Construction and antibacterial activities of walnut green husk polysaccharide based silver nanoparticles (AgNPs).

In this paper, the size-controllable nano­silver particles (AgNPs) were synthesized from walnut green husk polysaccharide, and its cytotoxicity and antibacterial activity were evaluated. Firstly, acidic polysaccharide WGHP2 was extracted from walnut green husk, and then the silver ion in AgNO3 was reduced in WGHP2 aqueous solution using NaBH4, so as to synthesize the nano­silver composite. The nano­silver composite was characterized by transmission electron microscope, Fourier infrared spectroscopy, ultraviolet-visible spectrometer, scanning electron microscope, inductively coupled plasma mass spectrometry and X-ray photoelectron spectroscopy. The results show that AgNPs stabilized by WGHP2 are mainly regular spheres with an average particle size distribution of 15.04-19.23 nm. The particle size distribution and morphology of AgNPs changed with the concentration of silver precursor, which is related to the dispersion of silver precursor in polysaccharide aqueous solution and the formation of AgO coordination bond between silver precursor and polysaccharide molecules. These coordination bonds changed the ability of nanoparticles to produce and release Ag+, and thus regulated their antibacterial activity and cytotoxicity, as evidenced by the experimental result of the cytotoxicity of the nano­silver particle against PC12 cells and the bacteriostatic effect on E.coli and S.aureus. Conclusively, WGHP2-Ag has good stability, antibacterial activity and low cytotoxicity.

Bacterial cellulose nanofibers used as nanofillers improved the fresh-keeping performance of gelatin-based edible coating for fresh-cut apples.

In this study, bacterial cellulose nanofibers (BCNs) (0%, 1%, 2%, and 3%) were used as nanofillers to prepare gelatin-based edible films, and their physical properties and fresh-keeping performance were investigated. The microstructure observation showed that the BCNs were well dispersed in the gelatin-based edible films and the surface roughness of the films increased with the increase of BCNs content. X-ray diffraction and thermogravimetric analysis showed that the crystallinity and thermal stability of the film were significantly increased with the increase of BCNs. Fourier-transform infrared spectroscopy analysis suggested that hydrogen bond interactions occurred between BCNs and gelatin polymers, leading to improved mechanical properties with the increase of BCNs content. Furthermore, the barrier performance was also improved with the increase of BCNs content, where gelatin-based edible films with 2% BCNs showed the best mechanical property. Meanwhile, the gelatin-based film-forming solutions (FFSs) containing different BCNs were coated on the fresh-cut apples and the corresponding fresh-keeping performance was investigated. The results showed that the fresh-keeping parameters of fresh-cut apples coated with FFSs containing BCNs were better as compared with those of pure gelatin FFSs. Moreover, the fresh-keeping parameters were improved with the increase of BCNs, especially the FFSs containing 2% BCNs that showed the best fresh-keeping parameters. Therefore, BCNs, used as nanofillers, are an excellent enhancer to improve the fresh-keeping performance of the gelatin-based edible coating, showing a promising potential application in the food preservation field.

Volatomics-assisted characterization of aroma and off-flavor contributors in fresh and thermally treated kiwifruit juice.

The distinctive aroma profile of kiwifruit juice was significantly changed during thermal treatment, however, the theoretical basis for clarifying and controlling the changes was deficient. In this study, we applied volatomics techniques to investigate the contributors of off-flavors in thermally treated kiwifruit juice. Sixteen aroma compounds were identified to be responsible for the typical "fruity", "grassy", and "cucumber-like" flavors of fresh kiwifruit by two different fused silica capillary columns coupled with chromatography-olfactometry/detection frequency (GC-O/DF) analysis and calculation of odor activity value (OAV). Thirty-one odor-active compounds were determined as important contributors to the sensory profile of thermally treated kiwifruit juice, 14 of which were common to all varieties investigated. The key aroma compounds on fresh kiwifruit significantly decreased after thermal treatment, while decanal, (E)-2-decenal, methional, ß-damascenone, 1-octen-3-one, DMHF, and dimethyl sulfide which presented undesirable cooked cabbage/potato, roasted fruit, and sulfurous odors, were accumulated in a large amount. By applying PLSR analysis, (E)-2-decenal, methional, ß-damascenone, DMHF, and dimethyl sulfide were further verified to have great contributions to the formation of the cooked off-flavor during thermal treatment. Moreover, XX was found to be more thermal-sensitive and more prone to forming cooked off-flavors after thermal treatment. This study could provide theoretical guidance for the regulation of thermal-induced off-flavors during the manufacturing of kiwifruit juice.

