects of DSS on the regulation of egg white protein (EWP) aggregation and gelation properties were investigated. The results found that the improvement of protein mechanical properties by MCS is closely related to the level of sulfate substitution. The higher the DSS, the more ordered protein aggregates and compact gel network formed during heating as compared to that of pure EWP. Lower DSS (0.33) shows little effect on the mechanical properties of EWP. Furthermore, all the MCSs could significantly destroy the tertiary structure of protein molecules during heating, while for the secondary structure, MCS with higher DSS (0.51 and 0.61) could effectively control the decreasing tendency of α-helix and increasing tendency of ß-sheet. Hydrophobic interactions were recognized as the major intermolecular force in the compact mixed gels (EWP/MCS2 and EWP/MCS3 gels, DSS was 0.51 and 0.61, respectively). These findings provide a vital understanding of the gelling mechanism of the protein-polysaccharide system and the application of sulfated polysaccharides in protein-based food products.

Potential Benefits of Egg White Proteins and Their Derived Peptides in the Regulation of the Intestinal Barrier and Gut Microbiota: A Comprehensive Review.

Impaired intestinal barrier function can impede the digestion and absorption of nutrients and cause a range of metabolic disorders, which are the main causes of intestinal disease. Evidence suggests that proper dietary protein intake can prevent and alleviate intestinal diseases. Egg white protein (EWP) has received considerable attention, because of its high protein digestibility and rich amino acid composition. Furthermore, bioactive peptides may have an increased repair effect due to their high degradation efficiency in the gut. In this study, we aimed to review the effects of EWP and its bioactive peptides on intestinal structural repair. The potential modulation mechanisms by which EWP and their peptides regulate the gut microbiota and intestinal barrier can be summarized as follows: (1) restoring the structure of the intestinal barrier to its intact form, (2) enhancing the intestinal immune system and alleviating the inflammatory response and oxidative damage, and (3) increasing the relative abundance of beneficial bacteria and metabolites. Further in-depth analysis of the coregulation of multiple signaling pathways by EWP is required, and the combined effects of these multiple mechanisms requires further evaluation in experimental models. Human trials can be considered to understand new directions for development.

Untargeted metabolomics revealed the regulatory effect of Maillard reaction products of ovalbumin with different degrees on the metabolism of colitis mice.

The Maillard reaction (MR) is inevitable in food processing and daily cooking, but whether the MR degree would affect the biological activity of the protein in vivo remains unknown. In this study, we used untargeted metabolomics techniques to explore the effects of two different levels of Maillard reaction products (MRPs) of ovalbumin (OVA) on metabolites in colitis mice. Studies have shown that MR could affect protein metabolites in vivo and MRPs of OVA could reduce the concentrations of IL-6 and IL-1ß and intestinal permeability. Metabolomics results showed that the degree of MR affected the abundance of oligopeptides and bile acids in vivo. This study revealed that MRPs could regulate the abundance of metabolites such as taurocholic acid and putrescine, and repair the intestinal barrier in colitis mice through signaling pathways such as secondary bile acid biosynthesis, bile secretion and ABC transporters. The investigation has significant implications for the digestion properties and metabolite regulation of MRPs in vivo, and also promotes the application of MRPs in functional foods.

Integrated Microbiome and Metabolomic Analysis Reveal the Repair Mechanisms of Ovalbumin on the Intestine Barrier of Colitis Mice.

The development and progression of colitis would detrimentally destroy the intestine barrier. However, there remains a paucity of evidence on whether ovalbumin (OVA) can be used as a nutritional food protein to repair the intestinal barrier. In this study, the repairing mechanism of OVA on intestinal barrier was thoroughly investigated by gut microbiota and untargeted metabolomics techniques. The findings demonstrated that OVA reduced intestinal permeability and restored mucin (0.75 ± 0.06) and tight junction (TJ) protein (0.67 ± 0.14) expression in colitis mice caused by 3% dextran sulfate sodium (DSS). In addition, the inflammation response and oxidative stress were also attenuated. The intake of OVA upregulated the abundance of Lactobacillaceae (7.60 ± 3.34%) and Akkermansiaceae (10.39 ± 5.97%). Furthermore, OVA upregulated the abundance of inosine (6.06 ± 0.36%), putrescine (4.14 ± 0.20%), and glycocholic acid (5.59 ± 0.23%) in colitis mice through ATP binding cassette (ABC) transporters and bile secretion pathways. In summary, our findings revealed that OVA could maintain intestinal health, which may provide crucial insights for preventing and treating intestinal diseases.

