Physicochemical properties and bioactivity of whey protein isolate-inulin conjugates obtained by Maillard reaction.

The functional properties and physiological functions of whey protein isolate (WPI) decreased near its isoelectric point (PI). The Maillard reaction covalently binding polysaccharides to proteins is an effective method to improve the functional activities of proteins. WPI-inulin conjugates were prepared by wet-heating method at 70 °C for 2 h, 4 h and 6 h, respectively. New bonds at higher molecular zone appearing at SDS-PAGE, decreased free amino acid content and new formed CN bonds in FT-IR of conjugates compared with WPI confirmed the formation of the covalent bonds between WPI and inulin. As the increase of the reaction time, both the brown intensity and fluorescence intensity of WPI-inulin conjugates became higher. Amino acid contents, Circular dichroism analysis and SEM analysis presented the primary structure, secondary structure and surface structure change of protein after covalent with inulin. Emulsion properties of emulsion activity (EAI) and emulsion stability (ES) of WPI-inulin conjugates were assessed and both showed significantly enhanced compared with WPI at range of pH 3 to pH 7. AAPH+ scavenging test and ORAC measurement also revealed that covalent binding with inulin enhanced the antioxidant activities of WPI. This work presented the conjugation with inulin successfully enhanced the functional properties of WPI.

Comparative assessment of phytochemical profiles and antioxidant and antiproliferative activities of kiwifruit (Actinidia deliciosa) cultivars.

The present study was designed to analyze and compare phytochemical activities of four different cultivars of kiwifruit. Among all investigated varieties, Hua You (HY) and Cui Xiang (CUX) displayed the maximum concentration of phytochemical content, and the highest total phenolic results were observed in HY and CUX cultivars with 220.20 ± 1.12 mg GAE/100 g and 218.04 ± 1.11 mg GAE/100 g FW, respectively. Likewise, the richest total flavonoids results were estimated in red kiwifruit (RKF) and CUX varieties with 49.082 ± 0.14 mg CE/100 g FW and 48.327 ± 0.14 mg CE/100 g FW, respectively. Moreover, tests for oxygen radical scavenging capacity (ORAC) and peroxyl radical scavenging capacity (PSC) were observed maximum in RKF cultivar showing 131.229 ± 5.91 µM Trolox equivalent/g FW and 85.957 ± 11.75 µM vitamin C equivalent/g FW, respectively. Furthermore, the highest cellular antioxidant activity (CAA) with No PBS wash protocol was depicted in RKF 237.544 ± 4.12 µM QE equivalent/g FW with the lowest EC50 0.0128 mg/ml. In addition, high-performance liquid chromatography (HPLC) analysis confirmed the presence of ferulic acid, naringin, gallic acid, syringic acid, caffeic acid, rutin, protocatechuic acid, salicylic acid, and catechin in kiwifruit. Catechin as one main content in our study is consistent with the recent reports. The result suggested that the phytochemical profile and bioactivities were significantly affected by the type of cultivars. PRACTICAL APPLICATIONS: Kiwifruit is widely consumed over the world for its rich nutritious and medicinal values. Currently, phytochemicals are considered as one of the main bioactive components of kiwifruits, which are responsible for lots of bioactivities, such as antitumor, anti-inflammatory, antioxidant, hypoglycemic, and hypolipidemic activities. There are varieties of kiwifruits, and the bioactive components and bioactivities are greatly affected by the cultivars. But there have been no comparative studies on the phytochemicals from different varieties. This study aimed to make a comprehensive assessments of the free, bound, and total phenolics and flavonoids, as well as the chemical-based and cell-based antioxidant activities of four different subspecies of kiwifruit. This work would be beneficial to elucidate the function differences of different kiwifruit phytochemicals, promote its further research, as well as provide a basis for selecting cultivars.

A comparison study on polysaccharides extracted from Fructus Mori using different methods: structural characterization and glucose entrapment.

