Search Results (76)

Faced with the challenge of reducing food waste, transforming discarded fruit into functional ingredients useful for the food industry is a valuable solution. Ingredients from fruit such as persimmons, which are rich in indigestible carbohydrates and bioactive compounds with antiradical capacity, could positively impact on the health of certain population groups due to their potential prebiotic effect. This study aimed to select the most suitable drying conditions and milling intensity for obtaining powdered persimmon ingredients with a prebiotic-like effects observed in vitro for postmenopausal women, and to evaluate this effect by considering the stimulation of health-promoting bacterial growth and short-chain fatty acids (SCFAs) production. First, the effect of the drying method (hot air drying at 60 and 70 °C, and freeze-drying) and grinding intensity on antiradical capacity, particle size, and the release of bioactive antiradical components into the intestinal lumen after an in vitro gastrointestinal digestion was determined. Next, the effect of these conditions on the microbiota composition of postmenopausal women was preliminary assessed in a batch colonic fermentation experiment for 24 h. The results showed that the ingredient dried with air at 70 °C had the highest phenol and flavonoid content, suffered the least degradation during in vitro gastrointestinal digestion and promoted the differential growth of fiber-degrader genera. Consequently, this was the ingredient selected as the most suitable. Lastly, the impact of this ingredient on the microbiota composition of 4 postmenopausal women has been evaluated in a long-term study using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME^®) coupled to high throughput sequencing. The growth stimulation of health-associated bacteria, such as Akkermansia muciniphila, Faecalibacterium prausnitzii or Phascolarctobacterium faecium, and the promotion of beneficial metabolic pathways, such as the sugar uptake-specific phosphotransferase system, sugar metabolism and propionate and isobutyrate production, were detected along 14 days of persimmon powder supplementation. A holistic framework for promoting human health while advancing environmental sustainability is represented by the combination of sustainable by-product valorization and microbiota-targeted functional food development. Full article

Recently, the role of mucosal intestinal microbiota in human health has received increasing attention. Nevertheless, data on the response of this microbiota to various interventions remain limited. Here, we have employed the Mucosal Simulator of Human Gastrointestinal Microbial Ecosystem (M-SHIME^®) and luminal SHIME^® (L-SHIME^®) to examine mucosal microbiota responses to interventions that are known to impact the intestinal microbial community in humans and study relationships between the responses of mucosal and luminal microbiota. Specifically, we evaluated the effects of varying macronutrient levels over a 28-day standard, balanced dietary intervention and a parallel 14-day administration of Lacticaseibacillus rhamnosus GG. Observed shifts in mucosal microbiota in response to interventions differed significantly from those observed in luminal microbiota (p < 0.05). In particular, we found that the mucosal microbiota compared to luminal microbiota was more stable and that the abundance of several genera (i.e., Subdoligranulum, Parabacteroides and Fusobacterium) in the M-SHIME^® correlated positively with the intake of dietary macronutrients, especially protein, which was in line with results reported in previous human studies. This study demonstrates the reliability of advanced in vitro models in capturing diet-induced dynamics of the human mucosal microbiota, a compartment that remains understudied despite its critical role in intestinal immune regulation. Full article

Background/Objectives: This study examines the application of the novel double emulsion gel system for the delivery and release of encapsulated cannabidiol (CBD) and the probiotic strain Lactiplantibacillus plantarum DSM 24624. Methods: During a six-week experimental period comprising stabilization, treatment, and wash-out phases, the dynamic Simulator of the Human Intestinal Microbial Ecosystem (SHIME^®) model was employed to assess a system. The evaluation focused on the delivery of CBD and probiotics, as well as the system’s effects on microbial composition, diversity, and metabolic activity throughout the digestion process using 16S rRNA gene sequencing and digital PCR methods. Results: Microbial community analysis revealed significant shifts in both mucosal and luminal microbiota following supplementation. The treatment increased beneficial bacterial families such as Lachnospiraceae and Clostridiaceae, demonstrated effective delivery, release, and persistence of the probiotic L. plantarum, as well as enhanced butyrate and lactate production. Diversity analyses highlighted a transient rise in alpha diversity within the mucin layer and a decrease in the lumen, with significant changes in beta diversity across experimental phases. Conclusions: Findings suggest that double emulsion gel can be employed for the delivery of probiotics and CBD to the gastrointestinal tract. In addition, an innovative CBD-probiotic formulation can modulate gut microbiota composition and metabolic activity, suggesting its potential as a functional food innovation for intestinal health. However, the results are based on an in vitro model, which lacks the complexity of the human host environment, and further clinical studies are necessary to confirm the biological relevance and therapeutic potential of such delivery systems for gastrointestinal health. Full article

