West Nile Virus inhibits type I interferon response via TRIM71 in human microglial cells.

T cells are critical to control Toxoplasma gondii (T. gondii) parasite infection. Yet, our understanding of T. gondii-specific CD8+ T cell responses in humans remains scarce. Here, we studied 56 HLA-A2+ healthy blood donors, including 40 latently infected subjects. In addition to routine T. gondii serodiagnosis, we quantified IgG specific for the bradyzoite-restricted Brain Cyst Load-associated Antigen (BCLA) and we enumerated blood-circulating parasite-specific CD8+ T cells by IFN-γ ELISPOT using a panel of 29 T. gondii peptides, previously reported to be HLA-A2 ligands. We identified a set of 16 epitopes that stimulates detectable CD8+ T cell responses in 50% routine T. gondii seropositive subjects, yet none was a universal immunodominant T. gondii epitope. Among individuals with either positive BCLA-specific serology and/or detectable T. gondii-specific T cell response, T. gondii-specific ELISPOT responses were inversely correlated with BCLA-specific IgG titers, suggesting that stronger CD8+ T cell responses may contribute to reduce chronic parasite burden or that certain long-standing infections may result in waning of antibody levels but persistence of CD8+ T cell memory. Furthermore, 2 out of 56 subjects displayed null T. gondii-specific humoral responses across 4 serological assays, but had measurable CD8+ T cell responses to 2 T. gondii peptides. This study sheds light on the complexity of T. gondii-specific immune responses in humans and rationalizes the cellular and serological toolkits for immune monitoring of latent T. gondii infection in humans.

Despite guideline-directed reperfusion and pharmacological therapies, myocardial infarction (MI) continues to pose substantial clinical challenges, with high incidence and mortality rates. Mesenchymal stromal cell (MSC) transplantation represents a promising therapeutic strategy, yet its efficacy remains limited by poor survival and unstable paracrine function of transplanted cells within the hostile infarct microenvironment. Here, we developed an integrated microcarrier system via digital light processing, which encapsulates MSCs within microgels functionalized with cardiac-derived decellularized extracellular matrix (dECM) particles (termed MSCs@dECMMG). By leveraging the native biochemical composition and structural cues of cardiac dECM, this bioinspired microcarrier was designed not only to enhance MSC viability but also to actively modulate their secretory activity. Conditioned medium from MSCs@dECMMG promoted angiogenic responses in human umbilical vein endothelial cells and attenuated oxidative stress‑induced apoptosis in H9c2 cells. In the rat MI model, local implantation of MSCs@dECMMG markedly improved the left ventricular ejection fraction from 32.03% to 60.77%, reduced the fibrotic area by 17.44%, and increased the left ventricular wall thickness by 2.6-fold. Collectively, this study demonstrates that a dECM‑integrated microgel can serve as a bioactive carrier that directs MSC secretory signaling, offering a novel strategy to advance cell‑based therapy for MI. STATEMENT OF SIGNIFICANCE: Low survival and unstable paracrine effects of transplanted MSCs in the hostile infarct microenvironment remain a major clinical challenge. Addressing how to specifically enhance MSC paracrine function is critical for effective cardiac repair. Here, we present a precision-engineered microcarrier system that integrates mechanically ground cardiac dECM particles into DLP-bioprinted microgels to deliver MSCs. Unlike prior studies focusing on proliferation or differentiation behaviors, we further uncovered and harnessed cardiac dECM to potentiate MSC paracrine signaling, markedly improving myocardial repair following MI. This introduces a new dimension to dECM application-from guiding differentiation to paracrine enhancement-while providing a translational-ready platform with scalability and controllability for future cell-based therapies.

Adding beta-tricalcium phosphate ceramic to additive manufactured 3D porous Ti6Al4V scaffolds enhances osteogenic activity of human mesenchymal stromal cells in vitro.

Individual cells of the same type and in the same position in the cell cycle show stochastic cell-to-cell variability (noise) in gene and protein expression, even under the same environmental conditions. The aim of the study was to test if radiation-induced DNA damage response of cells shows noise, on top of the randomly distributed DNA damage. This was tested by comparing the frequency of DNA repair foci in sister nuclei of binucleated cells (intra BNC variability) and nuclei of two BNC (inter BNC variability). Inter cell variability was also measured for mononucleated cells (MNC). Binucleation was induced in U2OS cells by cytochalasin B. Cells were exposed to 2Gy of photons, 53BP1 and γH2AX foci were visualised and counted in nuclei of binucleated and mononucleated cells 30, 60, 120 and 180min post exposure. Coefficients of variation were calculated as indicators of variability. For both types of foci, the frequencies observed in sister nuclei within BNC were positively correlated and exhibited significantly lower variation compared to the variability measured between single nuclei from randomly paired BNC. The contribution of noise to the total variability was 28% for 53BP1 foci and 50% for γH2AX foci. The results demonstrate that the variability of focus frequency observed in nuclei of BNC results from a combination of noise and random damage variability.

