Impact of semolina-barley mixture on the volatolomic profile of dough and pasta: characterization by a multivariate chemometric approach.

BACKGROUND: Barley flour, known to be rich in various phytochemicals, has been demonstrated to improve the technological and nutritional properties of pasta; however, its volatile profile, on which its aromatic properties depend, also plays an important role in the acceptance of barley-enriched pasta. In the present work, volatile organic compounds (VOCs) of semolina doughs enriched with different percentages of barley and of the related pasta were characterized by solid phase micro-extraction (HS-SPME) coupled to gas-chromatography/mass spectrometry (GC-MS), and evaluated using a multivariate statistical approach, including principal component analysis (PCA), cluster heatmaps, Pearson's and Spearman's correlations, and partial least squares correlation (PLSC). RESULTS: The effects of single raw materials, and their interactions, were studied to establish their importance in the volatile profile of the samples, and the correlation between the dough VOCs and the processed product VOCs was assessed. The presence of barley flour markedly affected the volatile profile in comparison with the dough obtained with only durum wheat. For alcohols, esters, terpenes, and some aldehydes there was a clear correlation with the percentage of barley. For some of the VOCs, on the other hand, a strong dependence on the ingredients interaction effect due to the mixing stage has been demonstrated. CONCLUSION: The heatmaps allowed a good graphical visualization of the relationship between molecules and barley percentage, offering the possibility to select the best one according to the desired volatolomic footprint. Pasta with 40% of barley was demonstrated to give pasta with the most complex volatile profile. © 2024 Society of Chemical Industry.

A New Intraepithelial γδ T-Lymphocyte Marker for Celiac Disease Classification in Formalin-Fixed Paraffin-Embedded (FFPE) Duodenal Biopsies.

BACKGROUND: The histopathologic diagnosis of celiac disease (CD) may be challenging when the duodenal biopsies mucosal injury is limited. Intraepithelial T-lymphocytes (IELs) can be useful to characterize the degree of mucosal inflammation. A small fraction of IELs expresses the γδ T-cell receptor (named γδ-IELs), whose density, determined by flow cytometry or frozen section immunohistochemistry (IHC), is a specific marker for CD. AIM: To establish a new IHC assay for γδ-IELs applicable to formalin-fixed paraffin-embedded (FFPE) duodenal biopsies. METHODS: We analyzed γδ-IELs using IHC in 138 duodenal biopsies using a standard IHC staining protocol with a new monoclonal antibody H-41. IELs were quantitated with digital image analysis. RESULTS: Compared to those in non-celiac controls (n = 51), γδ-IEL density was significantly increased in newly diagnosed celiac disease patients (n = 22, p < 0.0001). In ROC-curve analysis, the cutoff of 6.5 γδ-IELs/100 enterocytes distinguished optimally active CD patients from non-celiac controls (sensitivity 96%, specificity 95%). γδ-IEL density in CD patients on a gluten-free diet (n = 53) were also higher than in controls (p < 0.0001), but lower than those in newly diagnosed CD (p < 0.0001). The diagnostic value of γδ-IELs outperformed that of CD3 + IELs in both patient groups. γδ-IELs were better than CD3 + IELs distinguishing between celiac disease and conditions histologically mimicking celiac disease (n = 12). CONCLUSIONS: Intraepithelial γδ T-lymphocytes can be stained and quantitated reliably in FFPE duodenal biopsies. The results showed excellent specificity and sensitivity for celiac disease. The new IHC method of detection of γδ-IELs is a promising addition to the routine histopathologic assessment methodology of celiac disease.

Effect of the Addition of Freeze-Dried Grape Pomace on Fresh Tagliatelle Gluten Network and Relationship to Sensory and Chemical Quality.

