Search Results (4,380)

Background: Tubular aggregate myopathy (TAM) is an autosomal dominant myopathy that results from gain-of-function mutations in the STIM1 and ORAI1 genes, which encode the two key proteins that coordinate store-operated Ca²⁺ entry in skeletal muscle and other cell types. Knock-in mice heterozygous for a glycine-to-serine point mutation in the ORAI1 pore (ORAI1^G100S/+ or GS mice) phenocopy several key aspects of TAM in humans with the analogous mutation including muscle weakness, exercise intolerance, elevated CK levels, hypocalcemia, and the presence of tubular aggregates. Methods: Since homozygous inheritance of the ORAI1-G100S mutation is embryonic lethal, we assessed the impact of homotypic ORAI1-G100S expression in skeletal muscle by crossing GS mice with constitutive, muscle-specific ORAI1 knock-in mice (cORAI1-KO). Results: Compound cORAI1-KO/GS mice exhibit only one active ORAI1 (GS) allele, and thus only express ORAI1-G100S monomers in skeletal muscle (‘homotypic’ GS mice). Homotypic GS mice exhibit an earlier onset and more severe muscle phenotype than age-matched heterotypic GS mice with both WT and GS alleles. Specifically, homotypic GS mice exhibit TAs at an earlier age, as well as significantly reduced in vivo muscle performance (grip strength, treadmill endurance, and rotarod endurance), maximal specific force production, and respiratory function, compared to those observed for both WT and heterotypic GS mice. Conclusions: These findings indicate that homotypic expression of the ORAI1-G100S mutation in skeletal muscle results in an earlier-onset and more severe muscle phenotype. Full article

Rapid changes in Arctic sea ice exert significant impacts on regional climate feedbacks and high-latitude maritime activities, increasing the demand for accurate short-term forecasting of key sea ice variables. This study proposes a GraphTransformer-based framework for joint forecasting of sea ice thickness (SIT) and sea ice concentration (SIC), designed to address their strong spatiotemporal coupling under irregular Arctic Ocean geometries. A static spatial graph is constructed over effective Arctic marine grid cells, where neighborhood aggregation is applied at each time step to explicitly encode spatial correlations. A shared-parameter temporal Transformer is subsequently employed to model node-level long-range temporal dependencies and to perform direct multi-step forecasting. The model generates 14-day daily forecasts of SIT and SIC in a single forward pass. Experiments are conducted using multi-source daily data spanning from 1 January 2019 to 15 May 2025, with evaluation restricted to valid marine grid nodes. Results indicate that the proposed GraphTransformer achieves either the best or second-best performance among the compared models in multi-step forecasting accuracy. Ablation experiments further confirm the critical role of graph-based spatial encoding in enhancing spatial coherence and mitigating error propagation. Full article

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease for which there is currently no cure. Dominant mutations in the TARDBP gene are causative of ALS. In particular, the p. G376D substitution in TDP-43 causes familial ALS and it is associated with TDP-43 mislocalization in the cytosol, increased presence of cytoplasmic aggregates, and lysosomal and mitochondrial dysfunction. We previously designed a small interfering RNA (siRNA) that specifically targets and silences the mutant allele and we demonstrated that, in patient-derived fibroblasts, it can reduce TDP-43 aggregation, decrease oxidative stress, and improve cell viability. Here, we investigated the ability of this siRNA to revert some ALS-associated pathological phenotypes in motor neurons derived from induced pluripotent stem cells (iPSCs), as motor neurons are the primary cells affected in ALS. siRNA treatment reduced TDP-43 mislocalization, enhanced lysosomal function and cell viability, and decreased oxidative stress. These findings indicate that this allele-specific siRNA effectively reverses key ALS-related cellular deficits in motor neurons, representing a promising candidate for targeted therapy in patients carrying the TDP-43 G376D mutation. Full article

Background: Oral iron supplementation or food fortification is essential for managing or preventing iron deficiency but often causes gastrointestinal side effects. While solubility has traditionally been considered a requirement for iron uptake via the DMT1 transporter, recent evidence shows that insoluble iron can also be absorbed through endocytosis, raising questions about particle size and epithelial responses. Methods: Human intestinal cell lines (Hutu-80 and Caco-2) were exposed to physiologically relevant but elevated iron levels (0.5 mM Fe, 48 h) as ferric pyrophosphate, ferrous fumarate (both prone to precipitation), and soluble ferric EDTA. Cell survival and COX-2 protein were quantified by ELISA, solubility by ICP-OES, and particle size in cell culture medium by dynamic light scattering analyses. Results: Ferric pyrophosphate (0.62–3.8 μm) markedly increased COX-2 expression in Hutu-80 cells (254% ± 37%, n = 3, p = 4.11 × 10⁻⁵) and in Caco-2 cells (78% ± 8%, n = 3, p = 0.01) compared to the control. Ferrous fumarate (237–866 nm) also induced COX-2, but only in Hutu-80 cells (62% ± 11%, n = 3, p = 0.04), whereas ferric EDTA showed no effect in either cell line. COX-2 induction was associated with larger particles in the medium (≥237 nm), whereas smaller particles (<146 nm) were not. Conclusions: Particle size appears to be a critical determinant of cell survival and iron-induced epithelial COX-2 expression. Iron compounds that present as both soluble and particulate forms may optimize bioavailability, but controlling aggregate size (<146 nm) could reduce inflammatory signaling. These findings may have important implications for cell culture systems and warrant in vivo validation in iron supplemental studies. Full article