Vascular endothelial cells (VECs) play a critical role in tissue and organ homeostasis, wound healing, and immune function. VECs are commonly utilized for in vitro cell culture and in vivo tissue engineering applications which experience intrinsic and extrinsic stressors. Severe stress can overwhelm VECs adaptive stress response mechanisms inducing damage and loss of cell viability. Improving VEC stress tolerance is therefore of increasing interest. In this study we assess the transgenic expression of tardigrade extremotolerance-associated proteins (TEAPs), specifically CAHS3, MAHS, LEAM, and Dsup for improved VEC tolerance to heat shock and hyperosmotic stress. In vitro studies of all TEAP-expressing VECs demonstrate up to 45% decrease in population metabolic activity in response to 24h incubation in hyperosmotic media solution without recovery. Interestingly, acute hyperosmotic stress treatments followed by a 24h recovery period showed significant increase in population metabolic activity for CAHS3 (186%), Dsup (202%), and LEAM (223%), while MAHS (161%) showed non-significant increases. Similarly, heat shock treatments followed by a recovery period showed significant increases for CAHS3 (198%), Dsup (182%), MAHS (195%), and LEAM (156%). Our results demonstrate that TEAP expression in VECs confer context dependent protective effects with respect to heat shock and hyperosmotic stress, motivating further exploration of TEAPs stress tolerance function and mechanism of action in mammalian cells.

Patients living with human immunodeficiency virus 1 (PLWH) develop accelerated liver fibrosis, but the exact mechanism remains unknown. Activation of hepatic stellate cells (HSCs)-a central driver of hepatic fibrogenesis-is influenced by various factors, including viral infection, hepatocellular injury, chronic immune activation, gut barrier dysfunction, and microbial translocation. The role of gram-positive microbial products in human immunodeficiency virus 1 (HIV-1) infection-associated liver inflammation and fibrosis remains poorly understood. This study investigates the effect of lipoteichoic acid (LTA), a major gram-positive bacterial component, on HSCs in the context of HIV-1 infection. Human primary HSCs (pHSCs) were isolated from liver tissues of HIV-1-infected and uninfected individuals undergoing hepatic resection. Inflammatory responses of HSCs to LTA stimulation were measured via ELISA in patient-derived HSCs, and in vitro Lx2 cells (a human HSC line) before and after HIV-1BaL exposure. Western blotting, ChIP-qPCR and RNA-seq were used on Lx2 cells to explore relevant gene associations. LTA modestly induced interleukin-8/CXCL8 (IL-8) production in HSCs, but this response was heightened in both patient-derived HIV-1 exposed pHSCs and in vitro HIV-1BaL exposed Lx2 cells. IL-8 mRNA levels were higher in HIV-1-infected tissues compared to uninfected controls. LTA exposure increased IL-8 mRNA in LX2 cells, and IL-8 treatment of Lx2 cells increased α-SMA and COL1A1 expression. Transcriptomic analysis on Lx2 cells co-exposed to LTA and HIV-1BaL suggested a link between histone acetylation and IL-8. HIV-1 exposure in patient-derived pHSCs and Lx2 cells was linked to increased IL-8 response of HSCs to LTA. In vitro IL-8 treatment of Lx2 cells supports a role of IL-8 in pro-fibrogenic signaling in HSCs. These findings link gut-derived gram-positive microbial products and IL-8 activation in HSCs, providing important insights to guide future mechanistic studies and potential therapeutic targets for managing HIV-associated liver disease.

