Revealing the regional distribution of soybean lunasin content in China and the effects of climate factors by sampling extensively.

BACKGROUND: Lunasin is a novel therapeutic peptide that was initially isolated from soybean. In this study, we quantified the variations in lunasin content in a total of 413 soybean samples that were collected from four major regions in China and harvested in 2014 and 2015 to reveal the regional distribution of soybean lunasin content in China and the effects of climate factors. RESULTS: The results showed that the cultivar Changmidou 30 collected from Jilin province and harvested in 2015 had the highest lunasin content (3.25 g kg-1 dry seeds). The data from both 2014 and 2015 showed that the lunasin content in soybean collected from northern China was significantly higher (P < 0.05) than that from south China. There was a positive correlation (P < 0.01) between lunasin content and hours of sunshine (HS) as well as diurnal temperature range (DTR); however, there was a negative correlation (P < 0.01) between lunasin content and rainfall (RF). In addition, combined analysis of data from 2014 and 2015 demonstrated that DTR was the dominant factor that affected lunasin content with a direct path-coefficient of 0.301. CONCLUSIONS: These results were anticipated to contribute to guiding the cultivation of soybean with high lunasin content. © 2018 Society of Chemical Industry.

Expression of Bioactive Lunasin Peptide in Transgenic Rice Grains for the Application in Functional Food.

Lunasin, a bioactive peptide initially isolated from soybean, has anticancer, anti-inflammatory, and antioxidant activity. Due its great application value, lunasin seems to be a candidate gene in improving the nutritional value of crops. In this study, lunasin was inserted into the rice genome to evaluate whether it was feasible to express lunasin using the rice expression system and improve the bioactivity of protein in rice for our needs. We generatedlunasin-overexpressing rice lines, and chose three independent transgenic rice lines for further study. The lunasin content in trans-lunasin rice detected by UPLC-MS/MS was 1.01 × 10-3 g·kg-1 dry rice flour with grease removal in the lunasin extracts. The antioxidant efficacy of LET (lunasin-enriched fraction from trans-lunasin rice) and PEW (peptide-enriched fraction from wild type rice) was compared. Due to the presence of lunasin, LET showed higher (p < 0.05) antioxidant activity than PEW. LET exhibited high DPPH radical scavenging activity (IC50 value, 8 g·L-1), strong ABTS⁺ radical scavenging activity (IC50 value, 1.18 g·L-1), and great oxygen radical scavenging activity (170 µmol·L-1 Trolox equivalents when the concentration reached 4 g·L-1). Moreover, LET presented significantly higher (p < 0.05) anti-inflammatory activity on macrophage cells, and the NO production and the release of pro-inflammatory cytokines (IL-6, MCP1, and TNF-α) were significantly inhibited by LET. However, because of the low purity, LET showed weaker antioxidant and anti-inflammatory activity when compared to the Lunasin standard. These results suggested that it is feasible to use the rice expression system to express the exogenous lunasin in rice, and lunasin-overexpressing rice seems to be a candidate resource for application in functional food. Rice rich in lunasin is beneficial for human health, and could be used as a functional food in the diets of cancer and obese patients in the future.

Antiproliferative Activity of Combined Biochanin A and Ginsenoside Rh2 on MDA-MB-231 and MCF-7 Human Breast Cancer Cells.

Breast cancer is the most frequently diagnosed cancer in women worldwide. The antiproliferative activities of biochanin A (BA) and ginsenoside Rh2 were determined by evaluating their inhibitory effect on MDA-MB-231 human breast cancer cell proliferation. The combination of BA with Rh2 was also assessed. In MDA cells, combination treatment led to a decrease in the EC50 values of BA and Rh2 to 25.20 µM and 22.75 µM, respectively. In MCF-7 cells, the EC50 values of combined BA and Rh2 decreased to 27.68 µM and 25.41 µM, respectively. BA combined with Rh2 also improved the inhibition of MDA-MB-231 and MCF-7 cell migration and invasion compared to the individual compounds. Western blot analysis demonstrated upregulation in p-p53, p-p38, and p-ASK1 proteins while levels of TRAF2 were downregulated. These results suggest that BA combined with Rh2 exhibits synergistic effects against MDA-MB-231 and MCF-7 cell proliferation.

