Effect of succinic acid deamidation-induced modification on wheat gluten

The effect of succinic acid deamidation-induced modification on wheat gluten was investigated in the present study. The changes of surface hydrophobicity, functional properties, secondary structure, and sensibility of proteolysis of modified samples were determined. The solubility of deamidated proteins increased in the isoelectric region of untreated wheat gluten. The isoelectric point of succinic acid deamidated wheat gluten was shifted to a basic pH and existed in the broad pH regions. Foaming property and molecular flexibility of wheat gluten were improved after the modification. The hydrolysis degree of the hydrolysates in proteolysis with flavorzyme and pancreatin increased after succinic acid deamidation. Moreover, succinic acid deamidation-induced modification resulted in little change in molecular weight and secondary structure of the protein. Thus, succinic acid could facilitate unfolding protein conformation. In addition, it could improve protein-water interactions, surface properties, and sensibility of the proteolysis of the deamidated wheat gluten.     

Effect of succinic acid deamidation-induced modification on wheat gluten

The effect of succinic acid deamidation-induced modification on wheat gluten was investigated in the present study. The changes of surface hydrophobicity, functional properties, secondary structure, and sensibility of proteolysis of modified samples were determined. The solubility of deamidated proteins increased in the isoelectric region of untreated wheat gluten. The isoelectric point of succinic acid deamidated wheat gluten was shifted to a basic pH and existed in the broad pH regions. Foaming property and molecular flexibility of wheat gluten were improved after the modification. The hydrolysis degree of the hydrolysates in proteolysis with flavorzyme and pancreatin increased after succinic acid deamidation. Moreover, succinic acid deamidation-induced modification resulted in little change in molecular weight and secondary structure of the protein. Thus, succinic acid could facilitate unfolding protein conformation. In addition, it could improve protein-water interactions, surface properties, and sensibility of the proteolysis of the deamidated wheat gluten.     

High-Molecular-Weight Glutenin Subunits: Genetics,Structures, and Relation to End Use Qualities

High-molecular-weight glutenin subunits (HMW-GSs) are storage proteins present in the starchy endosperm cells of wheat grain. Encoding the synthesis of HMW-GS, the Glu-1 loci located on the long arms of group 1 chromosomes of the hexaploid wheat (1A, 1B, and 1D) present multiple allelism. In hexaploid wheat cultivars, almost all of them express 3 to 5 HMW-GSs and the 1Ay gene is always silent. Though HMW-GSs are the minor components in gluten, they are crucial for dough properties, and certain HMW-GSs make more positive contributions than others. The HMW-GS acts as a “chain extender” and provides a disulfide-bonded backbone in gluten network. Hydrogen bonds mediated by glutamine side chains are also crucial for stabilizing the gluten structure. In most cases, HMW-GSs with additional or less cysteines are related to the formation of relatively more or less interchain disulfide bonds and HMW-GSs also affect the gluten secondary structures, which in turn impact the end use qualities of dough.     

Differential protein expression of Caco-2 cells treated with selenium nanoparticles compared with sodium selenite and selenomethionine

The study was designed to determine the differential protein expression of Caco-2 cells treated with different forms of selenium including sodium selenite, selenomethionine (Se-Met), and selenium nanoparticles (nano-Se). Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS) were used to identify the differentially expressed proteins. The results indicated that seven protein spots, ubiquitin-conjugating enzyme E2 (E2), glutathione synthetases (GS), triosephosphate isomerase (TSP), T-complex protein 1 subunit zeta (TCPZ), lamin-B1, heterogeneous nuclear ribonucleoprotein F (hnRNP F), and superoxide dismutase [Cu-Zn] (Cu, Zn-SOD) were significantly different among all the groups. According to the order of control, sodium selenite, Se-Met, and Nano-Se, the expression levels of two proteins (E2 and GS) increased and the other differential proteins were reverse. Except for E2, there were no significant differences in other protein expressions between the groups treated with nano-Se and Se-Met.     