Fine particulate matter 2.5 (PM2.5) is linked to rising cases of respiratory, cardiovascular, and immune-mediated diseases and increased hospitalizations in non-respiratory patients. Chiang Mai Province in Northern Thailand, faces seasonal severe PM2.5 pollution, but its impact on immune responses in human remains poorly characterized. This study aimed to investigate the association between air pollution and non-communicable diseases in Chiang Mai and to assess how PM2.5 collected in Chiang Mai (CM-PM2.5) during the peak of poor air quality in 2020 affected the innate immune responses of primary human monocytes. PM2.5, PM10, NO2, and O3 air quality index (AQI) levels concurrently increased throughout the year. Notably, PM2.5 and NO2 AQI levels were positively correlated with the incidence of chronic obstructive pulmonary disease (COPD) in Chiang Mai. Transcriptomic profiling of primary human monocytes stimulated with CM-PM2.5 (5 and 20μg/ml) for 24h indicated a dose-dependent effect on gene expression profiles. Alterations in pyrimidine ribonucleotide metabolism were observed at both CM-PM2.5 concentrations. Upregulation of key metabolic genes (CYP1B1, HK2, and XYLT1) and suppression of glycolytic activity were observed with treatment of 20μg/ml of CM-PM2.5. This study provides strong evidence that exposure to PM2.5 disrupts metabolism and alters responses of human innate immune cells which may contribute to increased severity of respiratory diseases.

Bioelectrical signaling, a collection of electrical currents driven mainly by ionic fluxes and transmembrane potential gradients, is recognized as a central regulator of cell behavior, tissue patterning, and regeneration, extending far beyond classical roles in excitable tissues. While electrically conductive materials are widely used to deliver exogenous stimulation, the specific contribution of intrinsic biomaterial charge (fixed charges, counterions, piezoelectric polarization, electric field) to bioelectrical signaling remains comparatively underexplored. This review focuses on how charged biomaterials influence bioelectrical signaling and downstream human cell responses. We first outline the fundamentals of bioelectricity, including electrochemical gradients, Nernst potentials, and resting membrane potential as key regulators of ion flux, metabolism, and cell fate. We then discuss major classes of charged biomaterials (charged polymers, ionically conductive hydrogels, piezoelectric systems, and charged nanoparticles) and how their charge density, polarity, and dynamic behavior shape protein adsorption, integrin-mediated signaling, or membrane potential. Mechanistic links between material charge and some signaling pathways are highlighted in the context of cell adhesion, viability, migration, proliferation, and differentiation. Finally, we examine tissue-specific applications in cardiac tissue engineering, bone regeneration, neural interfaces, and skin wound healing and immunomodulation. Charged biomaterials have the potential to modulate bioelectrical signaling not only through electronic conductivity but also via electrostatic fields, Donnan effects, and piezoelectric or ionic charge transport. In many regenerative systems, the effects of charge are confounded with mechanical and biochemical cues; mechanistic studies that isolate charge-driven bioelectric phenomena remain limited. Although it is known that surface charge affects the adaptive and innate immune systems, such as through macrophage polarization, the broader consequences of charge-mediated immune modulation remain inadequately explored. Therefore, a more comprehensive discussion and studies on the cell-biomaterial charge interactions are warranted to advance the field and guide future biomaterial design.

Malignant pleural mesothelioma (MPM) is a rare cancer of the pleura, frequently related to asbestos exposure. It is characterized by a grim prognosis, as few therapeutic options are available. Typically, MPM exhibits high molecular heterogeneity, providing opportunities for exploiting therapeutic vulnerabilities in well-defined subgroups of patients. Nevertheless, these options have been insufficiently explored. We found that inactivation of Protein Phosphatase 2A (PP2A) is a recurrent event in MPM, affecting the cancer cell phenotype and chemotherapy response. Specifically, in silico analysis showed that heterozygous loss of PTPA (PPP2R4), encoding the PP2A activator PTPA, occurred in 26% of MPM patients (TCGA, n = 87), correlating with significantly decreased overall survival. Re-expression of PTPA in PTPA-low human MPM cells decreased 2D colony formation and anchorage-independent cell growth, without affecting cell migration. At enzyme level, PTPA restoration increased PP2A catalytic subunit expression and activity (methylation), without favoring specific PP2A holoenzymes. Comparative RNAseq and Gene Set Enrichment Analysis revealed increased RAS pathway activation, activated p53 signaling and induction of senescence (SA-β-Galactosidase, SASP, NF-κB activation) upon PTPA restoration, presumably by increased PP2A-dependent dephosphorylation of Kinase Suppressor of Ras 1. Suppressing Ras signaling by Trametinib (MEK-inhibitor) prevented senescence induction, and pretreatment with senolytic agents restored impaired growth. ATUX-792, a pharmacologic PP2A activator, did not phenocopy senescence induction. Finally, PTPA restoration sensitized MPM cells to carboplatin as the likely consequence of lethal stress induction. PTPA promotes oncogene-induced senescence (OIS) in MPM. By preventing OIS, heterozygous PTPA loss may contribute to mesothelial transformation and carboplatin resistance.

