Pickering Emulsion Stabilized by Hordein-Whey Protein Isolate Complex: Delivery System of Quercetin.

As a lipophilic flavonol, quercetin has low bioavailability, which limits its application in foods. This work aimed to prepare a hordein-based system to deliver quercetin. We constructed hordein-whey isolate protein fibril (WPIF) complexes (H-Ws) by anti-solvent precipitation method at pH 2.5. The TEM results of the complexes showed that spherical-like hordein particles were wrapped in WPIF clusters to form an interconnected network structure. FTIR spectra revealed that hydrogen bonds and hydrophobic interactions were the main driving forces for the complex formation. H-W1 (the mass ratio of hordein to WPIF was 1:1) with a three-phase contact angle of 70.2° was chosen to stabilize Pickering emulsions with oil volume fractions (φ) of 40-70%. CLSM images confirmed that the oil droplets were gradually embedded in the three-dimensional network structure of H-W1 with the increase in oil volume fraction. The emulsion with φ = 70% showed a tight gel structure. Furthermore, this emulsion exhibited high encapsulation efficiency (97.8%) and a loading capacity of 0.2%, demonstrating the potential to deliver hydrophobic bioactive substances. Compared with free quercetin, the bioaccessibility of the encapsulated quercetin (35%) was significantly improved. This study effectively promoted the application of hordein-based delivery systems in the food industry.

The intracellular fate and transport mechanism of shape, size and rigidity varied nanocarriers for understanding their oral delivery efficiency.

Nanocarriers have become an effective strategy to overcome epithelial absorption barriers. During the absorption process, the endocytosis mechanisms, cell internalization pathways, and transport efficiency of nanocarriers are greatly impacted by their physical properties. To understand the relationship between physical properties of nanocarriers and their abilities overcoming multiple absorption barriers, nanocarriers with variable physical properties were prepared via self-assembly of hydrolyzed α-lactalbumin peptide fragments. The impacts of size, shape, and rigidity of nanocarriers on epithelial cells endocytosis mechanisms, internalization pathways, transport efficiency, and bioavailability were studied systematically. The results showed that nanospheres were mainly internalized via clathrin-mediated endocytosis, which was then locked in lysosomes and degraded enzymatically in cytoplasm. While macropinocytosis was the primary pathway of nanotubes and transported to the endoplasmic reticulum and Golgi apparatus, resulting in a high drug concentration and sustained release in cytoplasm. Besides, nanotubes can overcome the multi-drug resistance by inhibiting the P-glycoprotein efflux. Furthermore, nanotubes can open intercellular tight-junctions instantaneously and reversibly, which promotes transport into blood circulation. The aqueous solubility of hydrophobic bioactive mangiferin (Mgf) was improved by nanocarriers. Most importantly, the bioavailability of Mgf was the highest for cross-linked short nanotube (CSNT) which outperformed free Mgf and other formulations by in vivo pharmacokinetic studies. Finally, Mgf-loaded CSNT showed an excellent therapeutic efficiency in vivo for the intervention of streptozotocin-induced diabetes. These results indicate that cross-linked α-lactalbumin nanotubes could be an effective nanocarrier delivery system for improving the epithelium cellular absorption and bioavailability of hydrophobic bioactive compounds.

Comparative study of Pd@Pt nanozyme improved colorimetric N-ELISA for the paper-output portable detection of Staphylococcus aureus.

Staphylococcus aureus is a frequent pathogenic bacterium that has a significant detrimental influence on the health of the human body. Therefore, developing a practical and portable detection platform is critical to ensuring food safety. Nanozymes are a kind of engineered nanomaterials with superior enzyme-like activities, providing infinite possibilities for the development of highly sensitive analytical assays. In this study, mesoporous core-shell palladium@platinum (Pd@Pt) nanozymes were synthesized and then applied as a signal amplifier in Staphylococcus aureus colorimetric immunoassay. At the same time, a careful comparative study of the catalytic performance with natural horseradish peroxidase (HRP), Pd@Pt nanozymes and their complexes Pd@Pt-HRP (HRP coupling with Pd@Pt nanozymes) were firstly performed, as well as clever using a colorimeter to achieve portable signal output. Pd@Pt-HRP bioprobes enable remarkable peroxidase-like catalytic activity, resulting in the highest sensitivity with a limit of detection (LOD) improved from 1 × 105 to 1 × 103 CFU/mL. In all, this work conducted a valuable comparative study and presented an improved strategy for the portable detection of pathogenic bacteria, which was expected to be used in a wide range of applications in food safety regulation and biomedicals arenas.

