Egg Yolk Selenopeptides: Preparation, Characterization, and Immunomodulatory Activity.

In this study, egg yolk selenium peptides (Se-EYP) were prepared using double-enzyme hydrolysis combined with a shearing pretreatment. The properties of the selenopeptides formed were then characterized, including their yield, composition, molecular weight distribution, antioxidant activity, in vitro digestion, and immunomodulatory activity. The peptide yield obtained after enzymatic hydrolysis using a combination of alkaline protease and neutral protease was 74.5%, of which 82.6% had a molecular weight <1000 Da. The selenium content of the lyophilized solid product was 4.01 µg/g. Chromatography-mass spectrometry analysis showed that 88.6% of selenium in Se-EYP was in the organic form, of which SeMet accounted for 60.3%, SeCys2 for 21.8%, and MeSeCys for 17.9%. After being exposed to in vitro simulated digestion, Se-EYP still had 65.1% of oligopeptides present, and the in vitro antioxidant activity was enhanced. Moreover, Se-EYP exhibited superior immune detection indices, including immune organ index, level of immune factors in the serum, histopathological changes in the spleen, and selenium content in the liver. Our results suggest that Se-EYP may be used as selenium-enriched ingredients in functional food products.

Identification and Immunomodulatory Effect on Immunosuppressed Mice of Selenium-Enriched Peptides of Egg White.

Selenium-enriched egg white peptides (Se-EWP) were prepared by pre-heat treatment and enzymatic hydrolysis in this study. In addition, their selenopeptide sequence identification and immunomodulatory effect were investigated. Results showed that the yield of Se-EWP obtained from alkaline-neutral protease treatment reached 76.90%, and peptides with a molecular weight of 200-1000 Da accounted for 98.33%. Four characteristic selenopeptides, including SeCys-Trp-Leu-Glu, Trp-Ser-SeCys, SeMet-Ala-Pro, and SeMet-Leu, were identified by HPLC-ESI-MS/MS, which were rich in hydrophobic and branched-chain amino acids. Se-EWP (750 mg/kg/d) could effectively retard the decrease of immune organ index in immunosuppressed mice induced by cyclophosphamide. Moreover, supplementation of Se-EWP could promote a higher content of Se in liver, the number of white blood cells, and the levels of serum cytokines (IL-6, IL-2, and TNF-α) as compared with EWP groups, indicating that Se-EWP could effectively alleviate immunosuppression induced by cyclophosphamide. These findings suggested that Se-EWP exhibited great potential as functional foods for immunomodulatory effect.

[Advances in separation and analysis of aromatic amino acids in food].

Amino acids are important building blocks of proteins in the human body, which are involved in many metabolic pathways. Patients with metabolic diseases such as phenylketonuria, tyrosinemia, and hepatic encephalopathy are genetically defective and cannot metabolize aromatic amino acids (AAA) in food; hence, a regular diet may lead to permanent physiological damage. For this reason, it is necessary to restrict the intake of AAA in their daily diet by limiting natural protein intake, while ensuring normal intake of low protein foods and supplementation with low-AAA protein equivalents. Sources of low-AAA protein equivalents currently rely on free amino acid complex mixtures and low-AAA peptides (also known as high-Fischer-ratio peptides), which have better absorption availability and palatability. AAA separation and analysis techniques are essential for the preparation and detection of low-AAA peptides. Researchers in this field have explored a variety of efficient adsorption materials to selectively remove AAA from complex protein hydrolysates and thus prepare low-AAA peptide foods, or to establish analysis strategies for AAA. Covering more than 70 publications on AAA removal and separation in the last decade from Web of Science Core Collection and China National Knowledge Infrastructure, this review analyzes the structural characteristics and physicochemical properties of AAA, and summarizes the technological progress of AAA removal based on adsorbents such as activated carbon and resin. The applications of two-dimensional nanomaterials, molecular imprinting, cyclodextrins, and metal-organic frameworks in AAA adsorption and analysis from three dimensions, i. e., sample pretreatment, chiral separation and adsorption sensing, are also reviewed. The mainstream adsorbents for AAA removal, such as activated carbon, still suffer from poor specificity and cause environmental pollution during post-use treatment. Existing AAA separating materials show impressive selective adsorption capability in food samples and chiral mixtures as well as high sensitivity in adsorption sensing. The development of an efficient detection technology for AAA may help in detecting trace AAA in food and in evaluating chiral AAA adulteration in food samples. By exploring the advantages and disadvantages of each type of technology, we provide support for the advancement of the removal and analysis techniques for AAA.

