Development of chitosan/rice protein hydrolysates/ZnO nanoparticles films reinforced with cellulose nanocrystals.

In the present work, the composite films were obtained by the solution casting method from chitosan and rice protein hydrolysates, reinforced with cellulose nanocrystals (CNC) of different contents (0 %, 3 %, 6 % and 9 %). The influence of different CNC loadings on the mechanical, barrier and thermal properties was discussed. SEM showed the formation of intramolecular interactions between the CNC and film matrices, leading to more compact and homogeneous films. These interactions had a positive influence on the mechanical strength properties, which was reflected in higher breaking force of 4.27 MPa. The elongation dwindled from 132.42 % to 79.37 % with increasing CNC levels. The linkages formed between the CNC and film matrices reduced the water affinity, leading to a reduction in their moisture content, water solubility and water vapor transmission. Thermal stability of the composite films was also improved in the presence of CNC, by increasing maximum degradation temperature from 311.21 to 325.67 °C with increasing CNC contents. The strongest DPPH inhibition of the film was 45.42 %. The composite films exhibited the highest inhibition zone diameter against E. coli (12.05 mm) and S. aureus (12.48 mm), and the hybrid of CNC and ZnO nanoparticles exhibited stronger antibacterial activity than their single existent forms. The present work shows the possibility of obtaining CNC-reinforced films with improved mechanical, thermal and barrier properties.

Chitosan/rice hydrolysate/curcumin composite film: Effect of chitosan molecular weight.

The composite films were prepared by adding rice protein hydrolysate (RH) into chitosan (CH). Effect of four different chitosan molecular weights (MW, 0.8- 1.0, 30, 100, 300 kDa) was evaluated and curcumin was further incorporated into the film to enhance its bioactivities. With increasing MW (<100 kDa), moisture, solubility and water vapor transmission marginally decreased while tensile strength increased. Rice hydrolysate improved the film solubility and extensibility. All films showed extraordinary extensibility ranging from 180.26% to 204.08%. DSC confirmed MW did affect the thermal stability. The lower MW, the higher antioxidant activity of the CH/RH films. The 30 kDa CH/RH film exhibited higher antibacterial activity against both E. coli and S. aureus. Curcumin effectively improved both antibacterial and antioxidant activity of the CH/RH films. Hence, chitosan molecular weight must be chosen carefully according to its application and incorporation of protein hydrolysate into film would be an efficient strategy to modify film performance.

Anti-Inflammatory Effects of Gynura procumbens on RAW264.7 Cells via Regulation of the PI3K/Akt and MAPK Signaling Pathways.

Gynura procumbens is a traditional herb and food extensively cultivated in China and Southeast Asian countries. In this work, the crude extract (CE) of G. procumbens was purified with macroporous resin to obtain the refined fraction, and its anti-inflammatory activity was compared with that of CE. Moreover, the detailed mechanisms of anti-inflammatory activity were also investigated for the first time. The results indicated that CE was more effective in anti-inflammatory activity and it could reduce the secretion of NO, TNF-α, and PGE2 via decreasing the iNOS, TNF-α, and COX-2 genes transcription and related proteins translation, which were associated with the inhibition of AP-1 and NF-κB nuclear translocation and downregulation of PI3K/Akt and MAPK signaling pathways. In conclusion, the extract of G. procumbens has a promising potential in inflammation-related disorders alleviation, and these findings could provide the basis for the comprehensive utilization of G. procumbens and the new functional food development.

A base-mediated aerobic oxidative synthesis of cyclopent-2-enol derivatives from doubly activated cyclopropanes and substituted acetonitriles.

We report here that polysubstituted cyclopent-2-enols can be constructed by the one-pot reaction of doubly activated cyclopropanes and α-EWG substituted acetonitriles under mild basic conditions via a domino-ring-opening-cyclization/deacylation/oxidation sequence. Moreover, the synthetic applications of these cyclopent-2-enols have been demonstrated in the late-stage derivatization into functionalized cyclopentapyrimidin-4-ones and 2-hydroxy cyclopentanones with good yields.

