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Background The poor regenerative capability and structural complexity make the reconstruction of meniscus particularly challenging in clinic. 3D printing of polymer scaffolds holds the promise of precisely constructing complex tissue architecture, however the resultant scaffolds usually lack of sufficient bioactivity to effectively generate new tissue. Results Herein, 3D printing-based strategy via the cryo-printing technology was employed to fabricate customized polyurethane (PU) porous scaffolds that mimic native meniscus. In order to enhance scaffold bioactivity for human mesenchymal stem cells (hMSCs) culture, scaffold surface modification through the physical absorption of collagen I and fibronectin (FN) were investigated by cell live/dead staining and cell viability assays. The results indicated that coating with fibronectin outperformed coating with collagen I in promoting multiple-aspect stem cell functions, and fibronectin favors long-term culture required for chondrogenesis on scaffolds. In situ chondrogenic differentiation of hMSCs resulted in a time-dependent upregulation of SOX9 and extracellular matrix (ECM) assessed by qRT-PCR analysis, and enhanced deposition of collagen II and aggrecan confirmed by immunostaining and western blot analysis. Gene expression data also revealed 3D porous scaffolds coupled with surface functionalization greatly facilitated chondrogenesis of hMSCs. In addition, the subcutaneous implantation of 3D porous PU scaffolds on SD rats did not induce local inflammation and integrated well with surrounding tissues, suggesting good in vivo biocompatibility. Conclusions Overall, this study presents an approach to fabricate biocompatible meniscus constructs that not only recapitulate the architecture and mechanical property of native meniscus, but also have desired bioactivity for hMSCs culture and cartilage regeneration. The generated 3D meniscus-mimicking scaffolds incorporated with hMSCs offer great promise in tissue engineering strategies for meniscus regeneration. Graphical Abstract

Human gut microbiota played a key role in maintaining and regulating host health. Gut microbiota composition could be altered by daily diet and related nutrients. Diet polysaccharide, an important dietary nutrient, was one kind of biological macromolecules linked by the glycosidic bonds. Galactans were widely used in foods due to their gelling, thickening and stabilizing properties. Recently, effects of different galactans on gut microbiota have attracted much attention. This review described the structural characteristics of 4 kinds of galactans, including porphyran, agarose, carrageenan, and arabinogalactan, along with the effects of different galactans on gut microbiota and production of short-chain fatty acids. The ability of gut microbiota to utilize galactans with different structural characteristics and related degradation mechanism were also summarized. All these four galactans could be used by gut

Background Phosphorylation of proteins is one of the most essential and pleiotropic modifications. Although the importance of phosphorylation in protein function is known, investigation of protein phosphorylation of duck egg yolk (DEY) is still very limited. This study aimed to conduct a detailed phosphoproteomic study of DEY using immobilized metal affinity chromatography and ultra-high liquid chromatography tandem mass spectrometry. Results A total of 253 phosphorylation sites assigned to 66 phosphoproteins were identified in DEY, of which VTG-1, VTG-2, and fibrinogen alpha chain were found to be the highly phosphorylated proteins in DEY. The biological functions of the identified phosphoproteins were illuminated through Gene Ontology analysis, which showed these were mainly involved in binding, catalytic, immune response, and metabolic activities. S-X-E and S-X-S were found the most conserved serine motifs of phosphorylation in DEY. The comparison of DEY phosphoproteins with those of chicken egg yolk (CEY) revealed differences mostly involved molecular functions and biological processes. The comparison also revealed a higher phosphorylation level in DEY proteins. Conclusion The higher phosphorylation level in DEY proteins proposing to enhance duck growth performance and biological activities (e.g. antibacterial and antioxidant ability) to adapt the humid living environment. This article is protected by copyright. All rights reserved.

Regeneration of epithelia is crucial for maintaining the intestinal barrier and homeostasis. Our previous work showed that exopolysaccharides from Lactobacillus plantarum NCU116 (EPS116) regulated the barrier function and homeostasis of the intestine; however, the relevant mechanisms remain obscure. Therefore, we sought to explore the role of EPS116 in promoting intestinal epithelial regeneration. Our data showed that the administration of EPS116 markedly ameliorated inflammatory bowel disease-related phenotypes and promoted the regeneration of crypts in the colon of colitis mice. The results of immunofluorescence and reverse transcription-quantitative polymerase chain reaction experiments indicated that EPS116 strikingly increased the number of intestinal stem cells (ISCs) and the expression of differentiation markers for goblet cells, enterocytes, and enteroendocrine cells in the mouse colon. Intestinal microbiota analysis showed that EPS116 increased microbial populations associated with intestinal regeneration and glycan metabolism. Therefore, the present study revealed a novel model that EPS116 promoted the intestinal homeostasis through modulating the proliferation and differentiation of ISCs and altering the gut microbiota profile.

The release of lysosomal hydrolase into the cytoplasm is accompanied by several systems of apoptosis signal transduction, and the imbalance between cell viability and apoptosis induces tumorigenesis. Tea polysaccharides (TPs) are the main bioactive components in green tea with hopeful anti-tumor efficacy, while their mechanism is still unclear. Here, TPs significantly promoted the death of colon cancer cell line CT26. RNA-seq results showed that the signal pathways up-regulated by TPs included lysosome pathways, apoptosis, the release of mitochondrial pigment c and programmed cell death. Among them, the results of AO-EB and annexin V-FITC/PI double staining indicated that TPs significantly up-regulated apoptosis. In addition, TPs significantly disrupted the function of lysosomes, which would cause mitochondrial damage. Intriguingly, TPs treatment increased the expression of Bak1, cleaved caspase-9 and cleaved caspase-3, but decreased the level of Bcl-2 and mitochondrial membrane potential, which indicated that TPs induced mitochondrial-mediated apoptosis. Moreover, TPs ameliorated the reduced lysosomal numbers by Baf A1 (lysosomal inhibitor). Therefore, our data indicated that TPs targeted lysosomes and induced apoptosis by a lysosomal-mitochondrial pathway mediated caspase cascade, thereby inhibiting the proliferation of CT26 cells. In short, the data would help the development of TPs as potential cancer drug therapeutics.

