Exploring the Gut Microbiome and Metabolome in Individuals with Alopecia Areata Disease.

In recent years, heightened attention has been devoted to unravelling the intricate interplay between genetic and environmental factors shaping the gut microbiota and its significance for human health. This study delves into exploring the plausible connection between Alopecia Areata (AA), an autoimmune disease, and the dynamics of the gut microbiome. Examining a cohort of healthy adults and individuals with AA, both the gut microbiota composition and volatile organic compound (VOC) metabolites from faeces and urine were analysed. While overall microbiota composition showed no significant differences, intra-individual variability revealed distinctions related to age, gender, and pathology status, with AA individuals exhibiting reduced species richness and evenness. Differential abundance analysis identified microbial biomarkers for AA, notably Firmicutes, Lachnospirales, and Blautia, while Coprococcus stood out for healthy individuals. The Data Integration Analysis for Biomarker discovery using Latent Components (DIABLO) method further supported these findings including metabolite biomarkers, such as esters of branched chain fatty acids and branched chain amino acids as predictors for AA, suggesting potential links to oxidative stress. Despite certain limitations, the study highlights the complexity of the gut microbiome and its metabolites in the context of AA, while the biomarkers identified could be useful starting points for upcoming studies.

The Effect of Hydrolyzed and Fermented Arabinoxylan-Oligo Saccharides (AXOS) Intake on the Middle-Term Gut Microbiome Modulation and Its Metabolic Answer.

Although fermentation and hydrolyzation are well-known processes to improve the bioavailability of nutrients and enable the fortification with dietary fibers, the effect of such pre-treatments on the prebiotic features of arabinoxylan-oligosaccharides (AXOS) had not been explored. The middle-term in vitro simulation through the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) demonstrated that the feeding with different formulations (namely oat bran, rye bran and wheat bran) containing hydrolyzed AXOS fermented by lactic acid bacteria significantly increased the synthesis of short-chain fatty acids (SCFA) by colon microbiota, with hydrolyzed and fermented rye bran displaying the highest effect. After two weeks from the interruption of intake, SCFA concentrations significantly decreased but remained still significantly higher compared to the original condition. The microbiome was also affected, with a significant abundance increase in Lactobacillaceae taxon after feeding with all fermented and hydrolyzed formulates. Hydrolyzed and fermented rye bran showed the highest changes. The fungal community, even if it had a lower variety compared to bacteria, was also modulated after feeding with AXOS formulations, with an increase in Candida relative abundance and a decrease in Issatchenkia. On the contrary, the intake of non-hydrolyzed and non-fermented wheat bran did not produce relevant changes of relative abundances. After two weeks from intake interruption (wash out period) such changes were mitigated, and the gut microbiome modulated again to a final structure that was more like the original condition. This finding suggests that hydrolyzed AXOS fermented by lactic acid bacteria could have a more powerful prebiotic effect compared to non-hydrolyzed and non-fermented wheat bran, shaping the colon microbiome and its metabolic answer. However, the intake should be continuous to assure persistent effects. Opening a window into the ecological evolutions and plausible underlying mechanisms, the findings reinforce the perspective to explore more in depth the use of hydrolyzed and fermented AXOS as additional ingredient for bread fortification.

How cereal flours, starters, enzymes, and process parameters affect the in vitro digestibility of sourdough bread.

Digestibility of leavened baked goods relies on multiple factors: starch bioavailability and protein hydrolysis during food processing and digestion, presence of antinutritional factors, and satiety and gastrointestinal symptoms after intake. Several studies highlighted that bread digestibility might be positively affected by long-time sourdough fermentation. Nevertheless, most research is focused on single factors and their effect on digestibility, excluding the potential complementary effects of more than one factor. In this work, a multitude of factors influencing the the in vitro starch and protein digestibility and predicted glycemic index were assessed simultaneously. Forty-six different breads made with various raw material/ingredients (flour, enzymes, lactic acid bacteria cytoplasmic extracts and gluten), type of sourdoughs (fresh or commercial liquid or dried), strains of lactic acid bacteria and yeasts, and time and temperature of fermentation, were preliminarily investigated. Further selection of optimal conditions was based on statistical analysis and final breads were further characterized for their peptide profiles, total free amino acids and quality indexes of the digestible protein fraction. Among the factors considered, results identified a claimed in vitro digestibility for breads made with whole wheat, spelt, and rye flours, obtained with lactic acid bacteria strains selected for the peptidase activity, added of fungal proteases, and fermented at the optimal temperature of 37 °C.