Synergistic Effect of Bioactive Anticarcinogens from Soybean on Anti-Proliferative Activity in MDA-MB-231 and MCF-7 Human Breast Cancer Cells In Vitro.

Consumption of soybean products has been implicated in the prevention of breast cancer. This study provides insights into the anti-proliferative activity of 12 anticarcinogens from soybean by single or two-way combination treatment against MCF-7 and MDA-MB-231 human breast cancer cells. Results showed that genistein, daidzein, glycitein, genistin and dainzin showed stronger anti-proliferative activity against MCF-7 cells with EC50 values of 66.98 ± 4.87 µM, 130.14 ± 2.10 µM, 190.67 ± 5.65 µM, 72.82 ± 2.66 µM and 179.21 ± 6.37 µM, respectively. There is a synergistic effect of combination treatment of genistin plus daidzin in MCF-7 cells with combination index at inhibition of 50% (CI50) of 0.89 ± 0.12. Genistein, glycitein, genistin and ß-sitosterol were demonstrated to have a stronger anti-proliferative activity against MDA-MB-231 cells with EC50 values of 93.75 ± 5.15 µM, 142.67 ± 5.88 µM, 127.82 ± 4.70 µM and 196.28 ± 4.45 µM. The synergistic effect was observed in the mixture of genistein plus genistin, genistein plus ß-sitosterol or ß-sitosterol plus genistin with CI50 values of 0.56 ± 0.13, 0.54 ± 0.20 and 0.45 ± 0.12, respectively. These bioactive anticarcinogens were able to inhibit invasion and migration of breast cancer cells and the combination treatments enhanced the inhibitory effect. Regulation of PI3K/Akt/mTORpathway seems to be the main mechanisms involved in the anticancer activity.

Detection of lunasin in quinoa (Chenopodium quinoa Willd.) and the in vitro evaluation of its antioxidant and anti-inflammatory activities.

BACKGROUND: Lunasin is a novel cancer-preventive peptide that has been detected in various plants. However, the presence and bioactivity of lunasin in quinoa have not been demonstrated. RESULTS: Lunasin was detected in quinoa using ultrahigh-pressure liquid chromatography-electrosparay ionization-mass spectrometry. The content in 15 quinoa samples ranged from 1.01 × 10-3 g kg-1 dry seed to 4.89 × 10-3 g kg-1 dry seed. Significant differences (P < 0.05) in lunasin content among different cultivars (yy22, xsg86, xsg59, yy30 and yy19) from the same area and the same cultivar from different areas (Hainan, Hebei and Shanxi provinces) were observed. The lunasin isolated and purified from quinoa (QLP) exhibited a weak DPPH radical scavenging activity (no IC50 value), but a strong ABTS+ radical scavenging activity (IC50 value, 1.45 g L-1 ) and oxygen radical scavenging activity (40.06 µmol L-1 Trolox equivalents/g QLP when the concentration was 3.20 g L-1 ). In addition, QLP inhibited the production of nitric oxide (NO), tumor necrosis factor-α and interleukin-6 on lipopolysaccharide-stimulated RAW264.7 macrophages by up to 44.77%, 39.81% and 33.50%, respectively, at a concentration of 0.40 g L-1 . CONCLUSION: Taken together, these findings indicate that lunasin presents in quinoa and is bioactive, which strengthens the recommendations for the development of quinoa-based functional foods. © 2017 Society of Chemical Industry.

Antioxidant and immunoregulatory activity of polysaccharides from quinoa (Chenopodium quinoa Willd.).

The water-extractable (QWP) and the alkali-extractable (QAP) polysaccharides from quinoa (named QWP and QAP, respectively) and their four polysaccharide sub-fractions (QWP-1, QWP-2, QAP-1 and QAP-2), were isolated and purified by anion-exchange and gel filtration chromatography. QWP-1 and QWP-2 were composed of Rha, Ara, Gal and GalA. QAP-1 and QAP-2 were composed of Rha, Ara, Man, Gal and GalA. Antioxidant and immunoregulatory activities of the polysaccharides were evaluated. The results showed that QWP-1, QWP-2, QAP-1 and QAP-2 had significant antioxidant and immunoregulatory activities. The results suggest that QWP-1, QWP-2, QAP-1 and QAP-2 could be used as potential antioxidants and immunomodulators.