A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro

Background: Glioblastoma multiforme (GBM) is the most frequent and aggressive primary brain tumor, and macrophages account for 30–40% of its composition. Most of these macrophages derive from bone marrow monocytes playing a crucial role in tumor progression. Unraveling the mechanisms of macrophages-GBM crosstalk in an appropriate model will contribute to the development of specific and more successful therapies. We investigated the interaction of U87MG human GBM cells with primary human CD14⁺ monocytes or the THP-1 cell line with the aim of establishing a physiologically relevant heterotypic culture model. Methods: primary monocytes and THP-1 cells were cultured in the presence of U87MG conditioned media or co-cultured together with previously formed GBM spheroids. Monocyte differentiation was determined by flow cytometry. Results: primary monocytes differentiate to M2 macrophages when incubated with U87MG conditioned media in 2-dimensional culture, as determined by the increased percentage of CD14⁺CD206⁺ and CD64⁺CD206⁺ populations in CD11b⁺ cells. Moreover, the mitochondrial protein p32/gC1qR is expressed in monocytes exposed to U87MG conditioned media. When primary CD14⁺ monocytes or THP-1 cells are added to previously formed GBM spheroids, both invade and establish within them. However, only primary monocytes differentiate and acquire a clear M2 phenotype characterized by the upregulation of CD206, CD163, and MERTK surface markers on the CD11b⁺CD14⁺ population and induce alterations in the sphericity of the cell cultures. Conclusion: our results present a new physiologically relevant model to study GBM/macrophage interactions in a human setting and suggest that both soluble GBM factors, as well as cell-contact dependent signals, are strong inducers of anti-inflammatory macrophages within the tumor niche.     

Effects of 3‐D microwell culture on initial fate specification in human embryonic stem cells

Several studies have demonstrated that three‐dimensional (3‐D) culture systems influence human embryonic stem cell (hESC) phenotypes and fate choices. However, the effect that these microenvironmental changes have on signaling pathways governing hESC behaviors is not well understood. Here, a 3‐D microwell array was used to investigate differences in activation of developmental pathways between 2‐D and 3‐D cultures of both undifferentiated hESCs and hESCs undergoing initial differentiation in embryoid bodies (EBs). An increased induction into mesoderm and endoderm and differences in expression of genes from multiple signaling pathways that regulate development, including Wnt/β‐catenin, TGF‐β superfamily, Notch, and FGF during EB‐mediated differentiation were observed in 3‐D microwells as compared with the 2‐D substrates. In undifferentiated hESCs, differences in epithelial‐mesenchymal transition phenotypes and the TGFβ/BMP pathway between cultures in 3‐D and 2‐D were also observed. These results illustrate that 3‐D culture influences multiple pathways that may regulate the differentiation trajectories of hESCs. © 2014 American Institute of Chemical Engineers AIChE J, 60: 1225–1235, 2014     

Food uses of wheat gluten

New separation methods for wheat gluten have increased production and made possible the use of mass wheat as a raw material. This widens the use of gluten in the fields where functional as well as baking properties are important. Solubility, swelling, viscosity and nutritional aspects of gluten are reviewed. The current applications of wheat gluten are mainly in baked products, breakfast foods, and meat analogs. Many chemical modifications of gluten have been developed. Modifications are not in use, but the increased gluten production will bring them into use.     

AGE-RAGE Axis Stimulates Oxidized LDL Uptake into Macrophages through Cyclin-Dependent Kinase 5-CD36 Pathway via Oxidative Stress Generation

Advanced glycation end products (AGEs) are localized in macrophage-derived foam cells within atherosclerotic lesions, which could be associated with the increased risk of atherosclerotic cardiovascular disease under diabetic conditions. Although foam cell formation of macrophages has been shown to be enhanced by AGEs, the underlying molecular mechanism remains unclear. Since cyclin-dependent kinase 5 (Cdk5) is reported to modulate inflammatory responses in macrophages, we investigated whether Cdk5 could be involved in AGE-induced CD36 gene expression and foam cell formation of macrophages. AGEs significantly increased Dil-oxidized low-density lipoprotein (ox-LDL) uptake, and Cdk5 and CD36 gene expression in U937 human macrophages, all of which were inhibited by DNA aptamer raised against RAGE (RAGE-aptamer). Cdk5 and CD36 gene expression levels were correlated with each other. An antioxidant, N-acetyl-l-cysteine, mimicked the effects of RAGE-aptamer on AGE-exposed U937 cells. A selective inhibitor of Cdk5, (R)-DRF053, attenuated the AGE-induced Dil-ox-LDL uptake and CD36 gene expression, whereas anti-CD36 antibody inhibited the Dil-ox-LDL uptake but not Cdk5 gene expression. The present study suggests that AGEs may stimulate ox-LDL uptake into macrophages through the Cdk5–CD36 pathway via RAGE-mediated oxidative stress.     