Feeding with Sustainably Sourdough Bread Has the Potential to Promote the Healthy Microbiota Metabolism at the Colon Level.

The contribution of sustainably food processing to healthy intestinal microbial functions is of recent acquisition. The sourdough fermentation fits well with the most sustainable bread making. We manufactured baker's yeast (BYB) and sourdough (t-SB30) breads, which first underwent to an in-depth characterization. According to nutritional questionnaires, we selected 40 volunteers adhering to the Mediterranean diet. Data on their fecal microbiota and metabolome allowed the selection of two highly representative fecal donors to separately run the Twin Mucosal-SHIME (Twin M-SHIME) under 2-week feeding with BYB and t-SB30. Bread feeding did not affect the microbial composition at phylum and family levels of both donors, in all Twin M-SHIME colon tracts, and lumen and mucosal compartments. The genus core microbiota showed few significant fluctuations, which regarded the relative abundances of Lactobacillus and Leuconostoc according to feeding with BYB and t-SB30, respectively. Compared with BYB, the content of all short chain fatty acids (SCFA), and isovaleric and 2-methylbutyric acids significantly increased with t-SB30 feeding. This was evident for all Twin M-SHIME colon tracts and both donors. The same was found for the content of Asp, Thr, Glu, GABA, and Orn. The bread characterization made possible to identify the main features responsible for this metabolic response. Compared with BYB, t-SB30 had much higher contents of resistant starch, peptides, and free amino acids, and an inhomogeneous microstructure. We used the most efficient approach to investigate a staple food component, excluding interferences from other dietary factors and attenuating human physiology overlaps. The daily consumption of sourdough bread may promote the healthy microbiota metabolism at colon level. IMPORTANCE Knowledge on environmental factors, which may compose the gut microbiota, and drive the host physiology and health is of paramount importance. Human dietary habits and food compositions are pivotal drivers to assemble the human gut microbiota, but, inevitably, unmapped for many diet components, which are poorly investigated individually. Approximately 30% of the human diet consists of fermented foods and beverages. Bread, a fermented/leavened food, is a basic component of the human diet. Its potential effect on gut microbiota composition and functionality is challenging. In this study, we industrially made baker's yeast and sourdough breads, which were used to feed the Twin Mucosal-SHIME, a worldwide scientifically validated gastrointestinal simulator. Only the consumption of sourdough bread has the potential to enhance the synthesis of short chain fatty acids and free amino acids at the colon level.

Role of Lactic Acid Bacteria Phospho-ß-Glucosidases during the Fermentation of Cereal by-Products.

Bioprocessing using lactic acid bacteria (LAB) is a powerful means to exploit plant-derived by-products as a food ingredient. LAB have the capability to metabolize a large variety of carbohydrates, but such metabolism only relies on few metabolic routes, conferring on them a high fermentation potential. One example of these pathways is that involving phospho-ß-glucosidase genes, which are present in high redundancy within LAB genomes. This enzymatic activity undertakes an ambivalent role during fermentation of plant-based foods related to the release of a wide range of phenolic compounds, from their ß-D-glycosylated precursors and the degradation of ß-glucopyranosyl derived carbohydrates. We proposed a novel phenomic approach to characterize the metabolism drift of Lactiplantibacillus plantarum and Leuconostoc pseudomesenteroides caused by a lignocellulosic by-product, such as the brewers' spent grain (BSG), in contrast to Rich De Man, Rogosa and Sharpe (MRS) broth. We observed an increased metabolic activity for gentiobiose, cellobiose and ß-glucoside conjugates of phenolic compounds during BSG fermentation. Gene expression analysis confirmed the importance of cellobiose metabolism while a release of lignin-derived aglycones was found during BSG fermentation. We provided a comprehensive view of the important role exerted by LAB 6-phospho-ß-glucosidases as well the major metabolic routes undertaken during plant-based fermentations. Further challenges will consider a controlled characterization of pbg gene expression correlated to the metabolism of ß-glucosides with different aglycone moieties.