Improved retention ratio and bioaccessibility of lutein loaded in emulsions stabilized by egg yolk granules-lecithin complex.

BACKGROUND: Egg yolk granules (EYGs)-soy lecithin (SL) complex is a newly developed delivery system that is effective for improving the storage stability of hydrophobic bioactive compounds. However, the formation mechanism of EYGs and SL complex and its effect on the gastrointestinal fate of lutein-loaded emulsions needs to be investigated further. RESULTS: Adding SL greatly improved the surface activity of the EYGs, as evidenced by reduced surface tension and an increased adsorption rate to the oil/water interface. Hydrophobic interaction was the dominant force in the formation of EYG-SL complex, with hydrogen and ionic bonds playing complementary roles. Using the EYG-SL complex, stable oil-in-water emulsions were formed and exhibited an enhanced retention ratio and bioaccessibility of lutein after simulated digestion. Correlation analysis demonstrated that the additional anti-oxidant activity as a result of EYGs was responsible for the high retention of lutein, whereas low surface tension facilitated the micellization of bioaccessible lutein. CONCLUSION: The present study shows that the EYG and SL have a synergistic effect with respect to improving the retention ratio and bioaccessibility of lutein in emulsions stabilized by EYG-SL complex after digestion and this will guide the development of value-added oil-in-water emulsion products using protein-lecithin complex as a promising nutrient delivery vehicle. © 2022 Society of Chemical Industry.

Human neutrophil membrane-derived nanovesicles as a drug delivery platform for improved therapy of infectious diseases.

Resolvins are a group of specialized proresolving lipid mediators (SPMs) enzymatically produced from omega-3 fatty acids during acute inflammation response to infections or tissue injury. Resolvin D1 (RvD1) is one of resolvins and is well studied in resolution of inflammation to treat inflammatory diseases. Resolution of inflammation includes the inhibition of polymorphonuclear leukocyte recruitment and reduced cytokine production. However, effective delivery of RvD1 to inflammatory tissues is challenging because of its lack of tissue targeting and poor physicochemical properties. Here, we proposed nanovesicles made from human neutrophil membrane which can specifically target inflamed tissues, and we loaded RvD1 on the surface of nanovesicles and antibiotic (ceftazidime, CEF) inside nanovesicles for improved treatment of bacterial infections. In a mouse model of bacterium-induced peritonitis, we demonstrated that human neutrophil cell membrane-formed vesicles (NMVs) enhanced inflammation resolution and bacterial killing after co-delivery of RvD1 and CEF. Our studies reveal that neutrophil nanovesicles may be critical for enhanced therapy to infectious diseases.

Enhancing the oxidative stability of algal oil powders stabilized by egg yolk granules/lecithin composites.

This study reported a powder formulation containing omega-3-rich algal oil emulsions stabilized by egg yolk granules (EYGs)/lecithin composites. The improved physical stability of the algal oil samples due to increasing pH and lecithin addition was beneficial to the oxidative stability through analysis of free radical scavenging activities, metal ion chelating activities, and the release of primary and secondary oxidation products during accelerated storage (12 days, 60 °C). In addition, the effect of three antioxidants, i.e. ascorbic acid (VC), ascorbyl palmitate (AP), and α-tocopherol (VE), on lipid oxidation was investigated. Results showed that antioxidant partitioning at different regions of the emulsion system influenced its ability to prevent oxidation with the effectiveness of AP (at the O/W interface) > VE (in the oil phase) > VC (in the aqueous phase). This study developed a new powder-based emulsion formulation for algal oils with superior oxidative stability as an alternative source of omega-3.

Transcriptomic profiles of 33 opium poppy samples in different tissues, growth phases, and cultivars.

