Search Results (418)

Disruption of the intestinal epithelial barrier is a key driver of gut-derived inflammation in various disorders, yet strategies to preserve or restore barrier integrity remain limited. To address this, we evaluated a four-strain Bifidobacterium mixture—selected for complementary anti-inflammatory potency and industrial scalability—in lipopolysaccharide (LPS)-challenged RAW 264.7 macrophages and a Caco-2/THP-1 transwell co-culture model. Pretreatment with the probiotic blend reduced nitric oxide (NO) release in a dose-dependent manner by 25.9–48.3% and significantly down-regulated the pro-inflammatory markers in macrophages. In the co-culture system, the formulation decreased these markers, increased transepithelial electrical resistance (TEER) by up to 31% at 10⁵ colony-forming unit (CFU)/mL after 48 h, and preserved the membrane localization of tight junction (TJ) proteins. Adhesion to Caco-2 cells (≈ 6%) matched that of the benchmark probiotic Lacticaseibacillus rhamnosus GG, suggesting direct epithelial engagement. These in vitro findings demonstrate that this probiotic mixture can attenuate LPS-driven inflammation and reinforce epithelial architecture, providing a mechanistic basis for its further evaluation in animal models and clinical studies of intestinal inflammatory disorders. Full article

Tricuspid regurgitation (TR) is a well-recognized factor contributing to adverse outcomes and mortality. Recent developments in transcatheter valve repair techniques, with the emergence of tricuspid transcatheter edge-to-edge repair (TEER) devices, have altered the treatment algorithm of TR and now offer a safe and feasible alternative for the effective management of the disease and an improvement in patient symptoms. Evidence from large studies and registries showcases the benefit of tricuspid interventions in terms of heart failure hospitalization and quality of life; however, most studies do not report a significant benefit in terms of hard outcomes. Even though longer-term follow-up may be needed to identify such differences, it is important to also identify distinct patient phenotypes that would benefit the most from such interventions, moving from pure anatomical criteria to an overall assessment of the patient’s clinical status. Therefore, the aim of this review is to provide updates on potential moderators of the effect of tricuspid TEER, focusing on novel anatomical criteria, right cardiac function, and renal physiology, in order to guide patient selection and provide an insightful discussion on the optimal patient phenotype for future trial design. Full article

Oxidative stress, driven by impaired antioxidant defence systems, is a major contributor to cognitive decline and neurodegenerative processes in brain ageing. This study investigates the neuroprotective effects of a natural compound mixture—composed of Hericium erinaceus, Palmitoylethanolamide, Bilberry extract, and Centella asiatica—using a multi-step in vitro strategy. An initial evaluation in a 3D intestinal epithelial model demonstrated that the formulation preserves barrier integrity and may be bioaccessible, as evidenced by transepithelial electrical resistance (TEER) and the expression of tight junctions. Subsequent analysis in an integrated gut–brain axis model under oxidative stress conditions revealed that the formulation significantly reduces inflammatory markers (NF-κB, TNF-α, IL-1β, and IL-6; about 1.5-fold vs. H₂O₂), reactive oxygen species (about 2-fold vs. H₂O₂), and nitric oxide levels (about 1.2-fold vs. H₂O₂). Additionally, it enhances mitochondrial activity while also improving antioxidant responses. In a co-culture of neuronal and astrocytic cells, the combination upregulates neurotrophic factors such as BDNF and NGF (about 2.3-fold and 1.9-fold vs. H₂O₂). Crucially, the formulation also modulates key biomarkers associated with cognitive decline, reducing APP and phosphorylated tau levels (about 98% and 1.6-fold vs. H₂O₂) while increasing Sirtuin 1 and Nrf2 expression (about 3.6-fold and 3-fold vs. H₂O₂). These findings suggest that this nutraceutical combination may support the cellular pathways involved in neuronal resilience and healthy brain ageing, offering potential as a functional food ingredient or dietary supplement. 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

