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Phenolic acids are antioxidant nutrients in cereals and affect the quality of wheat products and the properties of gluten protein. Gallic acid (GA), caffeic acid (CA), syringic acid (SA), and p-coumaric acid (p-CA) were selected to study the interaction mechanism between cereal phenolic acids and gluten protein.The results showed that adding GA significantly (P 0.05) decreased the content of free sulfhydryl in gluten proteins by 70-87.26% compared with the control group. The aggregates' behavior of gluten protein induced by adding the phenolic acids would produce oversized particles (5000 nm). Adding the selected phenolic acids changed the hydrogen-bond linkage of protein secondary structure. Zeta potential values of gluten protein increased significantly (P 0.05) by 14.41%, 26.49%, 30.77%, and 57.93% for CA, p-CA, GA, and SA respectively added at 0.03 g kgThe results demonstrated that the selected phenolic acids could interact with gluten protein via covalent cross-linking as well as by hydrogen bonding, thereby changing the structure of the gluten protein. This exploration is expected to provide theoretical support for the development and utilization of whole-grain foods. © 2022 Society of Chemical Industry.

The Maillard reaction (MR) has been known to modify proteins and optimize their physicochemical properties by conjugating with reducing sugars. The structure and physicochemical properties of wheat gliadin and maize amylopectin conjugates induced by MR were investigated under different gliadin-amylopectin ratios (2:1, 1:1, 1:2, 1:4, and 1:8). The formation of conjugates was indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, degree of conjugation, and browning development analyses. The Fourier transform infrared and fluorescence spectroscopy analyses suggested changes in the structures of conjugates and the microenvironment of amino acids. A remarkable decrease in the β-turn structure content and an increase in the free sulfhydryl group content were observed at a ratio of 1:8, leading to decreased allergenicity. The reaction process was commendably controlled at a ratio of 1:1 with a 59.7% degree of conjugation in this group, contributing to the amelioration of solubility and foaming properties. Meanwhile, improvements in the oil holding capacity, surface hydrophobicity, and emulsifying properties were observed at a ratio of 1:4. PRACTICAL APPLICATION: The study revealed that the conjugates produced by MR might have various degrees of improved functional properties and reduced allergenicity at different ratios of substrates. Our study might be helpful for conjugates to assist in improving the texture of products and its potential in expanding the industrial application of products with gliadin.

The impact of different ultrasonic power on the structure and functional properties of wheat gliadin (WG) and green wheat gliadin (GG) was investigated and compared. Ultrasound had no obvious effect on subunit composition and bands of WG and GG, and there were more small molecular weight bands in GG. The results of Fourier transform-infrared spectroscopy, intrinsic fluorescence spectroscopy, and scanning electron microscopy analyses demonstrated that ultrasonic treatment had a significant effect on the structure of WG and GG, inducing the transformation from order structure to disorder structure. The dispersion and uniformity were better at 400 and 300 W, respectively. Under proper ultrasonic treatment, the particle size of WG and GG was significantly reduced, and the free sulfhydryl groups and surface hydrophobicity were significantly increased (p 0.05). Furthermore, the functional properties of WG and GG such as solubility, emulsification properties, water holding and oil holding properties, thermal stability, and digestibility were enhanced. The better functional properties of WG and GG were obtained at 400 and 300 W, respectively. These results indicated that ultrasonic treatment with appropriate power was a valuable method for improving functional characteristics of WG and GG. PRACTICAL APPLICATION: Ultrasonic treatment could cause structural changes of wheat gliadin (WG) and green wheat gliadin (GG), and their functional properties are improved under appropriate power. This study compares the effects of ultrasound on WG and GG, and the results will provide theoretical guidance for the development of GG in the food industry.

Proteins, one of the vital nutritional compounds sensitive to the environment, can be modified by interaction with polyphenols. Ultrasonication has been applied for enhancing the functional properties of proteins. In this study, the interactions of gliadin (G) and rutin (R) in the absence and presence of ultrasonication (0, 150, 300, 450, and 600 W) for 20 min were investigated, with a focus on the properties of emulsions prepared by G-R complexes. Ultrasonication improved the interaction, which increased the content of β-type secondary structure. Ultrasonication at 450 W increased the particle size of the conjugates. For Pickering emulsions, treating the covering of R on G with ultrasonication improves the stability of the G-based emulsion significantly, owing to the strong films formed on the oil-water interfaces. The G-R complexes treated at 450 W ultrasonication formed emulsions that showed higher potential and storage modulus (G') and denser microstructures than those of the untreated emulsions. Nevertheless, ultrasound treatment at 600 W weakened the emulsion properties that were stabilized by the conjugates. Ultrasound combined R was shown to be a potential processing technology for changing the protein structure and producing stable emulsions. PRACTICAL APPLICATION: The interactions between proteins and polyphenols are able to preserve the stability of the functional compounds, allow targeted and controlled release, and improve the texture of these complexes employed in the food industry. Improvements in the functional characteristics of the protein-polyphenol complexes so that they possess high emulsifying stability during food processing is a crucial factor for employing them in the food industry. Therefore, the aim of this research is using a soluble complex of gliadin-rutin for the development of its functional characteristics.

Objective To perform a meta-analysis that investigates the prevalence and seroprevalence of celiac disease (CD) in Chinese general population and high-risk populations, and to summarize data from a serological survey of high-risk populations in Guangdong, China. Methods We collected data from the serological survey of high-risk populations in Guangdong, China (n = 1390) by testing serum tissue transglutaminase immunoglobulin A (tTG-IgA), deamidated gliadin peptides immunoglobulin A (DGP-IgA) and deamidated gliadin peptides immunoglobulin G (DGP-IgG), and also searched Medline, PubMed, Embase, Cochrane and 3 Chinese databases up to December 20, 2020 to estimate the pooled prevalence and seroprevalence of CD in China. Results In the serological survey, 0.94% (13/1390) individuals were positive for CD antibodies. In a meta-analysis of 18 studies, the seroprevalence of CD in Chinese general population was 0.27% (95% CI, 0.02-0.71). The seroprevalence of CD in Chinese high-risk populations was 8.34% (95% CI, 4.90-12.54), significantly higher than that in general population (OR 7.27, 95% CI, 4.06-13.04). The prevalence of biopsied-proven CD in high-risk Chinese populations was 4.44% (95% CI, 1.53-8.58). The seroprevalence of CD was varying with geographic origin, higher in Northern China than that in Southern China. Conclusions The basis for diagnosis of CD in Chinese patients should be through early case-finding by serological screening in high-risk groups; generous serological testing in patients with vague symptoms, especially in Northern Chinese.