Isolation of B-constituent through selective complex-induced precipitation of hordein with ι-carrageenan.

The complex behavior of the crude hordein with ι-carrageenan (Car) as a function of pH (11.0-3.0) and Hordein/Car mass ratios (20:1-1:1, w/w) was studied through zeta potential analysis, turbidimetric titration and SDS-PAGE. By preferential binding with Car, B-hordein was isolated from the crude hordein at pHmax (6.2) and Hordein/Car mass ratio of 15:1, which was further confirmed by LC-MS/MS analysis. The results of zeta potential and size of separated B-hordein and C-hordein suggested that the difference in charge density was the main driving force of selective complexation between hordein and Car. Simultaneously, Fourier transform infrared spectroscopy also confirmed the existence of strong electrostatic interaction between B-hordein and Car. Additionally, the more ordered secondary structure of B-hordein at pHmax might be beneficial to its preferential binding with Car. This study further promotes the application of B-hordein in food industry.

A sandwich-based evanescent wave fluorescent biosensor for simple, real-time exosome detection†.

Exosomes are regarded as a promising biomarker for the noninvasive diagnosis and treatment of diseases. The value of exosomes for medical research has promoted the search for a fast, efficient, and sensitive detection method. This study reported a sandwich-based evanescent wave fluorescent biosensor (S-EWFB) for exosome detection. A two-step strategy was implemented to take advantages of the simple binding of fluorescent probes with exosomes via the hydrophobic interaction between the cholesteryl and phospholipid bilayer membrane, as well as real-time detection on an evanescent wave liquid-solid interface based on CD63 aptamer-specific capture to form an exosome@fluorescence probe/aptamer sandwich structure. The one-to-many connection between exosomes and signal molecules and the aptamer-modified evanescent wave optical fiber detection platform reduced the detection limit of exosomes to 7.66 particles/mL, with a linear range of 47.5-4.75 × 106 particles/mL. The entire detection process was simple, rapid, and real-time and lasted about 1 h while requiring no separation and purification. Additionally, this platform showed excellent surface regeneration capability and exhibited good performance during the analysis of tumor and non-tumor-derived exosomes.

Preparation and characterization of phytosterol-loaded nanoparticles with sodium caseinate/dextran conjugates.

Sodium caseinate (SC)/dextran conjugates were prepared via Maillard reaction under controlled dry-heating conditions. Moreover, the nanoparticles of phytosterols (PS) encapsulated by SC or SC/dextran were produced using the emulsion evaporation method. The encapsulation efficiency (78.81 ± 5.22%) of PS in SC/dextran nanoparticles was higher than that (73.5 ± 2.78%) in SC nanoparticles. Compared with the compact and dense structure of SC nanoparticles, SC/dextran nanoparticles existed as relatively loose aggregates. The result of differential scanning calorimetry demonstrated that the encapsulation of PS greatly decreased its crystallinity. The released rates of PS from SC and SC/dextran nanoparticles under acidic gastric conditions were 8.59% and 4.73%, respectively. After 7 h of intestinal digestion, the released rate (52.19%) of PS from SC/dextran nanoparticles was significantly higher than that from SC (32.67%) nanoparticles. Therefore, SC/dextran conjugates prepared by the Maillard reaction are more suitable to be used as wall material for the nano-encapsulation of PS.

Benefit-risk assessment of dietary selenium and its associated metals intake in China (2017-2019): Is current selenium-rich agro-food safe enough?