Opium poppy is one of the most important medicinal plants and remains the only commercial resource of morphinan-based painkillers. However, little is known about the regulatory mechanisms involved in benzylisoquinoline alkaloids (BIAs) biosynthesis in opium poppy. Herein, the full-length transcriptome dataset of opium poppy was constructed for the first time in accompanied with the 33 samples of Illumina transcriptome data from different tissues, growth phases and cultivars. The long-read sequencing produced 902,140 raw reads with 55,114 high-quality transcripts, and short-read sequencing produced 1,923,679,864 clean reads with an average Q30 rate of 93%. The high-quality transcripts were subsequently quantified using the short reads, and the expression of each unigene among different samples was calculated as reads per kilobase per million mapped reads (RPKM). These data provide a foundation for opium poppy transcriptomic analysis, which may aid in capturing splice variants and some non-coding RNAs involved in the regulation of BIAs biosynthesis. It can also be used for genome assembly and annotation which will favor in new transcript identification.

Beneficial effects of dietary ß-glucan on growth and health status of Pacific white shrimp Litopenaeus vannamei at low salinity.

An 8-week trial was conducted to evaluate the effect of dietary ß-glucan supplement (0, 0.01%, 0.02%, or 0.04%) on growth and health of Pacific white shrimp Litopenaeus vannamei at low salinity of 3 practical salinity unit (psu). The L. vannamei fed 0.02% and 0.04% ß-glucan gained more weight and showed higher activities of protease, amylase, superoxide dismutase, and glutathione peroxidase in the intestine than in the control (0% ß-glucan). The L. vannamei fed 0.04% ß-glucan had a higher condition factor than those fed the control diet. Amylase activity in the hepatopancreas of L. vannamei fed 0.02% ß-glucan was higher than those fed the control diet. Dietary ß-glucan supplement increased the mRNA expressions of Toll-like receptor, myostatin, immune deficiency or heat shock protein 70, but decreased the mRNA expressions of tumor necrosis factor-α and C-type lectin 3 in both hepatopancreas and intestine. The response of intestine microbiota in L. vannamei fed 0.04% ß-glucan was further compared to the control. The 0.04% ß-glucan supplement reduced richness and diversity of the intestinal microbial community as indicated by the low values of Chao1 estimator, ACE estimator, Simpson index and Shannon diversity index. Abundances of Bacillus, Chitinibacter, Geobacillus and Vibrio in the intestine increased, while Flavobacterium, Microbacterium and Mycobacterium decreased significantly in L. vannamei fed 0.04% ß-glucan compared to the control. This study indicates that dietary ß-glucan supplement at 0.02%-0.04% can significantly improve digestibility, antioxidant capacity and immunity in L. vannamei, and thus improve growth performance and survival at low salinity. These beneficial effects of ß-glucan probably are related to the dominance of probiotics over potential pathogens in the intestine.

On-Demand Versatile Prodrug Nanomicelle for Tumor-Specific Bioimaging and Photothermal-Chemo Synergistic Cancer Therapy.

Photothermal therapy is a promising approach for antitumor application although regrettably restricted by available photothermal agents. Physical entrapment of organic near-infrared dyes into nanosystems was extensively studied to reverse the dilemma. However, problems still remained, such as drug bursting and leakage. We developed here an amphiphilic prodrug conjugate by chemically modifying indocyanine green derivative (ICG-COOH) and paclitaxel (PTX) to hyaluronic acid (HA) backbone for integration of photothermal-chemotherapy and specific tumor imaging. The prepared ICG-HA-PTX conjugates could self-assemble into nanomicelles to improve the stability and reduce systemic toxicity of the therapeutic agents. The high local concentration of ICG-COOH in nanomicelles resulted in fluorescence self-quenching, leading to no fluorescence signal being detected in circulation. When the nanomicelles reached the tumor site via electron paramagnetic resonance effect and HA-mediated active targeting, the overexpressed esterase in tumor cells ruptured the ester linkage between drugs and HA, achieving tumor-targeted therapy and specific imaging. A series of in vitro and in vivo experiments demonstrated that the easily prepared ICG- HA-PTX nanomicelles with high stability, smart release behavio r, and excellent tumor targeting ability showed formidable synergy in tumor inhibition, which provided new thoughts in developing an organic near-infrared-dye-based multifunctional delivery system for tumor theranostics.

A dual-targeting reconstituted high density lipoprotein leveraging the synergy of sorafenib and antimiRNA21 for enhanced hepatocellular carcinoma therapy.