Accurate in vitro models of intestinal permeability are essential for predicting oral drug absorption. Standard models like Caco-2 cells have well-known limitations, including lack of segment-specific physiology, but are widely used. Emerging models such as organoid-derived monolayers and microphysiological systems (MPS) offer enhanced physiological relevance but require comparative validation. We performed a head-to-head evaluation of Caco-2 cells, human jejunal (J2) and duodenal (D109) enteroid-derived cells, and EpiIntestinal^TM tissues cultured on either static Transwell and flow-based MPS platforms. We assessed tissue morphology, barrier function (TEER, dextran leakage), and permeability of three model small molecules (caffeine, propranolol, and indomethacin), integrating the data into a physiologically based gut absorption model (PECAT) to predict human oral bioavailability. J2 and D109 cells demonstrated more physiologically relevant morphology and higher TEER than Caco-2 cells, while the EpiIntestinal^TM model exhibited thicker and more uneven tissue structures with lower TEER and higher passive permeability. MPS cultures offered modest improvements in epithelial architecture but introduced greater variability, especially with enteroid-derived cells. Predictions of human fraction absorbed (F_abs) were most accurate when using static Caco-2 data with segment-specific corrections based on enteroid-derived values, highlighting the utility of combining traditional and advanced in vitro gut models to optimize predictive performance for F_abs. While MPS and enteroid-based systems provide physiological advantages, standard static models remain robust and predictive when used with in silico modeling. Our findings support the need for further refinement of enteroid-MPS integration and advocate for standardized benchmarking across gut model systems to improve translational relevance in drug development and regulatory reviews. Full article

Mitral regurgitation (MR) is a common valvular heart disease associated with significant morbidity and mortality. For patients at high or prohibitive surgical risk, mitral transcatheter edge-to-edge repair (M-TEER) offers a less invasive alternative to surgery. This review outlines key aspects of patient selection and procedural planning for M-TEER, with a focus on clinical and echocardiographic criteria essential for success. Comprehensive imaging—especially 2D and 3D transesophageal echocardiography—is critical to assess leaflet anatomy, coaptation geometry, and mitral valve area. Selection criteria differ between primary and secondary MR and are guided by trials such as COAPT and MITRA-FR. Optimal outcomes rely on careful screening, anatomical suitability, and multidisciplinary evaluation. With growing experience and advancing technology, M-TEER has become a transformative option for treating severe MR in non-surgical candidates. Full article

Chickpeas are used as alternative protein sources in healthy snacks due to their bioactive compounds beneficial for gut health. Combining chickpeas with dark chocolate improves palatability and may enhance biological functionality, although mechanistic evidence is still limited. In this explorative research, we evaluate the nutrigenomic, antioxidant and anti-inflammatory properties of a chickpea and chocolate snack using in vitro Caco-2 (colon adenocarcinoma cells) and THP-1 (monocyte-derived macrophages) models. The total polyphenol content and antioxidant activity were measured after in vitro digestion (30.30 mg/mL to 1.9 mg/mL). Caco-2 epithelia and THP-1 were pre-treated for 4 days (2 h/day) with high (15.1 mg/mL) or low (3.8 mg/mL) concentrations of digests. Inflammation was induced for 3 h by LPS (Lipopolysaccharides) and IL-1β (Interleukin-1β). Transepithelial electrical resistance (TEER) was measured to assess barrier integrity. Gene expression related to tight junctions and inflammation was analysed using qPCR (quantitative polymerase chain reaction). Chocolate and snack digests showed the highest total polyphenol content and 2,2-diphenyl-1-picrylhydrazyl activity. Barrier integrity improved with all treatments. Chickpea upregulated tight junction gene expression. Chickpea and chocolate reduced IL-1β expression in both cell types. In THP-1, the chocolate and the snack upregulated CD206 (mannose receptor C-type 1) expression. IL-10 increased with all treatments. These results pave the way for future research that may support the potential use of this snack as a functional food with antioxidant, gut-protective and anti-inflammatory effects. Full article

