Search Results (2,269)

The purpose of this study was to characterise the effect of cranberry (Vaccinium macrocarpon) juice on unicellular and multicellular systems, specifically food spoilage yeasts (Wickerhamomyces anomalus, Dekkera bruxellensis and Rhodotorula mucilaginosa) and intestinal cells (IEC-6 and Caco-2 cells). The effects of both raw cranberry juice and juice digested in vitro were investigated. The juices were analysed for polyphenol content using high-performance liquid chromatography coupled with mass spectrometry. The cranberry juice was evaluated for its impact on yeast surface hydrophobicity and anti-adhesive action using the MATH test and luminometry/microscopy, respectively. We also assessed the effects of raw and digested cranberry juices on IEC-6 and Caco-2 cells by measuring cell viability, metabolic modulation, genotoxicity, and antioxidant activity. Chromatographic analysis of the raw cranberry juice revealed the presence of diverse bioactive compounds, identified as hydroxybenzoic and hydroxycinnamic acids, flavonols, and anthocyanins. After digestion, the cranberry juice remained a rich source of phenolic acids. The yeast strain R. mucilaginosa was characterised by the highest hydrophobicity and adhesive abilities, but cell adhesion in the presence of raw cranberry juice was several times lower for all the tested strains. Both tested cranberry juices reduced ROS levels and were well tolerated by intestinal epithelial cells, without significant cytotoxic or genotoxic effects. Our findings provide new insights into the safety of using cranberry juice across unicellular and multicellular systems. However, further validation in real-world settings is necessary before practical applications. Full article

Background/Objectives: Ulcerative colitis (UC) is an inflammatory bowel disease and a major cause of ulcers and chronic inflammation in the colon and rectum. Recurring symptoms include abdominal pain, rectal bleeding, and diarrhoea, and patients with UC are at a higher risk of developing comorbidities such as colorectal cancer and poor mental health. In UC, the decreased diversity and changed metabolic profile of gut microbiota, along with a diminished mucus layer, leads to disruption of the underlying epithelial barrier, with an ensuing excessive and detrimental inflammatory response. Treatment options currently rely on drugs that reduce the inflammation, but less emphasis has been placed on improving the resilience of the epithelial barrier. Macrolide antibiotics exhibit epithelial barrier-enhancing capacities unrelated to their antibacterial properties. Methods: We investigated two novel barriolides, macrolides with reduced antibacterial effects in common bacterial strains. Gut epithelial cell barrier resistance in the Caco-2 cell line, with and without co-culture with mucus-producing HT-29 cells, was increased when treated with barriolides. Using ^AXGut-on-Chip technology with inflammatory cytokine-stimulated Caco-2/HT-29 co-cultures, the effectiveness of the barriolides was confirmed. Lastly, we reveal the barrier-enhancing and inflammation-reducing effects of the barriolides in a dextran-sulphate sodium (DSS)-induced colitis mouse model. Results: We show the predictive power of the novel ^AXGut-on-Chip system and the effectiveness of the novel barriolides. Indications include reduced inflammatory response, increased epithelial barrier and decreased overall clinical score. Conclusions: The results of this study indicate the notion that barriolides could be used as a treatment option for UC. Full article

Background/Objectives: Assessing the bioavailability of nutrients and bioactive compounds in vitro commonly relies on coupling standardized gastrointestinal digestion models with intestinal epithelial cell systems. However, digests produced using static digestion protocols such as INFOGEST often impair epithelial barrier integrity, limiting their direct application to intestinal models and reducing reproducibility across studies. Methods: This work systematically compared five commonly used digest conditioning strategies, including acidification, centrifugation, rapid freezing, and ultrafiltration using 10 kDa and 3 kDa molecular weight cut-off membranes, to identify the approach that best preserves intestinal epithelial viability and barrier function while enabling exposure at physiologically relevant concentrations. INFOGEST digests of yogurt were initially evaluated, followed by validation using biscuit and canned mackerel digests. Cell viability and monolayer integrity were assessed in differentiated Caco-2 cells using MTT assay and transepithelial electrical resistance (TEER) measurements. Results: Among the tested approaches, ultrafiltration using 3 kDa membranes consistently preserved epithelial viability and barrier integrity at a 1:10 dilution across all food matrices, whereas other conditioning methods failed to maintain TEER despite acceptable cell viability. At lower dilutions, food-dependent effects emerged, highlighting the importance of matrix-specific evaluation. Conclusions: These findings identify 3 kDa ultrafiltration as an effective and minimally invasive strategy to improve the compatibility of INFOGEST digests with intestinal cell models. By enabling reproducible exposure conditions that preserve epithelial integrity, this approach supports more reliable in vitro assessment of nutrient bioavailability and contributes to methodological standardization in nutrition research. Full article