Sorafenib (So) is a multi-target kinase inhibitor extensively used in clinic for hepatocellular carcinoma therapy. It demonstrated strong inhibition both in tumor proliferation and tumor angiogenesis, while hampered by associated cutaneous side-effect and drug resistance. The knockdown of miR-21 with antisense oligonucleotides (antimiRNA21) was regarded as an efficient strategy for increasing tumor sensibility to chemotherapy, which could be employed to appreciate the efficacy of So. Herein, we successfully formulated a dual-targeting delivery system for enhanced hepatocellular carcinoma therapy by encapsulating So and antimiRNA21 in RGD pentapeptide-modified reconstituted high-density lipoprotein (RGD-rHDL/So/antimiRNA21). The RGD and apolipoprotein A-I (ApoA-I) on nanoparticles (NPs) could drive the system simultaneously to tumor neovascular and parenchyma by binding to the overexpressed ανß3-integrin and SR-B1 receptors, achieving precise delivery of therapeutics to maximize the efficacy. A series in vitro and in vivo experiments revealed that co-delivery of So and antimiRNA21 by RGD-rHDL significantly strengthened the anti-tumor and anti-angiogenic effect of So with negligible toxicity towards major organs, reversed drug-resistance and was capable of remodeling tumor environments. The constructed RGD-rHDL/So/antimiRNA21 with improved efficacy and excellent tumor targeting ability provided new idea for chemo-gene combined therapy in hepatocellular carcinoma. STATEMENT OF SIGNIFICANCE: Sorafenib (So) is a multi-target kinase inhibitor which was approved by FDA as first-line drug for hepatocellular carcinoma (HCC) therapy. However, long term application of So in clinic was hampered by serious dermal toxicity and drug resistance. Although numerous researchers were devoted to finding alternatives or therapies as combination treatments with So to reach more desired therapeutic efficacy, the therapeutic options were still limited. The present study prepares RGD pentapeptide decorated biomimic reconstituted high-density lipoprotein (rHDL) loaded with So and antimiRNA21 (RGD-rHDL/So/antimiRNA21) for enhanced HCC therapy. The RGD-rHDL/So/antimiRNA21 NPs offer an effective platform for anti-tumor and anti-angiogenesis therapy in HCC and provide new approach to reverse drug-resistance of So for feasible clinical application.

Protection of ß-carotene from chemical degradation in emulsion-based delivery systems using antioxidant interfacial complexes: Catechin-egg white protein conjugates.

The aim of the present study was to fabricate catechin-egg white protein (CT-EWP) conjugates as novel food-grade antioxidant emulsifiers designed to improve the physicochemical stability of ß-carotene (BC) emulsions. CT-EWP conjugates were synthesized using free radical grafting, and the formation of conjugates was confirmed by electrophoresis and liquid chromatography-mass spectrometry. The physicochemical stability of BC emulsions was characterized by measuring alterations in particle size, ζ-potential and BC retention. The particle size and ζ-potential changed more rapidly at 37°C than at 4 or 25°C, however no creaming or oiling-off were observed at any of the storage temperatures, suggesting all emulsions were physically stable throughout the 30-day storage period. Compared to emulsions stabilized by EWP or CT+EWP physical mixtures (no conjugation), CT-EWP conjugate-stabilized emulsions had better resistance to environmental stresses, such as thermal processing and high ionic strengths, which was attributed to a stronger steric repulsion between the oil droplets. CT-EWP conjugates also significantly reduced the degradation rate of BC in emulsions during storage (p<0.05), which was attributed to their strong antioxidant and interfacial activities. These results indicate that CT-EWP conjugates can be utilized to develop food-grade delivery systems to protect chemically labile lipophilic bioactive compounds.

Physical and antimicrobial properties of thyme oil emulsions stabilized by ovalbumin and gum arabic.

