Search Results (1,880)

Liquid infant formula has garnered increasing attention due to its mild thermal processing and superior retention of bioactive nutrients. Within such matrices, the lipid source is a critical determinant of protein digestion behavior, yet its influence on peptide bioavailability and intestinal homeostasis remains undefined. Given that efficient peptide absorption is vital for the systemic delivery of bioactivity in infants, understanding the lipid–protein synergy is essential for formula optimization. Moreover, excessive oxidative stress is closely associated with impaired intestinal health and developmental disorders in infants, making the regulation of oxidative stress crucial for maintaining intestinal function. The present study evaluated the effects of three distinct lipid sources—soybean oil (SM), bovine milk fat (BM), and goat milk fat (GM)—on the physicochemical stability, proteolytic digestion, peptide release, intestinal absorption, and oxidative stress modulation of goat-milk-based infant formula. An integrated approach combining physicochemical characterization, in vitro simulated infant digestion, and a Caco-2 intestinal epithelial cell model was employed. we demonstrate that all three lipids (3% w/w) formed stable emulsions with uniform spherical structures and mean particle diameters of 117–300 nm, as visualized by laser confocal microscopy. Following in vitro simulation of infant gastrointestinal digestion, the SM group exhibited the most extensive protein hydrolysis, yielding the highest total peptide content (4.28 ± 0.10 mg/mL) and generated the highest number of peptides identified by LC-MS/MS (474 types). Bioinformatic analysis predicted that peptides from all groups possess potential antihypertensive, hypoglycemic, and immunomodulatory activities. The Caco-2 monolayer cell model demonstrated that although the GM group produced fewer identified peptide species than the SM group (365 types), it achieved significantly higher intestinal peptide absorption rate (55.34 ± 1.05%). Furthermore, the GM digests provided superior protection against H₂O₂-induced oxidative stress in Caco-2 cells, markedly reducing reactive oxygen species levels and suppressing the expression of pro-inflammatory cytokines TNF-α and IL-6. Collectively, these findings reveal that while soybean oil promotes more extensive proteolysis, the use of homologous goat milk lipid enhances peptide bioaccessibility and confers potential cytoprotective effects on intestinal epithelial cells, underscoring its potential as a preferred lipid source in infant formula formulations. Full article

Osteoporosis is characterized by reductions in bone mineral density (BMD) and bone quality. While gut-derived signaling has been increasingly studied in relation to BMD, its contribution to molecular factors associated with bone quality remains less defined. Here, we investigated whether a heat-inactivated, freeze-dried, non-viable preparation of Levilactobacillus brevis AS-1 modulates intestinal epithelial-like cells and thereby promotes lysyl oxidase (LOX), a key enzyme involved in collagen cross-linking. Caco-2 cells were treated using 1 mM sodium butyrate and subsequently stimulated with 100 μg/mL L. brevis AS-1. Supernatants were collected and applied to MG63 cells. Cytokine mRNA expression in Caco-2 cells and LOX responses in MG63 cells were analyzed by qRT-PCR, and bone morphogenetic protein (BMP-1) and transforming growth factor-β (TGF-β)1 protein levels in the supernatant were measured by ELISA. L. brevis AS-1 stimulation up-regulated BMP-1 and TGF-β1 mRNA expression in SB-treated Caco-2 cells and increased BMP-1 protein secretion into the supernatant. LOX mRNA expression and total LOX activity were increased in MG63 cells treated with the conditioned supernatant, and inhibition of BMP-1/procollagen C-proteinase activity (UK383367) attenuated LOX mRNA induction. Collectively, these results suggest that L. brevis AS-1 stimulates intestinal epithelial-like cells to secrete BMP-1, which in turn promotes LOX mRNA expression in osteoblast precursor cells. This in vitro mechanism supports the concept of gut–bone crosstalk regulating molecular factors associated with bone quality. Full article

The intestinal epithelial barrier regulates paracellular transport, and its dysfunction is associated with inflammatory and metabolic diseases. Among dietary fibers, inulin has attracted considerable attention due to its beneficial effects on intestinal health. Inulin’s actions have been attributed to protecting the structure and function of gut barrier components against inflammatory-associated damage. This review integrates preclinical and clinical studies evaluating the impact of inulin on intestinal permeability. Evidence from in vitro and in vivo models shows that inulin regulates the expression of tight junction proteins (TJPs), Paneth cell proliferation, and antimicrobial peptides, and modulates inflammatory signaling pathways. In addition, inulin prebiotic activity, via microbiota, stimulates the production of short-chain fatty acids (SCFAs) as butyrate that reinforces the barrier function. Understanding these pathways highlights the therapeutic potential of inulin as a nutritional strategy for treating barrier dysfunction. Clinical studies in obesity, metabolic disorders and inflammatory intestinal disease have associated inulin supplementation with improvements in biomarkers of intestinal permeability. Future studies are needed to test inulin’s safety in order to prevent potential risks and hazards. Full article