Background/Objectives: Magnesium (Mg²⁺) supplements can contain different types of Mg²⁺ salts, which influence their bioavailability. A highly bioavailable and bioaccessible Mg²⁺ source is essential to meet requirements for many physiological processes that are fundamental to human health. The objective of this study was to compare the bioavailability of Mg²⁺ from different sources, with different composition and chemical structure, namely, Aquamin Mg Soluble (seawater), magnesium oxide, commercial magnesium bisglycinate 1, and analytical grade magnesium bisglycinate 2. In addition, the influence of the different Mg²⁺ sources on transported Mg²⁺ and expression of TRPM6 and TRPM7 genes in Caco-2 cell monolayers was also evaluated to estimate bioavailability. TRPM6 and TRPM7 are members of the transient receptor potential melastatin subfamily characterized as Mg²⁺ permeable channels. Method: The study involved analyzing bioavailability of the Mg²⁺ sources predigested with and without food using the Infogest model prior to application to a Caco-2 cell monolayer in transwells for assessing transport. Mg²⁺ concentration on the basolateral side was analyzed by ICP-MS, and expression of TRPM6 and TRPM7 genes in the monolayer was analyzed using real-time qPCR. Results: Aquamin Mg Soluble showed significantly higher bioavailability compared to magnesium bisglycinate 2 (p = 0.016) when digested with food prior to application to the Caco-2 monolayer. In the digestion without food prior to the Caco-2 monolayer, there was no significant difference between Mg²⁺ bioavailability among the tested supplements. The TRPM6 gene was significantly downregulated in Caco-2 monolayers exposed to Aquamin Mg Soluble compared to untreated Caco-2 cells (p < 0.001). Conclusions: The INFOGEST digestion model showed that Aquamin Mg Soluble provides a highly bioavailable form of Mg²⁺, while the Caco-2 monolayer model also demonstrated its increased bioavailability by the modulation of TRPM6 gene expression. Full article

Background/Objectives: Essential amino acids’ (EAAs) biological effects depend on both gastrointestinal stability and intestinal bioavailability. A commercially available EAA blend has previously shown to be highly bioaccessible and able to inhibit the DPP-IV enzyme both directly and at a cellular level following simulated digestion in vitro. In light with this consideration, the present study aimed to evaluate the intestinal in vitro bioavailability of GAF subjected to INFOGEST digestion (iGAF) and to investigate the metabolic effects of its bioavailable fraction on muscle cells using an integrated Caco-2/C2C12 co-culture model. Methods: Differentiated Caco-2 cell lines were treated with iGAF, and amino acid transport was quantified by ion-exchange chromatography. The basolateral fraction containing bioavailable EAAs was used to treat differentiated C2C12 myotubes for 24 h. Western blot analyses were performed to assess the activation of anabolic and metabolic pathways, including mTOR, Akt, GSK3, AMPK and GLUT-4. Results: More than 50% of each EAA present in iGAF crossed the Caco-2 monolayer, with BCAAs and phenylalanine particularly enriched in the basolateral fraction. Exposure of C2C12 myotubes to the bioavailable iGAF stimulated mTORC1 activation and increased the phosphorylation of Akt and GSK3, indicating an enhanced anabolic response. At a cellular level, iGAF also elevated the p-AMPK/AMPK ratio, suggesting activation of energy-sensing pathways. Moreover, GLUT4 protein levels and glucose uptake were significantly increased. Conclusions: The study focuses exclusively on a cellular model, and results suggested that iGAF is highly bioavailable in vitro and that its absorbed fraction activates key anabolic and metabolic pathways of skeletal muscle cells, enhancing both protein synthesis signaling and glucose utilization in vitro. Full article

