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Environmental pollutants used as plasticizers in food packaging and in thousands of everyday products have become harmful for their impact on human health. Among them, phthalates, recognized as emerging endocrine disruptors (EDs) can induce toxic effects leading to different health disorders. Only few studies evaluated the effects of di-n-hexyl phthalate (DnHP) in in vivo models and no studies have been conducted to investigate the effect of DnHP on the endocannabinoid system (ECS), one of the majors signaling pathways involved in the microbiota-gut-brain axis. Due to the current relevance of probiotic bacteria as beneficial dietary interventions, the present study was aimed at evaluating the potential neuroprotective impact of daily administration of Lactiplantibacillus (Lpb.) plantarum IMC513 on zebrafish adults exposed to DnHP, with a focus on ECS modulation. Gene expression analysis revealed a promising protective role of probiotic through the restoration of ECS genes expression to the control level, in the brain of zebrafish daily exposed to DnHP. In addition, the levels of main endocannabinoids were also modulated. These findings confirm the potential ability of probiotics to interact at central level by modulating the ECS, suggesting the use of probiotics as innovative dietary strategy to counteract alterations by EDs exposure.
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Endocanabinoides , Probióticos , Peixe-Zebra , Animais , Probióticos/administração & dosagem , Probióticos/farmacologia , Endocanabinoides/metabolismo , Ácidos Ftálicos/toxicidade , Encéfalo/metabolismo , Encéfalo/efeitos dos fármacosRESUMO
Paneth cells are versatile secretory cells located in the crypts of Lieberkühn of the small intestine. In normal conditions, they function as the cornerstones of intestinal health by preserving homeostasis. They perform this function by providing niche factors to the intestinal stem cell compartment, regulating the composition of the microbiome through the production and secretion of antimicrobial peptides, performing phagocytosis and efferocytosis, taking up heavy metals, and preserving barrier integrity. Disturbances in one or more of these functions can lead to intestinal as well as systemic inflammatory and infectious diseases. This review discusses the multiple functions of Paneth cells, and the mechanisms and consequences of Paneth cell dysfunction. It also provides an overview of the tools available for studying Paneth cells.
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Microbiota , Celulas de Paneth , Intestinos , Intestino Delgado , Microbiota/fisiologiaRESUMO
The mechanisms by which early microbial colonizers of the neonate influence gut development are poorly understood. Bacterial bile salt hydrolase (BSH) acts as a putative colonization factor that influences bile acid signatures and microbe-host signaling pathways and we considered whether this activity can influence infant gut development. In silico analysis of the human neonatal gut metagenome confirmed that BSH enzyme sequences are present as early as one day postpartum. Gastrointestinal delivery of cloned BSH to immature gnotobiotic mice accelerated shortening of the colon and regularized gene expression profiles, with monocolonised mice more closely resembling conventionally raised animals. In situ expression of BSH decreased markers of cell proliferation (Ki67, Hes2 and Ascl2) and strongly increased expression of ALPI, a marker of cell differentiation and barrier function. These data suggest an evolutionary paradigm whereby microbial BSH activity potentially influences bacterial colonization and in-turn benefits host gastrointestinal maturation.
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Microbioma Gastrointestinal , Transcriptoma , Feminino , Humanos , Camundongos , Animais , Amidoidrolases/genética , Amidoidrolases/metabolismo , Trato Gastrointestinal/microbiologia , Bactérias/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismoRESUMO
Systemic inflammatory response syndrome (SIRS) is a severe condition characterized by systemic inflammation, which may lead to multiple organ failure, shock and death. SIRS is common in burn patients, pancreatitis and sepsis. SIRS is often accompanied by intestinal dysbiosis. However, the mechanism, role and details of microbiome alterations during the early phase of acute SIRS are not completely understood. The current study aimed to characterize the dynamic alterations of both the intestinal and respiratory microbiome at two timepoints during the early phase of acute SIRS (4 and 8 h after LPS) and link these to the host response in a mouse model of a LPS-induced lethal SIRS. Acute SIRS had no effect on the microbiome in the large intestine but induced a rapid dysbiosis in the small intestine, which resembled the microbiome alterations commonly observed in SIRS patients. Later in the disease progression, a dysbiosis of the respiratory microbiome was observed, which was associated with the MMP9 expression in the lungs. Although similar bacteria were increased in both the lung and the small intestine, no evidence for a gut-lung translocation was observed. Gut dysbiosis is commonly observed in diseases involving inflammation in the gut. However, whether the inflammatory response associated with SIRS and sepsis can directly cause gut dysbiosis was still unclear. In the current study we provide evidence that a LPS-induced SIRS can directly cause dysbiosis of the small intestinal and respiratory microbiome.