Chronic enteropathies (CEs) in dogs describe a group of idiopathic disorders characterized by chronic persistent or recurrent gastrointestinal (GI) signs. Three major subgroups of CE can be identified by their response to treatment: Food-responsive disease (FRD), antibiotic-responsive disease (ARD), and steroid-responsive disease (SRD). The clinical diagnosis of CE is made by exclusion of all other possible causes of chronic diarrhea and includes histologic assessment of intestinal biopsies. The process of diagnosing canine CE can therefore be very time-consuming and expensive, and in most cases, does not help to identify dogs that will respond to a specific treatment. The development of novel diagnostic tests for canine CE has therefore focused on the accuracy of such tests to predict treatment responses. In this article, several novel assays that have the potential to become commercially available will be discussed, such as genetic tests, perinuclear anti-neutrophil cytoplasmic antibodies (pANCA), antibodies against transglutaminase/gliadin, antibodies against E coli OmpC/flagellin, and micro RNAs.

So far, limited studies have focused on the role of Macrophages (MQs) in the development or progression of celiac disease (CD). Researchers believe that increasing knowledge about the function of MQs in inflammatory disorders plays a critical role in finding a new treatment for these kinds of diseases.CD is a permanent autoimmune intestinal disorder triggered by gluten exposure in predisposed individuals. This disorder happens due to the loss of intestinal epithelial barrier integrity characterized by dysregulated innate and adaptive immune responses. MQs are known as key players of the innate immune system that link innate and adaptive immunity. MQs of human intestinal lamina propria participate in maintaining tissue homeostasis, and also intestinal inflammation development. Previous studies suggested that gliadin triggers a proinflammatory phenotype (M1 MQ) in human primary MQs. Moreover, M2-related immunosuppressive mediators are also present in CD. In fact, CD patients present an impaired transition from pro-inflammatory to anti-inflammatory responses due to inappropriate responses to gliadin peptides.The M1/M2 MQs polarization balancing regulators can be considered novel therapeutic targets for celiac disease.

While individuals diagnosed with autism spectrum disorders (ASD) have higher levels of antibodies directed towards gliadin, a component of wheat gluten, no study has examined anti-gliadin antibodies (AGA) in etiologically relevant periods before diagnosis. The objective of this study was to investigate if maternal levels of AGA, during pregnancy and at the time of birth, are associated with ASD in offspring. We analyzed AGA in archived neonatal dried blood spots (NDBS) for 921 ASD cases and 1090 controls, and in paired maternal sera collected earlier in pregnancy for a subset of 547 cases and 428 controls. We examined associations with ASD diagnoses as a group and considering common comorbidities (intellectual disability [ID] and attention-deficit/hyperactivity disorder). We compared 206 cases to their unaffected siblings to examine the potential for confounding by shared familial factors. Odds of ASD tended to be lower among those with the highest levels (≥90th percentile) of AGA compared to those with low levels (

BACKGROUND Sowing date, seeding rate, and nitrogen (N) topdressing ratio have strong effects on grain yield (GY) and bread-making quality (BQ) in bread wheat. Simultaneous improvement in GY and BQ in bread wheat has long been a challenge due to the inverse relationship between GY and grain protein concentration (GPC). In this study, we investigated whether the GY and BQ of bread wheat sown on different dates could be improved simultaneously by optimizing the seeding rate and the N topdressing ratio. RESULTS Delaying sowing beyond a certain period led to decreases in both GY and BQ. Optimizing the seeding rate and N topdressing ratio enhanced the N uptake during pre- and post-anthesis, as well as N remobilization during grain filling for all wheat plants sown on different dates, thereby increasing the GPC and the total N per grain (Ntot ). Consequently, grain protein composition was improved, resulting in an increased glutenin/gliadin ratio, sodium dodecyl sulfate-insoluble glutenin/total glutenin (i.e., glutenin polymerization index), and high-molecular-weight glutenin subunit/ low-molecular-weight glutenin subunit (HMW-GS/LMW-GS) ratio. Increased GPC and improved grain protein composition enhanced BQ. CONCLUSION The mechanism underlying simultaneous improvement in GY and GPC as well as Ntot was the greater increase in N accumulation in grains per unit area relative to increases in GY, or total grain number per unit area. The GY and BQ can be improved simultaneously regardless of sowing date by optimizing the seeding rate and N topdressing ratio via enhanced N uptake and N remobilization into grains. © 2021 Society of Chemical Industry.

This chapter discusses the acclaimed adverse effects of some constituents of whole grain cereals and their bran, including allergenic proteins and proteins that provoke food sensitivity; toxic effects of the heavy metal cadmium; phytate and phenolic compounds for their chelation of iron; and acrylamide, which is a heat- generated class II carcinogen. The possible adverse effects related to whole grain consumption are important to consider with increased consumption of whole grains and especially bran-rich fractions. Some of the cereal proteins may provoke immune responses causing allergies or trigger autoimmune responses leading to food sensitivity in genetically predisposed individuals. Gluten proteins, which is a collective name for glutamin and prolamin-rich proteins, such as gliadin in wheat, hordeins in barley and secalins in rye, may trigger celiac disease in genetically predisposed individuals carrying, for example, the HLA-DQ8 and/or HLA-DQ2 genotypes.