Morin,a Flavonoid from Moraceae,Induces Apoptosis by Induction of BAD Protein in Human Leukemic Cells

Evidence suggests that phytochemicals can safely modulate cancer cell biology and induce apoptosis. Here, we investigated the anti-cancer activity of morin, a flavone originally isolated from members of the Moraceae family in human leukemic cells, focusing on apoptosis. An anti-cancer effect of morin was screened with several human leukemic cell lines. U937 cells were most sensitive to morin, where it induced caspase-dependent apoptosis in a dose-dependent manner. It also induced loss of MMP (ΔΨ_m) along with cytochrome c release, down-regulated Bcl-2 protein, and up-regulated BAX proteins. The apoptotic activity of morin was significantly attenuated by Bcl-2 augmentation. In conclusion, morin induced caspase-dependent apoptosis through an intrinsic pathway by upregulating BAD proteins. In addition, Bcl-2 protein expression is also important in morin-induced apoptosis of U937 cells. This study provides evidence that morin might have anticancer properties in human leukemic cells.     

Use of lignin strengthened with modified wheat gluten in biodegradable composites

In this study, composites from alkali lignin and wheat gluten, modified with different percentages of sodium silicate, were prepared and characterized. Moreover, the addition of silica to the aforementioned composites was studied with the aim of improving the thermal and mechanical properties. The effect of wheat gluten percent and the extent of its modification on the blends properties were investigated via diametric tensile strength, thermomechanical analysis (TMA), scanning electron microscope (SEM), thickness swelling and thermogravimetric analysis (TGA). The results showed significant improvement in the diametric tensile strength, thickness swelling, uniformity in the fracture surface, and the shift of glass transition temperature (T_g) toward higher values with increasing wheat gluten percent and its modification extent. These results reflect the enhancement of interaction between alkali lignin and wheat gluten. Alkali lignin/wheat gluten blends filled with silica possessed distinguishable characteristics and improved diametric tensile strength, low thermal expansion, and high T_g. Interestingly, TMA results showed that high dimensional stability against heating (thermal expansion percent) could be obtained using 60% wheat gluten modified with 15% sodium silicate and filled with 10% silica. This sample showed the highest T_g and the lowest thickness swelling in addition to smooth, uniform, and glossy surface as shown from the SEM images and TMA charts. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Chemical lipophilization of soy protein isolates and wheat gluten

Chemical modification of soy protein isolate and wheat gluten can be used to investigate the influence of the chemical composition and the structure of the proteins on the final product properties and to adjust these properties to meet specific demands. Proteins were lipophilized by adding different levels of lauroyl chloride to aqueous protein dispersions. At optimum reaction conditions, incorporation rates of lauroyl chains to the protein were 400 and 300 μmol/g proteins for soy protein isolate and wheat gluten, respectively. The study showed that it was possible to lipophilize proteins in an aqueous medium, even with the protein significantly high in concentration.     

Novel cinnamyl hydroxyamides and 2-aminoanilides as histone deacetylase inhibitors: apoptotic induction and cytodifferentiation activity

Four novel series of cinnamyl-containing histone deacetylase (HDAC) inhibitors 1-4 are described, containing hydroxamate (1 and 3) or 2-aminoanilide (2 and 4) derivatives. When screened against class?I (maize HD1-B and human HDAC1) and class?II (maize HD1-A and human HDAC4) HDACs, most hydroxamates and 2-aminoanilides displayed potent and selective inhibition toward class?I enzymes. Immunoblotting analyses performed in U937 leukemia cells generally revealed high acetyl-H3 and low acetyl-α-tubulin levels. Exceptions are compounds 3?f-i, 3?m-o, and 4?k, which showed higher tubulin acetylation than SAHA. In U937 cells, cell-cycle blockade in either the G?/M or G?/S phase was observed with 1-4. Five hydroxamates (compounds 1?h-l) effected a two- to greater than threefold greater percent apoptosis than SAHA, and in the CD11c cytodifferentiation test some 2-aminoanilides belonging to both series 2 and 4 were more active than MS-275. The highest-scoring derivatives in terms of apoptosis (1?k, 1?l) or cytodifferentiation (2?c, 4?n) also showed antiproliferative activity in U937 cells, thus representing valuable tools for study in other cancer contexts.     