Promoting adsorption performance and mechanical strength in composite porous gel film.

Starch is widely used to prepare biodegradable films due to its superior biocompatibility, low immunogenicity, and renewability. In this work, a novel K+/carrageenan porous-starch/casein gel film with high oil absorption was prepared using modified porous starch. Optimal gel stability and uniformity were obtained when adding 10 mg/mL k-carrageenan and 2 mg/mL K+ to 2 mg/mL microgels, with significantly reduced crystallinity and elasticity and increased tensile strength. The concentration of k-carrageenan was the main factor affecting gel strength and the hydrophilic and mechanical properties of the film. In addition, the film-forming solution showed excellent fluidity and spreading typical of non-Newtonian fluids. The film also exhibited a highly porous structure, as visualized by SEM and AFM, in line with a cumulative oil absorption rate of 87.5 % within 20 min, which was significantly higher than that obtained with glutinous rice starch. In conclusion, reinforcement of starch-based microgels as described in this study can maximize the film's adsorption performance and mechanical properties, with promising applications in skin care and beauty products.

Effect of Juglone against Pseudomonas syringae pv Actinidiae Planktonic Growth and Biofilm Formation.

Pseudomonas syringaepv Actinidiae (P. syringae) is a common pathogen causing plant diseases. Limoli proved that its strong pathogenicity is closely related to biofilm state. As a natural bacteriostatic agent with broad-spectrum bactericidal properties, juglone can be used as a substitute for synthetic bacteriostatic agents. To explore the antibacterial mechanism, this study was carried out to examine the inhibitory effect of juglone on cell membrane destruction, abnormal oxidative stress, DNA insertion and biofilm prevention of P. syringae. Results showed that juglone at 20 µg/mL can act against planktogenic P. syringae (107 CFU/mL). Specially, the application of juglone significantly damaged the permeability and integrity of the cell membrane of P. syringae. Additionally, juglone caused abnormal intracellular oxidative stress, and also embedded in genomic DNA, which affected the normal function of the DNA of P. syringae. In addition, environmental scanning electron microscope (ESEM) and other methods showed that juglone effectively restricted the production of extracellular polymers, and then affected the formation of the cell membrane. This study provided a possibility for the development and utilization of natural juglone in plants, especially P. syringae.

Rapid Detection of Dimethoate in Soybean Samples by Microfluidic Paper Chips Based on Oil-Soluble CdSe Quantum Dots.

Given the imperative of monitoring organophosphorus pesticides (OPs) residues in the ecosystem, here a novel, facile and sensitive fluorescence sensor is presented for the rapid detection of dimethoate. In this work, surface molecularly imprinted polymer (SMIP) and microfluidic technology had been introduced to enhance the selectivity and portability of the described methodology. Oil-soluble CdSe quantum dots (QDs) synthesized in a green way were used as fluorescent material for the selective detection of dimethoate on the basis of static quenching and photoinduced electron transfer mechanism. Among many kinds of paper materials, glass fiber paper was used as the novel substrate of paper chip due to low pristine fluorescence and better performance when combining CdSe QDs. In the process of molecular imprinting, the interaction between several functional monomers and dimethoate molecule was investigated and simulated theoretically by software to improve the selectivity of the sensor. Consequently, the fabricated novel detection platform could effectively respond to dimethoate in 10 min with the concentration range of 0.45-80 µmol/L and detection limit of 0.13 µmol/L. The recovery in the spiked experiment soybean sample was in an acceptable range (97.6-104.1%) and the accuracy was verified by gas chromatography-mass spectrometry, which signified the feasibility and potential in food sampling.

Juglone Inactivates Pseudomonas aeruginosa through Cell Membrane Damage, Biofilm Blockage, and Inhibition of Gene Expression.

