Search Results (679)

Candida biofilms play a critical role in clinical settings, contributing to persistent and device-associated infections and conferring resistance to antifungal agents, particularly in immunocompromised or hospitalized patients. Biofilm formation varies among Candida species, including C. albicans and non-albicans species, such as C. glabrata, C. tropicalis, C. parapsilosis, and C. auris, due to species-specific transcriptional networks that regulate modes of biofilm development, extracellular matrix composition, and metabolic reprogramming. These differences influence biofilm responses to treatment and the severity of infections, which can be further complicated in polymicrobial biofilms that modulate colonization and virulence. Understanding the mechanisms driving biofilm formation and interspecies interactions is essential for developing effective therapies and requires appropriate experimental models. Available models range from simplified in vitro systems to more complex ex vivo and in vivo approaches. Static in vitro models remain widely used due to their simplicity and reproducibility, but they poorly mimic physiological conditions and require careful standardization. Ex vivo tissue models offer a balance between practicality and biological relevance, enabling the study of biofilm physiology, host–microbe interactions and immune responses. In vivo models, primarily in mice, remain the gold standard for testing antifungal therapies, while alternative systems such as Galleria mellonella larvae provide simpler, cost-effective approaches. Advanced in vitro platforms, including organ-on-chip systems, bridge the gap between simplified tests and physiological relevance by simulating fluid dynamics, tissue architecture, and immune complexity. This review aims to examine Candida biofilms across species, highlighting differences in structural diversity and clinical implications, and to provide a guide to the most widely used experimental models supporting studies on Candida biofilm biology for the development of new therapeutic targets or drug testing. Full article

The role of Signal Transducer and Activator of Transcription 2 (STAT2) in cancer remains poorly understood. STAT2 is a key mediator of type I interferon (IFN) signaling, activating the expression of IFN-stimulated genes with antiviral and antiproliferative effects. However, emerging evidence suggests that STAT2 can also promote tumor growth. Here, we show that high STAT2 mRNA expression in colon cancer tumors correlates with reduced overall survival in patients. In preclinical models, deletion of STAT2 in tumor cells suppressed tumor growth, whereas STAT2 overexpression enhanced tumor growth, supporting its pro-tumorigenic role. To determine whether this function depends on type I IFN receptor (IFNAR1) signaling, we generated IFNAR1 knockout (IFNAR1 KO) colon carcinoma cells and compared their growth with parental and STAT2-deficient (STAT2 KO) tumor cells. Loss of type I IFN signaling was confirmed by western blot and qPCR analyses. In vitro, IFNAR1 KO and STAT2 KO tumor cells proliferated at similar rates. However, in xenograft tumor transplantation models, IFNAR1 KO cells formed larger tumors while STAT2 KO tumor cells formed smaller ones compared to parental tumor cells. These findings indicate that STAT2 promotes colorectal cancer growth through mechanisms independent of IFNAR1 signaling. Full article

Polyphenols, as natural compounds abundant in plant-derived foods, have been recognised for their human health benefits. This study evaluates the multifunctional properties of Biombalance^TM (BB), a grape seed extract rich in oligomeric procyanidins, in various in vitro and in vivo models. BB was studied to assess (i) its antimicrobial effects in different bacterial species; (ii) its protective effects against oxidative and inflammatory stress in Caco-2 cells; and (iii) its effects in mice, which were fed a standard diet with or without BB at two different doses (BB1X and BB2X) to understand the impacts of BB on microbiota and gut homeostasis. BB selectively inhibited several bacterial species, including Staphylococcus aureus, Helicobacter pylori, and Blautia coccoides. In addition, BB protected Caco-2 cells against hydrogen peroxide (H₂O₂)-induced oxidative damage and lipopolysaccharide (LPS)-induced oxidative and inflammatory stress. In vivo, BB supplementation upregulated the expression of antioxidant and homeostasis genes in the colon, ileum, and liver, accompanied by dose-dependent changes in the gut microbiota composition. Functional predictions indicated favourable modulation of microbial metabolic pathways, including those involved in antioxidant capacity and glutamate degradation. Furthermore, BB positively influenced key gut–brain axis mediators, including GLP-1, the GLP-1 receptor, and NPY. These findings highlight the potential of Biombalance^TM to support health and gut–brain communication and to protect against oxidative and inflammatory stress in the gut. Full article