Dietary consumption of selenium-rich agro-food is an effective way to avoid selenium deficiency diseases, however, over consumption of selenium-rich agro-food will result in potential risk of selenosis and problems with associated metals. In this study, we measured the concentrations of selenium and its associated metals in 2756 common and 4894 selenium-rich agro-food samples in 10 regions of China. We found that selenium-rich rice, flour, edible fungi and algae, meat, and tea contain higher levels of associated metals than other selenium-rich agro-food samples. Increasing the consumption of selenium-rich agro-food could make the actual intakes (AIs) of selenium for all population to meet respective recommended daily intakes (RDIs). Benefit-risk assessment results indicated that increasing the consumption of selenium-rich agro-food make AIs of selenium for all populations meet RDIs, chromium intakes for people under 18 years old exceed provisional tolerated daily intake (PTDIs), while arsenic and cadmium intakes are close to PTDIs. The main dietary contributors of selenium, chromium, arsenic and cadmium were meat, edible fungi and algae, rice, and rice, respectively. The study supported the consumption of selenium-rich agro-food for effective selenium supplement, but also emphasized potential risk from associated metals in selenium-rich agro-food, especially chromium.

Determination of Selenium in Common and Selenium-Rich Rice from Different Areas in China and Assessment of Their Dietary Intake.

In this study, 41 common rice varieties and 211 selenium-rich rice varieties from ten representative areas in China were collected in 2017-2019. The selenium contents of rice were analyzed with optimized inductively coupled plasma mass spectrometry (ICP-MS). Selenium concentrations of common rice and selenium-rich rice ranges were 0.81-7.26 and 0.76-180.73 µg/100 g, respectively. The selenium contents in selenium-rich rice from different areas were significantly different (p < 0.001) while those in common rice from different areas were not. The selenium-rich rice in Harbin and Keshan showed the lowest selenium level and those from selenium-rich areas (Enshi and Ankang) were highest. Based on the estimation of the risk assessment software @risk7.0 (Palisade Corporation, New York, NY, USA), the consumption of selenium-rich rice can effectively increase dietary selenium intake for the population. However, the risk index of P95 (Percentile 95) selenium exposure at the tolerable upper intake level for children at 2-14 years old exceeded 100%, with potential risk currently. Therefore, the consumption of selenium-rich rice should be properly monitored for young children and adolescents.

Phenolic acid profiles of common food and estimated natural intake with different structures and forms in five regions of China.

Phenolic acids have been reported to have many biological activities, but daily intake information is scarce. In this study, the phenolic acid contents of 116 commonly consumed food in five regions of China (Beijing, Hangzhou, Wuhan, Chongqing and Guangzhou) were analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS) and dietary intakes estimated. Tea had the highest total phenolic acids (TPA) content in all regions investigated. Phenolic acids were in esterified and bound forms, and hydroxycinnamic acids accounted for more than half of TPA content, except in tea and legumes. Average TPA intake was 193.50 mg/ day at the national level but ranged from 157.09 to 263.01 mg/ day among the regions. Rice, tea, and fruits were the main contributors to dietary intakes of TPA. Also, TPA intake in the period 2009-2013 increased 31.65 mg/ day compared with 2002, largely due to increased intakes of fruits, vegetables, and legumes.

Correction: Mercury nanoladders: a new method for DNA amplification, signal identification and their application in the detection of Hg(ii) ions.

Correction for 'Mercury nanoladders: a new method for DNA amplification, signal identification and their application in the detection of Hg(ii) ions' by Yuxiang Feng et al., Chem. Commun., 2018, 54, 8036-8039.

Anti-obesity and hypolipidemic effect of water extract from Pleurotus citrinopileatus in C57BL/6J mice.

Obesity has become one of the most important health problems worldwide requiring urgent need for efficient control. Pleurotus citrinopileatus (P. citrinopileatus)-a type of edible mushroom with abundant bioactive molecules-is a promising source for achieving this goal. In the present study, we evaluated the anti-obesity and hypolipidemic effect of P. citrinopileatus water extract (PWE) using a series of biochemical assays in randomized high-fat diet-induced obese (DIO) C57BL/6J mice, which were gavaged daily with low or high levels of PWE (400 or 800 mg/kg of body weight, respectively) in addition to high-fat diet for 12 weeks. Results showed that PWE significantly reduced the weight gain, fat accumulation, and food intake of DIO mice within 12 weeks. PWE also decreased the serum triglycerides, cholesterol and low-density lipoprotein, aspartate transaminase, nonesterified fatty acid, and creatinine, but increased high-density lipoprotein. Additionally, PWE improved the glucose tolerance of mice fed with high fat. From above, we conclude that PWE has great potential as functional foods for management of obesity and/or associated metabolic disorders.