Viral infections with gastrointestinal involvement remain a significant global health burden with limited therapeutic options. While probiotics show antiviral potential, their impact on primary human intestinal epithelial defenses is poorly defined. This study utilized human intestinal organoid-derived monolayers (ODMs), generated from the non-inflamed mucosa of patients with inflammatory bowel disease, to examine how Bifidobacterium animalis ssp. lactis BB-12 (BB-12) and Lacticaseibacillus rhamnosus GG (LGG) modulate mucosal antiviral pathways. Unlike conventional Caco-2 cells, ODMs preserved physiological cellular diversity and intact innate signaling. Expression of viral receptors and interferon (IFN)-stimulated genes (ISGs) was quantified by RT-qPCR, while the effector 2′-5′-oligoadenylate synthetase 1 (OAS1) was also assessed by immunofluorescence and flow cytometry. Both probiotic strains modulated IFN-associated pathways; however, BB-12 induced a markedly stronger antiviral transcriptional response than LGG. Notably, OAS1 exhibited cell type-specific regulation; while goblet cells showed high basal levels, both probiotics enhanced OAS1 expression selectively in ileal enterocytes. Despite this shared effect, only BB-12 pretreatment significantly restricted Influenza A (H1N1) replication in ileal ODMs, whereas LGG did not significantly affect viral replication. These findings establish human ODMs as a superior platform for probiotic immunology, suggesting that BB-12 more effectively shapes epithelial antiviral “set-points” and highlighting OAS1 as a sensitive component of a broader antiviral program. Full article

The vitamin D receptor (VDR) acts as both a nuclear transcription factor and a non-genomic mediator that regulates a broad spectrum of physiological processes beyond calcium and phosphate homeostasis. VDR plays an important role in the modulation of ion channels across multiple tissues, including osteoblasts, renal and intestinal epithelial cells, neurons, and vascular smooth muscle. These regulatory mechanisms encompass genomic actions through vitamin D response elements in target genes—such as TRPV5, TRPV6, KCNK3, and KCNH1—as well as rapid, non-genomic actions at the plasma membrane involving protein disulfide isomerase A3 and associated signaling cascades. VDR-mediated transcriptional control of calcium, potassium, and chloride channels contributes to the fine-tuning of cellular excitability, calcium transport, and mitochondrial function. Evidence also implicates VDR–ion channel crosstalk in various pathological contexts, including renal cell carcinoma, breast and cervical cancers, pulmonary arterial hypertension, and osteoporosis. Understanding the molecular interplay between VDR and ion channels provides new perspectives on the pleiotropic effects of vitamin D and offers promising therapeutic opportunities in oncology, cardiovascular disease, and skeletal disorders. This review synthesizes previous and current evidence on the genomic and non-genomic mechanisms underlying VDR–ion channel regulation and highlights novel frontiers in vitamin D signaling relevant to human health and disease. Full article

Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea, dehydration and death in piglets, resulting in significant economic losses in the pig industry. It is crucial to identify the pathogenesis and mechanism between host–PEDV interactions. In our study, we performed transcriptomic and metabolomic analyses in PEDV-infected Large White (LW) pigs. PEDV infection caused blunted and fused intestinal villi, necrosis of the intestinal mucosal epithelial cells and atrophy of intestinal glands. Transcriptomic and metabolomic analyses revealed 692 differentially expressed genes and 1485 differential metabolites, respectively. Among them, differentially expressed genes were enriched in virion assembly, lipoprotein metabolic process and PPAR signaling pathway. Differential metabolites were enriched in primary bile acid biosynthesis and lipoic acid metabolism. An integrated analysis of the transcriptome and metabolome revealed that differentially expressed genes and metabolites were co-enriched in steroid hormone biosynthesis and bile secretion. In addition, key metabolites Dehydroepiandrosterone (DHEA) and Estriol in steroid hormone biosynthesis both inhibited PEDV infection and alleviated the excessive inflammatory response in vitro. Collectively, our study constructed a multi-omics landscape of PEDV infection in LW pigs, providing potential targets for developing metabolic-targeted antiviral interventions. Full article