Microplastics (MPs) have become widespread environmental contaminants, with increasing evidence of their harmful impacts on human health. MPs generally enter the human body via ingestion, inhalation, or dermal exposure, with the gastrointestinal tract acting as a crucial entrance route. This work utilized the SHIME system to evaluate the effects of polystyrene (PS) MPs on gut microbiota and short-chain fatty acid (SCFA) metabolism in distinct colonic areas. The results demonstrated regional and individual-specific variations in microbial diversity, significant shifts in Firmicutes/Bacteroidetes (F/B) ratio, and declines in beneficial bacteria, such as Bifidobacteriaceae. Moreover, SHIME supernatants were then tested with a co-cultured cell model (Caco-2/HT29-MTX-E12). Results indicated a deteriorative effect on the intestinal model, characterized by enhanced oxidative stress and mitochondrial malfunction. No significant effect on intestinal barrier integrity or mucus secretion was detected. These findings highlight the potential systemic toxicity of PS-MPs on human gut microbiota-mediated mechanisms, emphasizing the necessity for immediate mitigation efforts. Full article

Background: Cystic fibrosis (CF) is a genetic disorder that disrupts gut microbiota composition, promoting dysbiosis associated with chronic inflammation, impaired nutrient absorption, and poor clinical outcomes. While modulation of the intestinal microbiota through prebiotics, probiotics, and synbiotics has been proposed as a therapeutic strategy, clinical evidence remains limited, especially in children. Objective: This study aimed to evaluate the impact of three supplementation strategies (a prebiotic (β-glucan), a probiotic (Lacticaseibacillus rhamnosus GG), and their synbiotic combination) on the gut microbiota and metabolic activity of a CF child faecal donor using a dynamic in vitro colonic fermentation model (SHIME^®). Methods: Microbial composition (16S rRNA gene sequencing), and metabolic activity (quantification of short-chain fatty acids (SCFAs), ammonia, and lactate) were analysed. Results: Results showed that the prebiotic increased alpha diversity; while both the prebiotic and probiotic treatments significantly reduced Bacillota and increased Bacteroidota, modulating the Bacillota/Bacteroidota ratio. The synbiotic treatment showed the most beneficial overall profile, including enhanced production of SCFAs, particularly butyrate and propionate, and increased abundance of Faecalibacterium and Agathobacter, which are two bacterial genera generally associated with gut health. Notably, the synbiotic also reduced the relative abundance of potentially pathogenic genera such as Veillonella, Megasphaera, and Stenotrophomonas, but paralleled with an increase in Clostridium ss 1. Although the probiotic alone showed some positive effects, it was less effective overall compared to the prebiotic and synbiotic approaches. Conclusions: These findings support the potential of synbiotic supplementation as a promising strategy to modulate gut dysbiosis in CF, though in vivo studies are needed to confirm the translational relevance of these results. Full article

Xanthohumol (XN), a polyphenol from hops (Humulus lupulus), exhibits antioxidant, anti-inflammatory, antihyperlipidemic, and chemo-preventive activity. Preclinical evidence suggests gut microbiota are critical to mediating some of these bioactivities. Nevertheless, its precise impact on human gut microbiota, particularly at supplemental doses, remains poorly characterized. We evaluated 200 mg/day XN for 3 weeks on human gut microbiota in a eubiotic and dysbiotic model using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME^®). Functional assessments of microbiota included quantification of XN metabolites, short-chain fatty acids (SCFAs), and untargeted metabolomics of the digestive metabolome. Bacterial composition was assessed by 16S rRNA gene sequencing. XN reduced alpha-diversity and short-chain fatty acid production in both models, as well as altered taxa abundance variably between models. XN disrupted bile acid metabolism through inhibition of microbial bile salt hydrolase (BSH). The modulation of bile acid metabolism has important implications for host-level bioactivity of XN. Full article

