The discovery of microRNAs encapsulated in milk-derived exosomes has revealed stability under extreme conditions reflecting the protection of membranes. We attempted to determine the variations in nanoparticles derived from milk after fermentation, and provide evidence to determine the effects of these exosomes on cells with potential bioactivity. Using scanning electron microscopy and dynamic light scattering, we compared the morphology and particle size distribution of exosomes from yogurt fermented with three different combinations of strains with those from raw milk. The protein content of the exosome was significantly reduced in fermented milk. The cycle threshold showed that the expression of miR-29b and miR-21 was relatively high in raw milk, indicating a loss of microRNA after fermentation. Milk-derived exosomes could promote cell growth and activate the mitogen-activated protein kinase pathway. These findings demonstrated biological functions in milk exosomes and provided new insight into the nutrient composition of dairy products.Keywords: biological function; exosomes; microRNAs; probiotic cultures;

•MiR-29b promoted NTE differentiation and inhibited NCC differentiation from ESCs•MiR-29b targeted Dnmt3a to regulate neural differentiation•MiR-29b mediated the function of Pou3f1•The Pou3f1-miR-29b-Dnmt3a axis regulated the cell fate determinationDuring gastrulation, the neuroectoderm cells form the neural tube and neural crest. The nervous system contains significantly more microRNAs than other tissues, but the role of microRNAs in controlling the differentiation of neuroectodermal cells into neural tube epithelial (NTE) cells and neural crest cells (NCCs) remains unknown. Using embryonic stem cell (ESC) neural differentiation systems, we found that miR-29b was upregulated in NTE cells and downregulated in NCCs. MiR-29b promoted the differentiation of ESCs into NTE cells and inhibited their differentiation into NCCs. Accordingly, the inhibition of miR-29b significantly inhibited the differentiation of NTE cells. A mechanistic study revealed that miR-29b targets DNA methyltransferase 3a (Dnmt3a) to regulate neural differentiation. Moreover, miR-29b mediated the function of Pou3f1, a critical neural transcription factor. Therefore, our study showed that the Pou3f1-miR-29b-Dnmt3a regulatory axis was active at the initial stage of neural differentiation and regulated the determination of cell fate.Download high-res image (170KB)Download full-size image

The antioxidant differences between two flavonoid extracts from fruiting body and fermentation broth of Phellinus igniarius were determined, and the effects of flavonoid addition on the sensory characteristics of sturgeon caviar were investigated. Both extracts exhibited moderate antioxidant activity with EC50 values of 4.98 and 2.39 mg/mL, the extract of fermentation broth exhibiting higher antioxidant efficiency. Flavonoids were added at 0.01% and 0.02% to sturgeon caviars, which were then stored at 4 or 20 °C for 12 days. Peroxide values (meq/kg) were determined after regular intervals. The 0.02% level of flavonoids from fermentation broth of P. igniarius had stronger antioxidant effect than vitamin C, very close to that of butylated hydroxyanisole (BHA). Flavonoid addition improved the sensory profile of caviar, and the flavonoid extract from the fermentation broth had a more desirable effect on flavour and aroma than that from fruiting bodies.Highlights► Submerged fermentation products from Phellinus igniarius strain were good natural sources for flavonoid antioxidants. ► Flavonoids had positive antioxidant effects on sturgeon caviar storage. ► Sensory characteristics of sturgeon caviar were improved with flavonoids addition.

Pure d/l-theanine enantiomers were synthesized separately, and SEM was used for their crystal-structure observation. The novel enantiomeric separating method by HPLC was established using the chiral selector of β-CD in the mobile phase. Green tea, white tea, oolong tea, black tea and Pu-erh tea were tested for theanine enantiomers by different degrees of fermentation. The significantly higher d-enantiomeric proportion of theanine was found in white tea than the others, which was probably due to its specific processing step of withering. The effect of electrolyzed reduced water (ERW) on enantiomeric theanine and polyphenols in tea was explored. There was no change of theanine, but rather a loss of ECG (epicatechin gallate) and an increasing amount of GA (gallic acid). ERW also reduced tea cream, which contains significant amount of polyphenols, indicating its potential application in the tea-beverage industry.Highlights► Crystal structures of theanine enantiomers under SEM. ► A novel method separating theanine enantiomers. ► White tea boasts higher amount of d-theanine than others. ► Electrolyzed reduced water decreases ECG, increases GA, and reduces tea cream in tea.