Colorectal cancer (CRC) is the third most common cancer globally and the second leading cause of cancer-related deaths. While intestinal microbiota dysbiosis is linked to CRC, the direct role of intratumoral bacteria in metastasis remains poorly understood. In this study, we isolated pathogenic bacteria from CRC tumor tissues, identified as Hungatella hathewayi (H. hathewayi), through the 16S rRNA gene and whole-genome sequencing. We developed specific primers (P48/P52) and polyclonal antibodies for detecting H. hathewayi in samples. Using quantitative real-time PCR (qPCR), we found significant enrichment of H. hathewayi in fecal samples from CRC patients compared to healthy controls, with mean fold changes of 137-fold and 142-fold for primers P48 and P52, respectively. Analysis of tissue samples revealed that H. hathewayi abundance was higher in CRC tumor tissues compared to normal tissues, with mean fold changes of 2.90 for P48 and 3.97 for P52. Fluorescence in situ hybridization (FISH), immunofluorescence (IF), and immunohistochemistry (IHC) confirmed its spatial distribution within tumor tissues. In vitro assays using CRC cell lines demonstrated that H. hathewayi-derived succinate upregulates HIF-1α and SUCNR1 expression and promotes cell metastasis by inducing epithelial–mesenchymal transition (EMT). Collectively, these findings identify H. hathewayi as a novel pro-metastatic bacterium and a potential non-invasive biomarker for CRC diagnosis, providing direct evidence for the role of intratumoral bacteria in CRC progression. Full article

The high incidence of Candida albicans infections and the limited efficacy of current antifungal therapies highlight the need for new antifungal agents. In this study, we present a bio-based hybridization strategy aimed at enhancing the antifungal activity of natural product scaffolds, with a particular focus on trans-ferulic acid. A library of twenty-nine hybrid molecules was rationally generated by grafting naturally occurring lipophilic moieties onto either the phenolic or carboxylic acid functions of ferulic acid. The antifungal activity of these molecules was then assessed against C. albicans. While the parent natural compounds exhibited weak activity (MIC > 500 µM), several hybrid derivatives (ATF19, ATF20, and MB22) demonstrated enhanced potency, with MIC values of <50 µM. Esters of the carboxylic acid or phenol group were essential for activity, with the most potent effects observed for short linear or mildly branched lipophilic chains. These active compounds exerted a multifaceted anti-virulence effect, including mitochondrial membrane depolarization, inhibition of hyphal morphogenesis, alterations in cell wall composition, and strong suppression of biofilm formation. Additionally, lead compounds showed no detectable cytotoxicity in human macrophages and intestinal epithelial cells and significantly improved host survival in a Caenorhabditis elegans model of C. albicans infection. Overall, the ferulic acid, citronellol, and sinapic hybrid molecules emerged as promising lead compounds for the development of antifungals against C. albicans. Full article

Background: Transmissible gastroenteritis virus (TGEV) is a highly pathogenic porcine coronavirus that causes severe gastrointestinal damage in piglets. However, how TGEV affects host iron homeostasis, oxidative stress, and the ferroptosis process remains unclear. This study aimed to investigate the effects of TGEV infection on cellular iron metabolism, oxidative damage, and lipid peroxidation-mediated ferroptosis, as well as to evaluate the potential therapeutic role of retinoic acid (RA). Methods: Using an intestinal epithelial cell model of TGEV infection, we assessed key regulators of iron handling, oxidative stress, lipid peroxidation, and ferroptosis. The expression of ferroportin (FPN) and ferritin (FTH/L) and the activity of the p62–NRF2–GPX4/HO-1 antioxidant axis were analyzed, and the effects of exogenous RA treatment on these endpoints were examined. Results: TGEV infection disrupted cellular iron homeostasis by downregulating the expression of ferroportin (FPN) and ferritin (FTH/L), leading to the accumulation of intracellular free iron, which in turn induced the generation of a large amount of reactive oxygen species (ROS) and ultimately triggered ferroptosis in intestinal epithelial cells. Additionally, TGEV infection significantly inhibited the p62-NRF2-GPX4/HO-1 antioxidant signaling pathway, further exacerbating the ferroptosis process. Conclusions: This study reveals that ferroptosis is a key pathological mechanism in TGEV-induced intestinal injury and demonstrates that RA exerts a therapeutic effect by regulating iron metabolism and activating the p62-NRF2-GPX4/HO-1 signaling pathway. These findings provide new theoretical insights for potential intervention strategies targeting virus infection-associated ferroptosis and intestinal damage. Full article