Background: Bile salt hydrolase (BSH) is a key probiotic trait, as it facilitates both host metabolism and bacterial survival into the gastrointestinal tract (GIT), through bile acid (BA) deconjugation, keeping intestinal homeostasis. Objectives: The present study aims to investigate the viability of the Lacticaseibacillus rhamnosus VB4 strain and its effects on bile acid deconjugation during the gastrointestinal tract (GIT) passage, under a fed condition, using the in vitro SHIME^® (Simulator of the Human Intestinal Microbial Ecosystem) model. Methods: Gastric, small intestinal and colonic fractions were monitored and a fecal slurry from a healthy donor was inoculated into the colonic compartment to establish the intestinal microbiota. Samples were collected at the end of stomach, duodenum, jejunum, ileum phases, and colon after 0, 16 and 24 h. Strain survival was assessed by culturing method, and bsh gene expression was revealed by quantitative PCR (qPCR). In addition, UHPLC/HR-MS was performed to reveal the hypothetical changes in BAs profile after strain administration. Results: Good survivability of the VB4 strain in the upper GIT was revealed. Furthermore, VB4-inculated sample showed sustained expression of bsh in both the stomach/small intestine and colon fractions at all sampling times. Analysis of the BAs profile shown that the VB4 strain reduced the levels of the main conjugated BAs in the small intestine under fed condition and improved the deconjugation efficiency during colonic transit compared with the control. Conclusions: These findings highlight the survivability of L. rhamnosus VB4 strain inside the gut and its potential as biotherapeutic BAs-mediator candidate, demonstrating that transcriptomic and metabolomic approaches coupled to a dynamic in vitro gut model represent a robust tool for selection of a BSH-positive probiotic candidate. Full article

Background/Objectives: With increasing knowledge of the role of the microbiota in health and disease, the need for the reliable simulation of its behavior in response to various factors, such as diet and probiotic administration in in vitro conditions, has emerged. Although many studies utilize developed systems, data on how accurately these systems represent individual microbiota responses are scarce. Methods: In the present study, the Simulator of Human Intestinal Microbial Ecosystem (SHIME^®) was exposed to experimental conditions mimicking the application of probiotics and dietary changes in the study participant. Next-generation 16S rRNA sequencing was used to reveal the structure of the microbial communities in the analyzed samples. Results: Analysis of 17 samples revealed that predominantly diet and, to a lesser extent, probiotics had a divergent effect on the microbiota’s fluctuations dependent on the culture environment. Despite this, results from both in vitro and in vivo conditions aligned well with previously published data on the expected impact of dietary changes on the intestinal microbial community. Conclusions: The anecdotal evidence presented in this study suggested that current in vitro technology enables the reproduction of some of the microbiota responses that are well known from in vivo research. However, further work is required to enable simulations of an individual microbiota. Full article

Background/Objectives: The probiotic Lacticaseibacillus paracasei DG (LpDG) has shown promising results for various gastrointestinal diseases. This study evaluated the survival, metabolic activity, and impact on colonic microbiota of LpDG in an in vitro gastrointestinal tract simulation. Methods: Encapsulated LpDG was tested under simulated fed, fasted, and shortened fasted conditions compared with a blank control in a modified Simulator of the Human Intestinal Microbial Ecosystem (SHIME^®) reactor. Capsule integrity, and cell culturability and viability were assessed at the end of each digestion phase. Metabolic activity (pH, total gas production, and concentrations of short-chain fatty acids, lactate, and ammonium) was assessed after a 24 h colonic incubation with a faecal inoculum. The impact of LpDG on the colonic microbial community was analysed by quantitative polymerase chain reaction and shallow shotgun sequencing. Results: The capsule was completely degraded at the end of the jejunum under all conditions. A low pH had a minimal impact on LpDG culturability and viability. Compared with blank control, LpDG remained metabolically active in the microbial community following a 24 h colonic incubation (LpDG [0–24 h] vs. blank control [0–24 h]: ΔpH, decreased [0.29–0.38 vs. 0.12–0.34]; Δlactic acid, decreased [1.52–1.69 mM vs. 0.13–0.21 mM]; and Δbutyrate, increased [7.49–10.52 mM vs. 5.19–7.76 mM]). Under fed conditions, treatment with LpDG compared with blank control significantly decreased levels of Escherichia coli and Blautia wexlerae and increased Clostridiaceae, Eubacteriaceae, and Lachnospiraceae. Conclusions: LpDG remains viable and metabolically active in the gastrointestinal tract, positively affecting intestinal microbiota and metabolite production. Full article