Due to the strong drug resistance of Pseudomonas aeruginosa (P. aeruginosa), the inhibition effects of conventional disinfectants and antibiotics are not obvious. Juglone extracted from discarded walnut husk, as a kind of plant-derived antimicrobial agent, has the advantages of naturalness, high efficiency, and low residue, with a potential role in the inhibition of P. aeruginosa. This study elucidated the inhibitory effect of juglone on the growth of plankton and the formation of P. aeruginosa biofilm. The results showed that juglone (35 µg/mL) had an irreversible inhibitory effect on P. aeruginosa colony formation (about 107 CFU/mL). The integrity and permeability of the cell membrane were effectively destroyed, accompanied by disorder of the membrane permeability, mass leakage of the cytoplasm, and ATP consumption. Further studies manifested that juglone could induce the abnormal accumulation of ROS in cells and block the formation of the cell membrane. In addition, RT-qPCR showed that juglone could effectively block the expression of five virulence genes and two genes involved in the production of extracellular polymers, thereby reducing the toxicity and infection of P. aeruginosa and preventing the production of extracellular polymers. This study can provide support for the innovation of antibacterial technology toward P. aeruginosa in food.

Load phycocyanin to achieve in vivo imaging of casein-porous starch microgels induced by ultra-high-pressure homogenization.

Traditional bioactive substances are often limited in practical application due to their poor stability and low solubility. Therefore, it is imperative to develop biocompatible high loading microgel carriers. In this study, a novel type of casein-porous starch microgel was prepared under ultra-high-pressure homogenization, by using porous starch with the honeycomb three-dimensional network porous structure. Molecular interaction force analysis and thermodynamic analysis showed that electrostatic interaction played a major role in the formation of microgels. Circular dichroism and Fourier transform infrared spectroscopy showed that homogenization and pH were the main factors, which affected the formation and structural stability of microgels. Compared with casein-glutinous rice starch microgels, the encapsulation efficiency and loading capacity of phycocyanin in casein-porous starch microgels were increased by 77.27% and 135.10%, respectively. Thus, casein-porous starch microgels could not only achieve a sustained release effect, but also effectively transport phycocyanin to the gastrointestinal tract of zebrafish, while achieving good fluorescence imaging in vivo. Ultimately, the prepared casein-porous starch microgels could enrich the nanocarriers material, and contribute to the research of safe and effective fluorescent imaging materials.

Effect of Walnut Meal Peptides on Hyperlipidemia and Hepatic Lipid Metabolism in Rats Fed a High-Fat Diet.

As a natural active substance that can effectively improve blood lipid balance in the body, hypolipidemic active peptides have attracted the attention of scholars. In this study, the effect of walnut meal peptides (WMP) on lipid metabolism was investigated in rats fed a high-fat diet (HFD). The experimental results show that feeding walnut meal peptides counteracted the high-fat diet-induced increase in body, liver and epididymal fat weight, and reduce the serum concentrations of total cholesterol, triglycerides, and LDL-cholesterol and hepatic cholesterol and triglyceride content. Walnut meal peptides also resulted in increased HDL-cholesterol while reducing the atherosclerosis index (AI). Additionally, the stained pathological sections of the liver showed that the walnut meal peptides reduced hepatic steatosis and damage caused by HFD. Furthermore, walnut meal peptide supplementation was associated with normalization of elevated apolipoprotein (Apo)-B and reduced Apo-A1 induced by the high-fat diet and with favorable changes in the expression of genes related to lipid metabolism (LCAT, CYP7A1, HMGR, FAS). The results indicate that walnut meal peptides can effectively prevent the harmful effects of a high-fat diet on body weight, lipid metabolism and liver fat content in rats, and provide, and provide a reference for the further development of walnut meal functional foods.

Walnut green husk polysaccharides prevent obesity, chronic inflammatory responses, nonalcoholic fatty liver disease and colonic tissue damage in high-fat diet fed rats.

High-fat (HF) diets cause obesity, gut microbial dysbiosis and associated disorders and inflammatory bowel disease (IBD) due to increased intestinal permeability, which is an important reason for chronic inflammation and oxidative stress. This study was to investigate the effects and mechanism by which walnut green husk polysaccharides (WGHP) prevents obesity, oxidative stress, inflammation, liver and colon damage in HF diet induced rats. We found that WGHP alleviated HF-induced abnormal weight gain, disordered lipid metabolism, inflammation, oxidative stress, colonic tissue injury and up-regulate the expression level of colonic tight junction protein in the rats. Besides, the administration of WGHP promoted browning of iWAT and thermogenesis in BAT of HF-fed rats, and improved gut microbiota dysbiosis by increasing the bacterial diversity and reducing the relative abundance of potential pathogenic bacteria in the colon of the rats. Furthermore, WGHP consumption not only increased the SCFAs content but also improved the relative abundance of Prevotellaceae and Allobaculum in the gut of rats. Our results suggest that the protective effect of WGHP on metabolic inflammation caused by HF may be due to the regulation of gut microbiota and SCFAs.