Mung bean decreases plasma cholesterol by up-regulation of CYP7A1.

Our results affirmed that supplementation of 1 or 2% mung bean could decrease plasma total cholesterol and triacylglycerol level. Mung bean increased mRNA 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase. Most importantly, mung bean increased not only the protein level of cholesterol-7α-hydroxylase (CYP7A1) but also mRNA CYP7A1. It was concluded that the hypocholesterolemic activity of mung bean was most probable mediated by enhancement of bile acid excretion and up-regulation of CYP7A1.

A method for obtaining object defocus information in the RC observation mode.

The micro-operation robot is widely used in micro-manipulations of biological cells in biological and medical experiments. It plans and controls micro-effector movement based on image feedback information to achieve micro-operations. However, the displacement information of the micro-effector on the x-y plane can be obtained from the image, but not the position information of the micro-effector in the z-axis direction. This makes the micro-effector movement in the z-axis direction discontinuous, which is time-consuming and reduces operational efficiency. In this study, starting from the optical imaging principle of Robert Hoffman modulation contrast method (RC), we propose a defocus detection method for the RC observation mode of an optical microscope. Our method can determine the direction of defocus, which is not available in previous defocusing detection methods. Utilizing this method, we achieve rapid focus for the micro-effector while it is moving along the z-axis direction.

Immunomodulatory activity of glycoproteins isolated from chickpea (Cicer arietinum L.).

Chickpea (Cicer arietinum L.) is a well-known legume widely used as traditional medicine. This study aimed to characterize the structure and evaluate the immunomodulatory activity of one glycoprotein [crude chickpea glycoprotein-1 (CAG-1)] isolated from chickpea. CAG-1 was extracted with hot alkaline water and purified with DEAE-Sepharose Fast Flow and Superdex-200 column chromatography. CAG-1, with a molecular weight of 8,106 Da, contained 57.12% polysaccharide and 35.41% protein. The polysaccharide part was mainly composed of glucose (Glc). The protein part was connected mainly by aspartic (Asp) and glutamic (Glu). The results of nuclear magnetic resonance (NMR) analysis indicated the presence of α-d-Glcp-(1 → 4)-α-d-Glcp-(1 → 4)-α-d-Glcp-(1 → . In addition, the sugar chains of the glycoprotein were not hydrolyzed under alkaline conditions, suggesting that the glycoprotein was N-glycosidic; thus, the sugar chain was linked to the protein chain by Asp. An immunological study showed that CAG-1 stimulated the production of nitric oxide (NO), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and monocyte chemotactic protein 1 (MCP-1) in RAW 264.7 macrophages in a dose-dependent manner.

Structural characterization and bioactivity evaluation of water-extractable polysaccharides from chickpeas (Cicer arietinum L.) seeds.

Two water-extractable polysaccharide fractions designated as CWP (7. 37 × 105 Da) and CWP-0.2 (1.58 × 104 Da) were isolated and purified from chickpea (Cicer arietinum L.) seeds. The chemical structure of the two polysaccharides was characterized by various methods. Monosaccharide composition and methylation analysis showed that CWP was composed of Man and Glc in a molar ratio of 44.6:55.4, and CWP-0.2 was composed of Rha, Ara, Man, Glc, and Gal in a molar ratio of 10.6:23.3:5.2:4.9:56. Further structural characterization indicated that the main chain connection of CWP was → (2-ß-d-Fruf-1) n →, and the main chain connection of CWP-0.2 was explored as → 2,4)-α-l-Rhap-(1 → 3)-α-d-Galp-(1 → with the branched chain of → 2,4)-α-l-Rhap-(1 → o-4. Besides, both CWP and CWP-0.2 had antioxidant and immunoregulatory activity in vitro, through scavenging DPPH· and ABTS·+ as well as stimulating production of NO, IL-6, TNF-α and MCP-1 in RAW 264.7 macrophages. CWP-0.2 revealed significantly higher bioactivity than CWP.