Age-related macular degeneration (AMD) is the main cause of blindness in Western nations. AMD models addressing specific pathological pathways are desired. Through this study, a best-practice protocol for polarized porcine single-eye retinal pigment epithelium (RPE) preparation for AMD-relevant models of RPE barrier and polarity is established. Single-eye porcine primary RPE cells (from one eye for one well) were prepared in 12-well plates including Transwell inserts. Different coatings (laminin (Lam), Poly-ᴅ-Lysine (PDL), fibronectin (Fn) and collagens) and varying serum contents (1%, 5% and 10%) were investigated to determine optimal culture parameters for this model. Success rates of cultures, cell number (trypan-blue exclusion assay), morphology/morphometry (light and fluorescence microscopy), protein secretion/expression (ELISA, Western blot), gene expression (qPCR), transepithelial electric resistance (TEER) and polar location of bestrophin 1 (BEST1) by cryosectioning (IHC-Fr) were assessed. Cells seeded on Lam exhibited the highest level of epithelial cells and confluence properties. Fn resulted in the highest cell number growth. Lam and Fn exhibited the highest culture success rates. TEER values and vascular endothelial growth factor secretion were highest when Lam was used. For the first time, polar (Transwell) porcine single-eye RPE morphometry parameters were determined. RPE on Lam showed bigger cells with a higher variety of cell shapes. CIV displayed the lowest claudin 19 expression. The highest basolateral expression of BEST1 was achieved with Lam coating. The higher the serum, the better the cell number increase and confluence success. A reduction in serum on Lam showed positive results for RPE morphology, while morphometry remained stable. A five percent serum on Lam showed the highest culture success rate and best barrier properties. RPE65 expression was reduced by using 10% serum. Altogether, the most suitable coating of Transwell inserts was Lam, and a reduction in serum to 5% is recommended, as well as a cultivation time of 28 days. A protocol for the use of polar porcine single-eye cultures with validated parameters was established and is provided herein. Full article

Secondary mitral regurgitation (sMR) is commonly understood to be secondary to heart failure (HF), left ventricular (LV) dilation, and altered coaptation of the mitral annulus. Three forms of sMR exist: non-ischemic sMR, ischemic sMR, and atrial functional sMR. In the past, there have been limited treatment options for this condition besides medication. Recently, the management of sMR has been revolutionized by the recent advances in percutaneous transcatheter edge-to-edge repair of the mitral valve (m-TEER). However, the major trials investigating this technology have shown that appropriate patient selection is of critical importance to achieve benefit. As such, there is a renewed interest in the accurate diagnosis of sMR. Herein, we review the etiology, management, and diagnosis of sMR in the era m-TEER. Full article

Background: The Tendyne™ transcatheter heart valve (THV) system is a promising option for high-risk patients with severe mitral regurgitation (MR) who are ineligible for surgery or transcatheter edge-to-edge repair (TEER). As most fatal complications occur within the first 90 days, this study aimed to identify anatomical predictors of in-hospital mortality after transcatheter mitral valve replacement (TMVR). Methods: In this subanalysis of the TENDER registry, data from 110 patients who underwent TMVR across 26 centers between January 2020 and June 2022 were evaluated. Preprocedural imaging parameters were analyzed, including transthoracic echocardiography (TTE), transesophageal echocardiography (TEE), and cardiac 4D computed tomography (CT). Results: We identified LVEDDi as a significant predictor of in-hospital mortality (p = 0.022), with lower values in non-survivors (26.42 ± 3.76 mm/m²) than in survivors (30.37 ± 5.58 mm/m²). Both indexed and absolute LVEDDi predicted in-hospital complications (p < 0.001 and p = 0.008). In multivariate analysis, LVEDDi (p = 0.048; OR = 0.856) and STS score (p = 0.038; OR = 1.114) remained independent predictors of in-hospital mortality. In an extended model, only LVEDDi persisted as a significant predictor (p = 0.007), highlighting its robustness. Conclusions: This analysis identified a small LVEDDi as a novel, clinically relevant risk factor in TMVR and showed its added value alongside conventional markers. Its easy calculation supports incorporating LVEDDi thresholds into screening to improve patient selection and outcomes. Full article