A 90-day subchronic toxicology screen of genetically modified rice Lac-3 and its effects on the gut microbiota in Sprague-Dawley rats.

A 90-day subchronic toxicology screen of genetically modified (GM) rice Lac-3 expressing human lactoferrin (hLF) and its effects on the gut microbiota were studied in comparison to non-GM rice fed to Sprague-Dawley (SD) rats. Three different dietary concentrations (17.5%, 35% and 70%, w/w) of the GM rice or its corresponding non-GM rice were used. Additionally, the phylotypes of gut microbiota in the control group, the 70% GM rice diet group and the 70% non-GM rice diet group on day 90 were determined by 16S rRNA sequencing. The results of the 90-day subchronic feeding study demonstrated that the GM rice Lac-3 containing human lactoferrin (LF) gene is considered as safe as the non-GM rice. The results of bacterial 16S rRNA sequencing showed that the structure of gut microbiota in the 70% GM group slightly changed when compared with the control group and the 70% non-GM group. There were no significant differences in the microbiota diversity among the three groups.

Mercury nanoladders: a new method for DNA amplification, signal identification and their application in the detection of Hg(ii) ions.

A biosensor based on Hg(ii) nanoladders integrated with graphene oxide (GO) for Hg(ii) detection was developed. Nanoladders were essentially DNA sandwich structures supported by T-Hg(ii)-T and were used for Hg(ii) trapping and signal translation. GO was used to differentiate the fluorescence generated by Hg(ii) nanoladders from a background signal.

Phytosterol Profiles of Common Foods and Estimated Natural Intake of Different Structures and Forms in China.

Phytosterols are well-known for their cholesterol-lowering effects, and the structures and forms of phytosterols affect their bioactivity. We aimed to illustrate the phytosterol profiles in common foods and estimate their natural intake in five geographical regions and among different age groups in China. In total, 12 phytosterols in free and esterified forms of 119 foods from five regions across China were examined using gas chromatography-mass spectrometry. Then, the dietary intake of phytosterols was calculated combined with the dietary foods intake data of Chinese people. The total phytosterol content was highest in vegetable oils (150.4-1230.9 mg/100 g), followed by legumes (129.6-275.6 mg/100 g), nuts (18.9-255.2 mg/100 g), and cereals (11.9-93.8 mg/100 g). Vegetables and fruits contained lower contents of total phytosterols. Phytosterols were mainly esterified in most common foods except in nuts. The predominant phytosterols were ß-sitosterol, campesterol, and stigmasterol, all of which belonged to plant sterols and 4-desmethylsterols. Total phytosterol intake varied across different regions, ranging between 257.7 and 473.7 mg/standard-person (sp)/day, with the highest intake in Beijing, followed by Hangzhou, Wuhan, Chongqing, and Guangzhou. However, phytosterol proportion was similar across regions, with ß-sitosterol accounting for 46.5-50.3% of the natural intake. Phytosterol intake was mainly constituted by plant sterols and 4-desmethylsterols in esterified form (61.9-74.6%). At the age of 2-70 years, phytosterol intake ranged from 154.3 mg/day to 348.0 mg/day in the national scale.

Potential subchronic food safety of the stacked trait transgenic maize GH5112E-117C in Sprague-Dawley rats.