Toxic shock syndrome toxin-1 (TSST-1) is a critical superantigen produced by certain staphylococcal strains, closely associated with menstrual toxic shock syndrome and mucosal inflammation. While the systemic superantigenic function of TSST-1 is well-established, its early cytotoxic and inflammatory effects on human vaginal epithelial cells (HVEC) remain poorly characterized. This study, therefore, sought to investigate the initial temporal dynamics of TSST-1-induced changes in cell viability, inflammatory cytokine responses, epithelial stress-related responses, and barrier-associated transcriptional changes in a human vaginal epithelial cell model, along with transcriptomic analysis. We evaluated the impact of recombinant TSST-1 on HVEC at different doses and treatment durations. Subsequent exposure assessments included cell viability, apoptosis, and cell cycle distribution via flow cytometry, alongside inflammatory gene expression and transcriptomic profiling. TSST-1 reduced cell viability and proliferation in a dose-and time-dependent manner due to increased apoptosis and G1-phase accumulation. Notably, early exposure did not result in a uniform upregulation of all inflammatory mediators; rather, multiple cytokines exhibited dose-dependent segmented response patterns instead of a linear increase. RNA sequencing analysis of samples treated with 125 or 1000 ng/mL TSST-1 for 48h revealed dose-associated transcriptional changes involving chemokine signaling, cytokine-receptor interactions, epithelial stress-related pathways, and glycolysis-related pathways. Notably the moderate-dose group showed a higher number of statistically defined differentially expressed genes than the high-dose group, suggesting a non-linear dose-associated transcriptional pattern. These findings demonstrate that TSST-1 can trigger cytotoxicity, apoptosis, cell cycle disruption, and a graded inflammatory response in vaginal epithelial cells, providing a systematic transcriptomic framework for understanding the early mucosal responses induced by TSST-1.

Lumbar degenerative disc disease (LDD) is a disabling condition which was associated with progressive degeneration of the intervertebral discs. The miR-766-3p and its modulation of SIRT6 remains unclear in LDD. This study aims to explored the regulated function of miR-766-3p and SIRT6 during LDD. Human nucleus pulposus cells (HNPCs) were treated with lipopolysaccharide (LPS) to construct a model of LDD to simulate inflammatory conditions. RT-qPCR and ELISA assays were employed to measure the expression of miR-766-3p and SIRT6. Additionally, we evaluated the levels of TNF‑α, IL‑1β, IL‑6, aggrecan, and collagen II by ELISA. Dual-luciferase reporter assays validated direct targeting between miR-766-3p and SIRT6. We observed that miR-766-3p was upregulated in LDD patients, and SIRT6 was downregulated. It is consistent with the changes in LPS-induced HNPCs. Additionally, when we inhibited the miR-766-3p and overexpressed SIRT6, inflammatory cytokines decreased, whereas aggrecan and collagen II increased. Moreover, the inhibition of miR-766-3p increased SIRT6, which was decreased by LPS. And the binding relationship between miR-766-3p and SIRT6 was confirmed by luciferase activity. MiR-766-3p mimic reversed the effects of elevated SIRT6 expression. The findings identify the miR-766-p/SIRT6 axis as a critical regulator mediating inflammatory responses and driving extracellular matrix degradation during LDD. Targeting this molecular pathway would offer novel therapeutic strategies for mitigating tissue damage of LDD.

Supercritical CO2\u2013Decellularized amniotic ECM hydrogel promotes immunomodulatory skin regeneration