Sweet tea (Lithocarpus polystachyus Rehd.) has been consumed as herbal tea to prevent and manage diabetes for a long time. Recent studies indicate that sweet tea is rich in a variety of bioactive compounds, especially a class of nonclassical flavonoids, dihydrochalcones. In order to provide a better understanding of sweet tea and its main dihydrochalcones on human health, this review mainly summarizes related literature in the recent ten years, with the potential molecular mechanisms emphatically discussed. Phlorizin, phloretin, and trilobatin, three natural sweeteners, are the main dihydrochalcones in sweet tea. In addition, sweet tea and its dihydrochalcones exhibit plenty of health benefits, such as antioxidant, anti-inflammatory, antimicrobial, cardioprotective, hepatoprotective, antidiabetic, and anticancer effects, which are associated with the regulation of different molecular targets and signaling pathways. Therefore, sweet tea, as a rare natural source of dihydrochalcones, can be processed and developed into nutraceuticals or functional foods, with the potential application in the prevention and management of certain chronic diseases.

Unlike adult mammalian heart, zebrafish heart has a remarkable capacity to regenerate after injury. Previous study has shown Notch signaling activation in the endocardium is essential for regeneration of the myocardium and this activation is mediated by hemodynamic alteration after injury, however, the molecular mechanism has not been fully explored. In this study we demonstrated that blood flow change could be perceived and transmitted in a primary cilia dependent manner to control the hemodynamic responsive klf2 gene expression and subsequent activation of Notch signaling in the endocardium. First we showed that both homologues of human gene KLF2 in zebrafish, klf2a and klf2b, could respond to hemodynamic alteration and both were required for Notch signaling activation and heart regeneration. Further experiments indicated that the upregulation of klf2 gene expression was mediated by endocardial primary cilia. Overall, our findings reveal a novel aspect of mechanical shear stress signal in activating Notch pathway and regulating cardiac regeneration.

Onion (Allium cepa L.) is a common vegetable, widely consumed all over the world. Onion contains diverse phytochemicals, including organosulfur compounds, phenolic compounds, polysaccharides, and saponins. The phenolic and sulfur-containing compounds, including onionin A, cysteine sulfoxides, quercetin, and quercetin glucosides, are the major bioactive constituents of onion. Accumulated studies have revealed that onion and its bioactive compounds possess various health functions, such as antioxidant, antimicrobial, anti-inflammatory, anti-obesity, anti-diabetic, anticancer, cardiovascular protective, neuroprotective, hepatorenal protective, respiratory protective, digestive system protective, reproductive protective, and immunomodulatory properties. Herein, the main bioactive compounds in onion are summarized, followed by intensively discussing its major health functions as well as relevant molecular mechanisms. Moreover, the potential safety concerns about onion contamination and the ways to mitigate these issues are also discussed. We hope that this paper can attract broader attention to onion and its bioactive compounds, which are promising ingredients in the development of functional foods and nutraceuticals for preventing and managing certain chronic diseases.

The seed of Tartary buckwheat (Fagopyrum tataricum) is rich in nutrients and functional ingredients and is recommended as a healthy cereal food. The total proteins of Tartary buckwheat seed (TBS) were extracted and digested; then, the phosphopeptides and glycopeptides were separately enriched and identified by nano liquid chromatography/tandem mass spectrometry. A total of 2613 phosphorylation sites from 1670 phosphoproteins and 404 N-glycosylation sites from 285 N-glycoproteins were identified in TBS. Function and pathway analyses showed that TBS phosphoproteins were significantly enriched in transport, energy metabolism, amino acids biosynthesis/metabolism, and signaling and TBS N-glycoproteins were significantly enriched in modification regulation. The present study reports the first profiles of the phosphoproteome and N-glycoproteome of TBS and provides important post-translational modifications information on the proteins in TBS. The results of this study will aid the understanding of the underlying mechanism of the germination of TBS during cultivation and edible quality changes during storage and processing.

Protein glycosylation is a ubiquitous posttranslational modification that modulates protein properties, thereby influencing bioactivities within a system. Duck egg white (DEW) proteins exhibit diverse biological properties compared with their chicken egg white (CEW) counterparts, which might be related to glycosylation. N-Glycoproteome analysis of DEW was conducted, and a total of 231 N-glycosites from 68 N-glycoproteins were identified. Gene ontology analysis was used to elucidate the biofunctions of DEW N-glycoproteins and compare them with those of CEW, which showed that the differences mostly involved molecular functions and biological processes. The biological functions of DEW N-glycoproteins were illuminated through bioinformatics analysis and comparison with CEW orthologues, which showed different allergenicities and antibacterial abilities. These divergences might be initiated by specific alterations in glycosylation, which can enhance the proteolysis resistance and protein steric hindrance. These results provide new insights for discovering the effects of N-glycosylation on biofunctions during the divergence of homologous proteins.

Morchella (morels) have been used as herbs throughout history, and current scientific research has confirmed their various health benefits. However, as an edible fungus, information regarding the chemical composition of morels is limited. In the present work, the metabolic and proteomic profiles of the cultured morel fruiting body (Morchella importuna) were investigated. The metabolic analysis, which was performed using a GC–MS protocol, identified 7 organic acids, 7 sugar alcohols, 6 monosaccharides, and 3 amino acids. In the shotgun proteome analysis based on the LC–MS/MS, a total of 4047 morel proteins were identified. Based on these omics data, an integrated analysis between the small molecule metabolites and the proteins (enzymes) in the morel fruiting body was performed. Results indicated that the taste of the morel was primarily due to polar metabolites, which were primarily related to respiratory enzymes. Besides, 139 carbohydrate-active enzymes and 10 browning related enzymes were found in morel, and would involve in the texture and color regulation of morel fruiting body. These results provide important information for the understanding of the edible qualities of the morel fruiting body. Moreover, the functional characterization of morel proteins may contribute to technologies for the preservation and processing of morels.

The zebrafish is a species of freshwater fish, popular in aquariums and laboratories. Several advantageous features have facilitated zebrafish to be extensively utilized as a valuable vertebrate model in the lab. It has been well-recognized that natural products possess multiple health benefits for humans. With the increasing demand for natural products in the development of functional foods, nutraceuticals, and natural cosmetics, the zebrafish has emerged as an unprecedented tool for rapidly and economically screening and identifying safe and effective substances from natural products. This review first summarized the key factors for the management of zebrafish in the laboratory, followed by highlighting the current progress on the establishment and applications of zebrafish models in the bioactivity evaluation of natural products. In addition, the zebrafish models used for assessing the potential toxicity or health risks of natural products were involved as well. Overall, this review indicates that zebrafish are promising animal models for the bioactivity and safety evaluation of natural products, and zebrafish models can accelerate the discovery of novel natural products with potential health functions.

