5-methylcytosine modification by Plasmodium NSUN2 stabilizes mRNA and mediates the development of gametocytes.

5-methylcytosine (m5C) is an important epitranscriptomic modification involved in messenger RNA (mRNA) stability and translation efficiency in various biological processes. However, it remains unclear if m5C modification contributes to the dynamic regulation of the transcriptome during the developmental cycles of Plasmodium parasites. Here, we characterize the landscape of m5C mRNA modifications at single nucleotide resolution in the asexual replication stages and gametocyte sexual stages of rodent (Plasmodium yoelii) and human (Plasmodium falciparum) malaria parasites. While different representations of m5C-modified mRNAs are associated with the different stages, the abundance of the m5C marker is strikingly enhanced in the transcriptomes of gametocytes. Our results show that m5C modifications confer stability to the Plasmodium transcripts and that a Plasmodium ortholog of NSUN2 is a major mRNA m5C methyltransferase in malaria parasites. Upon knockout of P. yoelii nsun2 (pynsun2), marked reductions of m5C modification were observed in a panel of gametocytogenesis-associated transcripts. These reductions correlated with impaired gametocyte production in the knockout rodent malaria parasites. Restoration of the nsun2 gene in the knockout parasites rescued the gametocyte production phenotype as well as m5C modification of the gametocytogenesis-associated transcripts. Together with the mRNA m5C profiles for two species of Plasmodium, our findings demonstrate a major role for NSUN2-mediated m5C modifications in mRNA transcript stability and sexual differentiation in malaria parasites.

Genome-wide landscape of ApiAP2 transcription factors reveals a heterochromatin-associated regulatory network during Plasmodium falciparum blood-stage development.

Heterochromatin-associated gene silencing controls multiple physiological processes in malaria parasites, however, little is known concerning the regulatory network and cis-acting sequences involved in the organization of heterochromatin and how they modulate heterochromatic gene expression. Based on systematic profiling of genome-wide occupancy of eighteen Apicomplexan AP2 transcription factors by ChIP-seq analysis, we identify and characterize eight heterochromatin-associated factors (PfAP2-HFs), which exhibit preferential enrichment within heterochromatic regions but with differential coverage profiles. Although these ApiAP2s target euchromatic gene loci via specific DNA motifs, they are likely integral components of heterochromatin independent of DNA motif recognition. Systematic knockout screenings of ApiAP2 factors coupled with RNA-seq transcriptomic profiling revealed three activators and three repressors of heterochromatic gene expression including four PfAP2-HFs. Notably, expression of virulence genes is either completely silenced or significantly reduced upon the depletion of PfAP2-HC. Integrated multi-omics analyses reveal autoregulation and feed-forward loops to be common features of the ApiAP2 regulatory network, in addition to the occurrence of dynamic interplay between local chromatin structure and ApiAP2s in transcriptional control. Collectively, this study provides a valuable resource describing the genome-wide landscape of the ApiAP2 family and insights into functional divergence and cooperation within this family during the blood-stage development of malaria parasites.

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.

A cascade of transcriptional repression determines sexual commitment and development in Plasmodium falciparum.

Gametocytogenesis, the process by which malaria parasites produce sexual forms that can infect mosquitoes, is essential for the transmission of malaria. A transcriptional switch of the pfap2-g gene triggers sexual commitment, but how the complex multi-step process is precisely programed remains largely unknown. Here, by systematic functional screening of a panel of ApiAP2 transcription factors, we identify six new ApiAP2 members associated with gametocytogenesis in Plasmodium falciparum. Among these, PfAP2-G5 (PF3D7_1139300) was found to be indispensable for gametocytogenesis. This factor suppresses the transcriptional activity of the pfap2-g gene via binding to both the upstream region and exonic gene body, the latter is linked to the maintenance of local heterochromatin structure, thereby preventing initiation of sexual commitment. Removal of this repressive effect through pfap2-g5 knockout disrupts the asexual replication cycle and promotes sexual commitment accompanied by upregulation of pfap2-g expression. However, the gametocytes produced fail to mature fully. Further analyses show that PfAP2-G5 is essential for gametocyte maturation, and causes the down-regulation of pfap2-g and a set of early gametocyte genes activated by PfAP2-G prior to gametocyte development. Collectively, our findings reveal a regulation cascade of gametocyte production in malaria parasites, and provide a new target for transmission blocking interventions.

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.

Advances in Lipid Extraction Methods-A Review.

Extraction of lipids from biological tissues is a crucial step in lipid analysis. The selection of appropriate solvent is the most critical factor in the efficient extraction of lipids. A mixture of polar (to disrupt the protein-lipid complexes) and nonpolar (to dissolve the neutral lipids) solvents are precisely selected to extract lipids efficiently. In addition, the disintegration of complex and rigid cell-wall of plants, fungi, and microalgal cells by various mechanical, chemical, and enzymatic treatments facilitate the solvent penetration and extraction of lipids. This review discusses the chloroform/methanol-based classical lipid extraction methods and modern modifications of these methods in terms of using healthy and environmentally safe solvents and rapid single-step extraction. At the same time, some adaptations were made to recover the specific lipids. In addition, the high throughput lipid extraction methodologies used for liquid chromatography-mass spectrometry (LC-MS)-based plant and animal lipidomics were discussed. The advantages and disadvantages of various pretreatments and extraction methods were also illustrated. Moreover, the emerging green solvents-based lipid extraction method, including supercritical CO2 extraction (SCE), is also discussed.

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).

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.