Postbiotics, defined as metabolites produced by probiotics, encompass both bacterial cells and their metabolic byproducts, and offer significant health benefits to the host. However, there are relatively few reports on their effects on intestinal microbiota. In this study, we investigated the components, total antioxidant capacity of Lactobacillus helveticus postbiotics (LHPs) and their impact on intestinal flora using the Simulator for Human Intestinal Microecology Simulation (SHIME). The results indicate that the primary components of postbiotics include polysaccharides, proteins, and organic acids. Furthermore, LHPs have a strong ability to inhibit the growth of harmful bacteria while promoting the growth of probiotics. Additionally, LHPs significantly increased the total antioxidant capacity in the intestine and regulated the balance of intestinal microbiota. Notably, there was also a significant increase in the content of short-chain fatty acids (SCFAs) in the intestine. Overall, LHPs have the potential to aid in the prevention and treatment of diseases by enhancing gut microbiology. Full article

Background/Objectives: Selective serotonin reuptake inhibitors (SSRIs), widely prescribed for anxiety disorders, may negatively impact the gut microbiota, contributing to dysbiosis. Considering the gut–brain axis’s importance in mental health, probiotics could represent an effective adjunctive strategy. This study evaluated the effects of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 on microbiota composition, metabolic activity, and immune markers in fecal samples from patients with anxiety on SSRIs, using the SHIME^® (Simulator of the Human Intestinal Microbial Ecosystem) model. Methods: The fecal microbiotas of four patients using sertraline or escitalopram were inoculated in SHIME^® reactors simulating the ascending colon. After stabilization, a 14-day probiotic intervention was performed. Microbial composition was assessed by 16S rRNA sequencing. Short-chain fatty acids (SCFAs), ammonia, and GABA were measured, along with the prebiotic index (PI). Intestinal barrier integrity was evaluated via transepithelial electrical resistance (TEER), and cytokine levels (IL-6, IL-8, IL-10, TNF-α) were analyzed using a Caco-2/THP-1 co-culture system. The statistical design employed in this study for the analysis of prebiotic index, metabolites, intestinal barrier integrity and cytokines levels was a repeated measures ANOVA, complemented by post hoc Tukey’s tests to assess differences across treatment groups. For the 16S rRNA sequencing data, alpha diversity was assessed using multiple metrics, including the Shannon, Simpson, and Fisher indices to evaluate species diversity, and the Chao1 and ACE indices to estimate species richness. Beta diversity, which measures microbiota similarity across groups, was analyzed using weighted and unweighted UniFrac distances. To assess significant differences in beta diversity between groups, a permutational multivariate analysis of variance (PERMANOVA) was performed using the Adonis test. Results: Probiotic supplementation increased Bifidobacterium and Lactobacillus, and decreased Klebsiella and Bacteroides. Beta diversity was significantly altered, while alpha diversity remained unchanged. SCFA levels increased after 7 days. Ammonia levels dropped, and PI values rose. TEER values indicated enhanced barrier integrity. IL-8 and TNF-α decreased, while IL-6 increased. GABA levels remained unchanged. Conclusions: The probiotic combination of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 modulated gut microbiota composition, metabolic activity, and inflammatory responses in samples from individuals with anxiety on SSRIs, supporting its potential as an adjunctive strategy to mitigate antidepressant-associated dysbiosis. However, limitations—including the small pooled-donor sample, the absence of a healthy control group, and a lack of significant GABA modulation—should be considered when interpreting the findings. Although the SHIME^® model is considered a gold standard for microbiota studies, further clinical trials are necessary to confirm these promising results. Full article

Fucoidans have demonstrated a wide range of bioactivities including immune modulation and benefits in gut health. To gain a deeper understanding on the effects of fucoidan from Undaria pinnatifida (UPF) on the colonic microbiome, the short-term Simulator of the Human Intestinal Microbial Ecosystem^®, a validated in vitro gut model, was applied. Following a three-week intervention period on adult faecal samples from three healthy donors, microbial community activity of the colonic microbiota was assessed by quantifying short-chain fatty acids while composition was analysed utilising 16S-targeted Illumina sequencing. Metagenomic data were used to describe changes in community structure. To assess the secretion of cytokines, co-culture experiments using Caco-2 and THP1-Blue™ cells were performed. UPF supplementation over a three-week period had a profound butyrogenic effect while also enriching colonic microbial diversity, consistently stimulating saccharolytic genera, and reducing genera linked with potentially negative health effects in both regions of the colon. Mild immune modulatory effects of UPF were also observed. Colonic fermentation of UPF showed anti-inflammatory properties by inducing the secretion of the anti-inflammatory cytokines IL-6 and IL-10 in two out of three donors in the proximal and distal colon. In conclusion, UPF supplementation may provide significant gut health benefits. Full article