Macrolepiota albuminosa (Berk.) Pegler is abundant in active polysaccharides, but little is known about their structures and solution properties. In this study, water-extracted polysaccharides from M. albuminosa (MAWP) were purified into three fractions with structural heterogeneity, which was attributed to the diversity in molecular weight, monosaccharide composition and linkage patterns, further affecting their solution properties. Methylation and NMR analysis revealed MAWP-60p and MAWP-70 were a 3-O-methylated glucomannogalactan and a previously unreported glucomannogalactan, whereas MAWP-80 was elucidated as a branched galactoglucan. Besides, three fractions exhibited random coil conformation in aqueous solution, while MAWP-60p had the highest viscosity due to its highest molecular weight, mean square radius of gyration (Rg) and O-methyl group attached to the backbone. The molecular weight, monosaccharide composition and glycosidic linkages might be the major contributors to the flexibility, molecular size and stereochemistry of mushroom polysaccharide chains.

Cardiac regenerative capacity varies widely among vertebrates. Zebrafish can robustly regenerate injured hearts and are excellent models to study the mechanisms of heart regeneration. Recent studies have shown that enhancers are able to respond to injury and regulate the regeneration process. However, the mechanisms to activate these regeneration-responsive enhancers (RREs) remain poorly understood. Here, we utilized transient and transgenic analysis combined with a larval zebrafish ventricle ablation model to explore the activation and regulation of a representative RRE. lepb-linked enhancer sequence (LEN) directed enhanced green fluorescent protein (EGFP) expression in response to larval ventricle regeneration and such activation was attenuated by hemodynamic force alteration and mechanosensation pathway modulation. Further analysis revealed that Notch signaling influenced the endocardial LEN activity as well as endogenous lepb expression. Altogether, our work has established zebrafish models for rapid characterization of cardiac RREs in vivo and provides novel insights on the regulation of LEN by hemodynamic forces and other signaling pathways during heart regeneration.

Wholegrains have been promoted for human consumption due to their various health benefits. However, different wholegrains vary in nutritional composition and their beneficial impact on health. In this study, we compared the in vitro starch and protein digestibility, as well as dietary fiber content of eight different wholegrains including barley, buckwheat, coix seed, foxtail millet, oat, proso millet, quinoa, and sorghum and their porridges. We found that boiling improved starch digestibility of all grains, and protein digestibility except proso millet and sorghum. Porridges made from oats, quinoa, or buckwheat are considered healthier than others due to their lower glycemic index and glycemic load, higher digestible protein content and amino acid bioaccessibility, and higher dietary fiber content (>12%). This study could provide a comprehensive nutritional composition and digestibility of the eight types of wholegrains and their porridges. Dietary recommendations were also given for different populations based on factor analysis.

Atherosclerotic cardiovascular disease is one of the leading causes of death worldwide. Establishing animal models of atherosclerosis is of great benefit for studying its complicated pathogenesis and screening and evaluating related drugs. Although researchers have generated a variety of models for atherosclerosis study in rabbits, mice and rats, the limitations of these models make it difficult to monitor the development of atherosclerosis, and these models are unsuitable for large scale screening of potential therapeutic targets. On the contrast, zebrafish can fulfill these purposes thanks to their fecundity, rapid development ex utero, embryonic transparency, and conserved lipid metabolism process. Thus, zebrafish have become a popular alternative animal model for atherosclerosis research. In this mini review, we summarize different zebrafish models used to study atherosclerosis, focusing on the latest applications of these models to the dynamic monitoring of atherosclerosis progression, mechanistic study of therapeutic intervention and drug screening, and assessment of the impacts of other risk factors.

The potential effects of tocopherols (100 μM in emulsions) on the physicochemical stability of whey protein isolate (WPI), soy lecithin (SL), or Tween 20 (TW) stabilized flaxseed oil (FO)-in-water emulsions were investigated. During the storage (18 days at 55 ℃), the particle size, microstructure and multiple light scattering results showed WPI-stabilized emulsions exhibited better physical stability when added tocopherols. The hydroperoxides and TBARS concentration in TW-stabilized emulsions were higher than those of SL or WPI, which were suppressed differently by tocopherols. Among homologues, δ-tocopherol was more effective in inhibiting lipid oxidation than α-tocopherol, which was related to the higher interface partitioning. Moreover, the increased interfacial tension indicated tocopherols, especially δ-tocopherol, were adsorbed on the interface and interacted with WPI or SL via hydrophobic or electrostatic interactions determined by isothermal titration calorimetry. Our results suggest tocopherols are more applicable in WPI emulsion systems to achieve steady-state delivery of ALA.

Type 2 diabetes (T2D) is a metabolic disease characterized by hyperglycemia. Intake of dietary fiber is inversely associated with risks of T2D. Here, metabolomics and 16S rRNA gene sequencing were employed to investigate the effects of arabinoxylan on gut microbiota and their metabolites in type 2 diabetic rats. T2D increased the abundance of opportunistic pathogens (such as Desulfovibrio and Klebsiella) and the levels of 12α-hydroxylated bile acids and acylcarnitines (C3) in diabetic rats, which eventually contribute to insulin resistance and hyperglycemia. Supplementation with arabinoxylan promoted the growth of fiber-degrading bacteria to increase short-chain fatty acids (SCFAs), as well as decreased the abundance of opportunistic pathogens. Arabinoxylan treatment also decreased the concentrations of 12α-hydroxylated bile acids, and increased the levels of equol, indolepropionate, and eicosadienoic acid. This study indicated that the beneficial effects of arabinoxylan on T2D may be partially attributed to the modification of gut microbiota and related metabolites.

