Structural characterization and high-efficiency prebiotic activity of the polysaccharide from Tremella aurantialba endophytic bacteria.

Herein, the low-molecular-weight heteropolysaccharide (designated as TABP), with a weight-average Mw of 5408 Da, was produced by the endophytic bacterium Bacillus sp. TAB, which was initially isolated from the fruiting bodies of the wild Tremella aurantialba. A relatively high TABP accumulation was obtained and enhanced to 6.94 g/L in 5 L fed-batch fermentation by high-density cultivation. Monosaccharide composition analysis showed that the TABP comprised arabinose, glucosamine, galactose, glucose, and mannose with a molar ratio of 0.073: 0.145: 0.406: 0.182: 0.195, respectively. Methylation and NMR analyses indicated that TABP contained 1,4-linked ß-d-Galp and 1,4-linked ß-d-Manp pyranosyl backbone, extensively substituted at the side chains to form a complex structure. Prebiotic potential analysis exhibited significant growth-promoting effects for various lactic acid bacteria by more than 90 %. Overall, this study initially provides valuable insights into the endophytic exopolysaccharides from T. aurantialba and their biological activity, which provides prospective sources of prebiotics for functional foods and aids in understanding the endophytes symbiosis mechanism in edible mushrooms.

Moisture content of tea dhool for the scenting process affects the aroma quality and volatile compounds of osmanthus black tea.

To improve the quality of osmanthus black tea, samples produced with different scenting methods were prepared. The sensory quality was assessed and the characteristic aromatic components were explored using solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry. According to the results, osmanthus black tea obtained by adding osmanthus scenting in the fermentation process had the strongest floral aroma. The major contributors to the aroma of osmanthus black tea were identified as ß-ionone, dihydro-ß-ionone, benzeneacetaldehyde, citral, geraniol, and linalool by calculating their relative odor activity values. An analysis of the causes revealed that the moisture content of tea dhool significantly affected the adsorption of fresh flower aroma by tea. The experimental results showed that osmanthus black tea produced using tea dhool containing 30% moisture content had the highest content of crucial aroma components, suggesting the tea dhool under this condition had the strongest adsorption capacity for osmanthus aroma.

The Effect of Guisangyou Tea on Abnormal Lipid Metabolism in Mice Induced by High-Fat Diet.

This study was aimed to investigate the effect of Guisangyou tea (GSY tea) in improving abnormal lipid metabolism in mice with obesity induced by a high-fat diet (HFD). The results showed that intervention of the water extract of GSY tea (WE) decreased serum levels of lipids, and positively regulated the related antioxidant enzyme activities and the inflammatory factors in the serum and liver. In the liver, the mRNA and protein expression levels of sterol regulatory element-binding proteins-1 (SREBP-1), stearoyl-CoA desaturase-1 (SCD-1), fatty acid synthase (FASN), and acetyl CoA carboxylase (ACC) related to lipid synthesis were downregulated, and the mRNA and protein expression levels of the farnesoid X receptor (FXR) and small heterodimer partner (SHP) related to bile acid production were upregulated. The results demonstrate that GSY tea can improve abnormal lipid metabolism in obese mice by improving the body's antioxidant capacity, regulating the inflammatory state, and reducing the synthesis of lipids and increasing the production of bile acids. GSY tea can be processed and utilized as a safe and effective resource for improving abnormal lipid metabolism.

HTR2A agonists play a therapeutic role by restricting ILC2 activation in papain-induced lung inflammation.

Group 2 innate lymphoid cells (ILC2s) are a category of heterogeneous cells that produce the cytokines IL-5 and IL-13, which mediate the type 2 immune response. However, specific drug targets on lung ILC2s have rarely been reported. Previous studies have shown that type 2 cytokines, such as IL-5 and IL-13, are related to depression. Here, we demonstrated the negative correlation between the depression-associated monoamine neurotransmitter serotonin and secretion of the cytokines IL-5 and IL-13 by ILC2s in individuals with depression. Interestingly, serotonin ameliorates papain-induced lung inflammation by suppressing ILC2 activation. Our data showed that the serotonin receptor HTR2A was highly expressed on ILC2s from mouse lungs and human PBMCs. Furthermore, an HTR2A selective agonist (DOI) impaired ILC2 activation and alleviated the type 2 immune response in vivo and in vitro. Mice with ILC2-specific depletion of HTR2A (Il5cre/+·Htr2aflox/flox mice) abolished the DOI-mediated inhibition of ILC2s in a papain-induced mouse model of inflammation. In conclusion, serotonin and DOI could restrict the type 2 lung immune response, indicating a potential treatment strategy for type 2 lung inflammation by targeting HTR2A on ST2+ ILC2s.