Kolbach index (KI) is an important index to evaluate the qualities of malt, which will affect protein molecular composition, enzyme activity, and other macromolecules degradation during wheat malting. In this paper, the relationship between wheat (Triticum aestivum L.) malts KI and the (i) characterization of albumins, globulins, gliadins, and glutenins and their hydrolysis and (ii) the enzymatic breakdown of starch and arabinoxylans during malting were studied. As malt KI values increased, all fractions of glutenins and gliadins were extensively hydrolyzed. The higher Mw globulins (36.6 to 70.8 kDa) were also increasingly degraded at higher KI values, but the concentration of smaller globulin fractions (14.9 to 35.0 kDa) had increased significantly. As for albumins, although their overall concentration had increased as KI increased, changes in the concentration of individual albumin fractions was more complex. While there were significant increases in the concentration of some new albumin proteins (43.8 and 84.4 kDa), the concentration of some albumins decreased (21.1 to 64.3 kDa), and some fractions had completely disappeared (28.8 and 64.3 kDa). Following mashing, the hydrophobicity of the worts had decreased significantly at higher KI values. At malt KI values between 39.5% and 42.7%, the enzymatic activity was at its highest, the degradation of starch was adequate and stable, and the concentration of water-soluble arabinoxylans was optimal. A KI value of about 39.5% to 42.7% was therefore considered optimal for the production of wheat malts with superior quality attributes. PRACTICAL APPLICATION: The findings from this study will be valuable to beer companies; a more precise control of the malting and brewing parameters, fundamental for the production of high-quality wheat malts and wheat beer, can be achieved.

Summary Most alpha‐gliadin genes of the Gli‐D2 locus on the D genome of hexaploid bread wheat (Triticum aestivum) encode for proteins with epitopes that can trigger coeliac disease (CD), and several contain a 33‐mer peptide with six partly overlapping copies of three epitopes, which is regarded as a remarkably potent T‐cell stimulator. To increase genetic diversity in the D genome, synthetic hexaploid wheat lines are being made by hybridising accessions of Triticum turgidum (AB genome) and Aegilops tauschii (the progenitor of the D genome). The diversity of alpha‐gliadins in A. tauschii has not been studied extensively. We analysed the alpha‐gliadin transcriptome of 51 A. tauschii accessions representative of the diversity in A. tauschii. We extracted RNA from developing seeds and performed 454 amplicon sequencing of the first part of the alpha‐gliadin genes. The expression profile of allelic variants of the alpha‐gliadins was different between accessions, and also between accessions of the Western and Eastern clades of A. tauschii. Generally, both clades expressed many allelic variants not found in bread wheat. In contrast to earlier studies, we detected the 33‐mer peptide in some A. tauschii accessions, indicating that it was introduced along with the D genome into bread wheat. In these accessions, transcripts with the 33‐mer peptide were present at lower frequencies than in bread wheat varieties. In most A. tauschii accessions, however, the alpha‐gliadins do not contain the epitope, and this may be exploited, through synthetic hexaploid wheats, to breed bread wheat varieties with fewer or no coeliac disease epitopes., Significance Statement We sequenced the expressed alpha‐gliadins in grains of Aegilops tauschii across its range. Some accessions contained the 33‐mer peptide with six overlapping coeliac disease epitopes, indicating that this was introduced into bread wheat from A. tauschii. We also found a large variation in the occurrence of coeliac disease epitopes, including accessions with small numbers of epitopes, suitable as basis for breeding bread wheat varieties with fewer or no coeliac disease epitopes.

ABSRACT The interaction between gluten and hydrocolloid additive, as well as the pH condition during dough formation is very important in making flour products. In this study, the influence of different pH conditions on the interactions between gluten proteins (including glutenin and gliadin) and carboxymethylcellulose (CMC), and on the rheological and microstructure changes of gluten proteins was investigated. The dynamic frequency sweep indicated CMC-gluten displayed more solid-like behavior under alkaline conditions than that under acidic conditions. The creep-recovery experiment suggested acidic conditions were not conducive to maintain the elasticity of CMC-gluten. Microstructural changes of various glutens with CMC showed that a higher ratio of β-sheets was observed in the CMC-gluten and CMC-glutenin under alkaline conditions. Total free sulfhydryl contents and changes in tryptophan microenvironment showed glutenin played a key role in the G polymerization with the addition of CMC. Lower surface hydrophobicity of CMC-gluten was displayed under acidic conditions. Scanning electron microscopy images showed that neutral and alkaline conditions were conducive to the network structure formation of CMC-gluten and CMC-glutenin. PRACTICAL APPLICATION This study investigated the interaction of CMC with gluten, gluten, and gliadin under different pH conditions, providing a basis for expanding dough quality improvement, and extending the in-depth application of CMC in the food industry.

Prolamins are a group of safe food additives that are biocompatible, biodegradable, and sustainable. Zein, gliadin, kafirin, and hordein are common prolamins that have been extensively studied, particularly as these form colloidal particles because of their amphiphilic properties. Prolamin-based binary/ternary complexes, which have stable physicochemical properties and superior functionality, are formed by combining prolamins with polysaccharides, polyphenols, water-soluble proteins, and surfactants. Although the combination of prolamins with other components has received attention, the relationship between the structural design of prolamin-based complexes and their functionalities remains uncertain. This review discusses the production methods of prolamin-based complexes, the factors influencing their structural characteristics, and their applications in the food industry. Further studies are needed to elucidate the structure-function relationships between prolamins and other biopolymers, as well as the toxicological effects of these complexes in food.