The aim of this study was to evaluate water-holding capacity, texture, rheological properties, interactions and microstructure of myofibrillar proteins (MP) gel with the addition of fibrin during the heat-induced process. Compared with the control, whiteness, water-holding capacity and hardness of MP gel with the addition of 2% fibrin increased by 9.70%, 79.36%, 323.07%, respectively. Storage modulus and loss modulus values in 2% fibrin addition group were higher than those of the control. The conformation of random coil gradually transformed to β-sheet and β-turn with the increase of fibrin content; nonspecific association and hydrophobic interactions increased from 14.27 g L−1 and 9.00 g L−1 in the control to 28.96 g L−1 and 12.90 g L−1 in the addition of 2% fibrin, respectively, indicating that these interactions played a key role in the improvement of gelling and rheological properties of MP. Scanning electron microscopy revealed that the incorporation of fibrin facilitated the formation of composited gel with a more ordered and homogeneous gel network structure. These results indicated that fibrin enhanced the functionality of MP, which could be beneficial to the development of gel-type meat products.

Clarification of the mechanism of heat-induced gel formation by proteins under natural food systems could provide important references for the regulation of food texture. In the present study, the proteins involved in the early stage (heating at 72 °C for 8 min) of egg-white thermal gel (EWG) formation were studied quantitatively through comparative proteomic analysis. We discovered that the abundance of ovalbumin and ovomucoid increased significantly (p 0.01), whereas that of ovotransferrin, lysozyme, ovomucin (mucin 5B and mucin 6) decreased significantly (p 0.01), in the supernatant of EWG. If the initial interaction of egg white proteins was altered by ultrasonic pretreatment, the abundance of ovomucin and lysozyme in the supernatant of EWG increased, and was accompanied by the change from a solid gel to a fluid gel. Based on these results, we hypothesize that ovomucin has a key role in the formation and regulation of EWG properties.

Plantago (Plantaginaceae) is an herbal plant, which is used in folk medicine, functional food, and dietary supplement products. Recent pharmacological and phytochemical studies have shown that polysaccharides isolated from Plantago have multiple medicinal and nutritional benefits, including improve intestinal health, hypoglycemic effect, immunomodulatory effect, etc. These health and pharmacological benefits are of great interest to the public, academia, and biotechnology industries. This paper provides an overview of recent advances in the physicochemical, structural features, and biological effects of Plantago polysaccharides and highlights the similarities and differences of the polysaccharides from different species and in different parts, including leaves, seeds, and husks. The scientific support for its use as a prebiotic is also addressed. The purpose of this review is to provide background as well as useful and up-to-date information for future research and applications of these polysaccharides.

Konjac flour can quickly swell and dissolve in a short time, leading to extremely high viscosity and easily agglomerative phenomenon. Such characters limit its application as ingredients in brewed foods and there is a high demand to restrict its swelling. Ethanol is usually adopted as a swelling inhibitor but it cannot be applied in the food industry. In this study, two types of milk (full-fat milk and low-fat milk) were selected and the swelling rates of konjac flour in these milk systems were measured. The results showed that the swelling rate of konjac flour significantly decreased. The konjac particles quickly cracked and dissolved in water within 3 min while only swelled in milk systems. To reveal which component in milk played a key role, we further studied the influence of the three components (lactose, whey protein isolate and sodium caseinate) on the swelling limitation of konjac flour. Sodium caseinate was found to be the strongest component than lactose and whey protein isolate in limiting the swelling of konjac flour. Our study revealed that casein was the crucial actor in inhibiting konjac flour swelling in milk and simulated milk system, indicating a new way to better control its swelling behavior in aqueous solution.

Background and objectives Dietary fibers have beneficial effects on human health through the interaction with gut microbiota. Larch wood arabinogalactan (LA-AG) is one kind of complex soluble dietary fibers that may be utilized by human gut microbiota. Methods and results In this study, the LA-AG degradation by gut microbiota were characterized by investigating the change of LA-AG, microbiota composition, and the production of short-chain fatty acids (SCFAs), lactic acid, succinic acid, as well as volatile organic metabolites. During the fermentation, pH decreased continuously, along with the organic acids (especially acetic acid and lactic acid) accumulating. LA-AG was degraded by gut microbiota then some beneficial metabolites were produced. In addition, LA-AG inhibited the proliferation of some gut microbiota (Unclassified_Enterobacteriaceae and Citrobacter) and the accumulation of some metabolites (Sulfide and indole) released by gut microbiota. Conclusion LA-AG was partly fermentable fibers with prebiotic potential for human gut health.

In order to explore the mechanism of nano eggshell calcium (NEC) enhancing the gel properties of surimi. The effects of NEC on the structure, physicochemical, and gel properties of myosin were investigated. The results revealed that the addition of NEC improved gel strength, storage modulus G′, and surface hydrophobicity of myosin. The maximum value was reached when the addition was 1.0 g/100 g. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the addition of NEC has a significant effect on the cross-linking of myosin heavy chain (MHC). FT-IR and fluorescence spectroscopy analysis indicated that NEC could induce the unfolding of myosin and promoted the conformational transition from α-helix to β-sheet. Scanning electron microscopy (SEM) observed that adding a moderate content of NEC (≤1.0 g/100 g) was conducive to the formation of homogeneous and dense three-dimensional network structure of myosin mixed gel. However, the addition of excessive NEC (≥1.25 g/100 g) weakened its structural properties. These results of the present study suggested that NEC has potential to be applied in surimi seafood products, and has potential applications in improving the gel structure and gel properties.

The quantitative comparisons of longissimus thoracis lipidomes among cattle-yak, yak, and cattle were carried out, and differentially abundant lipids (DALs) related to post-mortem physiology and meat quality were examined and discussed. Multivariate statistical analyses showed that the muscle lipidomes of cattle-yak, yak, and cattle were significantly different. Quantitative analysis results showed that there were 108 (cattle-yak vs. cattle), 106 (cattle-yak vs. yak), and 77 (yak vs. cattle) DALs between different muscles. Phospholipids containing long-chain polyunsaturated fatty acids were more abundant in the muscles of plateau cattle (cattle-yak and yak) than plain cattle (cattle), which is a bonus for the nutritional value of plateau cattle meat. Long-chain acylcarnitines accumulated in yak muscle, suggesting different levels of fatty acid β-oxidation and a potential influence on the post-mortem physiology of yak meat. The results show that the muscle lipid profiles of cattle-yak, yak, and cattle were significantly different, and could clearly distinguish by the OPLS-DA analysis. DALs reveal the difference in energy metabolism and lipid nutrition quality between plateau cattle (cattle-yak and yak) muscles and cattle muscle.