Improved Stability and In Vitro Anti-Arthritis Bioactivity of Curcumin-Casein Nanoparticles by Ultrasound-Driven Encapsulation.

Curcumin possesses beneficial biological functions, namely anti-inflammation and anti-diabetic functions. However, due to its low solubility and crystallinity, its applications are limited. In this work, curcumin was encapsulated in casein micelles in order to form curcumin-casein nanoparticles by ultrasound treatment (5 min). The ultrasound treatment induced the entry of the hydrophobic groups to the inner micelles and the polar sulfydryl groups to the surface of the micelles in order to form compact curcumin-casein nanoparticles of an appropriate size (100-120 nm) for cellular endocytosis. The product exhibited excellent stability during 8 months of cold storage, 6 days at room temperature, and 2 days at body temperature. Advanced in vitro experiments demonstrated that curcumin-casein nanoparticles displayed significantly greater inhibitory activity against the proliferation and proinflammatory cytokines of human fibroblast-like synoviocyte-osteo arthritis (HFLS-OA) cells and HFLS-rheumatoid (RA) cells than native curcumin due to better cellular uptake as a result of the low crystallinity and the appropriate nano-size of the nano-form. The results provide a reference for the use of ultrasound treatment to encapsulate other drug molecules and curcumin-casein nanoparticles as potential treatment for arthritis.

Hyaluronan Oligosaccharides-Coated Paclitaxel-Casein Nanoparticles with Enhanced Stability and Antitumor Activity.

This study aims to develop specific-molecular-weight hyaluronic acid oligosaccharides-coated paclitaxel-loaded casein nanoparticles (HA-PT-Cas NPs) via chemical conjugation to increase the stability and antitumor effects. Optimized HA-PT-Cas NPs (HA/casein of 3:1) were obtained with a mean size of 235.3 nm and entrapment efficiency of 93.1%. HA-PT-Cas exhibited satisfactory stability at 4 °C for 12 days and 37 °C for 3 h; paclitaxel was retained at rates of 81.4% and 64.7%, respectively, significantly higher than those of PT-Cas (only 27.8% at 4 °C after 16 h and 20.3% at 37 °C after 3 h). HA-PT-Cas exhibited high efficiency (61.3%) in inhibiting A375 tumor owing to the enhanced stability of HA oligosaccharides barrier, which was comparable with that of 10 µg/mL cis-platinum (64.9%). Mice experiments showed the 74.6% tumor inhibition of HA-PT-Cas by intravenously administration, significantly higher than that of PT-casein (39.8%). Therefore, this work provides an effective carrier for drug delivery via HA oligomers-coated modification.

The Effect of Microwave Radiation on the Green Color Loss of Green Tea Powder.

Microwave radiation is one of the main heating methods for food processing, especially affecting the color quality of colorful foods. This work presents the effect of microwave radiation on the green color loss of green tea powder (GTP) by the color description (L*, a*, b*, and Ha of green tea powder, L*:whiteness/darkness, a*: redness/greenness, and b*: yellowness/blueness; Ha derived from Hunter a and b could visually describe the color space) of the Hunter color system. First, the L*, a*, and b* were determined from the GTP samples treated with various microwave powers with the change of time to investigate the kinetic of color loss. Then, the L*, a*, and b*and temperature of GTP samples with serious thickness treated with constant microwave power (700 W) for a different time were determined to study the effect of sample thickness on the color loss. Finally, the chemicals that contributed to color change in the GTP samples treated with mild, moderate, and severe radiation were analyzed. The results showed that L*, |a*| (|a*|was the absolute value of a*), b*, and Ha decreased with the power increase in microwave radiation, and their changes conformed to the first-order kinetics. The activation energies (Ea) of different thickness GTP for change of L*, a*, b*, and Ha values could be predicted with the fitting models, and Ea for 20 mm-thick GTP were approximately 1/5, 1/8, 1/8, and 1/13 of those for 4 mm-thick GTP. The color loss was mainly caused by the Mg2+ loss of chlorophylls and the formation of derivates under mild radiation, the degradation of chlorophylls and the formation of theaflavin from catechins under moderate radiation, and the degradation of chlorophylls and their derivates accompanied by Maillard reaction between reducing sugar and amino acids under severe radiation. The results indicate that sample thickness and radiation time are two key parameters to keeping the color of GTP in food processing and microwave pasteurization.