The present study examined the wheat protein gliadin-induced oxidative and nitrosative stress and its downstream responses in human intestinal HCT-116 and HT-29 cells. The beneficial role of dietary phytochemical curcumin and role of multifunctional enzyme Apurinic/aprymidinic endonuclease 1 (APE1) a major player involved in the base excision repair (BER)-pathway in gliadin intolerant intestinal HCT-116 and HT-29 cell lines were evaluated as an in vitro model study. The cultured cells were exposed to gliadin protein, H

Background Glutenin macropolymer (GMP), glutenin and gliadin proteins are important indicators of the baking quality of dough. This study investigated the impacts of wheat bran and a mixture of ferulic acid (FA) and dietary fiber (DF) on the constitution of gluten proteins. Addition of wheat bran (100 and 150 g kg-1 ) into gluten decreased the gliadin/glutenin ratio, while the addition of different amounts of FA + DF (C1 group: 20 g kg-1 FA and 60 g kg-1 DF; C2 group: 30 g kg-1 FA and 90 g kg-1 DF) had the opposite effect. Results The GMP contents of wheat bran groups (B1 group: 100 g kg-1 ; B2 group: 150 g kg-1 ) were similar to that of the control group, and disulfide bond contents were increased. However, both GMP and disulfide bond contents of FA + DF groups significantly decreased. The GMP gel properties and microstructures were destroyed after addition of wheat bran and FA + DF. The wheat bran and FA + DF additives induced different effects on the thermal properties and secondary structures of glutenin, gliadin and GMP. Conclusions The results suggest that the interaction mechanism of bran fractions and gluten proteins is not only related to the physicochemical properties of the additives, but also to interactions between the additives and the components of gluten protein. © 2020 Society of Chemical Industry.

Human Leukocyte Antigen (HLA)-DQ2 and HLA-DQ8 are genetic risk factors for Type 1 Diabetes Mellitus (T1DM) and Celiac disease (CD) in Caucasians, but their association with Taiwanese Han population is unknown. We screened 532 Taiwanese T1DM patients for CD biomarkers including anti-tissue transglutaminase (TGM2), anti-gliadin and anti-neoepitope antibodies (Abs), sequencing DQB1 genotypes, and characterized the TGM2 Abs. We report that 3.76% of Taiwanese patients had TGM2-Abs and all had no CD's symptoms. In contrast to Caucasian's CD patients, DQ2/DQ8 only constituted ~4/5 of TGM2-Abs positive patients, while the other ~1/5 patients belonged to different HLA genotypes. Either anti-gliadin or anti-neoepitope Abs coexisted with ~3/4 of TGM2-Abs positive patients that were likely due to gluten-ingestion, while the cause of TGM2-Abs production for other ~1/4 of patients was unknown. Purified anti-TGM2 IgA (TGA) and anti-TGM2 IgG (TGG) could bind on endothelial cells surface, recognized native better than denatured forms of TGM2, and TGA inhibited TGM2's transamidation activity by up to 80% but TGG had no effects. Epitope mapping of all TGM2-Abs positive sera demonstrated that TGM2-Abs had heterogeneity in specificities. This is the first study on the differences between Taiwanese Han group and Caucasian in HLA genotypes and properties of TGM2-Abs.

Previously we detected increased levels of IgA to the enamel matrix protein amelogenin in children with untreated coeliac disease (CeD). The biological impact of autoantibodies to amelogenin may depend on which part of the amelogenin (epitopes) they bind. Sera or blood samples from 146 untreated children with CeD from two different cohorts and 25 non-CeD control children were tested for IgA anti-amelogenin (Emdogain) reactivity. The 32 CeD children and six controls with the highest reactivity were selected for detailed IgA anti-amelogenin (AMELX) epitope mapping using 31 overlapping, 10-22mer peptides in ELISA. The dominating reactivity were to six peptides in a 75-amino-acid (aa)-long central segment (aa 75-150) containing enzymatic cleavage sites for matrix metalloproteinase 20 (MMP-20) and kallikrein-related peptidase 4 (KLK-4) and to two N-terminal peptides (aa 13-41) included in the tyrosine-rich amelogenin peptide fragment, which is important for self-assembly. Only two of the six control children reacted to single, but different peptides. Solid-phase extraction with gliadin- and Emdogain-coated Sepharose revealed a gliadin cross-reactive central region and a putative conformation-dependent, apparently non-cross-reactive N-terminal region. High IgA anti-amelogenin reactivity may interfere with normal amelogenesis by inhibiting amelogenin self-assembly, amelogenin-hydroxyapatite interaction, and/or enzymatic degradation.

Although the impact of heat on molecular properties of wheat gluten is well understood, changes in its microstructure have rarely been studied. Here, formation of the thermoset gluten network in a model system relevant for bread baking was studied with confocal laser scanning microscopy and protein network analysis. From 65 °C onwards, gluten converts from thick aligned protein strands in a highly branched and homogeneous network of small thin protein threads. Neither gliadin incorporation in the network nor application of aqualysin 1, the thermo-active serine peptidase from Thermus aquaticus which recently has been reported to hydrolyze gluten proteins in dough only at temperatures exceeding 80 °C, impacts on the gluten microstructure. As starch causes structure setting itself and thereby decreases protein mobility, molecular scale changes in the gluten network at temperatures exceeding 80 °C brought about by aqualysin 1 do not impact its microstructure. ispartof: Journal of Food Science and Technology vol:55 issue:2 pages:891-898 status: Published online

The quality traits of beer, which include flavor, texture, foam stability, gushing, and haze formation, rely on contributions from beer proteins and peptides. Large-scale proteomic analysis of beer is gaining importance, not only with respect to authenticity of raw material in beer but also to improve quality control during beer production. In this work, foam proteins were first isolated from beer by virtue of their high hydrophobicity. Then sequential filter-aided sample preparation coupled with liquid chromatography and tandem mass spectrometry was used to analyze both beer protein and foam protein. Finally, 4692 proteins were identified as beer proteins, and 3906 proteins were identified as foam proteins. In total, 7113 proteins were identified in the beer sample. Several proteins contributing to beer quality traits, including lipid transfer protein, serpin, hordein, gliadin, and glutenin, were detected in our proteins list. This work constructed a comprehensive beer proteome map that may help to evaluate potential health risks related to beer consumption in celiac patients.