The interaction between lipase and quercetin 3-rhamnoside was studied by fluorescence spectroscopy, enzyme kinetics, and molecular dynamics simulation. The results showed that quercetin 3-rhamnoside had a strong quenching effect on the intrinsic fluorescence of lipase. The binding constant decreased with increasing temperature, and the number of binding sites approached 1. Thermodynamic parameters indicated that hydrogen bonding and van der Waals forces are the dominant forces when the interaction occurs. Circular dichroism spectroscopy and infrared spectroscopy proved that the ligand perturbed the structure of lipase. Enzyme kinetics results showed that quercetin 3-rhamnoside inhibited lipase, and the inhibitory effect was dose-dependent. Molecular dynamics simulation further explained the interaction mechanism and inhibitory effect. This study confirmed the inhibitory effect of quercetin 3-rhamnoside on lipase explained their binding mechanism, which will contribute to guiding the development of fat-reducing functional foods.

1,7-bis(4-hydroxy-3-methoxyphenyl)heptane-3,5-dione (tetrahydrocurcumin, THC) is a major bioactive metabolite of curcumin, demonstrating the potential anti-inflammatory, antioxidant and neuroprotective properties, etc. In this study, it was found that Aβ induced decreased cell viability, cell cycle arrest and apoptosis in BV-2 cells, which were ameliorated by THC. In vivo, THC administration rescued learning and memory, and reduced Aβ burden in the hippocampus of APP/PS1 mice. By proteomic analysis of the hippocampus of mice, 157 differentially expressed proteins were identified in APP/PS1 mice treated with THC (comparing with APP/PS1 mice), which also suggested that the effects of THC on the cell cycle and apoptosis were mostly related to the “Ras signaling pathway”, etc. In APP/PS1 mice, the down-regulation of Gab2 and K-Ras, and the up-regulation of caspase-3, TGF-β1 and TNF-ɑ were observed; THC attenuated the abnormal expression of Gab2, K-Ras, caspase-3 and TNF-ɑ, and up-regulated TGF-β1 and Bag1 expression. In BV-2 cells, Aβ induced the down-regulation of Gab2, K-Ras and TGF-β1, and the overexpression of caspase-3, PARP1, cleaved-PARP1 and TNF-ɑ, which were restored by THC. Moreover, THC up-regulated Bag1 expression in Aβ-treated BV-2 cells. The decreased transcriptional expression of Ccnd2 and Cdkn1a were also observed in Aβ-treated BV-2 cells, and THC alleviated the down-regulation of Ccnd2. For the first time, we identified that the action of THC in preventing AD was associated with inhibition of cell cycle arrest and apoptosis of microglia via the Ras/ERK signaling pathway, shedding new light on the role of THC in alleviating the progression of AD.

Senile plaque accumulation and neurofibrillary tangles are primary characteristics of Alzheimer's disease. We aimed to assess the protective functions of naringenin against β-amyloid protein fragment 25-35 (Aβ25-35)-caused nerve damage in differentiated PC12 cells, and study the potential mechanisms. We evaluated cell viability and apoptosis using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) test and flow cytometry, respectively. Moreover, we measured protein kinase B (Akt), glycogen synthase kinase-3β (GSK-3β), and caspase-3 activity via western blotting and RT-PCR. We found that naringenin protected cell against Aβ25-35-caused nerve damage by increasing cell viability, promoting Akt and GSK3β activation, and inhibiting cell apoptosis and caspase-3 activity. However, treatment with the estrogen receptor (ER) antagonist ICI182, 780 or phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002 suppressed the effects of naringenin. Our results suggested that naringenin could effectively suppress Aβ25-35-caused nerve damage in PC12 cells by regulating the ER and PI3K/Akt pathways.

Egg yolk phosvitin, one of the most highly phosphorylated extracellular matrix proteins known in nature, has a strong calcium binding and reducing capacity. Here, we investigated the effects of phosvitin on osteoblast differentiation and osteogenic gene expression in cultured mouse osteoblastic MC3T3-E1 cells by using alkaline phosphatase activity analysis, alizarin red S staining and real-time PCR assay.; Results: Alkaline phosphatase activity and alizarin red S staining analyses demonstrated no significant difference between differentiating MC3T3-E1 cells cultured in the presence of phosvitin and those cultured in the presence of ascorbic acid after 21 days of differentiation. Our real-time PCR assay also indicated the two groups were similar in the expression of the osteogenic gene markers, collagen type I, osteocalcin, runt-related transcription factor 2, and bone morphogenetic protein-2.; Conclusion: Our findings indicate that phosvitin plays a similar role to that of ascorbic acid in osteoblast differentiation and mineralisation. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Egg yolk contains multifunctional glycoproteins, which modulate embryogenesis and indicate species-specific adaptation. Here, we conducted an N-glycoproteomic study of duck egg yolk (DEY) and identified 359 N-glycosites corresponding to 113 glycoproteins. The comparison of DEY N-glycoproteins with those of chicken egg yolk (CEY) revealed a higher N-glycosylation level in DEY proteins, suggesting a biological adaption to meet the requirements for embryo development by modulating protein structure and binding ability, including immune response and ligand transport. This variation may be correlated to the strong resistance to the influenza virus disease of duck. A large set of corresponding glycoproteins between DEY and CEY were detected with similar molecular functions, which suggested a relatively lower evolution speed of avian egg yolk proteins than that of egg white proteins.

In this study, immobilized metal affinity chromatography and ultra-high liquid chromatography tandem mass spectrometry were used to establish detailed phosphoproteomes of fertilized egg, and further quantitative compared those from Tibetan chicken and lowland chicken. A total of 138 phosphosites from 42 phosphoproteins were identified. Specifically, 103 phosphosites representing 41 phosphoproteins were detected in Tibetan chicken egg, and 125 sites in 40 phosphoproteins were found in lowland chicken egg; 47 phosphosites in 26 phosphoproteins were found in fertilized egg white, while 109 phosphosites in 24 phosphoproteins were identified in fertilized egg yolk. Thirty-one phosphoproteins carrying 83 phosphosites (12 phosphosites from 10 phosphoproteins in egg white and 52 sites from 17 proteins in egg yolk) showed significantly different phosphorylation between Tibetan and lowland chicken eggs, which were mainly involved in binding, catalysis, allergenic and cryoprotectant activities. Besides, several amino acid polymorphisms in ovalbumin and vittelligion-2 were detected, which might alter 3D structures and biofunctions. Our data recorded the egg phosphoprotein abundances and biological activities of two chicken species. These results provide a solid foundation to better understand the egg phosphoprotein characteristics, especially to explore the impacts of cross-species alteration of phosphorylation status on the structure, function and nutritional properties.

