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This work implemented a non-invasive volatile organic compounds (VOCs) monitoring approach to study how food components are metabolised by the gut microbiota in-vitro. The fermentability of a model food matrix rich in dietary fibre (oat bran), and a pure prebiotic (inulin), added to a minimal gut medium was compared by looking at global changes in the volatilome. The substrates were incubated with a stabilised human faecal inoculum over a 24-h period, and VOCs were monitored without interfering with biological processes. The fermentation was performed in nitrogen-filled vials, with controlled temperature, and tracked by automated headspace-solid-phase microextraction coupled with gas chromatography-mass spectrometry. To understand the molecular patterns over time, we applied a multivariate longitudinal statistical framework: repeated measurements-ANOVA simultaneous component analysis. The methodology was able to discriminate the studied groups by looking at VOCs temporal profiles. The volatilome showed a time-dependency that was more distinct after 12 h. Short to medium-chain fatty acids showed increased peak intensities, mainly for oat bran and for inulin, but with different kinetics. At the same time, alcohols, aldehydes, and esters showed distinct trends with discriminatory power. The proposed approach can be applied to study the intertwined pathways of gut microbiota food components interaction in-vitro.
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Fezes , Microbioma Gastrointestinal , Compostos Orgânicos Voláteis , Compostos Orgânicos Voláteis/análise , Compostos Orgânicos Voláteis/metabolismo , Humanos , Microbioma Gastrointestinal/fisiologia , Fezes/microbiologia , Fezes/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Fermentação , Fibras na Dieta/metabolismo , Inulina/metabolismo , Microextração em Fase Sólida/métodosRESUMO
The popularity of plant-based meat alternatives (PBMAs) has sparked a contentious debate about their influence on intestinal homeostasis compared to traditional animal-based meats. This study aims to explore the changes in gut microbial metabolites (GMMs) induced by the gut microbiota on different digested patties: beef meat and pea-protein PBMA. After digesting in vitro, untargeted metabolomics revealed 32 annotated metabolites, such as carnitine and acylcarnitines correlated with beef meat, and 45 annotated metabolites, like triterpenoids and lignans, linked to our PBMA. Secondly, (un)targeted approaches highlighted differences in GMM patterns during colonic fermentations. Our findings underscore significant differences in amino acids and their derivatives. Beef protein fermentation resulted in higher production of methyl-histidine, gamma-glutamyl amino acids, indoles, isobutyric and isovaleric acids. In contrast, PBMAs exhibit a significant release of N-acyl amino acids and unique dipeptides, like phenylalanine-arginine. This research offers valuable insights into how PBMAs and animal-based proteins differently modulate intestinal microenvironments.
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Microbioma Gastrointestinal , Metabolômica , Animais , Bovinos/metabolismo , Bactérias/metabolismo , Bactérias/classificação , Bactérias/genética , Pisum sativum/metabolismo , Pisum sativum/química , Pisum sativum/microbiologia , Fermentação , Proteínas de Plantas/metabolismo , Humanos , Aminoácidos/metabolismo , Aminoácidos/análise , Modelos Biológicos , Carne/análiseRESUMO
This study explored differences in microbial lipid metabolites among sunflower seeds, soybeans, and walnuts. The matrices were subjected to in vitro digestion and colonic fermentation. Defatted digested materials and fiber/phenolics extracted therefrom were added to sunflower oil (SO) and also fermented. Targeted and untargeted lipidomics were employed to monitor and tentatively identify linoleic acid (LA) metabolites. Walnut fermentation produced the highest free fatty acids (FFAs), LA, and conjugated LAs (CLAs). Defatted digested walnuts added to SO boosted FFAs and CLAs production; the addition of fibre boosted CLAs, whereas the addition of phenolics only increased 9e,11z-CLA and 10e,12z-CLA. Several di-/tri-hydroxy-C18-FAs, reported as microbial LA metabolites for the first time, were annotated. Permutational multivariate analysis of variance indicated significant impacts of food matrix presence and type on lipidomics and C18-FAs. Our findings highlight how the food matrices affect CLA production from dietary lipids, emphasizing the role of food context in microbial lipid metabolism.