Novel Angiotensin-Converting Enzyme Inhibitory Peptides Identified from Walnut Glutelin-1 Hydrolysates: Molecular Interaction, Stability, and Antihypertensive Effects.

In recent years, angiotensin-converting enzyme (ACE) inhibitory peptide has become a research hotspot because of its essential role in maintaining human blood pressure balance. In this study, two novel ACE inhibitory peptides of Val-Glu-Arg-Gly-Arg-Arg-lle-Thr-Ser-Val (Valine-Glutamate-Arginine-Glycine-Arginine-Arginine-Isoleucine-Threonine-Serine-Valine, VERGRRITSV) and Phe-Val-Ile-Glu-Pro-Asn-Ile-Thr-Pro-Ala (Phenylalanine-Valine-Isoleucine-Glutamate-Proline-Asparagine-Isoleucine-Threonine-Proline-Alanine, FVIEPNITPA) were isolated and purified from defatted walnut meal hydrolysates through a series of preparation processes including ultrafiltration, Sephadex G-15 gel chromatography, and reverse high performance liquid chromatography (RP-HPLC). Both peptides showed high ACE inhibitory activities. The molecular docking study revealed that VERGRRITSV and FVIEPNITPA were primarily attributed to the formation of strong hydrogen bonds with the active pockets of ACE. The binding free energies of VERGRRITSV and FVIEPNITPA with ACE were -14.99 and -14.69 kcal/mol, respectively. Moreover, these ACE inhibitory peptides showed good stability against gastrointestinal enzymes digestion and common food processing conditions (e.g., temperature and pH, sugar, and salt treatments). Furthermore, animal experiment results indicated that the administration of VERGRRITSV or FVIEPNITPA exhibited antihypertensive effects in spontaneously hypertensive rats. Our results demonstrated that walnut could be a potential source of bioactive peptides with ACE inhibitory activity.

Characterization of structural, functional and antioxidant properties and amino acid composition of pepsin-derived glutelin-1 hydrolysate from walnut processing by-products.

Glutelin-1 of defatted walnut meal protein (DWPG-1) was modified by pepsin enzymatic hydrolysis to improve its functional properties and antioxidant activities. The amino acid composition, structural characteristics, physicochemical and functional properties as well as antioxidant activities of the hydrolysate were compared with those of unmodified DWPG-1. The analysis of X-ray diffraction patterns, surface microstructure and particle size distribution indicated that enzymatic hydrolysis changed the structures of DWPG-1. Compared with the natural unhydrolyzed protein, the hydrolysate showed better physicochemical properties, such as surface hydrophobicity, solubility, emulsifying properties, foaming properties and water absorption capacity. In addition, the hydrolysate also exhibited significantly stronger antioxidant activities than DWPG-1. In conclusion, the results of this study prove that pepsin-mediated hydrolysis of walnut glutelin-1 can effectively modify the structure, function and antioxidant activity of DWPG-1, and could be used as an effective technology to produce bioactive multifunctional hydrolysates.

Boronate affinity material-based sensors for recognition and detection of glycoproteins.

Glycoproteins are closely linked to the occurrence and development of many diseases. Therefore, it is of great importance to develop highly selective, sensitive, efficient detection methods for glycoproteins. To overcome the problems with traditional detections methods, such as mass spectrometry, chromatography-mass spectrometry, and enzyme-linked immunosorbent assay, boronate affinity material (BAM)-based sensors have developed rapidly for the specific recognition and detection of glycoproteins because of the advantages of pH-controlled binding/release, reversibility of the reaction, high specificity, and high selectivity, showing their wide application prospects. In recent years, there have been many significant leaps in the use of BAMs for sensing and detecting glycoproteins, but there are still many challenges and room for development. Therefore, this review critically investigates and summarizes recent advances with BAM-based sensors for glycoprotein detection. We focus on the common boronate affinity ligands of BAMs and their grafting methods, functional materials utilized in the synthesis of BAM-based sensors, advanced technologies, and applications. Finally, we propose the remaining challenges and future perspectives to accelerate the development of BAMs, and to utilize it for further developing versatile BAMs with a variety of promising applications.