De novo transcriptome assembly and shotgun proteome analysis of Dictyophora indusiata fruiting bodies were performed. A total of 19,704 unigenes were sequenced, and 4380 proteins were identified. Annotation and functional analysis of the identified proteins were significantly enriched in small molecule synthetic and metabolic processes, protein modification regulation (phosphorylation and ubiquitination), and vesicle transport. Furthermore, quantitative developmental transcriptome analysis was performed between the peach-shaped and mature fruiting bodies, and the results revealed that the metabolism and transport activities were upregulated in the mature stage, while protein translation was downregulated; this regulation is likely the main reason for the significant changes in the nutrients of fruiting bodies. Furthermore, the cell wall stress-dependent MAPK sub-pathway was activated in the mature stage, and fungal cell wall degradation-related genes were upregulated, which could promote reconstruction of the cell wall and might play a key role in the morphological development of D. indusiata fruiting bodies.

Avian egg yolk provides essential nutrients and physiological support for the developing embryo. To reveal the dynamics of yolk proteins during the entire period of incubation, we analyzed the alterations of egg yolk plasma proteins at different times during incubation (day 0, 7, 14, and 18). Day 14 (D14) of incubation was considered as the key point on the alteration of egg yolk proteins according to the SDS-PAGE analysis. The two-dimensional electrophoresis-based comparative proteomic analysis was conducted, and 26 spots representing 13 proteins were detected with significant changes in abundance. An accelerating transfer of ovalbumin from egg white into egg yolk was observed at D14 in fertilized eggs but not detected in unfertilized eggs, indicating an unrevealed absorbing pathway for the egg proteins/peptides particularly in fertilized eggs. Meanwhile, the abundance of yolk riboflavin-binding protein constantly decreased after D14, which might meet the need for essential riboflavin during embryo development. PRACTICAL APPLICATIONS: These findings provide fundamental insight into the effects of incubation on egg yolk plasma protein alterations during the entire embryonic development for a better understanding of plasma protein biological functions, especially in nutrient transportations and the formation of embryonic organs.

Theaflavin (TF), which is the key pigment in black tea, is a health-promoting food component with beneficial effects on humans. However, the interactions by which these effects are transferred and exerted into protein-rich foods are unclear. Here, egg ovalbumin (OVA) was selected as a representative dietary protein to ascertain their binding mechanism. Steady-state, time-resolved fluorescence and isothermal titration calorimetric results showed that TF can interact well with OVA with an affinity magnitude of 104. The noncovalent binding was mainly driven by hydrophobic interaction and hydrogen bonds. Structural analysis displayed that the TF binding pocket significantly overlapped with one of the surrounding specific IgE-binding epitopes, thereby causing a subtle structural adjustment on the secondary conformation of OVA. The biological complexation model that was delineated here will help understand how black tea dyes egg white in tea egg products and for the development of protein-rich carriers in functional foods.

Cysteine and glycine-rich protein 3 (CSRP3) is a striated muscle-specific cytoskeleton protein which participates in cardiac stretch sensing. Mutations in CSRP3 gene cause cardiomyopathies and deregulation of CSRP3 has been found in patients with heart failure and several skeletal muscle diseases. However, the mechanism underneath these disorders still remains poorly understood. Here we generated the first csrp3 knockout zebrafish. csrp3−/− embryos showed no gross morphological defects but csrp3 deficient skeletal muscle fibers were prone to lesions upon prolonged stretching force. Further studies revealed csrp3 cooperatively interacted with ilk to maintain skeletal muscle mechanical stability and regulated tcap activation. Thus, our work has established a zebrafish model to investigate the function of csrp3 gene, and provides novel insights towards how csrp3 defects may lead to skeletal myopathies by a mechanistic link between Csrp3 and force stimuli.

The whole seed of tartary buckwheat (Fagopyrum tataricum) is considered as a healthy and functional food, which is rich in kinds of flavonoids and with potential antioxidant effect. An in-depth analysis of tartary buckwheat seed (TBS) proteome was performed using a shotgun proteomics strategy. Total protein of TBS was extracted and digested, then the peptides were separated by offline two-dimensional liquid chromatography and identified by tandem mass spectrometry. Total of 3,363 high-confidence proteins were identified from 13,730 matched peptides, in which, 2,499 proteins were annotated by the Gene Ontology (GO) analysis with 1,720 involved in "biological process," 2,241 in "molecular function," and 693 in "cellular components." Based on the GO functional enrichment and Kyoto Encyclopedia of Genes and Genomes pathway enrichment results, buckwheat seed proteins were mostly enriched in metabolism of nucleic acid, respiration and energy metabolism, as well as synthesis and metabolism of protein. PRACTICAL APPLICATIONS: This study characterized the tartary buckwheat seed proteome on a scale of 3,000+ proteins and provide important information and clues for future research, especially in the mechanism of seed germination, nutrient composition changes, and metabolite production seed germination and material metabolism.

The use of probiotics poses challenges represented in loss of viability during harsh conditions, hence this study aims at exploring a simple green method to increase the survival of the encapsulated Lactobacillus salivarius during gastrointestinal conditions, thermal treatment, and storage. Single and dual biopolymer beads which consisted of whey protein, alginate, carboxymethyl cellulose, and chitosan were fabricated and then characterized by fourier transform infrared (FTIR) spectra. The morphology characteristics were investigated using scanning electron microscopy (SEM) and fluorescence microscopy. The encapsulation efficiency was reached up to 96% in all treatments. A biopolymer coated by a layer of carboxymethyl cellulose or chitosan showed a great protection for probiotics upon exposure to gastrointestinal fluid. Also, a high protection was observed after thermal treatment and storage when compared with free cells. Approximately the release of cells in colon solution was noted after 18 h for all treatments.