Background: Inflammatory bowel diseases (IBD) rely on invasive methods for detecting intestinal inflammation, with the needs for non-invasive molecular imaging tools being unmet. Integrin α4β7 is a key target in IBD pathogenesis due to its role in the recruitment of T cells. This study aimed to develop a novel ⁶⁸Ga-labeled integrin α4β7-targeted radiopharmaceutical (⁶⁸Ga-A2) and evaluate its feasibility for non-invasive PET/CT imaging of IBD inflammation in a dextran sulfate sodium (DSS)-induced murine colitis model. Methods: ⁶⁸Ga-A2 was synthesized via radiolabeling DOTA-A2 with ⁶⁸Ga. In vitro properties (radiochemical purity, stability, binding specificity, and affinity) of ⁶⁸Ga-A2 were validated. The DSS-induced colitis model was established and confirmed in C57BL/6J mice, followed by in vivo PET/CT imaging, ex vivo biodistribution studies, and histological (HE and IHC) analyses to evaluate the targeting efficacy of ⁶⁸Ga-A2. Results: ⁶⁸Ga-A2 was prepared efficiently (20 min) with a radiochemical purity of >95% and demonstrated good in vitro stability. It exhibited specific binding to integrin α4β7 with a Kd of 68.48 ± 6.55 nM. While whole-body PET/CT showed no visible inflammatory focus uptake, ex vivo imaging and biodistribution of colon tissue revealed significantly higher uptake in DSS-treated mice compared to that in healthy/blocking groups, which was consistent with histological evidence of inflammation. Conclusions: ⁶⁸Ga-A2 demonstrated specific targeting of IBD inflammatory foci in vitro and ex vivo. Despite whole-body imaging limitations, further optimization of its structure may enable it to become a promising non-invasive PET agent for IBD. These findings support future clinical investigations to validate its utility in IBD diagnosis and monitoring. Full article

Human exposure to Di(2-ethylhexyl) Phthalate (DEHP) occurs through ingestion of contaminated food. Yet, the effects of DEHP on gastrointestinal toxicity at the cellular level are poorly understood and studies conducted to date have used malignant cell lines, limiting our understanding of molecular signaling in intestinal epithelia of otherwise healthy individuals. The objective of our study was to use a non-transformed, colonic epithelial cell line (Young Adult Mouse Colonocytes; YAMCs) to characterize the in vitro effects of DEHP on non-malignant colonic epithelia. A 72 h DEHP exposure significantly reduced cell number and proliferation while short-term exposure increased: cellular apoptosis, BAX expression, Reactive Oxygen Species (ROS) production, gene expression linked to oxidative stress (NRF2, GCLC, HO-1, CHOP). Antioxidant pretreatment prior to DEHP exposure attenuated the phthalate’s apoptotic effect, suggesting a link between oxidative stress and apoptosis. Using YAMCs with a CRISPR-deleted Aryl Hydrocarbon Receptor (AhR) we further showed that the apoptotic and pro-oxidative effects of the phthalate are partially mediated through AhR. In conclusion, we have demonstrated that DEHP-induced toxicity in non-malignant colonocytes is due to ROS-induced oxidative stress and subsequently, apoptosis. We have further demonstrated that these effects are partly mediated by the AhR, a mechanism that deserves further investigation. Future studies should build on these findings by (a) characterizing the specific mechanisms linking ROS production to apoptosis demonstrated in our model of exposure, (b) measuring the dynamics of the receptor following DEHP exposure and (c) examining these effects over a longer exposure period. Full article

Background/Objectives: Consuming apples regularly has positive effects on human health due to their anti-inflammatory and antioxidant properties, which have been associated with their phenolic composition. To enhance the bioactive properties of apple phenolic compounds, high-pressure processing (HPP) has been studied as a tool to improve their extraction during gastrointestinal digestion with the aim of increasing their bioaccessibility and the amount that reaches the colon unchanged, which can serve as substrates for bacterial fermentation. This study aimed to analyze the impact of an HPP-apple ingredient on the metabolism of human gut microbiota using an in vitro dynamic simulator of gastrointestinal digestion and colonic fermentation (GID-CF) that allowed us to study the three colon regions separately (ascending—AC; transverse—TC; and descending—DC). Methods: Apples were HPP-treated (400 MPa/5 min) and lyophilized to obtain an HPP-apple ingredient in powder form. A GID-CF was employed to study the continuous intake of the HPP-apple ingredient for 14 days at 37.5 g/day. Results: The HPP-apple ingredient produced a significant accumulation of phenolic metabolites mainly in the DC, with benefits on human health. The main phenolic metabolites formed were phloroglucinol, 4-hydroxyphenylacetic acid, 4-hydroxy-3-methoxyphenylacetic acid, 3-(3-hydroxyphenyl)-propionic acid, and 3-(4-hydroxyphenyl)-propionic acid. A PCA revealed a perfect separation of the three colon regions based on the phenolic precursors and metabolites. The microbiota-modulatory effects were attributed to the increase in Bifidobacterium spp. and Lactobacillus spp. populations and the butyric acid (SCFA) concentration. Conclusions: The results obtained highlight the health benefits and potential prebiotic-like effect of the HPP-apple ingredient on the gut microbiota. 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