Deleterious crosstalk between the gut and distant organs is a key factor behind disease progression. Currently, the molecular signals mediating this communication remain elusive. We hypothesized that systemic oxidative stress and oxidatively modified serum proteins transmit injury signals from extraintestinal sites to the gut. In various murine models of organ injury, primary damage was consistently associated with systemic oxidative stress and intestinal damage. Specifically, ischemia/reperfusion (I/R)-induced acute kidney injury caused profound colonic barrier defects. Depleting the microbiota with antibiotics markedly improved survival and attenuated both renal and colonic injury, implicating translocated microbes in exacerbating pathology. Mechanistically, these changes were linked to systemic oxidative stress and were largely prevented by the antioxidant N-acetylcysteine. Furthermore, serum from I/R mice disrupted epithelial barrier integrity and induced cell death in vitro, effects that were recapitulated by exposure to oxidized serum proteins. Characterization of serum components identified albumin as the predominantly oxidized protein, which displayed potent cytotoxicity toward cultured intestinal epithelial cells. Our findings establish oxidative stress and oxidized serum albumin as key pathogenic factors mediating the detrimental interaction between remote organs and the gut. These data suggest that targeting oxidative modifications offers a promising therapeutic strategy to disrupt this pathological loop in critical illness. Full article

Physiologically relevant in vitro intestinal models are essential for studying key physiological processes, including barrier function, drug screening and gut-microbiota interactions. However, conventional 2D culture systems often fail to recapitulate structural and functional complexity. Here, we aimed to validate a 3D bioelectronic transmembrane platform, previously used for monitoring human intestinal epithelium and vascular endothelium, for modeling the rat small intestinal barrier in vitro. The device integrates a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) scaffold supporting co-cultures of rat intestinal epithelial cells (IEC-6) and rat fibroblasts (208F), enabling real-time monitoring of barrier formation through electrical measurements using electrochemical impedance spectroscopy (EIS). Barrier formation was monitored over 21 days and exhibited a time-dependent increase in barrier resistance. The 3D platform was compared with traditional 2D insert-based cultures and ex vivo rat tissue using an Ethylene Glycol Tetraacetic Acid (EGTA)-induced calcium switch assay to evaluate barrier disruption and recovery. EGTA treatment and removal induced reversible barrier disruption in the 3D in vitro and ex vivo models, whereas 2D in vitro cultures showed limited recovery. These findings demonstrate that the 3D platform more faithfully recapitulates native tissue architecture and function, closely paralleling ex vivo responses. Our study highlights the importance of validating advanced 3D in vitro models and establishes this bioelectronic platform as a robust tool for drug screening, barrier studies, and preclinical gastrointestinal research. Full article

Biochar, a carbon-rich material traditionally used to improve soil health and as a feed additive, has recently attracted attention for its potential biological activity. This study examined the effects of an aqueous biochar extract (BC-AE) on human intestinal epithelial cells (Caco-2), focusing on its influence on cell viability and apoptosis. The metabolic activity of Caco-2 cells exposed to BC-AE was first evaluated using an MTS assay. A concentration of 3 mg/mL, which promoted Caco-2 metabolic activity, was selected for further testing at 24 and 72 h. The effect of BC-AE on cell viability was assessed by epifluorescence microscopy (morphology) and flow cytometry (apoptosis profiling). The transcriptional response of cell viability-related genes (BAX, BAD, BCL-2, BCL-xL, MCL-1, P21, and P53) and microRNAs (miR-15b, miR-19, miR-21, miR-33a, miR-155, and miR-486) was analyzed by RT-qPCR. In parallel, selected proteins (BAD, BAX, BCL-2, and MCL-1) were examined by Western blotting. We showed that BC-AE decreased cell viability after 24 h via late apoptosis, while 72 h exposure increased necrosis without further viability loss. Both BAX and MCL-1 protein levels increased in Caco-2 cells after 72 h of BC-AE treatment, and miR-15b and miR-21 were upregulated, suggesting the involvement of a regulatory mechanism controlling cell survival. The obtained findings highlight the importance of considering both concentration and exposure duration when assessing biochar bioactivity and represent an additional contribution to the ongoing effort to better understand the biological role. Full article