The incorporation of 5 and 10% freeze-dried grape pomace powder (GPP) in fresh tagliatelle pasta preparation was evaluated for its effect on chemical composition, gluten protein structure, and sensory properties. The addition of the freeze-dried GPP led to a significant increase (p < 0.05) in polyphenol content in the raw and cooked fortified pasta samples with respect to 100% semolina pasta, although the phenolic content decreased after the cooking process. The opposite phenomenon was observed with the antioxidant activity, which increased significantly (p < 0.05) when switching from raw to cooked pasta samples fortified with GPP. The formation of a proper gluten structure was found in the fortified raw pasta, even if a change in the protein arrangement was shown in the fortified cooked samples, confirmed by a significant reduction (p < 0.05) in both the unextractable polymeric protein percentage (% UPP) and disulfide bond (S-S) formation. These results suggest a possible interaction between the protein sulfhydryl groups (-Cys) and polyphenols of grape pomace during cooking through non-disulfide covalent bonds, which was confirmed by the significant (p < 0.05) decrease in the -SH groups when comparing 100% semolina pasta with fortified pasta sample. Finally, a sensory analysis showed that the highest significant score (p < 0.05) was achieved by the 5% GP-fresh pasta sample.

Artificial aging of Uva di Troia and Primitivo wines using oak chips inoculated with Penicillium purpurogenum.

BACKGROUND: Two red wines (Primitivo and Uva di Troia) treated with oak chips inoculated with Penicillium purpurogenum were analysed in order to assess their contents of furfural, cis-ß-methyl-γ-octalactone, syringol, eugenol, vanillin and 4-vinylguaiacol. Two different sizes of oak chips (small and big, of length 2 and 8 mm respectively) and two different degrees of toasting (low and high) were used in the study. Aroma compounds were analysed by gas chromatography/mass spectrometry to determine differences among samples after 15 days of chip contact time. RESULTS: Big oak chips inoculated with P. purpurogenum increased the level of 4-vinylguaiacol, while small oak chips inoculated with P. purpurogenum, in some conditions, increased the level of eugenol. Chip size and degree of toasting also played an important role in the content of eugenol. CONCLUSION: The use of oak chips inoculated with mould might be a promising alternative to barrel aging. Moreover, different fungal inocula could contribute to the enrichment of wine with specific compounds (e.g. 4-vinylguaiacol and eugenol).

Adherence to Gluten-Free Diet Restores Alpha Diversity in Celiac People but the Microbiome Composition Is Different to Healthy People.

Celiac disease (CD) is an autoimmune disease with the destruction of small intestinal villi, which occurs in genetically predisposed individuals. At the present moment, a gluten-free diet (GFD) is the only way to restore the functionality of gut mucosa. However, there is an open debate on the effects of long-term supplementation through a GFD, because some authors report an unbalance in microbial taxa composition. METHODS: For microbiome analysis, fecal specimens were collected from 46 CD individuals in GFD for at least 2 years and 30 specimens from the healthy controls (HC). Data were analyzed using an ensemble of software packages: QIIME2, Coda-lasso, Clr-lasso, Selbal, PICRUSt2, ALDEx2, dissimilarity-overlap analysis, and dysbiosis detection tests. RESULTS: The adherence to GFD restored the alpha biodiversity of the gut microbiota in celiac people but microbial composition at beta diversity resulted as different to HC. The microbial composition of the CD subjects was decreased in a number of taxa, namely Bifidobacterium longum and several belonging to Lachnospiraceae family, whereas Bacteroides genus was found to be more abundant. Predicted metabolic pathways among the CD bacterial communities revealed an important role in tetrapyrrole biosynthesis. CONCLUSIONS: CD patients in GFD had a non-dysbiotic microbial composition for the crude alpha diversity metrics. We found significant differences in beta diversity, in certain taxon, and pathways between subjects with inactive CD in GFD and controls. Collectively, our data may suggest the development of new GFD products by modulating the gut microbiota through diet, supplements of vitamins, and the addition of specific prebiotics.

Healthy and pro-inflammatory gut ecology plays a crucial role in the digestion and tolerance of a novel Gluten Friendly™ bread in celiac subjects: a randomized, double blind, placebo control in vivo study.