Biotechnological re-cycling of apple by-products: A reservoir model to produce a dietary supplement fortified with biogenic phenolic compounds.

This study is an example of apple by-products (AP) recycling through a designed fermentation by selected autochthonous Lactobacillus plantarum AFI5 and Lactobacillus fabifermentans ALI6 used singly or as binary cultures with the selected Saccharomyces cerevisiae AYI7. Compared to Raw-, Unstarted- and Chemically Acidified-AP, Fermented-AP promoted the highest levels of total and insoluble dietary fibers, DPPH scavenging capacity, and free phenolics. The binary culture of L. plantarum AFI5 and S. cerevisiae AYI7 had the best effect on the bioavailability phenolic compounds as resulted by the LC-MS/MS validated method. The accumulation of phenolic acids derivatives highlighted the microbial metabolism during AP fermentation. Bio-converted phenolics were likely responsible for the increased DPPH scavenging capacity. The potential health-promoting effects of Fermented-AP were highlighted using Caco-2 cells. With variations among single and binary cultures, fermented-AP counteracted the inflammatory processes and the effects of oxidative stress in Caco-2 cells, and preserved the integrity of tight junctions.

LC-MS untargeted approach showed that methyl jasmonate application on Vitis labrusca L. grapes increases phenolics at subtropical Brazilian regions.

INTRODUCTION: Vitis labrusca L. grapes are largely cultivated in Brazil, but the tropical climate negatively affects the phenols content, especially anthocyanin. According to the projections of the incoming climatic changes, the climate of several viticulture zone might change to tropical. Therefore, researches are focusing on increasing grape phenols content; with methyl jasmonate application (MeJa) is considered a good alternative. OBJECTIVES: The aim was to investigate with an untargeted approach the metabolic changes caused by the MeJa pre-harvest application on two Vitis labrusca L. cultivars grapes, both of them grown in two Brazilian regions. METHODS: Isabel Precoce and Concord grapes cultivated under subtropical climate, in the south and southeast of Brazil, received MeJa pre-harvest treatment. Grape metabolome was extracted and analyzed with a MS based metabolomics protocol by UPLC-HRMS-QTOF. RESULTS: Unsupervised data analysis revealed a clear separation between the two regions and the two cultivars, while supervised data analysis revealed biomarkers between the MeJa treatment group and the control group. Among the metabolites positively affected by MeJa were (a) flavonoids with a high degree of methylation at the B-ring (malvidin and peonidin derivatives and isorhamentin) for Isabel Precoce grapes; (b) glucosides of hydroxycinnamates, gallocatechin, epigallocatechin and cis-piceid for Concord grapes; and (c) hydroxycinnamates esters with tartaric acid, and procyanidins for the Southeast region grapes. CONCLUSION: These results suggest that MeJa can be used as elicitor to secondary metabolism in grapes grown even under subtropical climate, affecting phenolic biosynthesis.

Lipid Profiling and Stable Isotopic Data Analysis for Differentiation of Extra Virgin Olive Oils Based on Their Origin.