Successful organ regeneration depends on coordinated cell-to-cell communication mediated by ligand–receptor interactions that regulate proliferation, differentiation, and axonal guidance. Sea cucumbers, particularly Holothuria glaberrima, exhibit remarkable regenerative capacity following evisceration, regenerating their complete intestinal system within weeks. To identify molecular signals orchestrating these events, we characterized five ligand–receptor groups of axonal guidance molecules (Netrin/UNC5-DSCAM, Ephrin/Eph receptors, Semaphorin/Plexin, RGMα/Neogenin, and SLIT/ROBO) using transcriptomic databases from regenerating intestines and the radial nerve cord. Comparative analyses confirmed these as highly conserved orthologs, retaining characteristic structural domains essential for guidance signaling. Multiple alternatively spliced isoforms were detected, with tissue-specific variants suggesting functional diversification. Differential gene expression analysis across intestinal regeneration stages (12 h to 21 days post-evisceration) revealed distinct temporal patterns: Netrin-1 showed significant upregulation at 7–14 days post-evisceration, coinciding with nerve fiber invasion into the intestinal anlage, while the Ephrin, Semaphorin, and SLIT–ROBO pathways exhibited late-stage expression associated with luminal tissue formation. Single-cell RNA sequencing from 9-dpe regenerating intestines localized Netrin to coelomic epithelial cells and UNC5B to differentiating epithelial cells, with CellChat analysis predicting strong epithelial-to-epithelial signaling. These findings strongly suggest that axonal guidance molecules play dual roles during intestinal regeneration: directing neural innervation in early-to-mid stages and orchestrating tissue boundary formation at later stages. Full article

Emerging evidence suggests that gut microbiota significantly influence female reproductive health by affecting hormonal, immune and metabolic processes. This research explored how a probiotic blend comprising Lactobacillus crispatus novaLCR6, Limosilactobacillus fermentum novaLF58 and Bifidobacterium bifidum novaBBF9 affects the gut–myometrium and gut–ovary axes. Intestinal epithelial cells were exposed to individual probiotics or their combination using a Transwell^® setup; their effects on barrier integrity, probiotic activity and short-chain fatty acid production were measured. Subsequently, basolateral metabolites were applied to myometrial and ovarian cells to assess viability, proliferation, oxidative stress, inflammation, signalling pathways and hormone production. All probiotics enhanced intestinal cell viability and barrier function. The combined probiotic showed synergistic effects, enhancing butyrate production by ~23–51%, improving myometrial proliferation by up to ~78%, decreasing ROS and TNF-α levels by ~49% and ~74% and modulating oxytocin signalling. In ovarian cells, the probiotic mixture activated ERK/MAPK and PI3K/AKT pathways, normalised PAK1, ERβ and PAX8 expressions and significantly increased LH and FSH secretion compared to single strains. These findings suggest that a multi-strain probiotic may modulate pathways involved in reproductive tissue homeostasis through gut–reproductive axis interactions, providing mechanistic insight from an in vitro study. Full article

The mycotoxin deoxynivalenol (DON) is a common contaminant found in swine diets, causing decreased growth performance and poor health. Additionally, F18 enterotoxigenic E. coli is a leading cause of post-weaning diarrhea. Nursery pigs are often exposed to each of them after weaning; however, it is unknown what impact the combination of these stressors has on gastrointestinal health. Therefore, the objective of this study was to investigate the effect of pre-exposure to DON on the response of intestinal porcine epithelial cells (IPEC-J2) to challenge with enterotoxigenic F18 E. coli. Four groups were compared: Control (untreated cells), DON (cells treated with 0.5 μM DON for 24 h), F18 E. coli (multiplicity of infection 5:1, varied duration) and DON + E. coli (DON treatment with subsequent E. coli infection). Gene expression of IL-8, IL-6 and TNFα was significantly increased in cells infected with E. coli for 3 h vs. uninfected cells (p < 0.0001, p < 0.0001 and p < 0.0001, respectively). There was an interactive effect between DON and E. coli on IL-8 gene expression; cells pretreated with DON before E. coli infection had a higher expression of IL-8 than those not pretreated (p < 0.05). The concentration of IL-8 protein was significantly increased by E. coli (p < 0.0001). Claudin 1 and Occludin protein abundance were reduced by E. coli as measured by Western blot. Cytotoxicity was increased by E. coli vs. Control (p < 0.05). Pretreatment with DON increased the amount of E. coli that adhered to IPEC-J2 cells (p < 0.01) 30 min post-infection. FITC-dextran passage was increased in the DON + E. coli treatment vs. E. coli alone (p < 0.0001). Transepithelial electrical resistance (TEER) was decreased by DON when compared to untreated cells at 0 h (p < 0.0001). Similarly, DON + E. coli exhibited lower TEER vs. E. coli alone at 2 h post-infection (p < 0.0001). Taken together, these results indicate that DON pre-exposure increased the severity of E. coli infection on endpoints such as barrier permeability and E. coli adhesion. Full article