In this study, the structure characteristics and the hypoglycemic and antioxidant activities of mulberry fruit polysaccharides obtained by the commonly used hot water (MFPh)-, ultrasonic (MFPu)-, acid (MFPc)- and alkali (MFPa)-assisted extraction methods were investigated. NMR analysis indicated that the four polysaccharides had similar glycosidic linkage patterns. Scanning electron microscopy analyses showed that the surface morphology of the polysaccharides was greatly affected by the extraction methods. The results of the bioactivity assays indicated that MFPh exhibited stronger antioxidant and α-amylase inhibitory activities than the other polysaccharides. Moreover, all the polysaccharides showed good α-glucosidase inhibitory activities except for MFPu with the lowest molecular weight. These results suggested that acid, alkali, and ultrasonic-assisted extractions have different effects on the degradation of polysaccharides without changing the main structure compared with hot water extraction. In addition, the molecular weight of polysaccharides plays a key role in the bioactivity of the mulberry fruit polysaccharides.

Comparative study on the physicochemical properties and bioactivities of polysaccharide fractions extracted from Fructus Mori at different temperatures.

This study examined the effects of different extraction temperatures (30 °C and 90 °C) on the physicochemical properties and bioactivities of polysaccharides (MFPs-30-60, MFPs-30-80, MFPs-90-40, MFPs-90-60 and MFPs-90-80) extracted at same ethanol gradation (40, 60 and 80%). The polysaccharides were extracted from Fructus Mori, and their antioxidant, hypoglycemic and hypolipidemic activities were evaluated. The results showed that all the polysaccharide fractions contained the same monosaccharides but at different molar ratios. MFPs-30-80 had the highest content of glucose (30.33%), and MFPs-90-40 had the highest content of uronic acid (43.46%). In addition, compared to the extraction temperature of 30 °C, the higher temperature (90 °C) caused the degradation of the extracted polysaccharides. Generally, MFPs-30-80 showed superior antioxidant activity, MFPs-90-40 displayed good bile acid-binding ability, and both MFPs-30-80 and MFPs-90-40 exhibited the strongest inhibitory effects on α-amylase and α-glucosidase activity. This study concludes that the bioactivities of mulberry fruit polysaccharides depend on a combination of structural factors that are greatly affected by the extraction temperature.

Modulation of gut microbiota by mulberry fruit polysaccharide treatment of obese diabetic db/db mice.

Increasing evidence indicates that gut microbiota is an important factor in mediating the development of metabolic disorders, especially type 2 diabetes. Herein, we investigated the protective effects of polysaccharides from mulberry fruit on diabetic db/db mice. We observed an inhibition in the body weight increase, a decrease in the blood glucose levels, and an improvement in glucose tolerance. Furthermore, our data showed increased enzymatic activities as evidenced by parameters such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT). In addition, high density lipoprotein cholesterol (HDL-C) increased, while total cholesterol (TC), triacylglycerol (TG), low density lipoprotein cholesterol (LDL-C), lipid peroxide content malonaldehyde (MDA), and free fatty acid (FFA) levels decreased, accompanied by the recovery from damage to the liver, kidneys and pancreas. Meanwhile, metformin and polysaccharide similarly shifted the abundance of the main gut microbiota, Bacteroidetes and Firmicutes, in diabetic mice toward levels observed in healthy mice. Especially at the genus level, the enrichment of some key bacteria like Bacteroidales, Lactobacillus, Allobaculum, Bacteroides, and Akkermansia was observed. Taken together, our findings suggest that polysaccharides from the mulberry fruit modulate gut microbiota, including enriching functional bacteria and reducing microbial diversity, which may contribute to their antidiabetic effects.

Chemical property and impacts of different polysaccharide fractions from Fructus Mori. on lipolysis with digestion model in vitro.