Helicobacter pylori urease (HpU) plays a central role in bacterial survival and virulence by hydrolyzing urea into ammonia and carbon dioxide, neutralizing gastric acidity, and facilitating host colonization. The increasing prevalence of antibiotic resistance underscores the need for alternative strategies targeting essential bacterial enzymes such as urease. In this study, a multistage computational pipeline integrating pharmacophore modeling, machine learning (ML), ensemble docking, and enhanced molecular dynamics simulations were applied to identify novel triazole-based HpU inhibitors. Starting from over seven million compounds in the ZINC15 database, pharmacophore- and ML-based filters progressively reduced the chemical space to 7062 candidates. Ensemble docking across 25 conformational frames of HpU, followed by quantum-polarized ligand docking (QPLD), identified seven promising ligands exhibiting strong binding energies and stable metal coordination. Molecular dynamics (MD) simulations under progressively relaxed restraints revealed three highly stable complexes (CA1, CA3, and CA6). Subsequent well-tempered metadynamics (WT-MetaD) simulations reconstructed free-energy landscapes showing deep, localized basins for CA3 and CA6, comparable to the potent reference inhibitor DJM, supporting their potential as strong urease binders. Finally, unsupervised chemical space mapping using the UMAP algorithm positioned these candidates within molecular regions associated with potent urease inhibitors, further validating their structural coherence and pharmacophoric relevance. An ADMET assessment confirmed that the selected candidates exhibit physicochemical and early safety properties compatible with subsequent in vitro evaluation. This multilevel screening strategy demonstrates the power of combining ML-driven classification, ensemble docking, and enhanced sampling simulations to discover non-hydroxamic urease inhibitors. Although the current findings are computational, they provide a rational foundation for future in vitro validation and for expanding the discovery of triazole-based scaffolds targeting ureolytic enzymes. Full article

Inflammatory bowel diseases (IBDs), including Crohn’s disease and ulcerative colitis, are chronic inflammatory disorders of the gastrointestinal tract, and current therapies are limited by adverse side effects. Combretum micranthum G. Don (kinkeliba), a medicinal plant traditionally used in West Africa, has been reported to possess pharmacological activities and a favorable safety profile. In this study, an ethanolic extract of Combretum micranthum (EECM) was characterized using HPLC-DAD-ESI-MS to identify its phenolic constituents. Acute colitis was induced in C57BL/6 mice using 3% DSS, while EECM (100 and 200 mg/kg) was administered orally for seven days. Disease Activity Index was monitored daily, and colonic injuries were evaluated through macroscopic and histological analyses, as well as hematological and biochemical assessments. In vitro, EECM contained 293.54 mg/g of total phenolic compounds and showed strong antioxidant activity in DPPH, ABTS, and FRAP assays. Furthermore, the extract exhibited antibacterial activity against Bacillus subtilis, Staphylococcus aureus, Salmonella enterica, and Streptococcus pyogenes at various concentrations. In contrast, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa were not affected at the tested concentrations. No antifungal activity was detected against the filamentous fungus Aspergillus brasiliensis and the yeasts Saccharomyces cerevisiae, Candida parapsilosis, and Candida albicans. In vivo, EECM alleviated the clinical signs of colitis, reduced histological damage, and modulated hematological and biochemical parameters. Overall, EECM exhibited significant antioxidant and anti-inflammatory activities and may represent a promising natural candidate for IBD management. Further investigations into chronic experimental models are necessary to establish their therapeutic relevance. Full article