High-throughput metabolomics using UPLC/Q-TOF-MS coupled with multivariate data analysis reveals the effect and mechanism of syringin against ovariectomized osteoporosis.

Osteoporosis is an increasing public health problem in the worldwide and has caused socioeconomic burden. Natural products as candidates have the potential to promote bone formation and suppress bone resorption for osteoporosis treatment. Previously, syringin has showed the potent anti-osteoporosis activity, however the detailed mechanism of syringin against osteoporosis is still unclear. This study aimed to reveal the pharmacological effect and mechanism of syringin through the high-throughput metabolomics. In this study, metabolomics techniques were used to explore the metabolic biomarkers and profiles provides deep insights into the pharmacological effects and mechanism of syringin against osteoporosis. The metabolite biomarkers were monitored based on the high-resolution mass spectrometry. By the integration analysis of metabolomics technology, a total of 23 metabolic biomarkers were discovered and we found the highly relevant pathway involved in glycine and serine metabolism, butyrate metabolism, methionine metabolism, catecholamine biosynthesis, tyrosine metabolism, etc. Interestingly, synthesis and degradation of ketone bodies, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, tyrosine metabolism, glycine, serine and threonine metabolism, butanoate metabolism, was related with efficacy of syringin. The present work showed that the metabolomics technology can provide novel strategies for revealing insights into the metabolic effects and action mechanism of drug.

Peptides Released from Extruded Adzuki Bean Protein through Simulated Gastrointestinal Digestion Exhibit Anti-inflammatory Activity.

Increasing attention has been focused on plant-derived peptides because of their potential bioactivities. In this study, bioactive peptides were released from extruded adzuki bean protein by simulated gastrointestinal digestion. A peptide (KQS-1) sequenced as KQSESHFVDAQPEQQQR was separated and identified using ultrafiltration, pre-high-performance liquid chromatography (HPLC), and ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). KQS-1 was shown to exert significant anti-inflammatory effects in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages by reducing the production of IL-1, IL-6, TNF-α, and MCP-1 to 38.31, 6.07, 43.96, and 41.74%, respectively. The involved signaling pathways were identified by transcriptome analysis. Overall, 5236 differentially expressed genes (DEGs) were identified. Gene ontology (GO) functions demonstrated that DEGs were significantly related to the NF-κB pathway. In conclusion, KQS-1 prevented the activation and expression of NF-κB/caspase-1 by upstream and downstream factors. These findings highlight the bioactivity of adzuki bean peptides.

Soy Protein Alleviates Malnutrition in Weaning Rats by Regulating Gut Microbiota Composition and Serum Metabolites.

Dietary intervention with plant protein is one of the main methods that is used to lessen the symptoms of malnutrition. Supplementary soy protein to undernourished weaning rats for 6 weeks significantly increased their body weight gain. After the intervention, the level of total short-chain fatty acids (SCFAs) was restored to 1,512.7 µg/g, while the level was only 637.1 µg/g in the 7% protein group. The amino acids (valine, isoleucine, phenylalanine, and tryptophan) increased in the colon, and vitamin B6 metabolism was significantly influenced in undernourished rats. The tryptophan and glycine-serine-threonine pathways were elevated, leading to an increase in the level of tryptophan and 5-hydroxytryptophan (5-HTP) in the serum. In addition, the relative abundance of Lachnospiraceae_NK4A136_group and Lactobacillus increased, while Enterococcus and Streptococcus decreased compared to undernourished rats. Overall, soy protein improved the growth of rats with malnutrition in early life by regulating gut microbiota and metabolites in the colon and serum.

Mung Bean Protein Suppresses Undernutrition-Induced Growth Deficits and Cognitive Dysfunction in Rats via Gut Microbiota-TLR4/NF-kB Pathway.