Three polysaccharides (MFP-1, MFP-2, and MFP-3) were fractioned from mulberry fruits with 30%, 60%, and 90% (v/v) ethanol, respectively. The MFP-1 had the largest molecular weight of 838kDa, followed by MFP-2 from 49.5 to 163kDa, and then MFP-3 from 1.41 to 15.30kDa. The three polysaccharides were composed of arabinose, galactose, glucose, mannose, xylose, and galacturonic acid but with different molar ratio. They also exhibited different surface morphology and thermogravimetric properties. In addition, the impact of the three polysaccharides on the digestion of lipid in a simulated saliva and gastrointestinal tract was investigated. The results indicated MFP-2 had the strongest ability to reduce the rate and extent of lipid digestion. These results have important implications for understanding the influence of polysaccharides on lipid digestion. The effect on retarding lipid digestion within the gastrointestinal tract might be important for the development functional food rich in oil to promote health.

Characterization of polysaccharide fractions in mulberry fruit and assessment of their antioxidant and hypoglycemic activities in vitro.

The present study aimed to characterize and investigate the polysaccharide fractions of mulberry fruit, and their antioxidant and hypoglycemic activities. Mulberry fruit polysaccharides (MFP) were extracted by using hot water as the solvent and fractioned by using a DEAE-Sepharose fast-flow column. In total four eluents of water (MFP-1), 0.1 M NaCl (MFP-2), 0.2 M NaCl (MFP-3) and 0.3 M NaCl (MFP-4) were fractionated. Arabinose, galactose, glucose, xylose and mannose were the main components present in MFP-1, while MFP-2 and MFP-3 were composed of arabinose and galactose, and MFP-4 of arabinose, galactose and glucose. High-performance gel permeation chromatography (HP-GPC) analysis indicated a narrower molecular weight distribution in the MFP-1 fraction. Scanning electron microscopy (SEM) exhibited a smooth surface for MFP-1 and MFP-3 fractions, whereas MFP-2 and MFP-4 have pore openings and flocculent fibers on a smooth surface. Tertiary structure analyses indicated that none of the fractions had a triple-helical conformation. On the whole, MFP-3 and MFP-4 showed better antioxidant activity and inhibitory effects on α-amylase and α-glucosidase compared to MFP-1 and MFP-2. These results show significant potential for the antioxidant and hypoglycemic activities of MFP-3 and MFP-4 indicating the need for their further exploration as potential antidiabetic agents.

Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro.

Single-factor experiment and Box-Behnken design (BBD) were applied to optimize the ultrasound-assisted extraction of mulberry fruits polysaccharides (MFP). Under optimum conditions: ratio of water to raw material 40.25, extraction temperature 69°C, ultrasonic power 190W and extraction time 75 min, the MFP yield was 3.13% (±0.07%), in accordance to the predicted value of 3.04%. The mulberry fruits polysaccharides fractions was obtained by deproteinization (MFP-1), followed by decolorization and deionization (MFP-2). Carbohydrate content in MFP, MFP-1 and MFP-2 was 58.61% (±1.47%), 69.98% (±0.91%), 81.18% (±1.29%), as well as proteins was estimated 16.50% (±0.86%), 1.57% (±0.63%), 1.02% (±0.18%), respectively. The FT-IR indicated that MFP, MFP-1 and MFP-2 were acidic polysaccharides. The MFP-1 exhibited the strongest antioxidant activity, while MFP-2 showed the strongest hyperglycemic activity in vitro. This may be caused by their different compositions and physical properties in the different mulberry fruit polysaccharides fractions.

Characterization, antioxidant and immunomodulatory activities of polysaccharides from Prunella vulgaris Linn.

Water-soluble polysaccharides from Prunella vulgaris Linn (P. vulgaris) were fractionated using DEAE-Sepharose fast-flow column to obtain several eluents of water (PV-P1), 0.1M NaCl (PV-P2) and 0.2M NaCl (PV-P3). Structural analyses showed that PV-P1 had a higher molecular weight and degree of branching as compared to PV-P2 and PV-P3. Tertiary structure analyses indicated that PV-P1, PV-P2 and PV-P3 did not have triple-helical conformation. PV-P2 and PV-P3 showed stronger antioxidant activities than PV-P1, as measured radical scavenging capacities. PV-P1 showed stronger immunomodulatory activities than PV-P2 and PV-P3 in term of stimulation of the production of pro-inflammatory cytokines, including nitric oxide (NO), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) in murine macrophage RAW 264.7 cells. PV-P1, PV-P2 and PV-P3 did not exhibit cytotoxicities against RAW 264.7 at the concentrations tested. These results suggest that P. vulgaris polysaccharides could be explored as potential antioxidant and immunomodulatory agents for the complementary medicine or functional foods.