Background and aim The aim of this study was to assess the prevalence and characteristics of celiac disease (CD) in all patients with type 1 diabetes mellitus attending a tertiary adult diabetes clinic in Durban, South Africa. Methods This was a cross-sectional observational study that screened 202 patients; of these, 56.4% were African (Black), 31.7% Asian Indian, 4.5% White, and 7.4% mixed race. Demographic data, symptoms, and anthropometry were documented. Blood tests included anti-tissue transglutaminase antibody (tTG), anti-endomysial antibody (EMA), and anti-gliadin antibody (AGA). Endoscopy and duodenal biopsy were performed in patients with celiac antibodies. Diagnosis of CD was based on the modified Marsh classification. Results Mean age and mean duration of diabetes were 26.4 ± 11.4 and 10.7 ± 9.1 years, respectively. Celiac antibodies were found in 65 (32.2%) patients: EMA 7.4%, tTG immunoglobulin A (IgA) 8.4%, tTG immunoglobulin G 1.9%, AGA IgA 18.3%, and AGA immunoglobulin G 21.8%. Histological evidence of CD was found in 5.9% (n = 12/202): 2.5% were classed as definite CD (Marsh 3) and 3.4% as potential CD (Marsh 1). None of the patients with CD were symptomatic. The sensitivity of AGA IgA, EMA, and tTG IgA antibodies for detecting histologically proven CD was 66.7%, 50.0%, and 41.7%, respectively. Conclusion The prevalence of CD was similar to reports from western countries. No ethnic specific differences were noted. CD was silent in all patients in this study. The sensitivity of EMA and tTG antibodies was poor and merits further evaluation as screening tools for CD in South African patients with type 1 diabetes mellitus.

Zonulin is a physiological modulator of intercellular tight junctions, which upregulation is involved in several diseases like celiac disease (CeD). The polyQ gliadin fragment binds to the CXCR3 chemokine receptor that activates zonulin upregulation, leading to increased intestinal permeability in humans. Here, we report a general hypothesis based on the structural connection between the polyQ sequence of the immunogenic CeD protein, gliadin, and enteric coccidian parasites proteins. Firstly, a novel interaction pathway between the parasites and the host is described based on the structural similarities between polyQ gliadin fragments and the parasite proteins. Secondly, a potential connection between coccidial infections as a novel environmental trigger of CeD is hypothesized. Therefore, this report represents a promising breakthrough for coccidian research and points out the potential role of coccidian parasites as a novel trigger of CeD that might define a preventive strategy for gluten-related disorders in general. Also see the video abstract here: https://youtu.be/oMaQasStcFI.

Background Gluten sensitivity among psoriasis patients and its association with gender, age, disease duration and severity of psoriasis are under studied in Indians. Objective To examine association among serum levels of anti-tTG and anti-gliadin antibodies and clinical features including gender, age, duration and severity of psoriasis. Methods Serum levels of anti-transglutaminase and anti-gliadin antibodies were measured quantitatively in 80 (M:F 57:23) psoriasis patients aged 15 to 83 years and matched healthy subjects. Results Forty-five (56.3%) patients were aged ≥41years, duration of disease was >5years in 43(53.8%) patients, and 22 (27.5%) patients had moderate-to-severe psoriasis. Two (2.5%) patients had arthritis and elevated serum anti-gliadin antibody. Significantly more patients than controls had elevated serum anti-gliadin antibody (67.5% vs. 2.5%) and anti-transglutaminase antibody levels (62.5% vs. 0%). Two patients, each with mild and moderate-to-severe psoriasis, had highly elevated serum anti-gliadin antibody and symptoms akin to coeliac disease. Except for a longer duration of psoriasis in patients with elevated anti-gliadin antibodies, there was no statistically significant difference in gender, age, and severity of psoriasis when compared with patients having normal levels. Conclusion Significant elevation of serum anti-transglutaminase and anti-gliadin antibodies levels is noted in psoriasis patients reflecting a possible link. However, results need careful interpretation for any significance of gluten sensitivity in pathogenesis of psoriasis/arthritis or as a stand-alone risk factor for chronicity/severity of psoriasis or whether gluten-free diet will be ameliorating. Small number of subjects, cross-sectional study design, lack of pathological/endoscopic diagnosis and follow-up are study limitations.

Background: The protein aggregation characteristics of three types of freshly milled wheat flour with high, medium or low gluten were investigated during 90 days of maturation. Changes in the content and particle size of the glutenin macropolymer (GMP), contents of sulfhydryl groups and disulfide bonds (SS), and secondary structure and molecular weight distribution of the protein were determined.; Results: For high, medium and low gluten flour, GMP content increased to 22.25, 13.72 and 10.32 g kg-1 ; free sulfhydryl group content decreased by 5.5%, 4.1% and 4.4%; and SS content increased by 1.6%, 1.8% and 2%, respectively. The proportion of β-sheet and random coil increased, and the proportion of α-helix and β-turns decreased. The polymeric protein content increased, whereas that of gliadin decreased.; Conclusion: Protein aggregation mediated by SS cross-linking helped develop a stronger gluten network. The findings provide theoretical support for the changes in protein structure during flour maturation and also help to predict the quality of wheat flour and its products. © 2018 Society of Chemical Industry.; © 2018 Society of Chemical Industry.

Water deficiency affects grain proteome dynamics and storage protein compositions, resulting in changes in gluten viscoelasticity. In this study, the effects of field water deficit on wheat breadmaking quality and grain storage proteins were investigated.; Results: Water deficiency produced a shorter grain-filling period, a decrease in grain number, grain weight and grain yield, a reduced starch granule size and increased protein content and glutenin macropolymer contents, resulting in superior dough properties and breadmaking quality. Reverse phase ultra-performance liquid chromatography analysis showed that the total gliadin and glutenin content and the accumulation of individual components were significantly increased by water deficiency. Two-dimensional gel electrophoresis detected 144 individual storage protein spots with significant accumulation changes in developing grains under water deficit. Comparative proteomic analysis revealed that water deficiency resulted in significant upregulation of 12 gliadins, 12 high-molecular-weight glutenin subunits and 46 low-molecular-weight glutenin subunits. Quantitative real-time polymerase chain reaction analysis revealed that the expression of storage protein biosynthesis-related transcription factors Dof and Spa was upregulated by water deficiency.; Conclusion: The present results illustrated that water deficiency leads to increased accumulation of storage protein components and upregulated expression of Dof and Spa, resulting in an improvement in glutenin strength and breadmaking quality. © 2018 Society of Chemical Industry.; © 2018 Society of Chemical Industry.