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Microbioma Gastrointestinal , Juglans , Fermentação , Nozes , Gorduras na Dieta , Ácidos Graxos não Esterificados , Ácido Linoleico , Fenóis , Óleo de Girassol , ColoRESUMO
The pathogenesis of non-alcoholic fatty liver disease (NAFLD) is associated with abnormalities of liver lipid metabolism. On the contrary, a diet enriched with n-3 polyunsaturated fatty acids (n-3-PUFAs) has been reported to ameliorate the progression of NAFLD. The aim of our study was to investigate the impact of dietary n-3-PUFA enrichment on the development of NAFLD and liver lipidome. Mice were fed for 6 weeks either a high-fat methionine choline-deficient diet (MCD) or standard chow with or without n-3-PUFAs. Liver histology, serum biochemistry, detailed plasma and liver lipidomic analyses, and genome-wide transcriptome analysis were performed. Mice fed an MCD developed histopathological changes characteristic of NAFLD, and these changes were ameliorated with n-3-PUFAs. Simultaneously, n-3-PUFAs decreased serum triacylglycerol and cholesterol concentrations as well as ALT and AST activities. N-3-PUFAs decreased serum concentrations of saturated and monounsaturated free fatty acids (FAs), while increasing serum concentrations of long-chain PUFAs. Furthermore, in the liver, the MCD significantly increased the hepatic triacylglycerol content, while the administration of n-3-PUFAs eliminated this effect. Administration of n-3-PUFAs led to significant beneficial differences in gene expression within biosynthetic pathways of cholesterol, FAs, and pro-inflammatory cytokines (IL-1 and TNF-α). To conclude, n-3-PUFA supplementation appears to represent a promising nutraceutical approach for the restoration of abnormalities in liver lipid metabolism and the prevention and treatment of NAFLD.
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Ácidos Graxos Ômega-3 , Hepatopatia Gordurosa não Alcoólica , Camundongos , Animais , Hepatopatia Gordurosa não Alcoólica/etiologia , Hepatopatia Gordurosa não Alcoólica/genética , Ácidos Graxos Ômega-3/farmacologia , Ácidos Graxos Ômega-3/uso terapêutico , Ácidos Graxos Ômega-3/metabolismo , Metionina/metabolismo , Colina/metabolismo , Fígado/metabolismo , Ácidos Graxos Insaturados/metabolismo , Racemetionina/metabolismo , Racemetionina/farmacologia , Ácidos Graxos não Esterificados/metabolismo , Dieta Hiperlipídica/efeitos adversos , Colesterol/metabolismo , Triglicerídeos/metabolismo , Camundongos Endogâmicos C57BLRESUMO
In this study, two rice varieties (RS4 and GZ93) with different amylose and lipid contents were studied, and their starch was used to prepare starch-palmitic acid complexes. The RS4 samples showed a significantly higher lipid content in their flour, starch, and complex samples compared to GZ93. The static in vitro digestion highlighted that RS4 samples had significantly lower digestibility than the GZ93 samples. The C∞ of the starch-lipid complex samples was found to be 17.7% and 18.5% lower than that of the starch samples in GZ93 and RS4, respectively. The INFOGEST undigested fractions were subsequently used for in vitro colonic fermentation. Short-chain fatty acids (SCFAs) concentrations, mainly acetate, and propionate were significantly higher in starch-lipid complexes compared to native flour or starch samples. Starch-lipid complexes produced a distinctive microbial composition, which resulted in different gene functions, mainly related to pyruvate, fructose, and mannose metabolism. Using Model-based Integration of Metabolite Observations and Species Abundances 2 (MIMOSA2), SCFA production was predicted and associated with the gut microbiota. These results indicated that incorporating lipids into rice starch promotes SCFA production by modulating the gut microbiota selectively.