In the present study, a tandem mass tag-labeled quantitative proteomic analysis was performed to compare the differences in protein profiles between Tibetan pig (TP) tenderloin and Yorkshire pig (YP) tenderloin. A total of 171 proteins were identified as differentially abundant proteins (DAPs) from 1448 quantified proteins. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) database pathway analyses revealed that upregulated DAPs in the TP tenderloin were mainly involved in energy production, muscle contraction, immunity and defense, while downregulated DAPs were mainly involved in glutathione metabolism. These DAPs and the pathways they participate in were related to meat characteristics. TP tenderloin has a deeper color and higher pH, but lower water content and cooking loss. This study provides insights into the mechanism underlying the differences in protein profiles between TP and YP tenderloins. The findings can provide a better understanding of the formation mechanism for Tibetan pork edible quality.

These studies were conducted to determine the most effective methods for increasing shoot elongation during the initial proliferation stage of micropropagation in two dwarfing apple, Malus ×domestica (Borkh.), rootstock cultivars. Several experiments were conducted to compare explant collection date, exposure to chilling (5 ± 1 °C) temperatures, and varying concentrations of plant growth regulators in Murashige and Skoog (MS) media. Microshoot growth of ‘Geneva 41’ (‘G.41’) was very low and unaffected by chilling duration from 0 to 8 weeks or by gibberellic acid (GA3) concentration from 0 to 1.0 mg·L−1, but was improved by an additional subculture which increased shoot length from 1 to 15 mm. In ‘Geneva 30’ (‘G.30’), shoot elongation was most affected by date, chilling explants, and by optimizing cytokinin concentration and type. Explant collection date in April increased shoot growth compared with August or November. Microshoot growth of ‘G.30’ was increased by chilling nodal explants for 4 and 6 weeks when explants were collected in August and November, but not in April. Eight weeks chilling was detrimental for explants collected in April, and generally had little or no effect with August and November. The cytokinin 6-benzylaminopurine (BA) increased shoot number to a greater extent than thidiazuron (TDZ) or zeatin (ZT), and was also more effective for increasing shoot elongation with concentrations of 0 to 2.0 mg·L−1. In ‘G.30’, GA3 increased shoot growth at the optimum concentration of BA, but not with lower concentrations. ‘G.30’ microshoots were fewer and shorter with 24-epi-brassinolide (EBR) at concentrations of 0.1 and 1.0 mg·L−1. Chemical names: N-phenyl-N’-(1,2,3-thiadiazol-5-yl)urea (TDZ), 6-(4-hydroxy-3-methylbut-2-enylamino)purine (ZT).

Fresh chicken egg whites can be easily categorized into two types, including the colloid-like egg white and the solution-like egg white, which are called thick egg white (TKEW) and thin egg white (TNEW), respectively. In the present study, the heat-induced gel properties of TKEW and TNEW were systematically analyzed and compared. The results showed that TKEW (72.51 °C) had higher temperature of heat denaturation than TNEW (67.01 °C). The texture profile analysis demonstrated that the TKEW gel appeared soft and tough (lower hardness and higher cohesiveness), while the TNEW gel appeared hard and brittle (higher hardness and springiness, but lower cohesiveness). SEM images showed that the TKEW gel exhibited a “mesh structure” with “mountain-like” protrusions and micropores, while the TNEW gel exhibited a “block structure” with a dense section and “rope-head” protrusions. Quantitative proteomic analysis revealed that a higher content of ovotransferrin in TNEW might be the main reason for its lower temperature of thermal denaturation, and a higher content of ovomucin in TKEW might regulate the microstructure of the gel during heating and cause the differences in the gel texture properties. Overall, this study provides insights into the mechanism for the differences of heat-induced gel properties between TKEW and TNEW from the comprehensive “composition-structure-function” perspective, and can give guidance for the regulation of egg white gel properties during its application.

As two major proteins of egg white, the interaction between lysozyme (LYZ) and ovalbumin (OVA) has been studied. In the present research, the mechanism and kinetics of the interaction between LYZ and OVA were studied using several methods, including cyclic voltammetry (CV), dual polarization interferometry (DPI) and surface plasmon resonance (SPR), and the effect of their interaction on the activity of LYZ was investigated using a bacteriolytic activity assay. Cyclic voltammograms showed that there was an interaction between these two proteins. DPI results indicated that the interaction is a type of chimeric mode, and the KA and KD of the interaction were 498 and 2.01 × 10−3 mol/L, respectively, calculated from the fitting of the surface plasmon resonance curve. The bacteriolytic activity assay showed that the interaction between these two proteins reduced the bacteriolytic activity of LYZ.

As the most abundant protein in chicken eggs, ovalbumin plays an important role in the processing of high value-added poultry products. The present study investigated the effects of hydroxyl radical-induced early stage oxidation on the physicochemical and interfacial properties of chicken egg white ovalbumin. Protein carbonyl content of ovalbumin increased (from 0.78 to 1.13 nmol/mg) with the oxidation time (0–5 h), while free sulfhydryl content (from 0.43 to 0.09 nmol/mg) and free amino group content (from 0.49 to 0.43 nmol/mg) decreased. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that the exposure of ovalbumin to hydroxyl radicals caused self-cross-linking and resulted in the formation of dimers and trimers. Accompanied by these changes, the surface hydrophobicity of ovalbumin was enhanced about 1.5-fold with the deepening of oxidation, and the value of zeta potential became more negative (from –7.15 to –20.51 mv). About 2 h of moderate oxidation improved the foaming and emulsifying properties of ovalbumin (1.2-fold to 1.8-fold), while excessive oxidation (3 h) decreased these interface properties. Hydroxyl radical-induced oxidation changed the surface chemical groups and structures of ovalbumin, thereby affecting the surface properties. The foaming and emulsifying properties of ovalbumin could be improved by oxidation, increasing the application possibilities of ovalbumin in the food interface system.