The enzymatic modification of wheat proteins during dough fermentation and its digestion as supported by peptidases of microbiological origin can result in the degradation of important peptides in the pathogenesis of coeliac disease. However, baking bread and the high temperature associated with this could change the physicochemical and immunological properties of proteins. Thermal changes in the spatial structure of proteins and their hydrolysis can lead to a masking or degrading of immunoreactive peptides.; Results: The addition of prolyl endopeptidase (PEP), comprising peptidases isolated from Lactobacillus acidophilus 5e2 (LA) or transglutaminase (TG) in the course of fermentation, decreases its immunoreactivity by 83.9%, 51.9% and 18.5%, respectively. An analysis of the fractional composition of gliadins revealed that γ- and ω-gliadins are the proteins most susceptible to enzymatic modification. Hydrolysis of wheat storage proteins with PEP and LA reduces the content of αβ-, γ- and ω-gliadins by 13.7%, 60.2% and 41.9% for PEP and by 22.1%, 43.5% and 36.9% for LA, respectively. Cross-linking of proteins with TG or their hydrolysis by PEP and LA peptidases during the process of forming wheat dough, followed by digesting bread samples with PEP and LA peptidases, decreases the immunoreactivity of bread hydrolysates from 2.4% to 0.02%. The content of peptide detected in polypeptide sequences is 263.4 ± 3.3, 30.9 ± 1.5 and 7.9 ± 0.4 mg kg-1 in samples of hydrolysates of bread digested with PEP, as produced from dough modified by TG, PEP and LA, respectively.; Conclusion: Enzymatic pre-modification of proteins during the process of dough fermentation decreases their immunoreactive potential, such that fewer peptides recognised by R5 antibodies are released during the digestion process from the bread matrix. Immunoreactive peptides are degraded more effectively when digestive enzymes are supported by the addition of PEP. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Monitoring of the molecular motions and secondary structures of gliadin (Gli) and glutenin (Glu) in blends with 10, 20, 30, and 40% glycerol was performed by solid-state (SS) and time domain (TD) NMR spectroscopy. Increasing the glycerol content increased the relative amount of β-sheets and disordered structures, while decreasing α-helices in Gli/Glu–glycerol blends studied by 13C CPMAS NMR. For ≥20% glycerol samples, the protein side-chain mobility increased similarly for Gli and Glu. A higher proportion of α-helices versus β-sheets was found in Gli-glycerol blends compared with Glu–glycerol blends. Glycerol acted as “immobilized” in 10–20% glycerol Gli samples and was found mainly “free” in 30 and 40% glycerol Gli/Glu samples. During temperature experiments, 30 and 40% glycerol amounts impacted the dynamic molecular behavior of the Gli and Glu proteins differently than lipids, as observed by TD-NMR. The combination of TD-NMR together with SS-NMR showed details of the dynamic molecular variations in Gli/Glu protein structure and are promising techniques to monitor the molecular dynamics of plasticized proteins. (Less)

BACKGROUND Paroxysmal gluten-sensitive dyskinesia (PGSD) in border terriers (BTs) results from an immunologic response directed against transglutaminase (TG)2 and gliadin. Recent evidence suggests that PGSD is only one aspect of a range of possible manifestations of gluten sensitivity in the breed. HYPOTHESIS/OBJECTIVES Gluten sensitivity in BTs is a heterogeneous disease process with a diverse clinical spectrum; to characterize the phenotype of PGSD using TG2 and gliadin autoantibodies as diagnostic markers. ANIMALS One hundred twenty-eight client-owned BTs with various disorders. METHODS Prospective study. BTs with paroxysmal episodes and a normal interictal examination were phenotyped using footage of a representative episode and assigned to 3 groups: idiopathic epilepsy (IE), paroxysmal dyskinesia (PD), or other. Owners of each dog completed a questionnaire to obtain information regarding clinical signs. Healthy BTs formed a control group. Serum antibodies against TG2 and AGA were measured in all dogs. RESULTS One hundred twenty-eight BTs were enrolled; 45 with PD, 28 with IE, 35 with other conditions, and 20 controls. Three overlapping phenotypes were identified; PD, signs suggestive of gastrointestinal disease, and dermatopathy. AGA-IgG concentrations were increased in PD, compared with IE (P = 0.012), controls (P

Coeliac disease is an autoimmune disorder triggered in genetically predisposed individuals by the ingestion of gluten proteins from wheat, barley and rye. The α‐gliadin gene family of wheat contains four highly stimulatory peptides, of which the 33‐mer is the main immunodominant peptide in patients with coeliac. We designed two sgRNAs to target a conserved region adjacent to the coding sequence for the 33‐mer in the α‐gliadin genes. Twenty‐one mutant lines were generated, all showing strong reduction in α‐gliadins. Up to 35 different genes were mutated in one of the lines of the 45 different genes identified in the wild type, while immunoreactivity was reduced by 85%. Transgene‐free lines were identified, and no off‐target mutations have been detected in any of the potential targets. The low‐gluten, transgene‐free wheat lines described here could be used to produce low‐gluten foodstuff and serve as source material to introgress this trait into elite wheat varieties., The Spanish Ministry of Economy and Competitiveness (Projects AGL2013‐48946‐C3‐1‐R and AGL2016‐80566‐P) and the European Regional Development Fund (FEDER) supported this work. Javier Gil‐Humanes acknowledges the Fundacion Alfonso Martín Escudero for its funding support.