Highly sensitive detection of target ssDNA based on SERS liquid chip using suspended magnetic nanospheres as capturing substrates.

A new approach for sensitive detection of a specific ssDNA (single-stranded DNA) sequence based on the surface enhanced Raman spectroscopy (SERS) liquid chip is demonstrated. In this method, the probe DNA (targeting to one part of target ssDNA) was attached to the nano-SERS-tags (poly(styrene-co-acrylic acid)/(silver nanoparticles)/silica composite nanospheres), and the capture DNA (targeting to the other part of target ssDNA) was attached to the Fe3O4/poly(acrylic acid) core/shell nanospheres. The nano-SERS-tags with probe DNA were first allowed to undergo hybridization with the target ssDNA in solution to achieve the best efficiency. Subsequently, the magnetic composite nanospheres with capture DNA were added as the capturing substrates of the target ssDNA combined with the nano-SERS-tags. Upon attraction with an external magnet, the nanospheres (including the nano-SERS-tags) were deposited together due to the hybridization, and the deposit sediment was then analyzed by SERS. Quantitative detection of target ssDNA was achieved based on the well-defined linear correlation between the SERS signal intensity and the target ssDNA quantity in the range of 10 nM to 10 pM, and the limit of detection was approximately 10 pM. Multiplexed detection of up to three different ssDNA targets in one sample was demonstrated using three different types of nano-SERS-tags under a single excitation laser. The experimental results indicated that the liquid-phase DNA sequencing method, thus named the SERS liquid chip (SLC) method, holds significant promises for specific detection of trace targets of organisms.

Ultrafast hydrothermal synthesis of high quality magnetic core phenol-formaldehyde shell composite microspheres using the microwave method.

An ultrafast, facile, and efficient microwave hydrothermal approach was designed to fabricate magnetic Fe(3)O(4)/phenol-formaldehyde (PF) core-shell microspheres for the first time. The structure of the Fe(3)O(4)/PF core-shell microspheres could be well controlled by the in situ polycondensation of phenol and formaldehyde with magnetic Fe(3)O(4) clusters as the seeds in an aqueous solution without any surfactants. The effect of synthetic parameters, such as the feeding amounts of phenol, the dosages of formaldehyde, the reaction temperatures, and the microwave heating time, on the morphologies and sizes of the Fe(3)O(4)/PF microspheres were investigated in details. The phenol-formaldehyde shell is found to be evenly coated on Fe(3)O(4) clusters within 10 min of the irradiation. The as-prepared microspheres were highly uniform in morphology, and the method was found to allow the shell thickness to be finely controlled in the range of 10-200 nm. The properties of the composite microspheres were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), thermogravimetic analysis (TGA), Fourier transform infrared (FT-IR) spectra, X-ray diffraction (XRD), and vibrating sample magnetometer (VSM). The as-prepared Fe(3)O(4)/PF microspheres were monodisperse and highly dispersible in water, ethanol, N,N-dimethyformamide, and acetone, a beneficial quality for the further functionalization and applications of the Fe(3)O(4)/PF microspheres.

Tailor-made magnetic Fe3O4@mTiO2 microspheres with a tunable mesoporous anatase shell for highly selective and effective enrichment of phosphopeptides.