The polysaccharide from Camellia oleifera fruit shell enhances immune responses via activating MAPKs and NF-κB signaling pathways in RAW264.7 macrophages.

Background: Macrophage plays an important role in innate immune responses by secreting immune molecules and phagocytosis. Camellia oleifera fruit shell, accounting for approximately 60% weight of the single C. oleifera fruit, is rich in polysaccharides and has several biological activities such as anti-oxidation, lipid regulation and anticancer. However, the immunomodulatory activity of the polysaccharide from C. oleifera fruit shells (CPS) has not been reported. Objective: This study aimed to investigate the immunomodulatory activities and mechanisms of CPS in RAW264.7 macrophages. Methods: The Methyl Thiazolyl Tetrazolium (MTT) method was used to evaluate the effect of CPS on the cell viability of RAW264.7 macrophages, and cell morphology was pictured using microscope. The production of immune-related molecules, including nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor α (TNFα), interleukin (IL)-1ß and IL-6, was detected by Griess assay and enzyme-linked immunosorbent assay (ELISA). The protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX2) and the phosphorylation level of mitogen-activated protein kinases (MAPKs) were analyzed through western blotting. The mRNA levels of related genes were tested using reverse transcription-polymerase chain reaction (RT-PCR). The nuclear translocation of nuclear factor-kappa B (NF-κB) was detected using immunofluorescence technology. Results: The results indicated that CPS treatment stimulated the production of NO and PGE2 and significantly enhanced the protein expression of iNOS and COX2 with little effect on the cell morphology and viability. Also, the secretion and mRNA levels of TNFα were increased by the treatment of CPS. In addition, CPS treatment markedly upregulated the phosphorylation level of MAPKs including Extracellular Signal Regulated Kinase (ERK), P38, and c-Jun N-terminal Kinase (JNK) at different time points and caused the activation and nuclear translocation of NF-κB. Conclusion: Our data implied that CPS exerts immunomodulatory activities by activating MAPKs and NF-κB signaling pathways in RAW264.7 macrophages.

Characteristics of rice dreg protein isolate treated by high-pressure microfluidization with and without proteolysis.

The characteristics of rice dreg protein isolate (RDPI) treated by microfluidization (0, 40, 80, 120, and 160 MPa) with or without proteolysis were investigated. Alcalase, Neutrase, and the combination of the two (Alcalcase:Neutrase = 1:1 [w/w]) were adopted for hydrolysis. The surface hydrophobicity and solubility of RDPI were increased. As pressure increased, different structures of RDPI exhibited disaggregation (<120 MPa) and reaggregation (160 MPa), and the effect on proteolysis was significant. The solubility of Neutrase and combined enzyme hydrolysates was improved after microfluidization. Additionally, the optimum choice of microfluidization (40 MPa) and Neutrase was efficient for improving the DPPH radical scavenging activity. The results indicate that both pressure level and enzyme type synergistically determine the functionality and antioxidant activities of products. This work may provide an alternative methodology for improving the utilization of RDPI in the food industry through desirable modifications.

Maillard conjugates of whey protein isolate-xylooligosaccharides for the microencapsulation of Lactobacillus rhamnosus: protective effects and stability during spray drying, storage and gastrointestinal digestion.

The Maillard reaction products (MRPs) of whey protein isolate (WPI) and xylooligosaccharides (XOS) were prepared by a moist heat method for use as protectants to encapsulate Lactobacillus rhamnosus via spray drying. The protective effects of MRPs on bacterial cells during drying, storage, and in vitro digestion were explored. FTIR results indicated that MRPs were successfully prepared. All MRPs showed good thermo-protective effect on the bacteria, and the survival ratio achieved with 1 : 2 XOS-WPI as a wall material reached 99.83 ± 8.44%, which was around 2 times as high as that of the WPI wall material and 1.5 times as high as that of the 1 : 2 XOS-WPI mixture. The dried lactobacilli showed similar growth curves to the fresh culture. After 10 weeks of storage at 4 °C, the decrease in the bacterial activity was less than 1 log CFU g-1 for all types of microcapsules, while the microcapsules composed of all MRPs had better storage stability. MRPs improved the stability of microcapsules during in vitro digestion. The number of viable bacteria in 1 : 2 XOS-WPI MRPs microcapsules was maintained at 4.09 ± 0.59 × 109 CFU g-1 after simulated gastrointestinal digestion for 4 hours, which only decreased by 0.20 log CFU g-1.