With the increasing demand for high-quality foodstuffs and concern for environmental sustainability, late-season nitrogen (N) foliar fertilization of common wheat is now an important and widespread practice. This study investigated the effects of late-season foliar versus soil N fertilization on yield and protein content of four varieties of durum wheat, Aureo, Ariosto, Biensur and Liberdur, in a three-year field trial in northern Italy.; Results: Variations in low-molecular-weight glutenins (LMW-GS), high-molecular-weight glutenins (HMW-GS) and gliadins were assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). It was found that N applied to the canopy did not improve protein rate compared with N application to the soil (general mean 138 mg g-1 ), but moderately increased productivity in the high-yielding varieties Liberdur and Biensur (three-year means 7.23 vs 7.13 and 7.53 vs 7.09 t ha-1 respectively). Technological quality was mainly related to variety choice, Aureo and Ariosto having higher protein rates and glutenin/gliadin ratios. Also found was a strong 'variety × N application method' interaction in the proportions of protein subunits within each class, particularly LMW-GS and gliadins. A promising result was the higher N uptake efficiency, although as apparent balance, combined with higher HMW/LMW-GS ratio in var. Biensur.; Conclusion: Late-season foliar N fertilization allows N fertilizer saving, potentially providing environmental benefits in the rainy climate of the northern Mediterranean area, and also leads to variety-dependent up-regulation of essential LMW-GS and gliadins. Variety choice is a key factor in obtaining high technological quality, although it is currently associated with modest grain yield. This study provides evidence of high quality in the specific high-yielding variety Biensur, suggesting its potential as a mono-varietal semolina for pasta production. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Summary Biodegradable food packaging promises a more sustainable future. Poly(lactic acid) (PLA) is a promising alternative for petroleum-derived polymers. However, PLA films suffer from poor barrier properties compared with petroleum-derived ones. To address this issue, we designed bilayer and sandwich-architectured trilayer films based on PLA and gliadin. We reported firstly the direct formation of PLA–gliadin bilayer/trilayer films without surface modification on PLA film. The films were compact and uniform, and double/triple layers were combined firmly, preventing delamination. This strategy enhanced mechanical resistance, ductility and moisture barrier of gliadin films and concomitantly enhanced the oxygen barrier for PLA films. Thymol loadings endowed bilayer/trilayer films with antimicrobial activity and antioxidant activity. The antimicrobial activity of the films depended on film types, and gliadin layer presented larger inhibition zone than PLA layer, hinting that the films possessed directional releasing role. This study opens a promising route to fabricate bilayer/trilayer architecture helping to create synergism of the biopolymers.

Wheat proteins are classified according to solubility into the so-called Osborne fractions. Because wheat flour contains both free thiol and disulfide groups, thiol–disulfide interchange reactions are possible during extraction. Osborne fractionation of 12 different wheat flour samples was performed in the presence of N-ethylmaleinimide (NEMI) to alkylate free thiol groups and without addition of NEMI (control). The addition of NEMI during extraction tended to decrease the content of gliadins (predominantly α-gliadins) and caused an increase of the content of glutenins in most flour samples. Thus, alkylation of free thiol groups during extraction led to a decline of the gliadin/glutenin ratio from 2 (control) to approximately 1.5 (NEMI). NEMI and control gliadins were separated by gel-permeation HPLC into an oligomeric subfraction (high-molecular-weight [HMW] gliadins) and two monomeric subfractions. In most flours (8 of 12), the addition of NEMI led to a significant increase of the content of HMW gliadin...

Thirty hard red spring (HRS) wheat cultivars released between 1910 and 2013 were analyzed to determine how they cluster in terms of parentage and HPLC analysis of gliadins and unreduced proteins. Markers for reduced height genes were assessed to determine associations between semidwarfism and protein composition, as well as quality characteristics. In terms of parentage, the clusters formed were grouped according to release year and semidwarfism. Farinograph development time and stability showed significant (P ≤ 0.05) differences between clusters, indicating improvement of mixing characteristics over time. Flour ash content was significantly different (P ≤ 0.05) between clusters, indicating improved milling quality. The clusters based on the gliadin reverse-phase HPLC peak binary data did not group cultivars according to release year and the semidwarf characteristic. However, cultivars were grouped according to year in the absorbance area based dendrogram. The clusters based on absorbance area of size-exclusion HPLC showed significant (P ≤ 0.05) differences for release year, the semidwarf characteristic, and farinograph stability. Overall, the results indicate that the introduction of reduced height genes accompanied the improvements of dough mixing and breadmaking quality characteristics, without altering the composition of gliadin proteins over the last 100 years of HRS wheat breeding.

Problem Celiac disease (CD) is an autoimmune intestinal inflammatory disease triggered by gluten in the diet. Untreated CD has been associated with pregnancy loss and infertility. The purpose of this study was to screen unselected women with recurrent pregnancy loss (RPL) for markers of CD to determine whether a correlation exists between RPL and CD serum markers. Method of study Frequencies of three serum markers of CD [tissue transglutaminase (TTG) IgA, endomysial (EMA) IgA, and deaminated gliadin peptide (DGP) IgA] were determined by enzyme-linked immunoassay (ELISA). Seven hundred and eight women who had two or more failed clinical pregnancies (cases) and one hundred women with at least one live birth and no miscarriages (controls) were included in this study. All cases had a full workup for RPL based on the American Society for Reproductive Medicine 2013 guidelines. Antiphospholipid antibodies (aPL) were correlated with CD markers based on their potential prothrombotic role. Results The results show no significant difference in the prevalence of CD autoantibodies when comparing the RPL patients with the controls. Over half of the patients who tested positive for serum markers for CD also had positive aPL. Conclusion Screening unselected women with RPL who are asymptomatic for CD is not supported based on these data. Women who test positive for CD may be candidates for aPL testing based on the association of adverse pregnancy outcomes.