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Cancer remains the second leading cause of mortality, with nearly 10 million deaths worldwide in 2020. In many cases, radiotherapy is used for its anticancer effects. However, radiation causes healthy tissue toxicity as a side effect. In intra-abdominal and pelvic malignancies, the healthy bowel is inevitably included in the radiation field, causing radiation-induced enteritis and dramatically affecting the gut microbiome. This condition is associated with significant morbidity and mortality that impairs cancer patients' and survivors' quality of life. This Review provides a critical overview of the main drivers in modulating the gut microenvironment in homeostasis, disease, and injury, focusing on gut microbial metabolites and microorganisms that influence epithelial regeneration upon radiation injury.
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Enterite , Microbioma Gastrointestinal , Neoplasias , Lesões por Radiação , Humanos , Qualidade de Vida , Enterite/etiologia , Lesões por Radiação/complicações , Neoplasias/complicações , Regeneração , Microambiente TumoralRESUMO
BACKGROUND: Advanced Glycation End products (AGEs) are a heterogeneous group of stable reaction products formed when amino acids, peptides, or proteins are glycated by the non-enzymatic Maillard Reaction. The formation and accumulation of these products in vivo are linked to many inflammation-based pathological outcomes and part of the pathophysiology of non-communicable diseases like eye cataracts and Alzheimer's disease. Since our diet contains high levels of the same compounds, it has been questioned whether their consumption is also detrimental to health. However, this is still under debate. In this context, the intestinal epithelium is an important target tissue since it is chronically exposed to relatively high concentrations of dietary AGEs. SCOPE OF REVIEW: This review summarizes the current evidence on the impact of dietary AGEs on the intestinal epithelium and critically reflects on its methodology. MAJOR CONCLUSIONS: In healthy rodent models, an inflammation-independent impaired intestinal barrier function is claimed; however, dietary AGEs showed anti-inflammatory activity in IBD models. In vitro studies could be a valuable tool to unravel the underlying mechanisms of these effects, however the available studies face some limitations, e.g. lack of the physicochemical characterization of the glycated proteins, the inclusion of the proper controls and the dose-dependency of the effect. In addition, studies using more advanced in vitro models like intestinal organoids and co-cultures with immune cells exposed to gut microbial metabolites derived from the fermentation of AGEs are still needed.
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Produtos Finais da Glicação Avançada em Alimentos , Produtos Finais de Glicação Avançada , Humanos , Produtos Finais de Glicação Avançada/metabolismo , Reação de Maillard , Inflamação , Mucosa Intestinal/metabolismoRESUMO
In this research, the physicochemical properties, sensory quality, and storage stability of mayonnaise prepared from egg yolk fermented for different times (0, 3, 6, and 9 h) have been investigated. Compared with control mayonnaise (3.50 µm and 92.88%), mayonnaise prepared from fermented egg yolk possessed significantly lower particle size (3.32-3.41 µm) and higher emulsion stability (97.26-98.72%). Meanwhile, texture, color, and gas chromatography-mass spectrometry (GC-MS) analysis revealed that the fermented egg yolk significantly enhanced the firmness, consistency and cohesiveness, lightness and redness, and flavor profile of mayonnaise. Sensory evaluation showed that mayonnaise with 3 h-fermented egg yolk exhibited the highest sensory scores. And the microscopic and appearance characteristics revealed that fermented egg yolk endowed mayonnaise with a more stable appearance after 30 days of storage. These results indicated that lactic acid fermentation of egg yolk is a feasible way to improve consumer acceptability and shelf life of mayonnaise.