Purification of Four Caffeoylquinic Acid Derivatives from the Flowers of Gynura Procumbens by HSCCC.

Four caffeoylquinic acid derivatives from the Gunura procumbens flowers (GPF) were successfully isolated and purified by high-speed counter-current chromatography (HSCCC). Ethyl acetate-methanol-water (3:1:3, v/v/v) was the optimum biphasic solvent system, which was selected by high-performance liquid chromatography (HPLC) and run on a preparative scale where the lower aqueous phase was used as the mobile phase with a head-to-tail elution mode. Chlorogenic acid (3.83 mg), Isochlorogenic acid A (6.51 mg), Isochlorogenic acid B (4.38 mg) and Isochlorogenic acid C (4.47 mg) were obtained for the first time in an one-step HSCCC separation from 800 mg of the crude extracts. The purities of four compounds were determined to be >95% by HPLC. Chemical structures of each isolated compounds were identified by nuclear magnetic resonance and electrospray ionization mass spectrometry methods. It is worth noting that all the four compounds were isolated here for the first time from GPF and this work confirms the effectiveness of HSCCC for the separation of compounds contained in complex samples, and provides a foundation for further exploitation of G. procumbens.

Novel synthesis of fused spiro piperidone-cyclopropanes from cyclopropyl amides and electron-deficient alkenes.

We report here that a series of bridged O,O-ketal fused spiro piperidone-cyclopropane derivatives 3 can be constructed with excellent yields and good diastereoselectivity by the one-pot reaction of 1-acylcyclopropanecarboxamides 1 with electron-deficient alkene 2a (EWG = CHO) via the domino process involving [4 + 2] annulation/intermolecular electrophilic addition/intramolecular cyclization. Furthermore, reactions of 1 with 2b/2c (EWG = CN, COOMe), leading to spiro piperidone-cyclopropane derivatives 4 or 5 by base catalyst selection, were also presented.

Effect of cold and hot enzyme deactivation on the structural and functional properties of rice dreg protein hydrolysates.

This study explored the effect of three different enzyme deactivation treatments: 4 °C slow cold deactivation (RDPH-(4 °C)), -18 °C rapid cold deactivation (RDPH-(-18 °C)) and 100 °C water bath (RDPH-(100 °C)), compared to that without enzyme deactivation (RDPH-(control)) on the structural and functional properties of rice dreg protein hydrolysates (RDPHs). The RDPHs from the different enzyme deactivation methods led to significant differences in the degree of hydrolysis, surface hydrophobicity, average particle size, intrinsic fluorescence and emulsion stability. FTIR analysis revealed that the strength of RDPH-(100 °C) spectrum peaks decreased significantly. All samples showed high solubility (>85%) and potent antioxidant capacity: DPPH (~90%), ABTS (~99%), and reducing power (0.86-1.03). Among the hydrolysates evaluated, the RDPH-(100 °C) led to the lowest reducing power and hydroxyl radical scavenging activity. Results reported here will be instrumental for the development of rice protein-based products and in the optimization and scale up of manufacturing process.

Formation of fibrils derived from whey protein isolate: structural characteristics and protease resistance.

The formation of lactoglobulin (ß-lg) fibrils from its native globular form has been investigated hitherto with pure lactoglobulin conditions. For commercially feasible production of ß-lg fibrils for food and therapeutic applications, the effect of impurities (albeit at very small fractions) found in industrial grade lactoglobulin proteins will need to be considered. In this work, food grade whey protein isolate (WPI), with its associated trace globulin and other impurities, was used to study the ß-lg fibril formation process. SDS-PAGE analysis supported by Thioflavin T measurement showed that the WPI fibril formation accompanied by hydrolysis limited the formation reaction rate; further analysis indicated that main impurities especially for α-lactalbumin may affect the intermolecular contacts among ß-lg. The increase of ß-sheet and α-helix conformation analyzed by circular dichroism revealed that the ß strand axis with an α-helix stabilized the linear structure of fibrils. The WPI fibrils exhibited a significant variation in lengths and differences in morphology when compared to some other protein fibrils reported in the literature. Two-stage pepsin followed by pancreatin digestion analysis revealed that the WPI fibrils produced will have the potential as a medium for active compound delivery to the intestinal environment.