The food safety of stacked trait genetically modified (GM) maize GH5112E-117C containing insect-resistance gene Cry1Ah and glyphosate-resistant gene G2-aroA was evaluated in comparison to non-GM Hi-II maize fed to Sprague-Dawley rats during a 90-day subchronic feeding study. Three different dietary concentrations (12.5, 25 and 50 %, w/w) of the GM maize were used or its corresponding non-GM maize. No biologically significant differences in the animals' clinical signs, body weights, food consumption, hematology, clinical chemistry, organ weights and histopathology were found between the stacked trait GM maize groups, and the non-GM maize groups. The results of the 90-day subchronic feeding study demonstrated that the stacked trait GM maize GH5112E-117C is as safe as the conventional non-GM maize Hi-II.

Genome-wide identification of cytosine-5 DNA methyltransferases and demethylases in Solanum lycopersicum.

Recent studies have reported that decreased level of DNA cytosine methylation in the global genome was closely related to the initiation of tomato (Solanum lycopersicum) fruit ripening. However, genome-scale analysis of cytosine-5 DNA methyltransferases (C5-MTases) and demethylases in tomato has not been engaged. In this study, 7 C5-MTases and 3 demethylases were identified in tomato genome, which probably contributed to DNA cytosine methylation level in tomato. The 7 C5-MTases were categorized into 4 subgroups, and the 3 demethylases were classified into 2 subgroups based on phylogenetic analyses. Comprehensive analysis of their structure and genomic localization was also performed in this paper. According to online RNA-seq data, 4 S. lycopersicum C5-MTase (SlC5-MTase) genes (SlMET, SlDRM1L1, SlDRM5, SlMET3L) were expressed higher than others, and one DNA demethylase gene (SlDML) was significantly changed during tomato fruit development and ripening. Furthermore, all these five gene expressions at breaker (BK) stage changed with 1-methylcyclopropene (1-MCP) treatment, indicating that they were regulated by ethylene directly or indirectly in tomato fruit. In addition, subcellular localization analysis indicated that SlDRM1L1 and SlDRM5 located in the nucleus might have responsibility for RNA-directed DNA methylation (RdDM). Collectively, this paper provided a framework for gene discovery and functional characterization of C5-MTases and DNA demethylases in other Solanaceae species.

Cloning, expression and characterization of recombinant elastase from Pseudomonas aeruginosa in Picha pastoris.

The gene lasB from Pseudomonas aeruginosa, which encoded elastase, was cloned and firstly successfully expressed in Pichia pastoris stain KM71 under the control of AOX promoter. The effects on the recombinant elastase activities of different pH, different temperatures and different metal ions were assayed. The full-length gene (1497 bp) encodes a preproenzyme including an N-terminal signal peptide (23 aa), a propeptide (197 aa) and mature elastase (301 aa). The recombinant elastase was secreted into culture supernatants using signal sequence from lasB and showed a single band at about 34 kDa by SDS-PAGE. The recombinant elastase expression hit the highest level of approximately 450 mg/L and the specific elastolytic activity of the recombinant elastase was 130 U/ml, which was approximately 26-fold higher than that of elastase obtained from P. aeruginosa. The optimal temperature and pH of the recombinant elastase was 28 degrees C and 7.4, respectively. The enzyme possessed high resistance to heat, and can be activated by Ca(2+). These enzyme properties suggested that it could be produced in an industrial scale and has the potential to be a commercial enzyme.

Expression, purification and characterization of pectin methylesterase inhibitor from kiwi fruit in Escherichia coli.

A significant problem in production of fruit juices for human consumption is auto-clarification, where enzyme catalyzes pectin demethylation resulting in loss of the ''natural" cloudy appearance of juices. To overcome this problem, a plant inhibitor protein which blocks the action of pectin methylesterase has been used. In this paper, expression of recombinant kiwi pectin methylesterase inhibitor (PMEI) was carried out in Escherichia coli, and the target protein was expressed in the form of inclusion bodies. The expression level reached 46% of total cell protein. Then the fusion protein was purified by nickel ion metal affinity chromatography, and the purity was finally up to 98%. After refolding in GSH/GSSG redox system, recombinant PMEI not only could efficiently inhibit PMEs from eight different plants, but could remain effective inhibitor activity in the pH 3.0-10.0 and 20-40 degrees C. Thus, recombinant PMEI has potential application in the production of fruit juices product industry.