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Condimentos , Gema de Ovo , Gema de Ovo/química , Emulsões/química , Tamanho da PartículaRESUMO
SCOPE: Epidemiological evidence suggests that a reduced risk of colorectal cancer (CRC) is correlated with high consumption of fruits and vegetables, which are major sources of fiber and phytochemicals, such as flavan-3-ols. However, it remains unknown how these phytochemicals and their specific gut-related metabolites may alter cancer cell behavior. METHODS AND RESULTS: A focused screening using native (poly)phenols and gut microbial metabolites (GMMs) on 3D HCT116 spheroids is carried out using a high-throughput imaging approach. Dose-responses, IC50 , and long-term exposure are calculated for the most promising native (poly)phenols and GMMs. As a result, this research shows that (poly)phenol catabolites may play a key role in preventing cancer propagation. Indeed, µM concentration levels of (4R)-5-(3',4'-dihydroxyphenyl)-γ-valerolactone significantly decrease spheroid size at early stages of spheroid aggregation and gene expression of matrix metalloproteinases. CONCLUSION: A chronic exposure to (4R)-5-(3',4'-dihydroxyphenyl)-γ-valerolactone may lead to a reduced CRC risk. Daily intake of monomeric, oligomeric, and polymeric flavan-3-ols may increase the colonic concentrations of this metabolite, and, in turn, this compound may act locally interacting with intestinal epithelial cells, precancerous and cancer cells.
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Microbioma Gastrointestinal , Fenóis , Flavonoides/metabolismo , PolifenóisRESUMO
Lactic acid bacteria (LAB) are involved in producing a considerable number of fermented products consumed worldwide. Many of those LAB fermented foods are recognized as beneficial for human health due to probiotic LAB or their metabolites produced during food fermentation or after food digestion. In this review, we aim to gather and discuss available information on the health-related effects of LAB-fermented foods. In particular, we focused on the most widely consumed LAB-fermented foods such as yoghurt, kefir, cheese, and plant-based products such as sauerkrauts and kimchi.
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Risk-benefit assessment is the comparison of the risk of a situation to its related benefits, i.e. a comparison of scenarios estimating the overall health impact. The risk-benefit analysis paradigm mirrors the classical risk analysis one: risk-benefit assessment goes hand-in-hand with risk-benefit management and risk-benefit communication. The various health effects associated with food consumption, together with the increasing demand for advice on healthy and safe diets, have led to the development of different research disciplines in food safety and nutrition. In this sense, there is a clear need for a holistic approach, including and comparing all of the relevant health risks and benefits. The risk-benefit assessment of foods is a valuable approach to estimate the overall impact of food on health. It aims to assess together the negative and positive health effects associated with food intake by integrating chemical and microbiological risk assessment with risk and benefit assessment in food safety and nutrition. The 2019 Parma Summer School on risk-benefit in food safety and nutrition had the objective was to provide an opportunity to learn from experts in the field of risk-benefit approach in food safety and nutrition, including theory, case studies, and communication of risk-benefit assessments plus identify challenges for the future. It was evident that whereas tools and approaches have been developed, more and more case studies have been performed which can form an inherent validation of the risk-benefit approach. Executed risk-benefit assessment case studies apply the steps and characteristics developed: a problem formulation (with at least 2 scenarios), a tiered approach until a decision can be made, one common currency to describe both beneficial and adverse effects (DALYs in most instances). It was concluded that risk-benefit assessment in food safety and nutrition is gaining more and more momentum, while also many challenges remain for the future. Risk-benefit is on the verge of really enrolling into the risk assessment and risk analysis paradigm. The interaction between risk-benefit assessors and risk-benefit managers is pivotal in this, as is the interaction with risk-benefit communicators.
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Inocuidade dos Alimentos , Estado Nutricional , Alimentos , Medição de Risco , Instituições AcadêmicasRESUMO
SCOPE: Several studies suggest that regular coffee consumption may help preventing chronic diseases, but the impact of daily intake and the contribution of coffee metabolites in disease prevention are still unclear. The present study aims at evaluating whether and how different patterns of coffee intake (one cup of espresso coffee/day, three cups of espresso coffee/day, and one cup of espresso coffee/day and two cocoa-based products containing coffee two times per day) may impact endogenous molecular pathways. METHODS AND RESULTS: A three-arm, randomized, crossover trial is performed in 21 healthy volunteers who consumed each treatment for one month. Urine samples are collected to perform untargeted metabolomics based on UHPLC-IMS-HRMS. A total of 153 discriminant metabolites are identified. Several molecular features are associated with coffee consumption, while others are linked with different metabolic pathways, such as phenylalanine, tyrosine, energy metabolism, steroid hormone biosynthesis, and arginine biosynthesis and metabolism. CONCLUSION: This information has provided new insights into the metabolic routes by which coffee and coffee-related metabolites may exert effects on human health.