The protective effects of walnut green husk polysaccharide on liver injury, vascular endothelial dysfunction and disorder of gut microbiota in high fructose-induced mice.

This study aimed to investigate the protective effects of walnut green husk polysaccharide (WGHP) on liver injury, vascular endothelial dysfunction and disorder of gut microbiota in mice induced by high fructose (HF) diet. The chemical analysis results show that the walnut green husk polysaccharide is a low molecular weight acidic heteropolysaccharide, composed mainly of glucuronic acid, arabinose and galactose. Biochemical analysis showed that WGHP significantly improved glucose metabolism and lipid metabolism and decreased oxidative stress in HF-diet induced obesity mice. Histopathological observation of liver and cardiovascular aorta confirmed the protective effects of WGHP on hepatic steatosis and vascular endothelial dysfunction. Furthermore, 16S rRNA sequencing results demonstrated that WGHP reversed the disorders of gut microbiota caused by HF, decreased the relative abundance of Verrucomicrobia and increased the relative abundance of Deferribacteres at the phylum level, decreased the relative abundance of Akkermansia, Lachnoclostridium and norank_f__Muribaculaceae and increased the relative abundance of Prevotellaceae_UCG-001, Helicobacter, Alloprevotella and Allobaculum at the genus levels. Our results indicate that WGHP may act as a functional polysaccharide for protecting liver and cardiovascular in HF-fed mice.

New insights into the alleviating role of Melaleuca alternifolia oil on metabolites pathway disorder of grapes caused by Aspergillus niger, verified by corresponding key genes expression.

Grape berries are susceptible to Aspergillus niger (A. niger) infection during storage, leading to a significant reduction in its nutritional quality. However, most alternations in nutrient contents and related gene expression during fungal infection or treated with antimycotics remain unexplored. This work aimed to monitor and verify the metabolic changes in berries caused by A. niger or Melaleuca alternifolia oil (MAO) by using UHPLC-ESI-MS2 and Quantitative Real-time PCR (RT-qPCR). Results showed that sucrose, glucose, fructose, trans-resveratrol and pterostilbene levels were down and pentose phosphate pathway, glycolysis pathway and phenylpropanoid pathway were significantly down-regulated compared with healthy berries due to A. niger infection, all of which were alleviated by MAO treated. A. niger also induced down-regulation of key genes expression associated with metabolic pathways and magnitude of down-regulation was reduced by MAO. These results provide a theoretical basis for MAO used to control the risk of A. niger-mediated diseases.

Influence of modified atmosphere treatment on post-harvest reactive oxygen metabolism of pomegranate peels.

Modified atmosphere storage can regulate the reactive oxygen metabolism of fruits and vegetables, reduce the accumulation of hazardous free radicals, and mitigate the peroxidation degree of fruit membrane lipids. In this study, different gas matching ratios were adopted for the modified atmosphere treatment of pomegranate fruits. Up to 120 d of storage, compared with the control treatment, the H2O2 and malonaldehyde (MDA) contents in treatment 2 decreased by 8.88% and 18.28%, respectively, when the activities of superoxide dismutase (SOD), catalase (CAT) and ascorbic acid peroxidase (APX) in treatment 2 increased by 21.44%, 117.38% and 114.95%, the ascorbic acid (ASA) and glutathione (GSH) contents in treatment 2 also increased by 116.83% and 50%, these results showed that treatment 2 (6.0% O2, 6.0% CO2) could effectively regulate various indexes of the reactive oxygen metabolism of pomegranate peels, maintain the normal physiological actions of the fruits, and postpone the ripening and senescence of histocytes. Under treatment 4 (10.0% O2, 10.0% CO2), H2O2 contents in the pomegranate peel significantly increased, and the activities of SOD, CAT and APX significantly reduced. ASA and GSH were degraded, the MDA content abruptly increased, the membrane lipid peroxidation accelerated, and the cytomembrane structure was destroyed.