Purification of Four Flavonoid Glycosides from Lotus (Nelumbo nucifera Gaertn) plumule by Macroporous Resin Combined with HSCCC.

High-speed counter-current chromatography (HSCCC) combined with macroporous resin (MR) column was successfully applied to the isolation and purification of four flavonoid glycosides from the medicinal herb Lotus plumule (LP). A polar two-phase solvent system composed of ethyl acetate-n-butanol-water (1:2:3, v/v/v) was selected by high-performance liquid chromatography (HPLC) and run on a preparative scale where the lower aqueous phase was used as the mobile phase with a head-to-tail elution mode. Quercetin-3-O-ß-D-glucopyranoside (15 mg), isorhamnetin-3-O-ß-D-glucopyranoside (13 mg), apigenin 6-C-ß-D-glucopyranosyl-8-C-α-L-arabinopyranoside (18 mg) and apigenin 6,8-di-C-ß-D-glucopyranoside (48 mg) were obtained in a one-step HSCCC separation from 240 mg of the sample. The purity of each compound was over 95% as determined by HPLC. Chemical structures of the isolated compounds were identified by electrospray ionization mass spectrometry (ESI-MS-MS) and nuclear magnetic resonance (NMR) methods. Moreover, the four compounds were isolated from LP for the first time.

Characteristics and Feasibility of Trans-Free Plastic Fats through Lipozyme TL IM-Catalyzed Interesterification of Palm Stearin and Akebia trifoliata Variety Australis Seed Oil.

Akebia trifoliata var. australis seed oil (ASO) was used as an edible oil in China. However, in-depth research studies on ASO have yet to be conducted for production of plastic fats in food industry. In this work, an immobilized lipase from Thermomyces lanuginosus (TL IM) was employed to catalyze palm stearin (PS) with different ratios of ASO in a laboratory-scale operation at 60 °C. The physical properties [e.g., fatty acid profile, slip melting point (SMP), solid fat content (SFC), polymorphic form, and microstructure] of physical blends (PBs) were analyzed and compared with those of the interesterified products (IPs). Results showed that SMPs of IPs (33.20-37.60 °C) decreased compared with those of PBs (48.03-49.30 °C). Meanwhile, IPs showed a good SFC range from 16.11% to 28.29% at 25 °C with mostly ß' polymorphic forms determined by X-ray diffraction analysis. It should be mentioned that no trans fatty acids (TFAs) were detected in any products, suggesting much more health-benefits of IPs. Texture tests showed that PBs (3318.19 ± 86.67 g) were markedly harder than IPs (557.02 ± 12.75 g). Conclusively, our study demonstrated that ASO can be utilized to produce trans-free plastic fats with good qualities through lipase-catalyzed interesterification.

Antioxidant activities of Se-SPI produced from soybean as accumulation and biotransformation reactor of natural selenium.

A study to compare the uptake, translocation, and distribution of selenium (Se) in soybean planted in natural seleniferous soil in Fengcheng city of China was conducted to clarify the relationship between the Se content levels of soybean proteins and their radical scavenging activity. The data showed that the total Se content in different parts of soybean plants varied with the growth periods. The selenoprotein (Se-SPI) content increased remarkably with the increase of Se content in seleniferous soils. The Se-SPI content obtained from the region with the highest Se level was almost 18 times higher than that of the control group, while antioxidant activities were about 4-fold compared to the control, suggesting that Se played a positive role in enhancing the antioxidant activity of Se-SPI. The increase in the Se level also led to changes in amino acids composition, but with nearly no effects on the subunit composition of soybean Se-SPI.