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Biomarcadores/urina , Café , Adulto , Aminoácidos/metabolismo , Cacau , Cafeína/urina , Relação Dose-Resposta a Droga , Feminino , Humanos , Masculino , Redes e Vias Metabólicas , Metabolômica/métodos , Esteroides/metabolismoRESUMO
Glioblastoma (GB) is the most frequent and aggressive type of glioma. The lack of reliable GB models, together with its considerable clinical heterogeneity, has impaired a comprehensive investigation of the mechanisms that lead to tumorigenesis, cancer progression, and response to treatments. Recently, 3D cultures have opened the possibility to overcome these challenges and cerebral organoids are emerging as a leading-edge tool in GB research. The opportunity to easily engineer brain organoids via gene editing and to perform co-cultures with patient-derived tumor spheroids has enabled the analysis of cancer development in a context that better mimics brain tissue architecture. Moreover, the establishment of biobanks from GB patient-derived organoids represents a crucial starting point to improve precision medicine therapies. This review exemplifies relevant aspects of 3D models of glioblastoma, with a specific focus on organoids and their involvement in basic and translational research.
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Dietary patterns, microbiome dysbiosis, and gut microbial metabolites (GMMs) have a pivotal role in the homeostasis of intestinal epithelial cells and in disease progression, such as that of colorectal cancer (CRC). Although GMMs and microorganisms have crucial roles in many biological activities, models for deciphering diet-microbiome-host relationships are largely limited to animal models. Thus, intestinal organoids (IOs) have provided unprecedented opportunities for the generation of in vitro platforms with the sufficient level of complexity to model physiological and pathological diet-microbiome-host conditions. Overall, IO responses to GMM metabolites and microorganisms can provide new insights into the mechanisms by which those agents may prevent or trigger diseases, significantly extending our knowledge of diet-microbiome-host interactions.
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Microbioma Gastrointestinal/fisiologia , Organoides/microbiologia , Animais , Humanos , Microbiota/fisiologia , Análise de Célula ÚnicaRESUMO
Tryptophan and tyrosine metabolism has a major effect on human health, and disorders have been associated with the development of several pathologies. Recently, gut microbial metabolism was found to be important for maintaining correct physiology. Here, we describe the development and validation of a UHPLC-ESI-MS/MS method for targeted quantification of 39 metabolites related to tryptophan and tyrosine metabolism, branched chain amino acids and gut-derived metabolites in human plasma and urine. Extraction from plasma was optimised using 96-well plates, shown to be effective in removing phospholipids. Urine was filtered and diluted ten-fold. Metabolites were separated with reverse phase chromatography and detected using triple quadrupole MS. Linear ranges (from ppb to ppm) and correlation coefficients (r2 > 0.990) were established for both matrices independently and the method was shown to be linear for all tested metabolites. At medium spiked concentration, recovery was over 80% in both matrices, while analytical precision was excellent (CV < 15%). Matrix effects were minimal and retention time stability was excellent. The applicability of the methods was tested on biological samples, and metabolite concentrations were found to be in agreement with available data. The method allows the analysis of up to 96 samples per day and was demonstrated to be stable for up to three weeks from acquisition.
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The transient receptor potential (TRP) channels superfamily are a large group of proteins that play crucial roles in cellular processes. For example, these cation channels act as sensors in the detection and transduction of stimuli of temperature, small molecules, voltage, pH, and mechanical constrains. Over the past decades, different members of the TRP channels have been identified in the human gastrointestinal (GI) tract playing multiple modulatory roles. Noteworthy, TRPs support critical functions related to the taste perception, mechanosensation, and pain. They also participate in the modulation of motility and secretions of the human gut. Last but not least, altered expression or activity and mutations in the TRP genes are often related to a wide range of disorders of the gut epithelium, including inflammatory bowel disease, fibrosis, visceral hyperalgesia, irritable bowel syndrome, and colorectal cancer. TRP channels could therefore be promising drug targets for the treatment of GI malignancies. This review aims at providing a comprehensive picture of the most recent advances highlighting the expression and function of TRP channels in the GI tract, and secondly, the description of the potential roles of TRPs in relevant disorders is discussed reporting our standpoint on GI tract-TRP channels interactions.