Designation and characterization of cold-set egg white protein/dextran sulfate hydrogel for curcumin entrapment.

This study aimed to design a cold-set hydrogel of egg white protein (EWP) with good mechanical properties for encapsulating curcumin. Dextran sulfate (DS) and transglutaminase (TGase) were used to control the aggregation and gelation behavior of EWP at preheating step and gelation step, respectively. The optimum soluble protein aggregate size was obtained in the EWP/DS mixture at a mass ratio of 10 under 85 °C preheated (HED10). The presence of TGase further enhanced the cross-linking degree between protein aggregates during the gelation step. The highest gel hardness was found in HED10 hydrogel with TGase, which is almost 10 times the pure EWP gel. Besides, the HED hydrogels effectively slowed down the release rate of curcumin in gastrointestinal digestion. This work provides a theoretical basis for the development of cold-set EWP hydrogel with good mechanical strength by sulfated polysaccharide addition and TGase cross-linking as encapsulation delivery systems.

Aggregation of egg white peptides (EWP) induced by proanthocyanidins: A promising fabrication strategy for EWP emulsion.

Egg-white peptides (EWP, <1 kDa) have been shown to possess various bioactive properties. However, poor emulsification of EWP limits its application in functional foods. In this study, EWP aggregation induced by proanthocyanidins (PC) contributed to the improvement of emulsion properties. The two-step binding process of PC-EWP-EWP was confirmed by isothermal titration calorimetry, fluorescence spectroscopy, surface hydrophobicity, and Fourier transform infrared spectroscopy. We found that first EWP combines with PC via hydrogen bonding and hydrophobic interactions. Next, more EWPs bind to the EWP in PC-EWP via hydrogen bonding, thereby forming PC-EWP-EWP aggregates. The aggregates (PC to EWP ratio of 1:4) reduced the surface tension (6 %) and improved the contact angle (53 %). The co-adsorption of EWP and aggregates at the O/W interface improved the contact angle, protein adsorption rate, and emulsion stability. This study establishes EWP aggregates induced by PC as an effective emulsifier, thereby expanding the application fields of EWP.

Soybean meal peptides regulated membrane phase of giant unilamellar vesicles: A key role for bilayer amphipathic region localization.

Membrane phase separation forms liquid-ordered (Lo) and liquid-disordered (Ld) phases and is involved in cellular processes and functions. Our previous study has confirmed that peptides can regulate phase separation by increasing the Lo phase. However, the specific mechanisms underlying the phase separation regulation of peptides remain poorly understood. This study aimed to explore the effect of soybean meal peptides on phase separation and illustrate the correlation between phase regulation and membrane localization of the peptides. Phase separation was studied by giant unilamellar vesicles (GUVs), and membrane localization of the peptides was detected by steady-state fluorescence quenching. Our results revealed that peptides YYK, CLA, and SLW enhanced the Lo phase while WLQ decreased the Lo phase. The localization in the membrane amphiphilic region of the peptides played a crucial role in their regulation of phase separation. The more localization of the peptides (YYK, CLA, and SLW) in the membrane amphiphilic region, the stronger the capacity to increase the Lo phase.

Protective effects and potential mechanisms of fermented egg-milk peptides on the damaged intestinal barrier.

Introduction: Fermented egg-milk peptides (FEMPs) could enhance the colon-intestinal barrier and upgrade the expression of zonula occludens-1 and mucin 2. Besides, the underlying biological mechanism and the targets FEMPs could regulate were analyzed in our study. Methods: Herein, the immunofluorescence technique and western blot were utilized to evaluate the repair of the intestinal barrier. Network pharmacology analysis and bioinformatics methods were performed to investigate the targets and pathways affected by FEMPs. Results and discussion: Animal experiments showed that FEMPs could restore intestinal damage and enhance the expression of two key proteins. The pharmacological results revealed that FEMPs could regulate targets related to kinase activity, such as AKT, CASP, RAF, and GSK. The above targets could interact with each other. GO analysis indicated that the targets regulated by FEMPs could participate in the kinase activity of the metabolic process. KEGG enrichment revealed that the core targets were enriched in pathways related to cell apoptosis and other important procedures. Molecular docking demonstrated that FEMPs could bind to the key target AKT via hydrogen bond interactions. Our study combined the experiment in vivo with the method in silico and investigated the interaction between peptides and targets in a pattern of multi-targets and multi-pathways, which offered a new perspective on the functional validation and potential application of bioactive peptides.