Early undernutrition has been found to be closely associated with subsequent neurodevelopment. However, studies examining crude growth in terms of body weight/tail length cannot clarify how diets might mediate associations between the gut microbiota and cognitive dysfunction. In the present study, Sprague-Dawley (SD) rats were fed a 7% protein diet and mung bean protein diet (MBPD) for 6 weeks to assess central nervous system functions. Bifidobacterium longum subsp, Alloprevotella, and Lactobacillus were significantly altered after supplementary MBPD. Additionally, tryptophan, tyrosine, and glycine significantly restored in the brain, and the choline system also improved. Moreover, mung bean supplementation also upregulated expression of the brain-derived neurotrophic factor, postsynaptic density 95 protein (PSD95), synaptosome-associated protein 25 (SNAP25), downregulated toll-like receptor 4 (TLR4), and nuclear factor kB (NF-kB). Metabolites in the serum also underwent changes. Together, these results showed that malnutrition perturbed neurodevelopment, while MBPD reversed this trend.

Structural Characterization of Quinoa Polysaccharide and Its Inhibitory Effects on 3T3-L1 Adipocyte Differentiation.

Quinoa is a kind of nutritious food crop with anti-obesity activity, however, the mechanism is not unclear. In this study, we separated and purified bioactive polysaccharide from quinoa (denoted SQWP-2). The chemical structural was characterized and its effect on 3T3-L1 pre-adipocyte differentiation was evaluated. The molecular weight of SQWP-2 was found to be 7.49 × 103 Da, and the polysaccharide consisted of fructose and glucose. The Glc-(1→, Fru-(2→, →4)-Glcp-(1→, and →4,6)-Glcp-(1→ glycosidic linkages were identified in SQWP-2 through gas chromatography-mass spectrometry. Nuclear magnetic resonance confirmed the monosaccharide composition and glycosidic linkage content, and a suggestion of the structural formula is provided. In Western Blotting and RT-PCR assays, treatment with SQWP-2 significantly inhibited 3T3-L1 differentiation by suppressing PPARγ, C/EBPα, C/EBPß, C/EBPδ, SREBP1C and AP2 expression. Quinoa polysaccharide isolated here could represent an anti-obesity agent once the structures and differentiation inhibition are definitively characterized.

Anti-obesity effects of α-amylase inhibitor enriched-extract from white common beans (Phaseolus vulgaris L.) associated with the modulation of gut microbiota composition in high-fat diet-induced obese rats.

α-Amylase inhibitors (α-AI) have great potential to treat obesity. In this study, an α-AI enriched extract (α-AIE) with a specific activity of 1027.1 ± 154.2 (U per mg protein) was prepared from white common bean (Phaseolus vulgaris L.) seeds. Its anti-obesity effect and gut microbiota modulation properties were verified in high-fat diet-induced obese rats. The intake of the α-AIE significantly reduced body weight gain and improved serum lipid levels (p < 0.05). In addition, rats fed the α-AIE diet exhibited higher total short-chain fatty-acid (SCFA) concentrations (p < 0.05) in their colonic contents. ß-Diversity analysis, principal component analysis and a Venn diagram showed that α-AIE administration changed the gut microbiota composition. At the phylum level, the relative abundances of Firmicutes and Proteobacteria decreased and the relative abundances of Bacteroidetes and Akkermansia increased. In addition, 89 operational taxonomic units (OTUs) significantly responding to the high-fat diet and 30 OTUs significantly responding to the α-AIE were identified. The OTUs enriched by the α-AIE were mainly assigned to putative SCFA-producing bacteria, including Bacteroides, Butyricoccus, Blautia and Eubacterium. Twenty-two OTUs were found to be significantly correlated with obesity indexes. Taken together, the present results suggest that the intake of the α-AIE attenuated obesity and modulated gut microbiota.

Functional properties and adipogenesis inhibitory activity of protein hydrolysates from quinoa (Chenopodium quinoa Willd.).