Wheat gluten as a protein ingredient

Wheat gluten is a unique cereal protein due to its property of high elasticity. This elasticity is only present in hydrated gluten, and it is destroyed by heating. The property permits the formation of gas cells in flour dough, and is the essential cause of the texture of our daily bread. Wheat gluten is produced industrially from flour rather than from wheat, although some progress has been made recently in the latter direct process. All the wet separation methods basically involve mixing flour and water and allowing the starch granules to leave the gluten matrix in a mixed aqueous phase. The various methods of wet separation give rise to essentially similar glutens, but the type and severity of the drying process can lead to major changes in degree of elasticity of the reconstituted gluten. In spite of a rather low level of lysine, methionine and tryptophane, the nutritive value of wheat gluten can be high, particularly when mixed with other proteins which supplement these particular amino acids. Although gluten can be used alose, as in some canned vegetarian dishes, generally it is used as an additive to cereal and meat or fish foods. In cereal applications the base material is usually flour or maize, rice or wheat. The market for gluten in the Western World is of the order of 90,000 tons per year, and it is expected that the industrial use of wheat gluten will continue to expand with our technical ability to make use of its unique properties.     

Dynamic mechanical analysis of the multiple glass transitions of plasticized wheat gluten biopolymer

Glass transition of thermo‐molded biomaterials made from wheat gluten and its main protein classes is studied by dynamic mechanical analysis (DMA). The materials are plasticized with variable contents of glycerol (30–40 wt %) and water (0–20 wt %). For all materials, three successive relaxation phases are systematically detected. Their positions shift to lower temperature as the plasticizer content of materials increases. Composition in gluten, glycerol and water of each relaxation phase is estimated using the Couchman‐Karasz model. Irrespective of the plasticizer content or composition, the relaxation phases shows rather constant plasticizer volume fractions. The low‐, middle‐, and high relaxation phases include respectively around 30, 60 and 80 vol % of gluten protein. These relaxations are assigned to the segmental motion of the surface amino‐acid side groups, to the collective motion of packed gluten proteins, and to the gain in protein conformational mobility as a 2D network of interacting plasticizer molecules forms. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43254.     

Role of Chain Entanglements in the Electrospinning of Wheat Protein-Poly(Vinyl Alcohol) Blends

The aim of this investigation was to determine if the rapid solvent removal evaporation that occurs during electrospinning enabled the gluten protein and poly(vinyl alcohol) (PVOH) chains to remain at least partially entangled in the final product. Natural and synthetic biopolymer blends are known to phase separate in the melt. Differential scanning calorimetry (DSC) was used to test our hypothesis, which we achieved by systematically comparing the thermal profiles of the nonwoven fibrous sheets comprising: 1) 100% commercial wheat gluten, 2) 100% PVOH, and 3) the (75/25) wheat gluten/PVOH blend. The DSC scans of the two PVOH-containing, nonwoven fibrous sheets exhibited differences in the characteristics and positions of the melting peaks (T_m) of the PVOH crystalline phase, while the DSC scans of the nonwoven fibrous sheets comprising either 100% commercial wheat gluten or the wheat gluten/PVOH blend yielded neither a measurable glass transition temperature (T_g) nor a T_m. Energy dispersive spectroscopy (EDS) was used to compare the elemental compositions of the individual fibers with the compositions of the spherical domains found in the nonwoven fibrous mats. These scans revealed that the mineral matter found in commercial wheat gluten (roughly 1% by weight) had phase-separated from the bulk gluten protein as a result of electrospinning.     

Novel biocomposites based on wheat gluten and rubber wood sawdust

Rubber wood sawdust (RWS) was used as a reinforcement for wheat gluten based bioplastics. The RWS content was varied from 0, 5, 10, 15–20 wt %. Effects of the RWS content on the morphology, water absorption, mechanical, thermal, and biodegradation properties of the wheat gluten based bioplastic were investigated. An addition of RWS caused an improvement of the tensile strength and water resistance of the wheat gluten based bioplastics. Scanning electron micrograph of the wheat gluten/RWS composites with a 10 wt % of RWS revealed a good dispersion and uniform embedding of the RWS within the wheat gluten matrix. Agglomeration of RWS was observed when the RWS loads were increased (15 and 20 wt %). The biodegradation process of the composites depended on the amount of RWS. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43705.     