Enhancing the antitumor activity of tea polyphenols encapsulated in biodegradable nanogels by macromolecular self-assembly.

Nanogels (NGs) with desirable stability have emerged as a promising platform for biomedical applications. Herein, a convenient approach was developed to encapsulate and protect tea polyphenols (TPs) by macromolecular self-assembly of lysozyme (Ly) and carboxymethyl cellulose (CMC) through a heating treatment. Biodegradable Ly-CMC NGs were formed on the basis of molecules driven by electrostatic interaction and hydrophobic forces. The particle size and morphology of the Ly-CMC NGs were analyzed using a Malvern particle size analyzer, fluorescence spectrophotometer, and scanning electron microscope. The results showed that the heated NGs were spherical with better stability and smaller particle size. The encapsulation efficiency of TP-loaded NGs was 89.05 ± 3.14%, and it indicated that the Ly-CMC NGs may have a strong binding force with TPs. Moreover, TP-loaded NGs showed a sustained release feature. The DPPH and ABTS-scavenging rates of the TP-loaded NGs were 76.5% and 86.1%, respectively. The antitumor activity of the TP-loaded NGs can effectively inhibit the proliferation of HepG2 cells. Furthermore, TP-loaded NGs were proven to significantly enhance the induction of apoptosis in hepatoma cells and exhibit obvious cell cycle arrest. Our results demonstrate that the Ly-CMC NGs have extensive application prospects as a biocompatible and biodegradable delivery carrier of food functional factors to improve their antitumor effects.

Apoptosis triggered by isoquercitrin in bladder cancer cells by activating the AMPK-activated protein kinase pathway.

Cancer cells are well known to require a constant supply of protein, lipid, RNA, and DNA via altered metabolism for accelerated cell proliferation. Targeting metabolic pathways is, therefore, a promising therapeutic strategy for cancers. Isoquercitrin (ISO) is widely distributed in dietary and medicinal plants and displays selective cytotoxicity to cancer cells, primarily by inducing apoptosis and cell cycle arrest. The aims of this study were to find out whether ISO could stabilize in a bladder-like acidic environment and inhibit bladder cancer cell proliferation by affecting their metabolism, and to investigate its molecular mechanism. In this study, the exposure of T24 bladder cancer cells to ISO (20-80 µM) decreased cell viability by causing ROS overproduction. This ROS change regulated the AMPK signaling pathway, and caused Caspase-dependent apoptosis as well as metabolism dysfunction. Metabolic alterations elevated metabolic pathway variation, which in turn destabilized lipid synthesis and altered anaerobic glycolysis. This linkage was proved by immunoblotting assay, and metabolomics as identified by UHPLC-QTOF-MS. Our findings provide comprehensive evidence that ISO influenced T24 bladder cancer cell metabolism, and that this process was mainly involved in activating the AMPK pathway. This study could lead to an understanding of how ISO suppresses bladder cancer cell growth, and whether the affected cancer metabolism is a common mechanism by which nutritional compounds suppress cancers.

Phenolic composition, DNA damage protective activity and hepatoprotective effect of free phenolic extract from Sphallerocarpus gracilis seeds.

The phenolic composition of the free phenolic extract from Sphallerocarpus gracilis seeds was analyzed by HPLC-MS and predominant compounds were chlorogenic acid, di-caffeoylquinic acid glucoside and luteolin-7-O-glucoside. The free phenolic extract was evaluated for DNA damage protective activity induced by ROO and OH radicals and hepatoprotective effect in vivo and in vitro. Results revealed that the free phenolic extract exhibited significant protective activity against both ROO and OH radical-induced DNA damage and the phenolic extract exerted more potent inhibitory activity against OH radical-induced damage than against that induced by ROO radicals. In vivo experimental results showed that the phenolic extract significantly prevented the increase of serum alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase activities and hepatic malondialdehyde level caused by CCl4 in rats, and markedly increased hepatic superoxide dismutase, catalase and glutathione peroxidase levels. Histopathological examinations further confirmed that the phenolic extract could protect the liver from CCl4-induced damage. In vitro experimental results showed that the phenolic extract could reduce BRL hepatocyte apoptosis and damage induced by CCl4. These findings indicate that the S. gracilis seed could be developed as a medicinal herb for the therapy and prevention of hepatic injury.