Excess serum Na level in rats administered with high doses of (-)-epigallocatechin gallate-casein nanoparticles prepared with sodium caseinate.

(-)-Epigallocatechin gallate (EGCG)-incorporated casein nanoparticles benefit from excellent antioxidant, anti-inflammatory and anti-cancer activities due to their synergistic efficiency, but few studies have evaluated their safety. In this study, the EGCG-casein nanoparticles (EGCG-NPs) formulated using caseinate by ultrasonic treatment were evaluated for their subacute toxicity. The subacute toxicity test of EGCG-NPs through 28-day oral administration in rats did not exhibit adverse effect, with a no-observed-adverse-effect level (NOAEL) of at least 5.0 g per kg body weight (BW) per day, which was equivalent to 500 mg per kg BW EGCG per day. However, the serum Na level in females and males treated with 10.0 g per kg BW EGCG-NPs increased significantly as compared to the control rats (P < 0.05). Similar indications appeared in rats treated with 10.0 g per kg BW pure casein nanoparticles without EGCG, which indicated that high doses of caseinate nanoparticles result in an excess serum Na level. Therefore, we should consider the safety of the nanoparticle formulation of caseinate when it is used as a loading nutrient and a functional substance in foods.

Enhanced antibacterial activity of eugenol-entrapped casein nanoparticles amended with lysozyme against gram-positive pathogens.

This study aimed to investigate the antibacterial efficiency and synergistic mechanisms of novel formulated eugenol-casein-lysozyme nanoparticles (ECL-NPs) against gram-positive bacteria. We obtained optimized ECL-NPs 151.9 nm in size and with an entrapment efficiency of 92.2%. ECL-NPs exhibited a satisfactory slow-release pattern, excellent storage stability (for 180 days at 4 °C), and freeze-drying stability. The synergy of low-dose lysozyme significantly enhanced the inhibitory efficiency of eugenol-casein nanoparticles against Staphylococcus aureus and Bacillus sp. by 5.83-fold and 5.53-fold, respectively; this resulted in a much lower minimum inhibitory concentration (3.75-fold and 4.16-fold) and minimum bacterial concentration (2.92-fold and 1.70-fold) values. Scanning electron microscopy clearly demonstrated that the entire cell morphological structure was broken into pieces after exposure to ECL-NPs. Furthermore, 100% microbial inhibition was observed in fresh fruits treated with ECL-NPs for 15 days. These findings suggest that ECL-NPs have an excellent potential for use in food industry against gram-positive bacteria.

Superior anti-infective potential of eugenol-casein nanoparticles combined with polyethylene glycol against Colletotrichum musae infections.

The aim of this study was to improve the stability of eugenol-casein nanoparticles (EL-CS-NPs) through polyethylene glycol (PEG) modification. The results show that modifying the EL-CS-NPs with PEG after loading with eugenol (EL) gives PEG-EL-CS-NPs, with increased stability. The NPs modified with higher-molecular-weight PEG showed better stability. A CS/PEG ratio of 200 : 1 (w/w) yielded the NPs with the best stability. A PEG20 K-EL-CS-NP dispersion remained stable in cold storage for over one year, and also exhibited stronger inhibitory effects against Colletotrichum musae inoculated on bananas than an EL-CS-NP dispersion, since it showed more prolonged sustained release of EL than the EL-CS-NP dispersion. Lyophilized PEG20 K-EL-CS-NP powder showed better effectiveness against mold on bread than lyophilized EL-CS-NPs powder. Using PEG to modify CS-NPs shows potential for improving the stability of CS-NPs loaded with hydrophobic substances for delivery in the fields of food and agriculture.