The functional properties and adipogenesis inhibitory activity of quinoa protein hydrolysates, prepared using papain, pepsin, and pancreatin for 0, 30, 60, 90, and 120 min, were studied. For these three kinds of proteases, the solubility of the hydrolysates significantly increased with the increasing DH in pH range of 3-8, while the EAI and ESI of these hydrolysates significantly decreased during hydrolysis. The anti-inflammatory activity of these protein hydrolysates was measured. All of these protein hydrolysates showed high anti-inflammatory activity. However, there was no significant difference in anti-inflammatory activity between protein hydrolysates and total protein from quinoa. These protein hydrolysates also inhibited lipid accumulation during differentiation within the range of concentrations of 0-1,600 µg/ml, which exerted no cytotoxicity toward 3T3-L1 cells. The protein hydrolysates from quinoa prepared using pepsin for 120 min (PEP-120) had the highest activity with an IC50 value of 786.58 µg/ml. Moreover, LC-MS/MS analysis of PEP-120 showed that five main bioactive peptides, which have been demonstrated to have ACE inhibitor, antioxidant, and antithrombotic activities, were present in PEP-120. In addition, gene expression and Western blot analysis revealed that PEP-120 suppressed the 3T3-L1 cell differentiation through the peroxisome proliferator-activated receptor γ (PPARγ) pathway.

Tandem repeated expression of lunasin gene in Pichia pastoris and its anti-inflammatory activity in vitro.

Lunasin is a novel promising health-beneficial peptide derived from soybean. However, the application of lunasin is limited by its high cost. In this study, we developed a successful protocol for expression of a dimer formation protein containing 4 tandem repeated lunasin analogs (lunasin-4) in Pichia pastoris. The expression level at the optimal condition (initial pH 7.0, 1.0% final methanol concentration and induction for 72 h at 26 °C) was 0.24 mg/mL cell-free broth. Lunasin analog, obtained from purified lunasin-4 protein through enterokinase digestion and ultrafiltration, significantly decreased (p < 0.05) the release of nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in a dose-dependent manner. In addition, intracellular signaling array analysis demonstrated down-regulated levels of phosphorylated Akt, mechanistic target of rapamycin (mTOR) and p70 s6 kinase (p70s6k) and an up-regulated level of glycogen synthase kinase-3ß (GSK-3ß) after lunasin analog treatment. These results suggest that lunasin analog exerted anti-inflammatory activities in LPS-stimulated RAW264.7 cells partly via inhibiting the activation of Akt/mTOR/p70s6k signaling pathway. In conclusion, this study provides a potential strategy for recombinant production of bioactive lunasin in industry.

Ultrasonic depolymerization of Li2O-Na2O-SiO2 type silicate melt.

Under the application of ultrasound, the organic polymers can be degraded for the reason of breakage of C-C or C-N bond induced by the cavitation effect, similar to organic polymers, the silicate melts are also one typical polymer connected with Si-O bonds. To verify the prediction of the depolymerization of silicate melt, the ultrasound was applied to one kind of silicate melt for the first time via a special designed experimental system. Results showed that, under the applying of the ultrasound, the Si-O (bridged O) bonds in the structure units of the silicate melt can be broken, which decreases the polymerization degree and the viscosity. Under the higher power of applied ultrasound, the polymerization degree and viscosity of the silicate melt further decrease. Unlike some organic polymers, once stopping applying ultrasound, the broken lower polymerized units reconnect to form higher polymerized units gradually, the degree of the polymerization and the viscosity reverse to its nature state of the silicate melts. According to these results, the potential applications in the steel continuous casting process and electrolytic aluminum process are predicted.

Interactions between wheat starch and cellulose derivatives in short-term retrogradation: Rheology and FTIR study.

The understanding of the interactions between starch and cellulose hydrocolloids is crucial for equipment design and definition of operation parameters in the food industry. In the present study, the possible interactions of wheat starch (WS) with different levels (2-10%) of carboxymethyl cellulose (CMC) or microcrystalline cellulose (MCC) in short-term retrogradation were explored by dynamic rheological and Fourier transform infrared spectrum (FTIR) measurements. The chains of water-soluble CMC could creep into the continuous phase during paste gelation. It was found the amylose network structure was broken and the inter- and intra-molecular hydrogen bond of WS were weaken after gelation. Moreover, there were interactions between the chains of CMC and chains of amylose at higher CMC concentration. On other hand, water connected with COO- groups blocked the interactions between CMC chains and amylose, and the interactions among CMC chains. The amorphous phases of high crystalline MCC were just swollen during paste gelation. It results that heterogeneous microstructure with amylose and MCC domains unevenly distributed was in WS/MCC paste.