Iron deficiency anemia remains a global nutritional challenge due to the low bioavailability and side effects of conventional inorganic iron supplements. A novel organic iron supplement, oyster peptide ferrous chelate (OP-Fe), was prepared using oyster peptides as ligands. Its preparation process was optimized via single-factor experiments and statistical methods with the optimal conditions identified as 1% peptide concentration, 35 °C chelation temperature, 3.91:1 peptide-to-iron ratio, 1.49% ascorbic acid concentration and pH 6.05. Under these conditions, the chelate’s iron content reached 15.44 ± 0.18 g/kg. Multi-dimensional characterization confirmed that Fe²⁺ formed stable complexes with oyster peptides through carboxyl, amino, and imidazole groups. In vitro Caco-2 cell experiments showed OP-Fe achieved a maximum iron absorption rate of 76.07%, significantly higher than ferrous sulfate (52.39%). In vivo pharmacokinetic studies in mice demonstrated higher iron accumulation in serum and small intestine for OP-Fe. Key iron transport-related genes (PEPT1, TFR1, DMT1) were upregulated, contributing to enhanced absorption. OP-Fe exhibits favorable structural stability and bioavailability, holding potential as an efficient iron supplement. Full article

Background/Objectives: Rotavirus remains a major cause of severe acute gastroenteritis in infants worldwide. The suboptimal efficacy of current vaccines underscores the need for alternative microbiome-based interventions, including postbiotics. Extracellular vesicles (EVs) from probiotic and commensal E. coli strains have been shown to mitigate diarrhea and enhance immune responses in a suckling-rat model of rotavirus infection. Here, we investigate the regulatory mechanisms activated by EVs in rotavirus-infected enterocytes. Methods: Polarized Caco-2 monolayers were used as a model of mature enterocytes. Cells were pre-incubated with EVs from the probiotic E. coli Nissle 1917 (EcN) or the commensal EcoR12 strain before rotavirus infection. Intracellular Ca²⁺ concentration, ROS levels, and the expression of immune- and barrier-related genes and proteins were assessed at multiple time points post-infection. Results: EVs from both strains exerted broad protective effects against rotavirus-induced cellular dysregulation, with several responses being strain-specific. EVs interfered with viral replication by counteracting host cellular processes essential for rotavirus propagation. Specifically, EV treatment significantly reduced rotavirus-induced intracellular Ca²⁺ mobilization, ROS production, and COX-2 expression. In addition, both EV types reduced virus-induced mucin secretion and preserved tight junction organization, thereby limiting viral access to basolateral coreceptors. Additionally, EVs enhanced innate antiviral defenses via distinct, strain-dependent pathways: EcN EVs amplified IL-8-mediated responses, whereas EcoR12 EVs preserved the expression of interferon-related signaling genes. Conclusions: EVs from EcN and EcoR12 act through multiple complementary mechanisms to restrict rotavirus replication, spread, and immune evasion. These findings support their potential as effective postbiotic candidates for preventing or treating rotavirus infection. Full article

To improve the oral bioavailability of oleanolic acid (OA), this study developed a menthol–fatty acid-based hydrophobic deep eutectic solvent (HDES) system. Through a comprehensive evaluation using in vitro simulated digestion and Caco-2 cell transport models, the short-chain HDES was found to increase the apparent in vitro bioavailability index of OA by 9.3-fold compared to conventional ethanol systems, with efficacy showing clear fatty acid chain-length dependence. The mechanism was systematically investigated through spectral characterization and cellular studies, revealing a two-stage enhancement process: during the digestion phase, HDES significantly improved OA bioaccessibility to 14.30% compared to 4.90% with ethanol; during the absorption phase, it markedly increased cellular uptake to 25.79% versus 4.71% with ethanol. Molecular analysis indicated that the optimal hydrophobicity and diffusion properties of HDES contributed to this enhancement. This study reveals a fatty acid chain-length-dependent mechanism in HDES-facilitated OA delivery, providing a tunable strategy for enhancing the absorption of hydrophobic bioactive compounds. Full article