Gluten Friendly™ (GF) is a new gluten achieved through a physicochemical process applied to wheat kernels. The goal of this research was to assess the in vivo effects of Gluten Friendly™ bread on celiac gut mucosa and microbiota. In a double-blind placebo-controlled intervention study, 48 celiac disease (CD) patients were randomized into 3 groups to eat 100 g of bread daily, containing different doses (0; 3 g; 6 g) of GF for 12 weeks. The small-bowel morphology (VH/CrD), intraepithelial densities of CD3+, celiac serology, MUC2, CB1, gut permeability, proinflammatory cytokines, gluten in stools, symptoms, and gut microbial composition were assessed. All 48 CD subjects experienced no symptoms. K-means analysis evidenced celiac subjects clustering around unknown parameters independent of GF dosage: K1 35%; K2 30%; K3 35%. VH/CrD significantly decreased in K1 and K2. VH/CrD did not correlate with IEL increase in K2. 33-mer was not detected in 47% and 73% of patients in both K1 and K2, respectively. VH/CrD and IEL did not change significantly and strongly correlated with the absence of 33-mer in K3. Inflammation and VH/CrD decrease are strongly related with the presence of proinflammatory species at the baseline. A boost in probiotic, butyrate-producing genera, is strongly related with GF tolerance at the end of the trial. Our research suggests that a healthy and proinflammatory ecology could play a crucial role in the digestion and tolerance of the new gluten molecule in celiac subjects. However, GF can be completely digested by gut microbiota of CD subjects and shapes it toward gut homeostasis by boosting healthy butyrate-producing populations. The clinical trial registry number is NCT03137862 (https://clinicaltrials.gov).

Characterizing the Rheological and Bread-Making Properties of Wheat Flour Treated by "Gluten FriendlyTM" Technology.

After discovering an innovative technology for the reshaping of gluten proteins-the "Gluten FriendlyTM" system-that confers to wheat flour some unprecedented characteristics, such as reduced epitope antigenicity and a positive modulation of the gut microbiota, its effects on the production and quality of bread have been studied. Mainly, we have investigated the chemical, rheological and pasting properties of Gluten Friendly Flour (GFF) and of control flour (CF) with the aim of analyzing and interpreting potential differences. Furthermore, the bread made from GFF and CF was evaluated in terms of microstructure properties and sensory quality. The experiments demonstrated that GFF became soluble in aqueous solution, making it unfeasible to isolate using the Glutomatic apparatus. Although the water absorption of GFF increased by 10% compared to CF, dough elasticity was reduced, and dough stability decreased from 5 to 2 min. A significant increase in the alveograph index (P/L) from 0.63 to 6.31 was detected, whereas pasting properties did not change from the control flour. Despite these profound modifications in the rheological properties, GFF exhibited a high ability to shape dough and to produce bread with high quality and negligible differences from the control bread in terms of appearance, taste, aroma, color and texture.

A Preliminary Report on the Use of the Design of Experiments for the Production of a Synbiotic Yogurt Supplemented With Gluten FriendlyTM Flour and Bifidobacterium infantis.

The main goal of this paper was to design a synbiotic yogurt containing Bifidobacterium infantis and Gluten Friendly FlourTM; the proposed approach relies upon milk fermentation through the classical starter of yogurt (Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus) to avoid a strong production of acetic acid by bifidobacterial and inoculum of B. infantis after the fermentation. The research was divided in 3 steps. The aim of the first step was the optimization of fermentation kinetic by L. delbureckii and S. thermophilus, by combining the amount of flour (either Gluten Friendly Flour-GF- or Control Flour-CF) in milk, temperature and inoculum level; the factors were combined through a mixture design. As a result of this step, the best combination was pointed out: flour at 2.5 g/l; L. delbrueckii subsp. bulgaricus at 6 log cfu/ml; temperature at 37-40°C. The goal of the second step was to study the effect of flour (2.5 g/l) on the viability of B. infantis. GF prolonged the viability of the probiotic for 14 days. In the last step, a synbiotic yogurt, supplemented with GF and fermented with L. delbureckii and S. thermophilus, and then inoculated with B. infantis, was produced. The product was stored at 8 and 15°C. A positive effect of GF was found at 15°C, with B. infantis at 7.0 log cfu/g in GF sample and 5.5.5.7 log cfu/g in CF sample.