Background: Chronic mitral regurgitation (MR) is categorized into primary and secondary MR (SMR). While primary MR arises from structural abnormalities of the mitral valve apparatus, SMR is a consequence of cardiac remodeling, typically due to heart failure or atrial fibrillation. Management strategies differ significantly, with primary MR requiring direct valvular intervention and SMR necessitating a comprehensive approach incorporating guideline-directed medical therapy (GDMT), revascularization, and resynchronization strategies. The MitraClip, a transcatheter edge-to-edge repair (TEER) device, has emerged as a recommended intervention for symptomatic severe SMR despite optimal GDMT. Objectives: This study aims to evaluate national trends in MitraClip placements in the U.S. from 2016 to 2021 and to assess 90-day readmission events following the procedure. Additionally, we analyze patient and socioeconomic factors associated with heart failure readmissions post-MitraClip placement to optimize patient selection criteria. Methods: The study utilized data from the National Inpatient Sample (NIS) for the years 2016–2021 and the National Readmissions Database (NRD) for 2021. Patients who underwent MitraClip placement were identified using ICD-10 code 02UG3JZ. We stratified the population based on demographics, hospital resource utilization, and comorbidities. Index admissions were classified based on the presence or absence of heart failure remissions within 90 days post-procedure. Statistical analyses, including ANOVA and logistic regression, were conducted to identify factors associated with readmissions. Results: MitraClip utilization demonstrated a rising trend from 2016 to 2021, with total annual procedures increasing from 869 to 2488. Mean patient age remained stable at 76–79 years, with a nearly equal sex distribution. In-hospital mortality remained low (1–3%) throughout the study period. A steady increase in hospital charges was observed, alongside a decline in the mean length of stay. Analysis of 4918 index admissions for MitraClip placement in 2021 identified 780 total readmissions within 90 days, with 206 (26.4%) attributed to heart failure. Factors significantly associated with increased risk of heart failure readmissions included atrial fibrillation (OR 3.77, CI 1.82–4.23), pulmonary hypertension (OR 3.96, CI 1.49–5.55), and chronic lung disease (OR 1.91, CI 1.32–2.77). Conclusions: The increasing adoption of MitraClip underscores its growing role in managing SMR. However, heart failure readmissions remain a significant concern. Identifying high-risk patient profiles can refine selection criteria and enhance post-procedural management strategies to improve clinical outcomes. Further research is needed to optimize patient selection and refine risk stratification for MitraClip interventions. Full article

Background: Intestinal epithelial barrier (IEB) dysfunction is related to multiple gastrointestinal disorders, notably inflammatory bowel disease (IBD). Betulinic acid (BA), a compound derived from birch bark, has demonstrated potential therapeutic benefits in IBD. Nevertheless, the impact of BA on IEB function has not been fully elucidated. Methods: The current study aimed to explore the potential underlying mechanisms of BA in dextran sodium sulfate (DSS)-induced IBD in mice and co-culture models involving Caco-2/HT29-MTX-E12 cell monolayers or mouse intestinal organoids (IOs) in conjunction with macrophages stimulated by lipopolysaccharide (LPS). Results: In vivo, BA treatment significantly improved body weight and colon length, alleviated disease activity index (DAI) scores, and reduced colonic histopathological injury in IBD mice. In vitro, BA reduced the flux of FITC-dextran; increased the TEER; and decreased the production of IL-6, IL-1β, and TNF-α while increasing IL-10 mRNA levels. Additionally, BA enhanced IEB formation by upregulating ZO-1, occludin (OCLN), and claudin-1 (CLDN1). Molecular docking studies revealed significant docking scores and interactions between BA and PPAR-γ. Moreover, BA significantly upregulated PPAR-γ protein expression, decreased NF-κB and MLC2 phosphorylation, and reduced MLCK protein expression. However, this effect was reversed by GW9662, an effective PPAR-γ antagonist. Conclusions: The findings reveal that BA mitigates IBD by safeguarding the intestinal barrier against dysfunction. This effect may be attributed to its ability to suppress inflammation and enhance the expression of tight junction proteins by modulating the PPAR-γ/NF-κB signaling pathway. Full article

Background: Transcatheter mitral valve edge-to-edge repair (M-TEER) reduces mitral regurgitation (MR) severity and improves symptoms, but early post-procedural assessment of left atrial pressure (LAP) remains challenging. Objectives: Investigating the impact of M-TEER on diastolic parameters and derived cutoff values to estimate post-procedural LAP. Methods: This retrospective study (n = 240) analyzed the effects of M-TEER on diastolic parameters. Cutoff values for predicting normal LAP were identified using classification tree analysis and validated with current methods for assessing LAP. Results: M-TEER increased E/e′ ratio in both normal and abnormal left ventricular ejection fraction (LVEF) groups. In normal LVEF, E wave velocity ≤ 85 cm/s at 30 days correlated with normal LAP (98% specificity, 90% positive predictive value). In abnormal LVEF, E/e′ ≤ 14 or E wave velocity ≤ 95 cm/s correlated with normal LAP (98%/90% specificity, 91%/83% positive predictive value). The validation of the proposed cutoff values with existing non-invasive methods showed 96% accuracy for normal LAP (24/25) and 90% for elevated LAP (27/30). Conclusions: M-TEER significantly alters diastolic parameters. Derived cutoff values based on easily obtainable diastolic measures show promise in estimating post-procedural LAP, but need further validation for clinical use. Full article