Biosynthesis of low-molecular-weight mannan using metabolically engineered Bacillus subtilis 168.

Mannans are functional polysaccharides with unique biological activities that have been employed widely in food, medicine and pharmaceutics. Recent breakthroughs in plant polysaccharide metabolism identified numerous genes involved in the biosynthesis of mannans. However, constructing highly efficient low-cost microbial cell factories to produce low-molecular-weight (LMW) mannans remains challenging. In this work, we designed a de novo mannan synthetic pathway in food-grade Bacillus subtilis, resulting in mannan accumulation of 0.97 g/L. By co-expressing the identified committed genes (manC, manB, manA and pgi), mannan production was significantly increased to 2.5 g/L. Furthermore, by redirecting the carbon flux using a glucose-repressed promoter to control pfkA expression, mannan production was substantially increased to 4.1 g/L. Production was further enhanced to 12.6 g/L (average MW 6370 Da) in 3-L fed-batch fermentation. This work provides alternative synthetic pathways for metabolic engineering of LMW mannans in B. subtilis, and a useful, optimisable approach to enhance mannans production.

Structural Modification of Nanomicelles through Phosphatidylcholine: The Enhanced Drug-Loading Capacity and Anticancer Activity of Celecoxib-Casein Nanoparticles for the Intravenous Delivery of Celecoxib.

This study aims to stabilize loaded celecoxib (CX) by modifying the structure of casein nanoparticles through phosphatidylcholine. The results show that Egg yolk phosphatidylcholine PC98T (PC) significantly increased the stability of CX-PC-casein nanoparticles (NPs) (192.6 nm) from 5 min (CX-ß-casein-NPs) to 2.5 h at 37 °C. In addition, the resuspended freeze-dried NPs (202.4 nm) remained stable for 2.5 h. Scanning electron microscopy indicated that PC may block the micropore structures in nanoparticles by ultrasonic treatment and hence improve the physicochemical stability of CX-PC-casein-NPs. The stability of the NPs was positively correlated with their inhibiting ability for human malignant melanoma A375 cells. The structural modification of CX-PC-casein-NPs resulted in an increased intracellular uptake of CX by 2.4 times than that of the unmodified ones. The pharmacokinetic study showed that the Area Under Curve (AUC) of the CX-PC-casein-NPs was 2.9-fold higher in rats than that of the original casein nanoparticles. When CX-PC-casein-NPs were intravenously administrated to mice implanted with A375 tumors (CX dose = 16 mg/kg bodyweight), the tumor inhibition rate reached 56.2%, which was comparable to that of paclitaxel (57.3%) at a dose of 4 mg/kg bodyweight. Our results confirm that the structural modification of CX-PC-casein-NPs can effectively prolong the remaining time of specific drugs, and may provide a potential strategy for cancer treatment.

Low-dose vardenafil potentiates the protective effect of (-)-epigallocatechin gallate on cardiomyocytes.

The major polyphenol (-)-epigallocatechin gallate (EGCG) of green tea shows well-known health benefits such as potential anti-cancer, anti-oxidation and ameliorating cardiovascular disease. This work aims to improve the bioactivity of EGCG on H9C2 cardiomyocytes by combination regimen of vardenafil and EGCG. The proliferative rates were significantly improved by 18.74%, 10.77% and 29.17% after 48 h with EGCG, vardenafil, and the combination of EGCG and low-dose vardenafil treatments, respectively. The treatments also increased the expression of the nitric oxide synthase (eNOS), and acutely stimulate production of vasodilators nitric oxide (NO) from 17.33µmol/L to 19.75, 20.87 and 24.47µmol/L in H9C2 cells. We further demonstrated that vardenafil also remarkably promoted EGCG to counteract H2O2-induced apoptotic damage in H9C2 by strengthening antioxidant defense systems and suppressing myocardial apoptosis. These results suggest that EGCG and low-dose vardenafil in combination may be a promising regimen to help prevent cardiovascular diseases.

Enhanced Antifungal Activities of Eugenol-Entrapped Casein Nanoparticles against Anthracnose in Postharvest Fruits.