To differentiate extra virgin olive oils (EVOO) according to the origin of purchase, such as monocultivar Italian EVOO with protected denomination of origin (PDO) and commercially-blended EVOO purchased in supermarkets, a number of samples was subjected to the analysis of various lipid species by liquid chromatography/mass spectrometry (LC-ESI-MS/MS, LC-ESI-IT-MS) and proton nuclear magnetic resonance analysis (1H-NMR). Many putative chemical markers were extracted as differentiators by uni- and multivariate statistical analysis. Commercially-blended EVOO contained higher concentrations of the majority of minor lipids, including free fatty acids, their alkyl (methyl and ethyl) esters, monoglycerides, and diglycerides, which may be indicative of a higher degree of triglyceride lipolysis in these than in monocultivar PDO EVOO. Triterpenoids and particular TAG species were also found in higher proportions in the samples from the commercially-blended EVOO class, suggesting a possible influence of factors such as the cultivar and geographical origin. The largest differences between the classes were determined for the concentrations of uvaol and oleanolic acid. The results of the analysis by isotopic ratio mass spectrometry (IRMS) were reasonably consistent with the information about the geographical origin declared on the labels of the investigated EVOOs, showing considerable variability, which possibly also contributed to the differences in lipid composition observed between the two investigated classes of EVOO.

LC-MS/MS analysis of free fatty acid composition and other lipids in skins and seeds of Vitis vinifera grape cultivars.

Lipids are important constituents of plant and animal cells, being essential due to their biological functions. Despite the significant role of these compounds in grape berries, knowledge of grape lipid composition is still limited. This study addresses the free fatty acid composition and other lipids in skins and seeds of several Vitis vinifera grape cultivars using LC-MS/MS. A different profile of free fatty acids was determined in the grape tissues, showing a higher proportion of unsaturated fatty acids in seeds (ca. 60%). Lignoceric acid was one of the main free saturated fatty acids found in grape skins, together with palmitic and stearic acids. Berry skins showed similar ranges of C18-unsaturated fatty acids, whereas linoleic acid was predominant in the seed fatty acid composition. Higher content of glycerophospholipids was determined in grape seeds. Uvaol and oleanolic acid were only quantified in skins (1.5-3.9 and 38.6-57.6 mg/kg fresh weight, respectively). These preliminary results suggest a certain diversity in grape lipids according to their location in the berry tissues and cultivar, providing useful information for the fermentation process due to their role in wine sensory profile and yeast growth.

Complementary Untargeted and Targeted Metabolomics for Differentiation of Extra Virgin Olive Oils of Different Origin of Purchase Based on Volatile and Phenolic Composition and Sensory Quality.

In order to differentiate the extra virgin olive oils (EVOO) of different origin of purchase, such as monovarietal Italian EVOO with protected denomination of origin (PDO) and commercial-blended EVOO purchased in supermarkets, a number of samples was subjected to the analysis of volatile aroma compounds by both targeted gas chromatography/mass spectrometry (GC-MS) and untargeted profiling by comprehensive two-dimensional gas chromatography/mass spectrometry (GC×GC-TOF-MS), analysis of phenols by targeted high-performance liquid chromatography/mass spectrometry (HPLC-DAD-ESI/MS), and quantitative descriptive sensory analysis. Monovarietal PDO EVOOs were characterized by notably higher amounts of positive LOX-derived C6 and C5 volatile compounds, which corresponded to the higher intensities of all the assessed positive fruity and green odor sensory attributes. Commercial-blended EVOOs had larger quantities of generally undesirable esters, alcohols, acids, and aldehydes, which coincided with the occurrence of sensory defects in many samples. Many minor volatile compounds that were identified by GC×GC-TOF-MS were found to differentiate each of the two investigated groups. The differences between the groups with respect to phenols and taste characteristics were evident, but less pronounced. The results that were obtained in this study have undoubtedly confirmed the existence of the large heterogeneity of oils that are sold declared as EVOO. It was shown that GC-MS, GC×GC-TOF-MS, and HPLC-DAD-ESI/MS analyses have complementary outputs, and that their use in combination has advantages in supporting the results of sensory analysis and objectively differentiating these groups of EVOO.