Hypoglycin A is a plant-derived protoxin that causes atypical myopathy in equids. In atypical myopathy-affected horses, metabolomic and microbiome studies have reported alterations in metabolic markers and faecal microbiota composition, pointing to a potential disruption of microbial homeostasis. However, in vivo observations are strongly confounded by host-related factors, underscoring the need for controlled in vitro approaches. To address this, we used an in vitro static batch fermentation model simulating the equine colon to investigate the direct effects of hypoglycin A on microbiota composition and activity. Faecal inocula from healthy horses were incubated in control and hypoglycin A-treated fermenters for 48 h, with serial analyses of hypoglycin A concentration, short-chain fatty acids, and 16S rRNA gene profiles. Hypoglycin A remained stable in the nutritive medium in the absence of microbiota, confirming that its degradation in inoculated fermenters was microbiota-dependent. The results showed significant microbial-associated hypoglycin A degradation without evidence of toxic metabolite formation. The analysis of α- and β-diversity revealed both an effect of incubation time, reflecting the natural temporal dynamics of microbial communities under batch fermentation, and a specific impact of hypoglycin A exposure, with certain taxa such as Paraclostridium being affected. This study provides the first in vitro evidence that the equine microbiota contributes to hypoglycin A degradation. Full article

The intensive development of livestock and poultry farming has heavily relied on antibiotics, leading to widespread antimicrobial resistance and posing serious threats to food safety and public health. As the industry transitions towards reduced antibiotic use and sustainable animal production, probiotics and their metabolites have garnered attention as functional alternatives. Probiotics are typically administered in the form of microecological preparations by mixing them into feed or water, offering advantages in cost-effectiveness and ease of use, with demonstrated efficacy in promoting animal health. Swine-derived probiotics, in particular, demonstrate host-specific advantages due to their natural adaptation to the porcine gastrointestinal environment, which improves intestinal colonization, pathogen inhibition, and immune modulation. Their metabolites, including short-chain fatty acids, bacteriocins, and exopolysaccharides, further contribute to these benefits through antimicrobial, anti-inflammatory, and barrier-strengthening effects. Recent studies have demonstrated improvements in average daily gain (18–22%) and feed conversion ratio (12–15%), along with a reduction in diarrhea incidence (up to 40–45%) in weaned piglets supplemented with certain probiotic consortia. It should be noted, however, that part of the supporting evidence is derived from in vitro or non-porcine models, and practical outcomes in swine may vary depending on husbandry conditions, probiotic strain, and husbandry conditions. This review systematically summarizes the isolation and identification of swine-derived probiotics, the active components and functions of their metabolites, and the mechanisms of action and application effects of these metabolites as antibiotic-alternative feed additives. It primarily focuses on innovative research advances in probiotic metabolites for enhancing antibacterial activity and improving pig growth performance. Furthermore, the review discusses the prospects for commercial applications and future research directions, aiming to provide theoretical foundations and technical references for green and healthy farming practices. Full article

This study characterized leaf extracts of Cymbopogon flexuosus (Ryukyu Lemongrass Corporation, Okinawa, Japan) and evaluated the bioaccessibility and bioactivities of phenolic compounds following a simulated in vitro gastrointestinal model of digestion (in vitro GID) of plant material. Undigested (controls, AqC, EtC) and digested aqueous (AqD) and ethanolic (EtD) extracts were analyzed. Control extracts contained higher total phenolics and flavonoids than digested ones, with EtC showing the highest values. UHPLC-QToF-MS (ultra-high-performance liquid chromatography system coupled to a quadrupole time-of-flight mass spectrometer) identified 32 compounds, including phenolic acids, flavone aglycones, C-glycosides, and derivatives. Hydroxybenzoic acids, coumaric acid, caffeic esters, flavones, tricin derivatives, vitexin, and isoorientin exhibited reduced recovery, while coumaric acid hexoside, ferulic acid hexoside, and isoschaftoside/schaftoside exceeded 100% recovery, suggesting release from the matrix. Some compounds were absent from AqD, and many were found in the pellet, indicating potential colonic metabolism. Antioxidant activity (DPPH, reducing power, β-carotene/linoleic acid) was stronger in controls but always weaker than BHT/ascorbic acid. Extracts mildly inhibited α-amylase but more strongly inhibited α-glucosidase as shown with applied enzyme inhibition assays, especially EtD (76.93% at a concentration of 10 mg/mL), which showed stronger activity than controls but remained below acarbose (87.74% at 1 mg/mL). All extracts promoted HaCaT keratinocyte growth and reduced HCT-116 colon cancer cell viability at 250 µg/mL, with the strongest effects in AqC and AqD. Overall, GID decreased antioxidant activity but enhanced antidiabetic potential, confirming the safety and selective anticancer effects of C. flexuosus extracts. Full article