This study aims to improve the antifungal effects of eugenol through low-energy self-assembly fabrication and optimization of eugenol-casein nanoparticles (EC-NPs). Optimized EC-NPs (eugenol/casein ratio of 1:5) were obtained with a mean size of 307.4 ± 2.5 nm and entrapment efficiency of 86.3% ± 0.2%, and showed high stability under incubated at 20 and 37 °C for 48 h. EC-NPs exhibited satisfactory sustained-release effect at 20 °C or 37 °C, with remaining eugenols amounts of 79.51% and 53.41% after 72 h incubation, respectively, which were significantly higher than that of native eugenol (only 26.40% and 19.82% after the first 12 h). EC-NPs exhibited a greater antifungal activity (>95.7%) against spore germination of fungus that was greater than that of native eugenol, showed 100% inhibition of the anthracnose incidence in postharvest pear after 7 d. EC-NPs is potential as an environmental-friendly preservatives in the food industry.

Enhanced Antiarthritic Efficacy by Nanoparticles of (-)-Epigallocatechin Gallate-Glucosamine-Casein.

This work aims to improve the antiarthritic activity of (-)-epigallocatechin gallate (EGCG) and glucosamine (GA) through fabrication and optimization of casein protein nanoparticles (EGC-NPs). Optimized EGC-NPs were obtained with a EGCG/GA/casein ratio of 1:2:8 (w/w/w). The EGC-NPs gave a mean size of 186 ± 3.5 nm and an entrapment efficiency of 86.8 ± 2.7%, and they exhibited a greater inhibitory activity against human fibroblast-like synoviocytes-osteoarthritis cells and human fibroblast-like synoviocytes-rheumatoid arthritis cells compared with that of the EGCG-GA mixture by 33.5% and 20.8%, respectively. Freeze-dried EGC-NPs stored at 25 °C during 12 months showed high dispersion stability. Moreover, the redispersion of the freeze-dried EGC-NPs produced almost no significant changes in their physicochemical properties and bioactivity. Rat experiments demonstrated that the antiarthritis effect of the EGC-NPs was significantly higher than that of EGCG-GA mixture, as assessed through an analysis of anti-inflammatory efficacy, radiographic images and histopathological assessments of paw joints, and immunohistochemical changes in serum cytokines. The enchanced antiarthritic activity in vivo was consistent with that in vitro. The EGC-NPs demonstrate potential as a food supplement for the treatment of arthritis.

Orange Fragrance with Sustained-Release Properties Prepared by Nanoethosomal Encapsulation of Natural Orange Essential Oil.

There is an upsurge of interest in improving the stability and prolong the scent holding time of fragrances in cosmetic industry. In this study, to encapsulate the orange essential oils (OEO), nanoethosomes were constructed with optimized proportions of ethanol, water, soybean phosphatidylcholine (SPC), Tween 80, and palm oil sucrose esters (PSE). The controlled-release behavior of nanoethosomes was then studied concerning physicochemical stability, microstructure, and olfactory sensation. The sustained-release effectiveness of the nanoethosomal fragrances was influenced by particle size and OEO amount of specific formula. Herein, there was a positive correlation between particle size and sustained-release effectiveness. In particular, the mean diameter of nanoethosomal orange fragrances (nano-OFs)-prepared by EO-ethanol-water-SPC-Tween 80-PSE (3-7:25:72-68:2:1.0:0.1) and 3%, 5%, and 7% OEO-was 68.6±3.6, 79.5±4.5 and 87.3±6.9 nm, respectively. The results of olfactory sensation and GC-MS analysis showed that these fragrances could sustainedly release the aromatic compounds to yield satisfactory smell longer than that of the conventional orange fragrance. Furthermore, the nano-OF made of 5% EO yield a satisfactory smell more than 3 h, which was 3 times of that of the conventional orange fragrance. This fragrance was stable when stored at 4 °C (>1 year) and 25 °C (>10 months). The knowledge gained from this study will be helpful to develop nanoethosomal fragrances or perfumes for commercial use.

New insight into the nitrogen metabolism of simultaneous heterotrophic nitrification-aerobic denitrification bacterium in mRNA expression.