Carotenoids: Dietary Sources, Extraction, Encapsulation, Bioavailability, and Health Benefits-A Review of Recent Advancements.

Natural carotenoids (CARs), viz. ß-carotene, lutein, astaxanthin, bixin, norbixin, capsanthin, lycopene, canthaxanthin, ß-Apo-8-carotenal, zeaxanthin, and ß-apo-8-carotenal-ester, are being studied as potential candidates in fields such as food, feed, nutraceuticals, and cosmeceuticals. CAR research is advancing in the following three major fields: (1) CAR production from natural sources and optimization of its downstream processing; (2) encapsulation for enhanced physical and chemical properties; and (3) preclinical, clinical, and epidemiological studies of CARs' health benefits. This review critically discusses the recent developments in studies of the chemistry and antioxidant activity, marketing trends, dietary sources, extraction, bioaccessibility and bioavailability, encapsulation methods, dietary intake, and health benefits of CARs. Preclinical, clinical, and epidemiological studies on cancer, obesity, type 2 diabetes (T2D), cardiovascular diseases (CVD), osteoporosis, neurodegenerative disease, mental health, eye, and skin health are also discussed.

Application of γ-cyclodextrin-lysozyme as host materials for encapsulation of curcumin: characterization, stability, and controlled release properties.

BACKGROUND: In this study, a safe and relatively stable γ-cyclodextrin-lysozyme (γ-CD-Lys) was synthesized using epichlorohydrin as the cross-linking agent, and curcumin was successfully encapsulated in γ-CD-Lys. RESULTS: The successful Lys grafting onto γ-CD can be demonstrated by a high grafting ratio (79.02%) and was further confirmed by Fourier transform infrared (FTIR) band shifts and the new signal obtained at δ 2.75 in proton nuclear magnetic resonance. The encapsulation efficiency value of γ-CD-Lys was 76.74%, and the successful encapsulation of curcumin into γ-CD-Lys was confirmed by crystal structure change, increased melting point, and FTIR band shifts. The intermolecular bonds results suggested that associative forces between curcumin and γ-CD-Lys were electrostatic interaction, hydrogen bonds interaction, and hydrophobic interaction. The designed nanoparticles had excellent stability at low pH and low salt concentration. The release rate of these nanoparticles was inhibited in simulated gastric conditions, whereas it increased significantly in intestinal media. Simulated gastrointestinal digestion experiments further confirmed that nanoparticles showed higher bioaccessibility (86.05%) compared with curcumin (58.82%). CONCLUSION: Overall, our study showed that the nanoparticles were highly promising for delivering curcumin because of their enhanced functional attributes and stabilization in acid or low salt environments. Also, it was an excellent wall material for targeting hydrophobic bioactive compounds in the intestinal tract via oral administration. © 2022 Society of Chemical Industry.

Stability of oil-in-water emulsions improved by ovalbumin-procyanidins mixture: A promising substrate with emulsifying and antioxidant activity.

The aim of this study is to develop a dual-functional ingredient with antioxidant activity and emulsification. The emulsion stability of ovalbumin (OVA) was improved by procyanidins (PC). The interactions between OVA and PC were investigated using multi-spectroscopy and molecular docking. Furthermore, the effect of the addition of the OVA-PC mixture on emulsion stability was evaluated as well. The fluorescence results showed that the quenching mechanism of PC to OVA's endogenous fluorescence was static quenching, and the binding ratio of OVA and PC was 1:1. Circular dichroism (CD) and Fourier Transform Infrared Spectrometer (FT-IR) showed that the addition of PC promoted the unfolding of OVA, and transformed the secondary structure of OVA from α-helix to ß-sheet. The main driving force of OVA and PC was hydrogen bonding, according to molecular docking analysis. Among all the samples, the stability of the emulsion of OVA-PC at a ratio of 1:30 exhibited extremely high stability and the smallest particle size. In comparison with individual OVA emulsions, the OVA-PC emulsions had excellent physical stabilities. Meanwhile, the oxidation degree of protein and oil for the OVA-PC emulsions was lower than that of the native OVA emulsion after 8-day storage. Our work provides important insights for understanding the interaction between OVA and expanding the application of OVA-PC.