Introduction: Drug development has traditionally used mathematical models to predict formulation behavior. Objective: Building artificial neural networks for the drug release evaluation of drug delivery systems using sustained-release metronidazole-coated colonic hydrophilic matrices as a model. Methods: The technological factors associated with the biopharmaceutical performance of hydrophilic metronidazole matrices were evaluated using a quality by design approach (QbD). The developed neural network includes variables related to the technological process for producing the matrices. These are related to the materials used, such as the type and viscosity of core polymers, the type of coating agent, or the matrix production process, such as the mixing time of core materials or the percentage of the coating agent. The output variables of the neural network were the percentages of drug released in vitro at 1, 6, 12, and 24 h and the mean dissolution time of the matrix. An iterative quasi-Newton method was used to train the artificial neural network. Results: A neural network with excellent prediction capacity allows selecting the technological variables with the greatest influence on the % of drug dissolved: the type of coating agent used and the percentage of the total weight increase after coating for 1 h and 6 h of drug release and also the viscosity of the HPMC for 12 and 24 h. Conclusions: The optimized neural network demonstrated an excellent predictive capacity for in vitro drug dissolution profiles, allowing the use of this type of methodology based on artificial intelligence methods in the optimization of drug delivery systems. Full article

This scoping review synthesized evidence from 2015 to 2025 on the anti-inflammatory and anticancer potential of pecan (Carya illinoinensis) kernel extracts, focusing on bioactive composition and cell signaling pathway modulation. Pecan kernels contain diverse phenolic compounds including gallic acid, catechin, epicatechin, and ellagic acid, along with tocopherols and unsaturated fatty acids, exhibiting significant cultivar-dependent variation influenced by ripening stage, processing conditions, and orchard management practices. In vitro studies demonstrate that kernel extracts possess substantial antioxidant capacity and exert antiproliferative and cytotoxic effects against various human cancer cell lines, including colon cancer cells, with evidence of apoptosis induction. Extraction methodologies significantly influence bioactive compound recovery and biological activity, with both lipid and phenolic fractions contributing to therapeutic potential. While current evidence highlights promising anti-inflammatory and anticancer properties mediated through modulation of apoptotic pathways, research remains predominantly limited to compositional analyses and in vitro models. Future investigations should elucidate specific molecular mechanisms, identify precise signaling pathway targets, conduct in vivo validation studies, and optimize processing conditions to maximize bioactive retention for potential therapeutic applications in cancer prevention and treatment. Full article

ADAM10 is a transmembrane metalloprotease that regulates diverse signalling functions via the shedding of membrane protein ectodomains, and is implicated in tumour development, including glioblastoma multiforme (GBM) and gastrointestinal (GI) cancers, where high ADAM10 expression is associated with poor prognosis. We assessed the role of ADAM10 by gene knockout (KO) in U251 GBM cells, and its effects on protein shedding and protein expression on cell proliferation and on the growth of tumour xenografts in mice. The growth of tumours was severely delayed, relative to modest effects on proliferation in vitro, suggesting roles particularly in the context of the tumour microenvironment (TME). Proteomics analysis of KO cell-conditioned medium showed decreased levels of known ADAM10 targets such as Notch and Eph receptors and ligands, as well as other proteins involved in cell–cell adhesion, migration, signalling, metabolism, differentiation, and development, including angiogenesis. KO cell and tumour lysate analysis also showed modulation of proteins associated with metabolic and catalytic activity, cell–matrix organisation and differentiation. Similar effects were also observed in the SW620 colon cancer model, indicating broader significance. Furthermore, expression of the associated protein sets also correlated with ADAM10 expression in human GBM and colon cancer specimens (TCGA datasets), indicating clinical relevance. Collagens and proteins associated with matrix deposition and fibril organisation were notably reduced in ADAM10 KO GBM tumours, and histology confirmed decreased collagen fibrils and blood vessels. Unexpectedly, increased chondrocyte differentiation was evident in ADAM10 KO U251 tumours, suggesting a role for ADAM10 in maintaining an undifferentiated phenotype in vivo. Together, our data indicate the importance of ADAM10 in diverse signalling mechanisms in tumours and the TME that promote tumour development. Full article