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Endotoxemia , Microbioma Gastrointestinal , Sepse , Animais , Disbiose/microbiologia , Endotoxemia/complicações , Inflamação/metabolismo , Lipopolissacarídeos/toxicidade , Metaloproteinase 9 da Matriz , Camundongos , Sepse/complicaçõesRESUMO
Lactiplantibacillus (Lpb.) plantarum is a versatile species commonly found in a wide variety of ecological niches including dairy products and vegetables, while it may also occur as a natural inhabitant of the human gastrointestinal tract. Although Lpb. plantarum strains have been suggested to exert beneficial properties on their host, the precise mechanisms underlying these microbe-host interactions are still obscure. In this context, the genome-scale in silico analysis of putative probiotic bacteria represents a bottom-up approach to identify probiotic biomarkers, predict desirable functional properties, and identify potentially detrimental antibiotic resistance genes. In this study, we characterized the bacterial genomes of three Lpb. plantarum strains isolated from three distinct environments [strain IMC513 (from the human GIT), C904 (from table olives), and LT52 (from raw-milk cheese)]. A whole-genome sequencing was performed combining Illumina short reads with Oxford Nanopore long reads. The phylogenomic analyses suggested the highest relatedness between IMC513 and C904 strains which were both clade 4 strains, with LT52 positioned within clade 5 within the Lpb. plantarum species. The comparative genome analysis performed across several Lpb. plantarum representatives highlighted the genes involved in the key metabolic pathways as well as those encoding potential probiotic features in these new isolates. In particular, our strains varied significantly in genes encoding exopolysaccharide biosynthesis and in contrast to strains IMC513 and C904, the LT52 strain does not encode a Mannose-binding adhesion protein. The LT52 strain is also deficient in genes encoding complete pentose phosphate and the Embden-Meyerhof pathways. Finally, analyses using the CARD and ResFinder databases revealed that none of the strains encode known antibiotic resistance loci. Ultimately, the results provide better insights into the probiotic potential and safety of these three strains and indicate avenues for further mechanistic studies using these isolates.
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We have previously isolated and characterized food-dwelling strains of Lactiplantibacillus (Lpb.) plantarum that are consumed naturally as part of the microbiota of table olives and raw milk cheeses. Despite being consumed at relatively high levels, the impact of such strains on the human gut microbiota is currently unclear. In the current study we evaluated the potential impact of food-dominant Lpb. plantarum strains on the human gut microbiota using a continuous fecal fermentation system. Daily inoculation of Lpb. plantarum strains led to significant, detectable levels in the fecal fermentation system. We examined the impact of the presence of Lpb. plantarum on the microbiota derived from two separate donors. For one donor, Lpb. plantarum increased alpha diversity and beta diversity. This was reflected in significant alterations in abundance of the unclassified genera, dominated by Enterobacteriaceae_unclass and Ruminococcaceae_unclass. The microbiota of the other donor was relatively unaffected following introduction of the Lpb. plantarum strains. Overall, the work describes the response of the human microbiota to the introduction of high levels of food-dominant Lpb. plantarum strains and indicates that the response may reflect interindividual differences between donor samples.