Chronic inflammation constitutes a well-established driver of colorectal carcinogenesis, yet the molecular circuitry linking inflammatory receptor signalling to tumour cell survival remains incompletely delineated. Here we demonstrate that the HMG-CoA reductase inhibitors atorvastatin and rosuvastatin modulate inflammatory survival pathways in colorectal cancer cells in a manner consistent with targeted interference with the protease-activated receptor 2 (PAR-2)–extracellular signal-regulated kinase (ERK)–tumour necrosis factor-α (TNF-α) signalling axis. Using lipopolysaccharide-stimulated HT-29 and Caco-2 cells as complementary models of inflammatory colorectal malignancy, we show that both statins selectively attenuate PAR-2 expression at the protein and transcript levels while leaving structurally related PAR-1 unaffected. This pattern of receptor modulation is accompanied by suppression of total ERK1/2 expression, ERK1/2 phosphorylation, and the transcriptional target DUSP6, together with attenuation of TNF-α secretion. Importantly, these signaling shifts are associated with dual apoptotic programs; the extrinsic pathway, reflected by transcriptional upregulation and proteolytic activation of caspase-8; and the intrinsic mitochondrial pathway, evidenced by reciprocal modulation of Bcl-2 family proteins favoring Bax over Bcl-2. Both pathways converge upon activation of executioner caspase-3 and an increase in Annexin V-defined apoptotic fractions, indicating re-engagement of programmed cell death under inflammatory stress. Notably, rosuvastatin consistently demonstrates superior potency across signaling endpoints, achieving comparable biological effects at lower concentrations than atorvastatin. Collectively, these data indicate that clinically deployed statins target the PAR-2–ERK axis and are associated with re-activation of apoptotic pathways in inflammatory colorectal cancer models, while leaving open the possibility that additional statin-responsive networks contribute to their pro-apoptotic effects. This mechanistic framework provides biological plausibility for epidemiologic observations linking statin use with reduced colorectal cancer risk and improved outcomes, and supports further translational evaluation of PAR-2-directed statin strategies in colorectal malignancy. Full article

This study examined whether co-fermentation with Lactobacillus casei CGMCC 15956 and Lactobacillus delbrueckii CGMCC 21287 could enhance the bioactivity of peptides derived from pecan nut cake (PNC) and clarify the underlying mechanisms. The fermented hydrolysate (PNCH) was compared with an unfermented control. PNCH showed higher antioxidant and α-glucosidase inhibitory activities. Total antioxidant capacity increased from 3.17 to 4.81 mM Trolox, and DPPH radical scavenging activity increased from 62.69% to 84.12%. In addition, the IC₅₀ value for α-glucosidase inhibition decreased from 7.549 to 4.509 mg/mL. In a mouse model of acute alcohol-induced liver injury, PNCH significantly alleviated liver damage through the synergistic enhancement of antioxidant and α-glucosidase inhibitory activities. Peptidomic analysis identified two representative bioactive peptides, FAGDDAPR (from actin) and LAGNPDDEFRPQ (from cupin domain–containing protein 1), both of which exhibited antioxidant and α-glucosidase inhibitory activities. Additionally, these peptides alleviated H₂O₂-induced oxidative stress in Caco-2 cells, significantly improving GSH and MDA levels, as well as SOD activity. Molecular docking suggested potential interactions of these peptides with superoxide dismutase, Keap1, and α-glucosidase. These findings support the high-value utilization of PNC and the development of functional peptide-based ingredients. Full article

Aflatoxin B1 (AFB1), deoxynivalenol (DON), and ochratoxin A (OTA) are common mycotoxins that frequently co-occur in cereals and pose potential risks to animal and human health. This study investigated the cytotoxic effects of AFB1, DON, and OTA, individually and in binary and ternary combinations, in four human-derived cell lines representing major target organs (Caco-2, HepG2, HK-2, and SK-N-SH). Individual toxin exposure revealed cell type–dependent sensitivity, with DON generally exhibiting the strongest cytostatic effect. Combined exposure analysis showed distinct interaction patterns across cell models, including antagonistic effects of AFB1 + OTA in most cell lines, dose-dependent interactions of DON + OTA, and low-dose synergistic effects in specific combinations. Overall, the results demonstrate that mycotoxin interactions are highly dependent on dose and target cell type, and that low-dose co-contamination may enhance toxicological risks, underscoring the importance of considering combined mycotoxin exposure in health risk assessment. Full article

Background: Gut barrier dysfunction and altered gut microbial metabolism are emerging signatures of chronic gut disorders. Considering growing interest in combining structurally and mechanistically distinct bioactives, we investigated the individual and combined effects of serum-derived bovine immunoglobulin (SBI) and N-acetylglucosamine (NAG) on the gut microbiome and barrier integrity. Methods: The validated ex vivo SIFR^® (Systemic Intestinal Fermentation Research) technology, using microbiota from healthy adults (n = 6), was combined with a co-culture of epithelial/immune (Caco-2/THP-1) cells. Results: While SBI and NAG already significantly improved gut barrier integrity (TEER, transepithelial electrical resistance, +21% and +29%, respectively), the strongest effect was observed for SBI_NAG (+36%). This potent combined effect related to the observation that SBI and NAG each induced distinct, complementary shifts in microbial composition and metabolite output. SBI most selectively increased propionate (~Bacteroidota families) and health-associated indole derivatives (e.g., indole-3-propionic acid), while NAG most specifically boosted acetate and butyrate (~Bifidobacteriaceae, Ruminococcaceae, and Lachnospiraceae). The combination of SBI_NAG displayed effects of the individual ingredients, thus, for instance, enhancing all three short-chain fatty acids (SCFA) and elevating microbial diversity (CMS, community modulation score). Conclusions: Overall, SBI and NAG exert complementary, metabolically balanced effects on the gut microbiota, supporting combined use, particularly in individuals with gut barrier impairment or dysbiosis linked to lifestyle or early-stage gastrointestinal disorders. Full article