Selective enrichment of phosphoproteins or phosphopeptides from complex mixtures is essential for MS-based phosphoproteomics, but still remains a challenge. In this article, we described an unprecedented approach to synthesize magnetic mesoporous Fe(3)O(4)@mTiO(2) microspheres with a well-defined core/shell structure, a pure and highly crystalline TiO(2) layer, high specific surface area (167.1 m(2)/g), large pore volume (0.45 cm(3)/g), appropriate and tunable pore size (8.6-16.4 nm), and high magnetic susceptibility. We investigated the applicability of Fe(3)O(4)@mTiO(2) microspheres in a study of the selective enrichment of phosphopeptides. The experiment results demonstrated that the Fe(3)O(4)@mTiO(2) possessed remarkable selectivity for phosphopeptides even at a very low molar ratio of phosphopeptides/non-phosphopeptides (1:1000), large enrichment capacity (as high as 225 mg/g, over 10 times as that of the Fe(3)O(4)@TiO(2) microspheres), extreme sensitivity (the detection limit was at the fmol level), excellent speed (the enrichment can be completed in less than 5 min), and high recovery of phosphopeptides (as high as 93%). In addition, the high magnetic susceptibility allowed convenient separation of the target peptides by magnetic separation. These outstanding features give the Fe(3)O(4)@mTiO(2) composite microspheres high benefit for mass spectrometric analysis of phosphopeptides.

Toward designer magnetite/polystyrene colloidal composite microspheres with controllable nanostructures and desirable surface functionalities.

An effective method was developed for synthesizing magnetite/polymer colloidal composite microspheres with controllable variations in size and shape of the nanostructures and desirable interfacial chemical functionalities, using surfactant-free seeded emulsion polymerization with magnetite (Fe(3)O(4)) colloidal nanocrystal clusters (CNCs) as the seed, styrene (St) as the monomer, and potassium persulfate (KPS) as the initiator. The sub-micrometer-sized citrate-acid-stabilized Fe(3)O(4) CNCs were first obtained via ethylene glycol (EG)-mediated solvothermal synthesis, followed by 3-(trimethoxysilyl)propyl methacrylate (MPS) modification to immobilize the active vinyl groups onto the surfaces, and then the hydrophobic St monomers were polymerized at the interfaces to form the polymer shells by seeded emulsion radical polymerization. The morphology of the composite microspheres could be controlled from raspberry- and flower-like shapes, to eccentric structures by simply adjusting the feeding weight ratio of the seed to the monomer (Fe(3)O(4)/St) and varying the amount of cross-linker divinyl benzene (DVB). The morphological transition was rationalized by considering the viscosity of monomer-swollen polymer matrix and interfacial tension between the seeds and polymer matrix. Functional groups, such as carboxyl, hydroxyl, and epoxy, can be facilely introduced onto the composite microspheres through copolymerization of St with other functional monomers. The resultant microspheres displayed a high saturation magnetization (46 emu/g), well-defined core-shell nanostructures, and surface chemical functionalities, as well as a sustained colloidal stability, promising for further biomedical applications.

Detecting trace melamine in solution by SERS using Ag nanoparticle coated poly(styrene-co-acrylic acid) nanospheres as novel active substrates.

A systematic study for the preparation of Ag nanoparticle (Ag-NP) coated poly(styrene-co-acrylic acid) (PSA) composite nanospheres by in situ chemical reduction is reported. The experimental results showed that the reaction temperature and the surface coverage of the -COOH determined the surface coverage and grain size of Ag nanoparticles on the PSA nanospheres. The surface enhanced Raman spectroscopy (SERS) sensitivity was investigated using 4-hydroxythiophenol (4-HBT) as the model probe in the solution of composite nanospheres stabilized by polyvinylpyrrolidone (PSA/Ag-NPs/PVP), with the detection limit of about 1 × 10(-6) M. Potential application of the new SERS substrate was demonstrated with the detection of melamine, and the detection limit was about 1 × 10(-3) M. Chemical noises from PVP and other impurities were observed and attributed mainly to the competitive adsorption of PVP on the surfaces of Ag-NPs. After tetrahydrofuran washing of the PSA/Ag-NPs/PVP substrates that removed the PVP and other residuals, the signal/noise levels of SERS were greatly improved and the detection limit of melamine was determined to be 1 × 10(-7) M. This result indicated that the new PSA/Ag-NPs system is highly effective and can be used as the SERS-active substrate for trace analysis of a variety of drugs and food additives.