Previously, we developed a porcine bronchial epithelial cell line designated as PBE cells and demonstrated that this cell line possesses functional Toll-like receptor 3 (TLR3), triggering the expressions of interferons (IFNs), antiviral factors, and inflammatory cytokines after its stimulation with the synthetic double-stranded ARN poly(I:C). In this work, we aimed to further characterize the PBE cell line as a reliable in vitro model for investigating swine influenza virus (SIV) infection and immunity. We evaluated the capacity of two SIV subtypes, H1N1 and H3N2, to replicate and induce cytopathic effects in PBE cells and to modulate the expressions of IFNs, antiviral factors, inflammatory cytokines, and negative regulators of the TLR signaling. We demonstrated that PBE cells are susceptible to both H1N1 and H3N2. SIV infected PBE cells inducing notable cytopathic effects as shown by the alteration of transepithelial electrical resistance (TEER) and cilia. Both SIV subtypes replicated in PBE cells in similar proportion and altered TEER values in comparable magnitudes. However, SIV H3N2 induced higher alterations of cilia than H1N1. SIV infection induced changes in all the immune factors evaluated in PBE cells. We detected quantitative differences when the subtypes H1N1 and H3N2 were compared. The fold expression changes of IFN-β, Mx1, Mx2, IFITM1, OAS1, OAS2, and OASL were higher in PBE cells infected with H3N2 than in cells challenged with H1N1. In addition, although both subtypes stimulated IL-8 expression, only the H3N2 induced IL-6 in infected PBE cells. SIV H1N1 and H3N2 also upregulated the expressions of the negative regulators A20, BCL-3, and MKP-1, while only H1N1 increased SIGIRR and Tollip. Immortalized respiratory cell lines from pigs can be useful in vitro systems for the study of viral infections and immune responses. These studies are of importance in the context of influenza infections not only for the agricultural field because pigs are natural hosts of these viruses but also because these animals serve as intermediate reservoirs of viruses that can threaten humans’ health. We demonstrated here that the PBE cell line can be a useful in vitro model to study SIV infection and immunity. Full article

Background and Objectives: MitraClip^® (MC) placement has been extensively used as an intervention for mitral transcatheter edge-to-edge repair (mTEER) for functional mitral regurgitation (FMR). The aim of our study is to analyze the association between the pre-procedural echocardiographic parameters of diastolic function (DF) and one-year outcomes after MC placement. Materials and Methods: The study was designed in a retrospective longitudinal cross-sectional format. In total, 224 patients who underwent MC placement between January of 2021 and March of 2024 were included in the study. The Primary Efficacy Endpoint (PEE) was determined by an absence of heart failure hospitalizations requiring Intravenous Diuretics or cardiac-related death in the one-year follow-up period. Multivariate regression analysis was carried out to identify the pre-procedural echocardiographic parameters of DF that had a significant association with a failure to reach the PEE. A two-tailed p-value < 0.05 was used to determine statistical significance. Results: Of the 224 patients included in the study, 85 patients (37.94%) failed to reach the PEE or had worsening symptoms. The mean mitral valve (MV) deceleration time was 176.88 ms (164.14–189.62) in the symptom-worsening group compared to 201.53 ms (186.01–217.07) in the symptom-improvement group. The mean of the E/A ratio (MV peak E velocity/A velocity) was noted to be 2.35 (1.97–2.74) in the symptom-worsening group compared to 1.90 (1.68–2.13) in the symptom-improvement group. After multivariate regression analysis, the E/A ratio was found to have a significant association with a failure to reach PEE: Odds Ratio (OR): 1.61 (1.13–2.29), p-value: 0.008. The area under the curve (AUC) analysis for the E/A ratio was calculated at 0.603 (0.50–0.69) for the symptom-worsening group. Conclusions: Patients that failed to reach the PEE had a lower pre-procedural MV deceleration time of 176.88 ms (164.14–189.62); however, no association was observed between MV deceleration time and a failure to reach the PEE in the multivariate regression analysis. The pre-procedural E/A ratio had a significant association with symptom worsening after multivariate regression analysis: OR: 1.61 (1.13–2.29). The AUC for the E/A ratio in the symptom-worsening group was 0.603, making it a more moderate predictor than random guessing for the failure to reach the PEE. Full article