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The glucocorticoid (GC) receptor (GR) is essential for normal development and in the initiation of inflammation. Healthy GRdim/dim mice with reduced dimerization propensity due to a point mutation (A465T) at the dimer interface of the GR DNA-binding domain (DBD) (here GRD/D) have previously helped to define the functions of GR monomers and dimers. Since GRD/D retains residual dimerization capacity, here we generated the dimer-nullifying double mutant GRD+L/D+L mice, featuring an additional mutation (I634A) in the ligand-binding domain (LBD) of GR. These mice are perinatally lethal, as are GRL/L mice (these mice have the I634A mutation but not the A465T mutation), displaying improper lung and skin formation. Using embryonic fibroblasts, high and low doses of dexamethasone (Dex), nuclear translocation assays, RNAseq, dimerization assays, and ligand-binding assays (and Kd values), we found that the lethal phenotype in these mice is due to insufficient ligand binding. These data suggest there is some correlation between GR dimerization potential and ligand affinity. We conclude that even a mutation as subtle as I634A, at a position not directly involved in ligand interactions sensu stricto, can still influence ligand binding and have a lethal outcome.
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Dexametasona , Mutação Puntual , Receptores de Glucocorticoides , Animais , Dexametasona/farmacologia , Glucocorticoides/farmacologia , Ligantes , Camundongos , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/metabolismoRESUMO
Fermentation processes have been used for centuries for food production and preservation. Besides the contribution of fermentation to food quality, recently, scientific interest in the beneficial nature of fermented foods as a reservoir of probiotic candidates is increasing. Fermented food microbes are gaining attention for their health-promoting potential and for being genetically related to human probiotic bacteria. Among them, Lactiplantibacillus (Lpb.) plantarum strains, with a long history in the food industry as starter cultures in the production of a wide variety of fermented foods, are being investigated for their beneficial properties which are similar to those of probiotic strains, and they are also applied in clinical interventions. Food-associated Lpb. plantarum showed a good adaptation and adhesion ability in the gastro-intestinal tract and the potential to affect host health through various beneficial activities, e.g., antimicrobial, antioxidative, antigenotoxic, anti-inflammatory and immunomodulatory, in several in vitro and in vivo studies. This review provides an overview of fermented-associated Lpb. plantarum health benefits with evidence from clinical studies. Probiotic criteria that fermented-associated microbes need to fulfil are also reported.
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Food-associated Lactiplantibacillus plantarum (Lpb. plantarum) strains, previously classified as Lactobacillus plantarum, are a promising strategy to face intestinal inflammatory diseases. Our study was aimed at clarifying the protective role of food-borne Lpb. plantarum against inflammatory damage by testing the scavenging microbial ability both in selected strains and in co-incubation with normal mucosa intestinal cells (NCM460). Here, we show that Lpb. plantarum endure high levels of induced oxidative stress through partially neutralizing reactive oxygen species (ROS), whereas they elicit their production when co-cultured with NCM460. Moreover, pre-treatment with food-borne Lpb. plantarum significantly reduce pro-inflammatory cytokines IL-17F and IL-23 levels in inflamed NCM460 cells. Our results suggest that food-vehicled Lpb. plantarum strains might reduce inflammatory response in intestinal cells by directly modulating local ROS production and by triggering the IL-23/IL-17 axis with future perspectives on health benefits in the gut derived by the consumption of functional foods enriched with selected strains.
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Inflamação/microbiologia , Interleucina-17/metabolismo , Interleucina-23/metabolismo , Mucosa Intestinal/microbiologia , Intestinos/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Linhagem Celular , Humanos , Inflamação/metabolismo , Mucosa Intestinal/citologia , Mucosa Intestinal/metabolismo , Intestinos/citologia , Lactobacillus plantarum/fisiologia , Estresse Oxidativo/fisiologiaRESUMO
Nowadays, the interest in the role of dietary components able to influence the composition and the activity of the intestinal microbiota and, consequently, to modulate the risk of genotoxicity and colon cancer is increasing in the scientific community. Within this topic, the microbial ability to have a protective role at gastrointestinal level by counteracting the biological activity of genotoxic compounds, and thus preventing the DNA damage, is deemed important in reducing gut pathologies and is considered a new tool for probiotics and functional foods. A variety of genotoxic compounds can be found in the gut and, besides food-related mutagens and other DNA-reacting compounds, there is a group of pollutants commonly used in food packaging and/or in thousands of everyday products called endocrine disruptors (EDs). EDs are exogenous substances that alter the functions of the endocrine system through estrogenic and anti-estrogenic activity, which interfere with normal hormonal function in human and wildlife. Thus, this paper summarizes the main applications of probiotics, mainly lactobacilli, as a bio-protective tool to counteract genotoxic and mutagenic agents, by biologically inhibiting the related DNA damage in the gut and highlights the emerging perspectives to enlarge and further investigate the microbial bio-protective role at intestinal level.