Chitosan was combined with nano-ZnO to increase its antimicrobial activity, using polyvinyl alcohol as a support, and then were electronspun to form composite nanofibres. Through SEM, EDX and XRD observations, chitosan was seen to be able to incorporate nano-ZnO in the composite nanofibres. Escherichia coli, expressing recombinant enhanced green fluorescent protein, and Candida albicans were used to test the antimicrobial efficacy of the newly synthesised chitosan/nano-ZnO antimicrobial composite. The CdTe quantum dots were used to rapidly detect the residual changes of C. albicans and determine the end point of using antimicrobial agents. Minimal minimum inhibitory concentration (MIC), post-antibiotic effect and continuous agent effect of the composite were determined. The MIC of chitosan/nano-ZnO against C. albicans was 160 μg/ml, close to the concentration of the treated composite with the lowest fluorescence intensity. The cell damage was observed by SEM, which indicated that nano-ZnO in the nanofibrous membranes played a cooperative role in the antimicrobial process of chitosan.Highlights► Chitosan and nano-ZnO were combined in nanofibrous membranes by electrospinning method for the first time. ► CdTe QDs coupled with Candida albicans were used for rapid fluorescence detection of antimicrobial agents. ► Chitosan/nano-ZnO nanofibrous membranes could be used as novel antimicrobial packaging in foods.

Terpinen-4-ol, an active component of tea tree oil, exhibits broad-spectrum antimicrobial activity. However, the high volatilization of terpinen-4-ol and its nonwettability property have limited its application. Our objective was to synthesize novel nanocarriers to deliver and protect terpinen-4-ol. The polyethylene glycol (PEG)-stabilized lipid nanoparticles were prepared and characterized by scanning electron microscope, Zetasizer, and differential scanning calorimetry. These nanoparticles had an average diameter of 397 nm and a Ζ-potential of 10 mV after being modified by glycine. Results showed that homogeneous particle size, high drug loading, stability, and targeting were obtained by the nanoparticles. Liquid chromatography/mass spectrometry showed a sustained release trend from nanoparticles for terpinen-4-ol. Minimum inhibitory concentration and minimum biofilm eradication concentration were tested against Candida albicans ATCC 11231. Studies on isolated mitochondria showed the blockage of biofilm respiration and inhibition of enzyme activity. The effects can be ascribed to localization of terpinen-4-ol on the membrane of mitochondria.

(−)-Epigallocatechin-3-gallate (EGCG) exhibited strong antimicrobial activity. However, the easy oxidation of EGCG has limited its application. To increase the antimicrobial activity and stability of EGCG, the EGCG−CuII complex was formed by chelating copper ions and then electronspun into polyvinyl alcohol (PVA) nanofibers. Electronspun nanofibrous membranes were investigated by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM), which showed that the average fiber diameter was 210 nm. Minimal inhibitory concentrations (MICs) of EGCG−CuII/PVA membranes were tested against the tested strains. The bactericidal activity of EGCG−CuII was suppressed by ethylenediaminetetraacetic acid (EDTA). Cell killing was accompanied by the leakage of intracellular proteins, indicating that the cytoplasmic membrane was badly damaged after exposure to the EGCG−CuII/PVA membrane. We observed the process of cell damage by SEM. On the basis of experimental evidence and theoretical analyses, the mechanism proposed that copper ions played a cooperative role in the bactericidal process of EGCG. To evaluate the antimicrobial activity of the EGCG−CuII/PVA membrane, we developed a rapid detection method by labeling cells with water-soluble CdTe quantum dots.

•Aminoacylase was immobilized on polyvinyl alcohol-based nanofibrous membranes.•The immobilized aminoacylase showed the optimal catalytic conditions at pH 8 and 52 °C.•Thermostability and storage stability of the immobilized aminoacylase were significantly improved.•The nanofibrous-immobilized aminoacylase could be used to produce pure l-theanine.The enzyme aminoacylase was used to produce l-theanine from its derivative N-acetyl-dl-theanine. For stabilization purposes the aminoacylase was immobilized on polyvinyl alcohol-based nanofibrous membranes generated by electrospinning. The immobilized aminoacylase exhibited better resistance to changes in temperature and pH than the free enzyme with optimal conditions being pH 8 and 52 °C. Under these conditions, Km values were two to three times higher than those of the free enzyme (3.6 mM). Thermostability was also significantly improved; the activity of the immobilized enzyme was retained at approximately 70% after 6 days at 52 °C. These results indicate potential applications of nanofibrous-immobilized aminoacylase for industrial production of pure l-theanine.Download full-size image