Here, the draft genome of simultaneous nitrification-denitrification strain (SND) Klebsiella sp. KSND revealed possible existence of genes involved in N-assimilation and -dissimilation pathways. The change levels of genes under defined N-sources were analyzed by Quantitative Real-Time PCR. It suggested that NH4+-assimilation via NADP-glutamate dehydrogenase pathway would occur preferentially. NirBD genes were tightly regulated in a lower level, so that nitrite was rapidly consumed for detoxication by denitrification. Three types of nitrate reductase homologues are surprisingly present in KSND, whereas the dominant nitrate reduction for assimilation and denitrification processes mediates by NapA-type nitrate reductase. Nitric oxide reductase homologues FlRd and FlRd-red provide an adequate capacity for NO detoxification. The recombinant hydroxylamine reductase showed high activity in hydroxylamine to generate ammonium, which might contribute to detoxification mechanism in nitrogen cycling. Overall, this study firstly provides valuable insights into the genes expression and enzyme action, which helps understanding the mechanism of SND processes.

Castanopsis lamontii Water Extract Shows Potential in Suppressing Pathogens, Lipopolysaccharide-Induced Inflammation and Oxidative Stress-Induced Cell Injury.

Castanopsis lamontii is traditionally used to prevent inflammatory diseases such as periodontitis and pharyngitis by residents in southwest China. However, little scientific evidence has been found to support this. In this research, the antibacterial activities of Castanopsis lamontii water extract (CLE) were assessed using the micro-dilution method. The anti-inflammatory and antioxidant activities of CLE were investigated in RAW264.7 cells. Key bioactive compounds in CLE were also explored. Results showed that CLE was capable of inhibiting the periodontitis pathogen Porphyromonas gingivalis and the pharyngitis pathogen ß-hemolytic Streptococcus. It suppressed lipopolysaccharide-induced inflammation in RAW 264.7 cells via inactivating the TLR4/NF-κB pathway. Besides, it reduced oxidative stress-induced cell injury via scavenging reactive oxygen species. Chemical composition analysis revealed that CLE was rich in epicatechin and procyanidin B2. Further studies confirmed that epicatechin predominantly contributed to the antibacterial activities of CLE, while procyanidin B2 was mainly responsible for the anti-inflammatory activities of CLE. Both compounds contributed to the antioxidant activities of CLE. Acute oral toxicity tests proved that CLE was practically non-toxic. These results provide experimental evidences of the health-beneficial effects of CLE and may help promote the application of CLE in the food and health industries.

Enhancement in Antibacterial Activities of Eugenol-Entrapped Ethosome Nanoparticles via Strengthening Its Permeability and Sustained Release.

The antibacterial efficiency and synergistic mechanisms of novel formulated eugenol entrapped ethosome nanoparticles (ELG-NPs) against fruit anthracnose were investigated. The results showed that concentrations of eugenol and ethanol significantly influenced the particle size and entrapment efficiency of nanoethosome, and the particle size significantly influenced the antibacterial effect. Superior ELG-NPs with optimized process (0.5% eugenol, 2% lecithin, and 30% ethanol) were obtained with a size of 44.21 nm and entrapment efficiency of 82%. ELG-NPs exhibited an antibacterial activity (>93%) against fruit pathogens that was greater than that of free eugenol and showed 100% inhibition of the anthracnose incidence in postharvest loquat after 6 d. The permeability study, first visualized in banana cortex with fluorescent indicators, demonstrated that eugenol delivered to the interior with ELG-NPs was 6-fold higher than that of free eugenol. ELG-NPs showed a satisfactory slow-release and prolonged antibacterial action. This work provides a promising strategy for disease controls in agricultural, food, cosmetic, and medical areas.

Improving the taste of autumn green tea with tannase.

Green tea processed from autumn leaves is more bitter and astringent than that from spring leaves, mainly due to gallated catechins. The present study aimed to improve the taste of autumn green tea and green tea infusion by using tannase to treat tea leaves and tea infusion. The results showed that, after hydrolysis, the sweet aftertaste and overall acceptability improved, and the ratio of gallated catechins decreased, as did the bitterness and astringency of the autumn green tea. The pH value was significantly correlated with the concentrations of gallated catechins (r = 0.930, p < 0.01), non-gallated catechins (r = -0.893, p < 0.01), and gallic acid (r = 0.915, p < 0.01), as well as with the intensities of bitterness, astringency, and sweet aftertaste during hydrolysis. Gallic acid contributed to the sweet aftertaste of green tea infusion. These results will help to improve autumn green tea products with tannase.