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Disruptores Endócrinos/metabolismo , Microbioma Gastrointestinal/fisiologia , Trato Gastrointestinal/química , Trato Gastrointestinal/microbiologia , Probióticos/metabolismo , Humanos , Lactobacillus/metabolismo , Mutagênicos/metabolismoRESUMO
Table olives are one of the oldest vegetable fermented foods in the Mediterranean area. Beside their economic impact, fermented table olives represent also an important healthy food in the Mediterranean diet, because of their high content of bioactive and health-promoting compounds. However, olive fermentation is still craft-based following traditional processes, which can lead to a not fully predictable final product with the risk of spontaneous alterations. Nowadays, food industries have to face consumer demands for safe and healthy products. This review offers an overview about the main technologies used for olive fermentation and the role of lactic acid bacteria and yeasts characterizing this niche during the fermentation. Particular attention is offered to the selection and use of microorganisms as starter cultures to fasten and improve the safety of table olives. The development and implementation of multifunctional starter cultures in order to obtain heath-oriented table olives is also discussed.
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[This corrects the article DOI: 10.3389/fmicb.2018.01287.].
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Lactobacillus plantarum species (recently re-named Lactiplantibacillus (Lpb.) plantarum subsp. plantarum) can be isolated from both either the mammalian gut or specific fermented foods where they may be present at high concentrations. Whilst Lpb. plantarum strains have been proposed as potential probiotic candidates, the ability of resident strains consumed in fermented foods to interact with the host is unclear. The main objective of this study was to investigate the cellular location and ability of three different food-borne Lpb. plantarum strains isolated from different sources (table olives and cheese) to modulate the immune response of a murine macrophage-like cell line (J774A.1). For that purpose, macrophages were exposed to the three different Lpb. plantarum strains for 24 h and the expression of a panel of genes involved in the immune response, including genes encoding pattern-recognition receptors (TLRs and NLRs) and cytokines was evaluated by qRT-PCR. We also utilized chemical inhibitors of intracellular pathways to gain some insight into potential signaling mechanisms. Results showed that the native food strains of Lpb. plantarum were able to modulate the response of J774A.1 murine macrophages through a predominately NOD signaling pathway that reflects the transient intracellular location of these strains within the macrophage. The data indicate the capacity of food-dwelling Lpb. plantarum strains to influence macrophage-mediated host responses if consumed in sufficient quantities.
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PURPOSE: Ewe's milk yogurt is richer in proteins, minerals and short- and medium-chain fatty acids compared to cow's milk yogurt. We aimed to evaluate the effects of cow's milk yogurt (CW, 3.0% fat), semi-skimmed (ES, 2.8% fat) and whole ewe's milk yogurts (EW, 5.8% fat) on inflammatory markers and gut microbiota in subjects with borderline-high plasma cholesterol. METHODOLOGY: 30 adults (16 women) were randomized into a crossover study to consume 250 g/yogurt/day during three 5-week periods (4-week washouts). Plasma insulin, leptin, adhesion molecules, cytokines and gut microbiota composition (qPCR) were analysed. Rates of change were used to assess treatment effects both in the whole group and in subgroups of subjects with different cholesterol/HDL-c ratio (Cho-I group A: the top 6 women and 4 men values; Cho-I group B: remaining subjects). RESULTS: The yogurts showed no different effects on the inflammatory biomarkers or the microbiota of the whole group. However, ICAM-1 and P-selectin rates of change were lower after EW compared to CW and ES, respectively, in subjects of the Cho-I group A (P = 0.047 and P = 0.020). Women of this group showed lower MCP-1 rates of change after EW compared to ES and CW (P = 0.028, both). Blautia coccoides-Eubacterium rectale decreased in women of the Cho-I group A during EW vs. ES (P = 0.028). CONCLUSION: Ewe's yogurt effects on inflammatory markers and microbiota were not different from those after cow's yogurt, but the attenuation of some inflammatory biomarkers with ewe's whole-milk yogurt in subjects with the highest TC/HDL-c deserves further study.