The Impact of Gluten Friendly Flour on the Functionality of an Active Drink: Viability of Lactobacillus acidophilus in a Fermented Milk.

The Gluten FriendlyTM Technology is an innovative method that induces structural changes in gluten proteins. In this paper a synbiotic fermented milk, containing Lactobacillus acidophilus La-5 and Gluten Friendly Flour (GFF), was proposed. A mixture design was used to combine flour, temperature and probiotic to study the effects of these variables on the acidification. The experiments were done on both GFF and control flour (CF). Thus, the following conditions were chosen to produce the fermented milk: L. acidophilus at 6.5 log cfu/ml; flour at 2.5 g/l; temperature at 37°C. Then, the fermented milk was produced and stored at 4°C for 90 days. The most important result was the positive effect of GFF on the viability of the probiotic, with a prolongation of the shoulder length to 20 days (12-13 days in the control). Moreover, GFF did not act on the sensory scores and on the physico-chemical parameters.

An In Vitro Fermentation Study on the Effects of Gluten FriendlyTM Bread on Microbiota and Short Chain Fatty Acids of Fecal Samples from Healthy and Celiac Subjects.

Recently, an innovative gluten detoxification method called Gluten FriendlyTM (GF) has been developed. It induces structural modifications, which abolish the antigenic capacity of gluten and reduce the in vitro immunogenicity of the most common epitopes involved in celiac disease, without compromising the nutritional and technological properties. This study investigated the in vitro effects of GF bread (GFB) on the fecal microbiota from healthy and celiac individuals by a three-stage continuous fermentative system, which simulates the colon (vessel 1, proximal colon; vessel 2, transverse colon; and vessel 3, distal colon), as well as on the production of short chain fatty acids (SCFA, acetate, propionate, butyrate). The system was fed with GFB and the changes in microbiota through fluorescence in situ hybridization and in SCFA content were assessed. GFB exerted beneficial modulations such as bifidogenic effects in each compartment of the model both with healthy- and celiac-derived samples, as well as growth in Clostridium clusters XIVa+b in celiac-derived samples. Furthermore, increased levels of acetic acid were found in vessel 1 inoculated with the fecal microbiota of healthy individuals, as well as acetic and propionic in vessel 1 and 2 with celiac-derived samples. In addition, the use of multivariate approaches showed that the supplementation of GFB could result in a different modulation of the fecal microbiota and SCFA, as a function of initial equilibrium.

Changes in wheat kernel proteins induced by microwave treatment.

Wheat kernels were subjected to microwave treatment, and the proteins were characterized by size exclusion high-performance liquid chromatography (SE-HPLC) and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Using this process, the proteins polymerize, forming intermolecular bonds among the same classes of proteins. Furthermore, the polymerization occurs only through disulphide bonds. Although SDS-PAGE did not show any differences for either the number or intensity of protein bands between flour samples before and after microwave treatment, gliadins from treated flours showed significantly reduced cross-reactivity with the R5 antibody. Moreover, the gluten became soluble in an aqueous saline solution, and it was not possible to isolate it using the Glutomatic apparatus. However, the treated flour, in the presence of water, was able to form dough and leaven and produce bread.

Impact of Gluten-Friendly Bread on the Metabolism and Function of In Vitro Gut Microbiota in Healthy Human and Coeliac Subjects.