Colorectal cancer is one of the leading causes of cancer-associated mortality, and multifactorial resistance remains one of the main challenges in its treatment. Essential oils and their main compounds show interesting anticancer properties, but their mechanism of action is yet to be defined. This study aims to assess the cytotoxic effects of eugenol (EU) and cinnamaldehyde (CN) on colorectal cancer (CRC) cells, highlighting possible mechanisms of action. These compounds were tested on normal immortalized colonocytes (NCM-460) and two CRC cell lines: Caco-2, a human colon epithelial adenocarcinoma cell line, and SW-620, colon cancer cells derived from a lymph node metastatic site. The efficacy of EU and CN was evaluated through CellTiter-Glo^® and clonogenic assays and by determining proinflammatory cytokine secretion. Transcriptome analysis was used to identify possible pathways affected by EU and CN treatments. The results confirmed that EU and CN were selectively cytotoxic and pro-apoptotic against CRC cells, with different putative mechanisms. While EU drove cytotoxicity through robust transcriptional remodeling, CN yielded a stronger anti-inflammatory action. We confirmed that EU and CN are promising natural candidates in CRC prevention and treatment, even in association with chemotherapeutic drugs. Full article

Fermentation, one of the oldest food processing techniques, is known to play a pivotal role in improving the nutritional and functional characteristics of cereals, with positive implications for gut health and overall well-being. The present study aims to examine the phenolic acids, peptides, and potential bioactive properties of 2 novel sorghum-based fermented beverages, Niselo and Delishe. A total of 48 phenolic compounds were identified through targeted and untargeted Ultra-High Performance Liquid Chromatography coupled with a Quadrupole Orbitrap High-Resolution Mass Spectrometer (UHPLC–Q-Orbitrap HRMS) analyses, revealing a higher content of phenolic acids in Niselo and a prevalence of flavonoids in Delishe. Niselo exhibited enhanced in vitro cytoprotective and reactive oxygen species (ROS)-scavenging activity and displayed a clear cytoprotective effect against hydrogen peroxide-induced oxidative stress in Caco-2 cells. Proteomic profiling using tryptic digestion revealed that Niselo has a substantial abundance of fragments of peptides matching several stress-related and antioxidant proteins, indicating a superior stress-response and/or defense capability. Overall, these findings highlight the functional potential of sorghum-based fermented beverages, supporting their role as health-promoting products. Full article

Chalcone-based derivatives were designed as dual-acting ligands targeting the histamine H₃ receptor (H₃R) and monoamine oxidase B (MAO-B), based on the lead compound DL76. Three series of compounds (1–18) were synthesised and characterised, including simple chalcones (1–9) and piperidinyl chalcones (10–18). All piperidinyl derivatives exhibited nanomolar affinity for human H₃R (hH₃R), with compounds 10–12 achieving K_i values ≤ 30 nM. Simple chalcones showed potent human MAO-B (hMAO-B) inhibition (IC₅₀: 0.85–337 nM), especially 3,4-dichloro derivatives. Compound 15 was the most active hybrid, with a K_i of 46.8 nM for hH₃R and an IC₅₀ of 212.5 nM for hMAO-B. Molecular docking and 250 ns simulations revealed stabilising interactions at both binding sites and clarified structural features behind dual activity. Preliminary ADMET profiling showed low Caco-2 permeability and rapid microsomal metabolism, mainly via hydroxylation. Compound 15 exhibited micromolar cytotoxicity in SH-SY5Y and HepG2 cells, induced G2/M arrest, disrupted mitochondrial homeostasis, and was genotoxic in Peripheral Blood Mononuclear Cells (PBMCs). Additionally, for H₃R ligands (15, DL76, pitolisant), the study reports the first use of Surface Plasmon Resonance Microscopy (SPRM) to assess their interactions with this receptor. Therefore, piperidinyl chalcones show promise as ligands with dual action on H₃R and MAO-B, useful in the treatment of neurodegeneration and/or CNS cancers. Full article