Stage-Specific Characterization of Physiological Response to Heat Stress in the Wheat Cultivar Norin 61

Bread wheat (Triticum aestivum) is less adaptable to high temperatures than other major cereals. Previous studies of the effects of high temperature on wheat focused on the reproductive stage. There are few reports on yield after high temperatures at other growth stages. Understanding growth-stage-specific responses to heat stress will contribute to the development of tolerant lines suited to high temperatures at various stages. We exposed wheat cultivar “Norin 61” to high temperature at three growth stages: seedling–tillering (GS1), tillering–flowering (GS2), and flowering–maturity (GS3). We compared each condition based on agronomical traits, seed maturity, and photosynthesis results. Heat at GS2 reduced plant height and number of grains, and heat at GS3 reduced the grain formation period and grain weight. However, heat at GS1 reduced senescence and prolonged grain formation, increasing grain weight without reducing yield. These data provide fundamental insights into the biochemical and molecular adaptations of bread wheat to high-temperature stresses and have implications for the development of wheat lines that can respond to high temperatures at various times of the year.     

Characterization of proteins from grain of different bread and durum wheat genotypes

The classical Osborne wheat protein fractions (albumins, globulins, gliadins, and glutenins), as well as several proteins from each of the four subunits of gliadin using SDS-PAGE analyses, were determined in the grain of five bread (T. aestivum L.) and five durum wheat (T. durum Desf.) genotypes. In addition, content of tryptophan and wet gluten were analyzed. Gliadins and glutenins comprise from 58.17% to 65.27% and 56.25% to 64.48% of total proteins and as such account for both quantity and quality of the bread and durum wheat grain proteins, respectively. The ratio of gliadin/total glutenin varied from 0.49 to 1.01 and 0.57 to 1.06 among the bread and durum genotypes, respectively. According to SDS-PAGE analysis, bread wheat genotypes had a higher concentration of α + β + γ-subunits of gliadin (on average 61.54% of extractable proteins) than durum wheat (on average 55.32% of extractable proteins). However, low concentration of ω-subunit was found in both bread (0.50% to 2.53% of extractable proteins) and durum (3.65% to 6.99% of extractable proteins) wheat genotypes. On average, durum wheat contained significantly higher amounts of tryptophan and wet gluten (0.163% dry weight (d.w.) and 26.96% d.w., respectively) than bread wheat (0.147% d.w. and 24.18% d.w., respectively).     

Role of Chain Entanglements in the Electrospinning of Wheat Protein-Poly(Vinyl Alcohol) Blends

The aim of this investigation was to determine if the rapid solvent removal evaporation that occurs during electrospinning enabled the gluten protein and poly(vinyl alcohol) (PVOH) chains to remain at least partially entangled in the final product. Natural and synthetic biopolymer blends are known to phase separate in the melt. Differential scanning calorimetry (DSC) was used to test our hypothesis, which we achieved by systematically comparing the thermal profiles of the nonwoven fibrous sheets comprising: 1) 100% commercial wheat gluten, 2) 100% PVOH, and 3) the (75/25) wheat gluten/PVOH blend. The DSC scans of the two PVOH-containing, nonwoven fibrous sheets exhibited differences in the characteristics and positions of the melting peaks (T_m) of the PVOH crystalline phase, while the DSC scans of the nonwoven fibrous sheets comprising either 100% commercial wheat gluten or the wheat gluten/PVOH blend yielded neither a measurable glass transition temperature (T_g) nor a T_m. Energy dispersive spectroscopy (EDS) was used to compare the elemental compositions of the individual fibers with the compositions of the spherical domains found in the nonwoven fibrous mats. These scans revealed that the mineral matter found in commercial wheat gluten (roughly 1% by weight) had phase-separated from the bulk gluten protein as a result of electrospinning.     

Aggregation and degradation of plasticized wheat gluten during thermo-mechanical treatments, as monitored by rheological and biochemical changes

Different thermo-mechanical treatments were carried out on plasticized gluten samples using the rubber process analyser (RPA 2000) which allows to record changes in viscoelastic properties during sample treatment. Temperatures ranging from 90 to 150 °C were tested at two oscillatory strains (7%: linear domain; 70%: high deformation domain). Biochemical changes of gluten proteins during treatments were followed by SE-HPLC. Rheological and biochemical changes were found to be in good accordance and allow us to characterize two major phenomena: aggregation and degradation of wheat gluten proteins upon heating. A kinetic approach of both phenomena was carried out in order to determine the temperature dependency of both reactions with respect to applied strain. Strain was found to decrease the temperature dependency of modulus values, to moderately accelerate the gluten protein aggregation reaction, and to greatly accelerate the gluten thermal degradation.