Natural biopolymer stabilized oil-in-water emulsions were formulated using ovalbumin (OVA), gum arabic (GA) solutions and their complexes. The influence of interfacial structure of emulsion (OVA-GA bilayer and OVA/GA complexes emulsions) on the physical properties and antimicrobial activity of thyme oil (TO) emulsion against Escherichia coli (E. coli) was evaluated. The results revealed that the two types of emulsions with different oil phase compositions remained stable during a long storage period. The oil phase composition had an appreciable influence on the mean particle diameter and retention of the TO emulsions. The stable emulsion showed a higher minimum inhibitory concentration (MIC), and the TO emulsions showed an improved long-term antimicrobial activity compared to the pure thyme oil, especially complexes emulsion at pH 4.0. These results provided useful information for developing protection and delivery systems for essential oil using biopolymer.

Nutritional composition of boletus mushrooms from Southwest China and their antihyperglycemic and antioxidant activities.

Thirteen samples representing five species were collected from different provinces of Southwest China, and their chemical composition, antihyperglycemic activity, and antioxidant activity were evaluated. These mushrooms had high crude protein (21.72-30.59g/100g dw) and total carbohydrate (49.18-62.58g/100g dw) contents, but low crude fat contents (1.96-7.87g/100g dw). They also accumulated notable quantities of potassium, zinc, sodium, magnesium and copper from the soil. The potassium content, in particular, was 18.75-39.21 times that found in the soil at the collection site. The natural habitat of these mushrooms, especially the mineral content of the soil, seems to have more influence on the mineral content of these mushrooms than their species. Most of the samples possessed antihyperglycemic and antioxidant activities. Suillellus luridus showed the highest antioxidant activity and antihyperglycemic activities, suggesting that S. luridus shows potential for development as a dietary nutritional supplement.

Artesunate ameliorates severe acute pancreatitis (SAP) in rats by inhibiting expression of pro-inflammatory cytokines and Toll-like receptor 4.

Severe acute pancreatitis (SAP) is a severe clinical condition with significant morbidity and mortality. Multiple organs dysfunction (MOD) is the leading cause of SAP-related death. The over-release of pro-inflammatory cytokines such as IL-1ß, IL-6, and TNF-α is the underlying mechanism of MOD; however, there is no effective agent against the inflammation. Herein, artesunate (AS) was found to increase the survival of SAP rats significantly when injected with 3.5% sodium taurocholate into the biliopancreatic duct in a retrograde direction, improving their pancreatic pathology and decreasing serum amylase and pancreatic lipase activities along with substantially reduced pancreatic IL-1ß and IL-6 release. In vitro, AS-pretreatment strongly inhibited IL-1ß and IL-6 release and their mRNA expressions in the pancreatic acinar cells treated with lipopolysaccharide (LPS) but exerted little effect on TNF-α release. Additionally, AS reduced the mRNA expressions of Toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) p65 as well as their protein expressions in the pancreatic acinar cells. In conclusion, our results demonstrated that AS could significantly protect SAP rats, and this protection was related to the reduction of digestive enzyme activities and pro-inflammatory cytokine expressions via inhibition of TLR4/NF-κB signaling pathway. Therefore, AS may be considered as a potential therapeutic agent against SAP.

Antidiabetic activity of mycelia selenium-polysaccharide from Catathelasma ventricosum in STZ-induced diabetic mice.

Se-polysaccharide from Catathelasma ventricosum (SPC-2) was purified by DEAE-52 and Sephadex G-100 column chromatography. The average size of SPC-2 was 1.6×10(5) Da, and it was mainly composed of glucose (87.4%) with the conformation of ß-pyran ring. The branched structure of SPC-2 was proved intuitively by atomic force microscope (AFM). The antidiabetic potential of SPC-2 was tested in STZ-induced diabetic mice. After STZ-induced diabetic mice being administered of SPC-2 for 30 days, SPC-2 treatment significantly reduced the levels of malondialdehyde (MDA) and low-density lipoprotein cholesterol (LDL-C) that were increased by the STZ treatment. Further, the SPC-2 treatment led to increased activity of antioxidant enzymes in liver and kidney and high-density lipoprotein cholesterol (HDL-C) that were decreased by the STZ. The results of histopathology also showed SPC-2 protected tissues (pancreas, liver and kidney) against peroxidation damage and maintained tissue integrity.

Effect of supplements Mn2+, Cu2+, and aromatic compounds and Penicillium decumbens on lignocellulosic enzyme activity and productivity of Catathelasma ventricosum.