Alpha herpesviruses (α-HV) initially infect mucosal epithelial cells and subsequently establish lifelong latency in the peripheral nervous system (PNS). Herpes simplex virus-1 (HSV-1), a human pathogen persisting in the majority of the adult population, shares neuroinvasive properties with Pseudorabies virus (PRV), a swine α-HV, commonly used as a model α-HV. Utilizing primary peripheral neuronal cultures, we previously showed that IFN-λ pre-treatment significantly reduced PRV yield. In this paper, we further characterized the early and late neuronal responses to IFN-λ by RNA-seq, and the antiviral potential of this response against HSV-1. Notably, HSV-1 exhibited neuron-specific resistance to IFN-λ mediated antiviral responses both in murine primary neurons and human neuronal cells. An ICP34.5-deficient HSV-1 (Δ34.5) mutant showed IFN-λ sensitivity in neurons, while replicating normally in untreated neurons showing that ICP34.5 is responsible for the neuron specific IFN-λ resistance of HSV-1. Our results further demonstrate that RSAD2 is strongly induced by IFN-λ in neurons, localizing to ER-associated membranes, and effectively restricting α-HV protein synthesis in the absence of ICP34.5. siRNA-mediated RSAD2 knockdown in IFN-λ-primed primary neurons largely restored replication of Δ34.5 HSV-1, highlighting the role of this IFN-λ induced host factor in neuronal infections. Together, neuronal IFN-λ-induced RSAD2 and HSV-1 ICP34.5 define a neuron-specific antagonistic mechanism that collectively determines the replication efficiency of HSV-1 in the PNS.

RETRACTION: Suppression of lncRNA MALAT1 Reduces LPS\u2010 or IL\u201017A\u2010Induced Inflammatory Response in Human Middle Ear Epithelial Cells via the NF\u2010\u03baB Signaling Pathway

Transcriptomic response of human mesenchymal stromal cells exposed to Boron- and Molybdenum-substituted mesoporous bioactive glass nanoparticles.

Exposure of cells or tissues to chemical compounds can be analyzed through transcriptomic signatures, which can be used to classify chemical agents. This information can also enrich Adverse Outcome Pathways (AOP). Transcriptional signatures have generally been obtained using "bulk" analysis, by which the global gene expression pattern of an entire tissue is determined. Although this approach has been useful in toxicology, some information is lost, especially when tissues containing multiple cell types are considered. With the advent of single-cell transcriptomics (scRNA-seq), it is now possible to obtain higher resolution, cell type-specific responses in complex tissues. The aim of the present study was to evaluate the added value of scRNA-seq in analysis of the acute response of human bronchial epithelial cells grown at the air/liquid interface (ALI) to a known toxic compound, CdCl2, with well described transcriptional signatures of exposure. Fully differentiated mucocilliary epithelia obtained from three independent donors were exposed to 10µM CdCl2 and scRNA-seq analysis was performed on a total of 18,255 cells to obtain cell type-specific signatures. Our results show that the contribution of each cell type to the overall transcriptomic bulk response varies. For example, the classical heavy metal detoxification response was only detected in multiciliated and secreting cells, while absent in basal cells. The data demonstrate that scRNA-seq provides high-resolution transcriptional signatures with unexpected features. This added information is likely to have implications for the refinement of AOPs and could serve as a basis for a new generation of tests in predictive toxicology.

PRGF eye drops mitigate oxidative stress and fibrosis in an in vitro glaucoma model: The role of platelet and plasma enrichment.

Background and Objectives:This study aimed to evaluate the anticancer potential of Lacticaseibacillus casei and Lacticaseibacillus rhamnosus cell-free supernatants (CFSs) against the PANC-1 human pancreatic cancer cell line, focusing on apoptosis, cell cycle modulation, and the expression of BAX and BCL-XL genes.Materials and Methods:PANC-1 cancer cells and adult human dermal fibroblast (HDFa) cells were treated with various concentrations of individual or combined CFSs. Cell viability was assessed using the MTT assay. Apoptosis was evaluated through Hoechst/PI staining and flow cytometry, while cell cycle distribution was analyzed via flow cytometry. Gene expression of BAX and BCL-XL was measured by quantitative real-time PCR.Results:At 20% (v/v), all CFS treatments significantly reduced PANC-1 cell viability while showing minimal effects on HDFa cells. Flow cytometry confirmed apoptotic rates of 39.33%, 42%, and 40.33% for L. casei, L. rhamnosus, and their combination, respectively, alongside a notable increase in S-phase cell population. Gene expression analysis showed a pro-apoptotic shift, characterized by BAX upregulation and BCL-XL downregulation.Conclusion:CFSs from L. casei and L. rhamnosus showed anticancer effects on PANC-1 cells, inducing apoptosis, S-phase arrest, and a favorable shift in apoptosis-related gene expression. These findings highlight their potential as promising adjuvant candidates for pancreatic cancer therapy.

Characterizing the osmotic response of human umbilical vein endothelial cells (HUVECs) to hypertonic PBS or sucrose exposure and demonstrating expected volume recovery upon re-exposure to isotonic conditions.