Background: Phenolic compounds, particularly anthocyanins and proanthocyanidins (PACs), are poorly absorbed in the upper digestive tract and reach the colon largely intact, where they may influence gut microbiota (GM) composition and, in turn, impact host health. We hypothesized that a PAC-rich aronia extract would beneficially modulate the GM, promote the growth of health-associated bacteria, and enhance short-chain fatty acid (SCFA) production across different colon sections, with partial reversion effects after supplementation ends. Methods: The Twin-M-SHIME^® system was used to simulate the digestion and colonic fermentation in two donors with contrasting microbiota profiles. The experimental design included four phases: stabilization (14 days), control (7 days), treatment with 500 mg/day PAC-rich aronia extract (21 days), and wash-out (10 days). SCFA production was monitored, and changes in microbiome composition were assessed using 16S rRNA gene sequencing. Results: PAC-rich aronia extract significantly modulated SCFA levels, increasing butyrate and reducing acetate, with some inter-donor variability. SCFA concentrations tended to return to baseline after the wash-out (WO) period. Metagenomic analysis revealed a decrease in Collinsella, Sutterella, Selenomonas, and Parabacteroides—genera linked to low-fiber diets and gut inflammation—while promoting Proteobacteria (e.g., Escherichia-Shigella, Klebsiella) and butyrate-associated Firmicutes such as Lactiplantibacillus. Although some microbial shifts partially reverted during the wash-out (e.g., Akkermansia, Bacteroides, and Bifidobacterium), other changes persisted. Conclusions: These findings suggest that PAC-rich aronia extract beneficially modulates GM and SCFA production, but continuous intake may be necessary to maintain these effects over time. Full article

The prolyl-hydroxylase inhibitor (PHI), used effectively in several countries for the treatment of renal anemia, activates the multifunctional hypoxia-inducible factors (HIFs). While hypoxic conditions in tumors are known to affect the response to radiation therapy, the effect of PHI on the radiation response of cancer cells has not been determined. Hypoxic pretreatment increased the radiation sensitivity of A549 lung adenocarcinoma cells, whereas hypoxic culture after irradiation decreased the radiation sensitivity of HSC2 oral squamous cell carcinoma cells. Treatment of PC9 lung adenocarcinoma and HSC2 cells with the PHI FG-4592 significantly increased radiation resistance, whereas A549 and TIG3 lung fibroblast cells tended to be sensitized, suggesting cell type-specific differential effects of PHI. Quantitative RT-PCR analyses revealed that the basal and radiation-inducible expressions of DEC2, BAX, and BCL2 may be related to PHI-mediated radiation responses. Knock-down experiments showed that silencing of DEC2 sensitized both A549 and PC9 cells under PHI-treated conditions. On the other hand, silencing of p53, which regulates BAX/BCL2, desensitized A549 cells expressing wild-type p53, but not PC9 cells, with mutant-type p53, to irradiation, regardless of whether PHI was treated or not. Taken together, PHI modifies radiation responses in a cell type-specific manner, possibly through DEC2 signaling. Full article

This study evaluated the impact of a proprietary blend of fruit and vegetable fibers rich in naturally occurring bound polyphenols (commercially marketed as NatureKnit^TM), compared to purified fibers (inulin and psyllium), on the human gut microbiome using the validated M-SHIME^® gastrointestinal model. A short-term single-stage colonic M-SHIME^® experiment (with fecal inoculum from three healthy human donors) was used to evaluate the test products compared to a negative control. Samples were assessed for pH, gas pressure, short-chain fatty acid (SCFA) production, lactate, and ammonium from 0 h to 48 h. Microbial community composition was assessed at 0 h (negative control only), 24 h, and 48 h (lumen) or 48 h (mucosal). All test products were fermented well in the colon as demonstrated by decreases in pH and increases in gas pressure over time; these changes occurred faster with the purified fibers, whereas NatureKnit™ demonstrated slow, steady changes, potentially indicating a gentler fermentation process. SCFA production significantly increased over the course of the 48 h experiment with all test products versus negative control. SCFA production was significantly greater with NatureKnit™ versus the purified fibers. Shifts in the microbial community composition were observed with all test products versus negative control. At the conclusion of the 48 h experiment, the absolute bacterial abundance and the richness of observed bacterial taxa in the lumen compartment was significantly greater with NatureKnit™ compared with inulin, psyllium, and negative control. Overall, NatureKnit™ demonstrated greater or similar prebiotic effects on study measures compared with established prebiotic fibers. Full article