This is the first report on using Catathelasma ventricosum for production of fruiting body and lignocellulosic enzymes. To improve the laccase activity and productivity of mushroom, the substrate was added with different supplements (eight aromatic compounds, Mn2+, and Cu2+). Based on the results, all these supplements can improve the laccase activity and productivity of C. ventricosum, and it seems that there is a critical value of laccase activity that affects the productivity of C. ventricosum. In addition, when Penicillium decumbens was inoculated into the substrate that had been cultivated with C. ventricosum for 20 days, the highest values of laccase activity, FPA activity, and productivity of C. ventricosum were obtained. Moreover, the laccase activity showed a positive correlation with the productivity of C. ventricosum. Finally, the effect of Mn2+, Cu2+, and P. decumbens on laccase activity was investigated by response surface methodology (RSM).

Chicken embryo extracts enhance spleen lymphocyte and peritoneal macrophages function.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditionally, chicken embryo eggs are widely considered as an immunomodulatory health product in China and southeast Asia. Besides that, Compendium of Materia Medica, the most famous Chinese pharmacopoeia, has recorded that chicken embryo eggs contributed to the appetite increase and immune function enhancement, especially for the pregnant and aged. AIM OF THE STUDY: Through assessing the immune efficacy of the chicken embryo extracts on immunocompetent cells, the immunomodulatory mechanism and components were explored preliminarily. MATERIALS AND METHODS: The raw material was subjected to in vitro gastric and pancreatic digestion successively, and then ligarine was added to separate the digested mixture to water-soluble fraction (WS) and lipid-soluble fraction (LS). Employing in vitro cell models, the immunomodulating activities of the extracts on immunosuppressed mouse were examined, at concentrations ranging from 10 to 500 µg/ml. The cell experiments consisted of the following experiments: spleen lymphocyte proliferation and IL-2 secretion, peritoneal macrophage phagocytosis and NO production. Before cell experiments, Kjeldal method and phenol-sulfuric acid method were employed respectively to determine the total protein and polysaccharide content of WS, and gas chromatography to analyze the fatty acid composition of LS. RESULTS: LS significantly enhanced the spleen lymphocyte proliferation and IL-2 secretion, while peritoneal macrophage phagocytosis and NO production activity were elevated by WS. Both extracts exhibited an immunomodulatory effect with a dose-effect relationship. CONCLUSION: The findings confirmed the chicken embryo eggs as a potent immunopotentiator on the immunosuppressed mouse, which would be useful for the fruitful application of the chicken embryo eggs to serve as a superior immunostimulatory tonic. Furthermore, the two extracts exhibited a different bioactivity tendency which makes it a significative work to screen the bioactive component for further study.

Chemical composition of five wild edible mushrooms collected from Southwest China and their antihyperglycemic and antioxidant activity.

Evaluation of the chemical composition and antihyperglycemic and antioxidant activity of five wild edible mushrooms (Clitocybe maxima, Catathelasma ventricosum, Stropharia rugoso-annulata, Craterellus cornucopioides and Laccaria amethystea) from Southwest China. The chemical composition assay includes proximate analysis (moisture, ash, crude protein, crude fat, total carbohydrates and total energy), bioactive compounds analysis (total phenolic, flavonoid, ascorbic acid, ergosterol, tocopherol), fatty acid analysis, amino acid analysis, phenolic compounds analysis and mineral analysis of these mushrooms. Furthermore, assays of α-glucosidase inhibitory and α-amylase inhibitory activity were used for evaluating antihyperglycemic activity of the mushrooms, and assays of reducing power, chelating effect on ferrous ions, scavenging effect on hydroxyl free radicals and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were used for evaluating antioxidant activity of the mushrooms. Based on the results, ethanolic and aqueous extract of these mushroom all showed antihyperglycemic and antioxidant potential. In particular, the aqueous extract of C. ventricosum revealed the highest α-glucosidase inhibitory activity (EC50 value 2.74 µg/mL), DPPH radical scavenging activity (EC50 value 2.86 mg/mL) and reducing power (EC50 value 0.96 mg/mL), while the aqueous extract of L. amethystea showed the highest α-amylase inhibitory activity (EC50 value 4.37 µg/mL) and metal chelating activity (EC50 value 2.13 mg/mL).