The main aim of this paper was to assess the in vitro response of healthy and coeliac human faecal microbiota to gluten-friendly bread (GFB). Thus, GFB and control bread (CB) were fermented with faecal microbiota in pH-controlled batch cultures. The effects on the major groups of microbiota were monitored over 48 h incubations by fluorescence in situ hybridisation. Short-chain fatty acids (SCFAs) were measured by high-performance liquid chromatography (HPLC). Furthermore, the death kinetics of Lactobacillus acidophilus, Bifidobacterium animalis subsp. lactis, Staphylococcus aureus, and Salmonella Typhimurium in a saline solution supplemented with GFB or CB were also assessed. The experiments in saline solution pinpointed that GFB prolonged the survival of L. acidophilus and exerted an antibacterial effect towards S. aureus and S. Typhimurium. Moreover, GFB modulated the intestinal microbiota in vitro, promoting changes in lactobacilli and bifidobacteria members in coeliac subjects. A final multivariate approach combining both viable counts and metabolites suggested that GFB could beneficially modulate the coeliac gut microbiome; however, human studies are needed to prove its efficacy.

A proteomic approach to study protein variation in GM durum wheat in relation to technological properties of semolina.

Genetic manipulation of durum wheats by tobacco rab-1 genes influence the trafficking of gluten proteins through the secretory system by up- or down-regulating the transport step from the ER to the Golgi apparatus which may in turn modify functional performance of the grain. Gluten proteins were extracted from two genetically manipulated lines - Svevo B730 1-1 and Ofanto B688 1-2 - and their control lines and were analyzed by two dimensional gel electrophoresis. When the two-dimensional maps were compared by image analysis no significant differences between the GM line with an up-regulated trafficking containing the wild type tobacco rab1 (Svevo B730 1-1) and its control (Svevo control). By contrast, significant differences were found between the GM line with a down-regulated trafficking due to the tobacco rab1 mutant form (Ofanto B688 1-2) and its control (Ofanto control). Of the new protein spots detected in the down-regulated Ofanto B688 1-2 map, only a beta-amylase was identified. The remaining spots were susceptible to chymotripsin action but not to trypsin one, as in the case of the gluten protein. Rheological measurements showed that gluten quality was enhanced in the down-regulated Ofanto B688 1-2 without an increase in the amount of gluten. Proteomics is a useful and powerful tool for investigating protein changes in GMOs and in understanding events in food science and technology.

Protein modifications in cooked pork products investigated by a proteomic approach.

To evaluate process-induced protein modifications in cooked ham and emulsion sausages, the proteomes of whole-cut (Parma and "Praga" cooked hams) and comminuted pork (mortadella and würstel) products were compared to raw pork using two-dimensional gel electrophoresis (2-DE) coupled to image analysis and mass spectrometry (MS). Other than heat-induced breakdown of part of the myosin heavy chains, the 2-DE pattern of cooked ham was substantially similar to that of raw pork. However, the MS-based analysis showed minor modifications, including the extensive oxidation of methionines. In contrast, likely due to emulsification, comminuted sausages were characterized by an abundant insoluble protein fraction (IPF). Interestingly, tropomyosin and myosin light chains in comminuted sausages were exclusively found in the IPF. Our results indicate that the protein aggregation systems of cooked hams and emulsion sausages reflect the processing conditions and are definitely different, the former being characterized mainly by disulphide bridges and the latter by additional covalent inter-protein links.

Cereal-based gluten-free food: how to reconcile nutritional and technological properties of wheat proteins with safety for celiac disease patients.

The gluten-free diet is, to date, the only efficacious treatment for patients with Celiac Disease. In recent years, the impressive rise of Celiac Disease incidence, dramatically prompted changes in the dietary habit of an increasingly large population, with a rise in demand of gluten-free products. The formulation of gluten-free bakery products presents a formidable challenge to cereal technologists. As wheat gluten contributes to the formation of a strong protein network, that confers visco-elasticity to the dough and allows the wheat flour to be processed into a wide range of products, the preparation of cereal-based gluten-free products is a somehow difficult process. This review focuses on nutritional and technological quality of products made with gluten-free cereals available on the market. The possibility of using flour from naturally low toxic ancient wheat species or detoxified wheat for the diet of celiacs is also discussed.