Parrotia subaequalis is an endangered palaeoendemic tree from disjunct montane sites in eastern China. Due to the lack of effective genomic resources, the genetic diversity and population structure of this endangered species are not clearly understood. In this study, we conducted paired-end shotgun sequencing (2 × 125 bp) of genomic DNA for two individuals of P. subaequalis on the Illumina HiSeq platform. Based on the resulting sequences, we have successfully assembled the complete chloroplast genome of P. subaequalis, as well as identified the polymorphic chloroplast microsatellites (cpSSRs), nuclear microsatellites (nSSRs) and mutational hotspots of chloroplast. Ten polymorphic cpSSR loci and 12 polymorphic nSSR loci were used to genotype 96 individuals of P. subaequalis from six populations to estimate genetic diversity and population structure. Our results revealed that P. subaequalis exhibited abundant genetic diversity (e.g., cpSSRs: Hcp = 0.862; nSSRs: HT = 0.559) and high genetic differentiation (e.g., cpSSRs: RST = 0.652; nSSRs: RST = 0.331), and characterized by a low pollen-to-seed migration ratio (r ≈ 1.78). These genetic patterns are attributable to its long evolutionary histories and low levels of contemporary inter-population gene flow by pollen and seed. In addition, lack of isolation-by-distance pattern and strong population genetic structuring in both marker systems, suggests that long-term isolation and/or habitat fragmentation as well as genetic drift may have also contributed to the geographic differentiation of P. subaequalis. Therefore, long-term habitat protection is the most important methods to prevent further loss of genetic variation and a decrease in effective population size. Furthermore, both cpSSRs and nSSRs revealed that P. subaequalis populations consisted of three genetic clusters, which should be considered as separated conservation units.

Background Almond (Prunus dulcis) is a popular tree nut worldwide and offers many benefits to human health. However, the importance of almond kernel proteins in the nutrition and function in human health requires further evaluation. The present study presents a systematic evaluation of the proteins in the almond kernel using proteomic analysis. Results The nutrient and amino acid content in almond kernels from Xinjiang is similar to that of American varieties; however, Xinjiang varieties have a higher protein content. Two-dimensional electrophoresis analysis demonstrated a wide distribution of molecular weights and isoelectric points of almond kernel proteins. A total of 434 proteins were identified by LC-MS/MS, and most were proteins that were experimentally confirmed for the first time. Gene ontology (GO) analysis of the 434 proteins indicated that proteins involved in primary biological processes including metabolic processes (67.5%), cellular processes (54.1%), and single-organism processes (43.4%), the main molecular function of almond kernel proteins are in catalytic activity (48.0%), binding (45.4%) and structural molecule activity (11.9%), and proteins are primarily distributed in cell (59.9%), organelle (44.9%), and membrane (22.8%). Conclusion Almond kernel is a source of a wide variety of proteins. This study provides important information contributing to the screening and identification of almond proteins, the understanding of almond protein function, and the development of almond protein products. © 2015 Society of Chemical Industry.

Background Hen egg white ovomacroglobulin (OVM) possesses a variety of bioactivities and could potentially be used as a pharmaceutical agent. It has been reported that OVM is involved in wound healing and cancer pathological processes, and previous results suggest that OVM plays a potential role in cell proliferation and migration; however, this has not yet been proven. In the present study, the effects of OVM on fibroblast proliferation and migration were evaluated. Results Results of cell counting, cell viability, and cell cycle indicated that proliferation of fibroblasts was not altered by OVM treatment. However, scratch assays showed that OVM could promote the migration of 3 T6 mouse embryonic fibroblasts and human skin fibroblasts (HSF). Also, the adhesion of HSF to the collagen matrix was also enhanced by OVM treatment. RT-qPCR and western blot analysis showed that β1 -integrin, β-tubulin, and β-actin were up-regulated while E-cadherin was down-regulated in OVM-treated HSF cells. The effect of OVM was silenced after forming a complex with trypsin, suggesting that the protease inhibitory ability of OVM is important for its effect on cell migration. Conclusion These results suggested that promotion of OVM on cell migration was achieved by enhancing cell adhesion to extracellular matrix, reducing intercellular aggregation, and strengthening cytoskeleton. The finding of the promotion effect of OVM on cell migration is important for understanding its role in wound healing and cancer pathological processes. © 2015 Society of Chemical Industry.

Two new alkaloids, plantadeprate A (1) and 1'-(4″-hydroxybutyl)plantagoguanidinic acid (2), along with three known compounds, were isolated from the seeds of Plantago depressa. Their structures were elucidated by physical data analyses including NMR, MS, and electronic circular dichroism (ECD) methods. Plantadeprate A (1), a monoterpene zwitterionic guanidium, possesses a unique 5/5/6-tricyclic ring system. Its absolute configuration was determined by X-ray crystallography and computational methods. Compound 1, plumbagine D (3), and plantagoguanidinic acid (4) exhibited potential antihyperglycemic properties attributed to suppression of hepatic gluconeogenesis with inhibitory rates of 8.2%, 18.5%, and 12.5% at 40 μM, respectively.

The influence of red and blue light wavelengths was tested to improve the initial in vitro multiplication of apple (Malus × domestica) rootstock cultivars Budagovsky 9 (B.9), Geneva 30 (G.30), and Geneva 41 (G.41). Single-node segments were established in semisolid Murashige and Skoog media and then transferred to proliferation media and cultured 40 days under white, red, or blue light irradiance. In a second experiment, G.30 was cultured under red, blue, or white light with and without gibberellic acid (GA3). The three rootstocks responded similarly under white light in terms of shoot number, length of the longest shoot, and the number of elongated shoots. Red light increased the number of shoots, length of the longest shoot, and the number of elongated shoots of B.9 and G.30 when compared with white or blue light. Red light increased the number of elongated B.9 and G.30 shoots to five per explant compared with one per explant under white light. In contrast, shoot growth of G.41 showed no difference under the three light quality treatments, and the number of elongated shoots per explant was less than one. When compared with an absence of GA3, a concentration of GA3 at 0.5 mg·L−1 promoted in vitro shoot growth of G.30 under red and blue light.

A novel aspartic protease gene (RmproA) was cloned from the thermophilic fungus Rhizomucor miehei CAU432 and expressed in Pichia pastoris. The RmproA was successfully expressed in P. pastoris as an active extracellular protease. High protease activity of 3480.4 U/mL was obtained by high cell-density fermentation. The protease was purified by the two step protocols to homogeneity. The molecular mass of the RmproA was estimated to be 52.4 kDa by SDS-PAGE and 50.6 kDa by gel filtration. The purified enzyme was optimally active at pH 5.5 and 55 °C, respectively. The enzyme exhibited a broad range of substrate specificity. RmproA-treated pork muscle showed lower shear force than papain-treated sample at a relative low concentration, suggesting its effectiveness on meat tenderization. Moreover, turtle hydrolysis by RmproA resulted in a large amount of small peptides, which exhibited high ACE-inhibitory activity. Thus, RmproA may be a potential candidate for several industrial applications.