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Gastroenteropatias/patologia , Canais de Potencial de Receptor Transitório/metabolismo , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Gastroenteropatias/metabolismo , Humanos , Hiperalgesia/metabolismo , Hiperalgesia/patologia , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Canais de Potencial de Receptor Transitório/genéticaRESUMO
Due to unique phytochemicals contained, Vaccinum berries are known to have a number of positive health effects. In this context, lingonberries (Vaccinium vitis-idaea) are considered to be the most effective, thus finding many uses. Recently, fraud suspicion on lingonberries-based products has been reported, partial or even total replacement by less valued cranberries (Vaccinium macrocarpon) was found. In this study, metabolomic fingerprinting employing instrumental platform consisting of U-HPLC-HRMS/MS was investigated for discrimination between the two Vaccinum berries species. Methanolic extracts of 33 authentic samples from two harvest years were analyzed and chemometric evaluation was performed to identify significant marker compounds, their stability during drying process was assessed, too. The characteristic markers most contributing to berries classification were representatives of polyphenols and phospholipids. Peonidin 3-O-arabinoside and myricetin 3-O-glucoside, not occurring in lingonberries, enabled to discover the presence of cranberries in prepared admixtures down to 1% (w/w).
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Análise de Alimentos/métodos , Metabolômica/métodos , Vaccinium macrocarpon/química , Vaccinium vitis-Idaea/química , Biomarcadores/análise , Cromatografia Líquida de Alta Pressão , Flavonoides/análise , Análise de Alimentos/estatística & dados numéricos , Contaminação de Alimentos , Glucosídeos/análise , Modelos Estatísticos , Fosfolipídeos/análise , Compostos Fitoquímicos/análise , Extratos Vegetais/análise , Polifenóis/análise , Espectrometria de Massas em Tandem , Vaccinium macrocarpon/metabolismo , Vaccinium vitis-Idaea/metabolismoRESUMO
The life sciences are currently being transformed by an unprecedented wave of developments in molecular analysis, which include important advances in instrumental analysis as well as biocomputing. In light of the central role played by metabolism in nutrition, metabolomics is rapidly being established as a key analytical tool in human nutritional studies. Consequently, an increasing number of nutritionists integrate metabolomics into their study designs. Within this dynamic landscape, the potential of nutritional metabolomics (nutrimetabolomics) to be translated into a science, which can impact on health policies, still needs to be realized. A key element to reach this goal is the ability of the research community to join, to collectively make the best use of the potential offered by nutritional metabolomics. This article, therefore, provides a methodological description of nutritional metabolomics that reflects on the state-of-the-art techniques used in the laboratories of the Food Biomarker Alliance (funded by the European Joint Programming Initiative "A Healthy Diet for a Healthy Life" (JPI HDHL)) as well as points of reflections to harmonize this field. It is not intended to be exhaustive but rather to present a pragmatic guidance on metabolomic methodologies, providing readers with useful "tips and tricks" along the analytical workflow.
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Biomarcadores/análise , Processamento Eletrônico de Dados/métodos , Metabolômica/métodos , Ciências da Nutrição/métodos , Cromatografia/métodos , Mineração de Dados , Ingestão de Alimentos , Prova Pericial , Análise de Alimentos , Humanos , Modelos Estatísticos , Análise Multivariada , Estado Nutricional , Reprodutibilidade dos TestesRESUMO
The authors should like to call your attention to the fact that Figure 5 of the article was taken from the book "Prebiotics and Probiotics in Human Milk" by Sarah S. Comstock and Sharon M. Donovan and printed with the permission from Elsevier. Unfortunately, the right referencing was missing in the original article.