SummaryBackground Although eosinophilic oesophagitis (EoE) is putatively mediated by an abnormal response to food antigen, the oesophagus is considered relatively impermeable to large molecules. Aim To assess whether food antigens penetrate the oesophageal mucosa in patients with EoE. Methods Anti-gliadin staining was performed in three groups: active EoE, inactive EoE and EoE patients on a low or gluten free diet. To appraise the specificity of our results, we also performed gliadin staining on six patients without oesophageal disease who were consuming gluten. The groups with EoE on gluten also underwent endoscopic infusion with gluten containing soy sauce and repeat biopsies during the endoscopy. We measured eosinophil density, dilated intercellular spaces (on a 0–4+ scale) and gliadin in oesophageal mucosa by immunofluorescence. Results Patients with active EoE had significantly greater epithelial density of anti-gliadin staining when compared to inactive EoE (P

Climate change bears the risk of more frequent drought stress in the northern hemisphere with more frequent early summer drought events affecting main grain crops. Winter wheat (Triticum aestivum L.) is susceptible for such drought events at the flowering and grain filling stages. After drought, the grain yield decrease of three hybrids was about 20% lower compared to three wheat lines analyzed. Wheat grain proteins are classified into four main components such as albumin and globulin, gliadin, and glutenin. The latter two are closely related to the baking quality of flour and might be affected by drought. However, detailed knowledge about the influence of drought on the synthesis of specific storage protein fractions is scarce. By analyzing the grain protein fractions by means of SDS-PAGE technique, we detected an increase in grain protein content as well as in HMW and some LMW glutenin sub-fractions. The glutenin fraction seems to be most variable in gene expression under different environmental scenarios such as drought. However, the protein yield as well as the grain yield may be strongly decreased, which might be not acceptable in practice.

We developed a new disomic addition line M11028-1-1-5 (2n = 44 = 21” + 1”) from a cross between wheat cv. ‘7182’ and octoploid Tritileymus M47 (2n = 8x = 56, AABBDDNsNs). Cytological observations demonstrated that M11028-1-1-5 contained 44 chromosomes and formed 22 bivalents during meiotic metaphase I. The genomic in situ hybridization (GISH) investigations showed this line contained 42 wheat chromosomes and a pair of L. mollis chromosomes. SSR, EST and PCR-based landmark unique gene (PLUG) markers were screened to determine the homoeologous relationships of the introduced L. mollis chromosomes in wheat background. Nine markers, i.e. Xwmc256, Xgpw312, Swes123, CD452568, BF483643, BQ169205, TNAC1748, TNAC1751 and TNAC1752, all of which were located on the homoeologous group 6 chromosomes of common wheat, amplified bands unique to L. mollis in M11028-1-1-5. Gliadin analysis also confirmed that the added chromosomes in M11028-1-1-5 were correlated with the sixth group chromosome. This indicated that M11028-1-1-5 contained a pair of introduced L. mollis chromosome belonging to homoeologous group 6, which we designated it as Lm#6Ns disomic addition line. This is the first report of a common wheat–L. mollis disomic addition line.

Globulin, albumin, gluten, and gliadin in oat bran were prepared by the Osborn method using oat bran as starting material. We characterized the secondary and tertiary structures of 4 proteins using circular dichroism, Fourier-transform infrared spectroscopy, and fluorescence spectroscopy in order to analyze the composition and functional mechanisms. The results showed that the amino acid composition in all the 4 proteins was relatively balanced, and the essential amino acid content in albumin and globulin was high. The molecular weights of albumin, globulin, gliadin, and gluten were 19 to 21, 15 to 53, 20 to 38, and 10 to 90 kDa, respectively. The composition of gluten was a little complex compared to those of the other oat bran proteins. The secondary structure distribution of the 4 proteins differed, and increase in the pH resulted in modification of the β-sheet structure to α-helical structure. Moreover, the α-helix content and surface hydrophobicity were negatively correlated (r = -0.988, P globulin > gluten > albumin, indicating that surface hydrophobicity of gliadin was the strongest and that of albumin was the weakest among the 4 proteins.

Coeliac disease (CD) is an inflammatory disorder of the small intestine. It includes aberrant adaptive immunity with presentation of CD toxic gluten peptides by HLA-DQ2 or DQ8 molecules to gluten-sensitive T cells. A ω-gliadin/C-hordein peptide (QPFPQPEQPFPW) and a rye-derived secalin peptide (QPFPQPQQPIPQ) were proposed to be toxic in CD, as they yielded positive responses when assessed with peripheral blood T-cell clones derived from individuals with CD. We sought to assess the immunogenicity of the candidate peptides using gluten-sensitive T-cell lines obtained from CD small intestinal biopsies. We also sought to investigate the potential cross-reactivity of wheat gluten-sensitive T-cell lines with peptic-tryptic digested barley hordein (PTH) and rye secalin (PTS). Synthesised candidate peptides were deamidated with tissue transglutaminase (tTG). Gluten-sensitive T-cell lines were generated by culturing small intestinal biopsies from CD patients with peptic-tryptic gluten (PTG), PTH or PTS, along with autologous PBMCs for antigen presentation. The stimulation indices were determined by measuring the relative cellular proliferation via incorporation of 3 H-thymidine. The majority of T-cell lines reacted to the peptides studied. There was also cross-reactivity between wheat gluten-sensitive T-cell lines and the hordein, gliadin and secalin peptides. PTH, PTS, barley hordein and rye secalin-derived CD antigen-sensitive T-cell lines showed positive stimulation with PTG. ω-gliadin/C-hordein peptide and rye-derived peptide are immunogenic to gluten-sensitive T-cell lines and potentially present in wheat, rye and barley. Additional CD toxic peptides may be shared.

Epitopes on the α-gliadins are known to give rise to immune responses that may lead to the development of celiac disease in genetically predisposed individuals. The reduction of epitope levels in wheat-based products would likely benefit this group of consumers and also consumers with non-celiac gluten sensitivity. Conventional breeding of wheats with lowered epitope levels will take time, but in this study we show for the first time that milling technology can be used to produce flour mill streams that are depleted in α-20 gliadin epitopes. Fifteen mill streams from two New Zealand wheat cultivars, Sapphire (a biscuit wheat) and Monad (a bread wheat), were tested with reversed-phase HPLC and an α-20 gliadin epitope ELISA kit. The level of α-20 epitope measured in Sapphire gliadins was significantly less than that found in Monad gliadins, even taking into account differences in total protein content. For both cultivars, compared with the straight-run flour, the break flours had similar or significantly hi...