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Colesterol/sangue , Microbioma Gastrointestinal/efeitos dos fármacos , Inflamação/sangue , Inflamação/dietoterapia , Iogurte , Adulto , Idoso , Animais , Biomarcadores/sangue , Estudos Cross-Over , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , OvinosRESUMO
Food-associated microbes can reach the gut as viable cells and interact with the human host providing potential health benefits. In this study, we evaluated the impact on cell viability and the adhesion ability of 22 Lactobacillus plantarum strains, mainly isolated from fermented foods, on a Normal-derived Colon Mucosa cell line. Furthermore, due to the presence of mucus layer on the gut epithelium, we also investigated whether the mucin could affect the microbial adhesion property. Our results demonstrated that all the strains displayed a strong ability to adhere to host cells, showing a strain-dependent behavior with preference for cell edges, that resulted not to be affected by the presence of mucin. Based on interleukin-8 release of intestinal cells induced by some Lb. plantarum strains, our data suggest a potential cross-talk with the host immune system as unconventional property of these food-associated microbes.
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Irrigation is widely used for the production of table olives because it increases fruit size and yield. However, irrigation also determines less accumulation of total phenols, an increase in water content, a decrease of firmness, lower concentrations of soluble sugars in the mesocarp, thus positively or negatively affecting the fermentation process for the production of table olives. In this study we tested the hypothesis that green fruits of cultivar Itrana obtained by different irrigation regimes had different phenolic concentration that responded differentially to spontaneous or inoculated fermentation. Fruits were harvested from two orchards in the Latina province of Latium, Italy, which had been irrigated with different volumes of water during the growing season to compare the evolution of spontaneous and inoculated fermentation processes. We measured fruit characteristics at harvest, changes in the concentrations of secoiridoids and lignans, and main microbial groups abundance during fermentation. At harvest and during fermentation the concentration of phenolic compounds was higher in fruits sampled from trees that had received less water in the field. Differences were observed between spontaneous and inoculated fermentations, with a prevalence of lactic acid bacteria (LAB) in inoculated samples. In particular, oleuropein concentration completely disappeared only from samples inoculated with the two selected strains used as starters. The inoculum with selected LAB positively influenced the fermentation process of green olives, whereas the irrigation regime previously experienced by trees did not alter fermentation.
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Yogurt is the most widely consumed fermented milk product worldwide. Studies have mainly used milk and dairy products from cow, which have a lower fat content than those from ewe and a different lipid profile. This study investigated the effect on the lipid profile of control subjects of three different set yogurts: (a) semi-skimmed ewe´s milk yogurt (2.8% milk fat); (b) whole ewe´s milk yogurt (5.8 % milk fat); (c) cow´s milk yogurt (3 % milk fat). A randomized crossover study included 30 healthy adults (16 women) to consume 250 g/yogurt/day during three consecutive 5-weeks periods separated by 4-week washouts. Blood samples were collected at the start and end of each period for the analysis of serum cholesterol (total, HDL-, LDL-) and triglycerides. We found no differences in the serum concentrations of lipid and lipoprotein fractions of the volunteers after the intake of any of the three types of yogurts. When the volunteers were grouped into two risk groups of risk according to their total cholesterol/HDL cholesterol ratio, the same differences between the groups in terms of the cholesterol (HDL-, LDL-) and triglyceride responses at baseline and after yogurt intake were found, with no effects due to the different types of yogurts. Moreover, we performed compositional analysis of the yogurts including determination of protein, fat, minerals and fatty acids (FA). Contents in protein, calcium, magnesium, non-protein nitrogen and some FA (mainly short-chain-FA) were higher for ewe's than for cow's milk yogurt. n6-n3 ratio was lower in the ewe's milk yogurt. In conclusion, yogurt intake, from ewe's and cow's milk, at levels of consumption compatible with a varied diet, neither decreases nor increases plasma lipoprotein cholesterol levels in apparently healthy individuals. As ewe's milk yogurt has a high content of macro- and micronutrients, certain target populations could benefit from its consumption.