The decline of honey bee health has intensified interest in microbiome-based strategies to support colony resilience and reduce reliance on chemical interventions. In this study, we performed an in vitro probiotic screening of five lactic acid bacteria (LAB) strains of honey bee origin and two multi-strain consortia for prospective application in apiculture. Two formulations were evaluated: LAB Mix 1 (Apilactobacillus kunkeei and Lactobacillus apis) and LAB Mix 2 (Lactiplantibacillus plantarum, Fructobacillus fructosus, and A. kunkeei). Functional and safety-related traits were investigated, including auto-aggregation, cell-surface hydrophobicity, inter-strain compatibility, organic acid production, oxidative detoxification capacity, antibiotic susceptibility, haemolytic activity, and growth dynamics in sugar-based feeding syrups. All strains exhibited time-dependent increases in aggregation and hydrophobicity, with A. kunkeei and F. fructosus showing particularly strong surface-associated properties. No mutual antagonism or haemolytic activity was observed. Organic acid profiling revealed strain-specific metabolic signatures, with high lactic and citric acid production by L. plantarum and LAB consortia. Several strains displayed peroxidase activity, suggesting a role in oxidative stress mitigation. Growth assays demonstrated that high sugar concentrations severely limited bacterial growth, whereas moderate dilution significantly improved growth. Under osmotic stress conditions, mixed cultures generally achieved higher optical density values than individual strains. Collectively, these findings support bee-associated LAB and multi-strain formulations as promising candidates for further probiotic development. Full article

Primary hepatocyte cultures serve as an ex vivo model of liver physiology. This study aims to employ poly(vinyl alcohol) (PVA) nanofiber membranes (NMs) to establish a three-dimensional (3D) culture system that supports the long-term functionality of primary hepatocytes. Primary hepatocytes were monocultured on a PVA NM or indirectly cocultured with NIH3T3 fibroblasts on a distinct polycaprolactone (PCL) NM layer. Monocultured and cocultured hepatocytes maintained prolonged survival without supplemental growth factors. Cocultured hepatocytes formed larger aggregates composed of cell clusters attached to untreated nanofibers than monocultured cells. However, most primary hepatocytes cultured on NaOH-treated PVA NM and Arg–Gly–Asp (RGD) peptide-blended PVA (RGD-PVA) NM, under monoculture and coculture conditions, formed non-aggregated cells in a single-cell layer. In a bioinert assay, unstimulated dendritic cells were activated on untreated but not NaOH-treated PVA NM. CYP3A4 activity was higher in cocultured cells on RGD-PVA NM with fibroblasts than in monocultured cells on PVA and RGD-PVA NM. Functional hepatocyte cultures were successfully maintained in a 3D single-cell layer on RGD-PVA NM, along with fibroblasts in a layer-by-layer coculture, for a prolonged period. The prolonged culture of hepatocytes in a 3D single-cell layer may facilitate further drug discovery, toxicity studies, and translational liver research. Full article

Prion diseases are fatal neurodegenerative disorders characterized by profound neuronal damage. Despite evidence supporting a neuroprotective role for β-synuclein (β-syn) in neurodegeneration, its potential functions and mechanisms in prion disease have not been elucidated. To investigate the role of β-syn, we systematically analyzed its alterations in the central nervous system of several prion-infected rodent models and cell models. A series of biochemical, cellular, and immunofluorescence assays were conducted to explore the relationship between β-syn and protein kinase B (Akt) signaling and between β-syn and prion protein (PrP), and its neuroprotective role in prion disease. Student’s t-test was used for statistics. At the terminal stage of prion disease, β-syn and Akt exhibited a parallel and remarkable decrease in rodent brains, contrasting with the slight but significant increase observed at early to middle stages. Dual-stained immunofluorescence assays confirmed that β-syn is localized within NeuN-positive neurons. Further structural and functional analyses revealed a high-affinity molecular interaction between β-syn and Akt, with the N-terminal region of β-syn being essential for binding to Akt1. In a cell model of PrP aggregation, β-syn overexpression suppressed cytochrome c-induced apoptosis, which was demonstrated by decreased levels of cleaved caspase-3. Notably, this anti-apoptotic effect was partially abolished upon Akt knockdown, indicating a dependence on Akt signaling. Moreover, colocalization of β-syn and PrP was observed in rodent brains. Consistently, in cellular models of prion infection and PrP aggregation, β-syn overexpression not only reduced PrP levels but also ameliorated its aberrant histological distribution. Our findings demonstrate that the anti-apoptotic activity of β-syn, mediated via Akt signaling, is severely lost during prion infection, thereby suggesting a mechanism of intrinsic neuronal vulnerability and revealing a novel therapeutic strategy. Full article

Prediction of fatigue failure in concrete structures remains a major challenge due to progressive material degradation. Reliable prediction, therefore, requires modeling the 3D heterogeneous microstructure of concrete to explain the underlying mechanisms governing fatigue failure. While such mesoscale models can reliably predict the fatigue-induced fracture mechanisms, the identification of the associated material parameters remains a significant challenge due to the high-dimensional parameter space introduced by the model. The key challenge addressed in this study is to capture microcrack initiation and coalescence under fatigue loading, using a model capable of representing fracture process: crack initiation, crack propagation, and final failure. Firstly, concrete domain is discretized into Voronoi cells, enabling explicit representation of aggregates and mortar by randomly assigning cohesive links connecting Voronoi cells as aggregates and mortar. After this, mortar links are modeled as coupled damage–plasticity 3D Timoshenko beam elements with nonlinear kinematic hardening and isotropic softening introduced using embedded discontinuity formulation, enabling fracture Modes I–III, whereas aggregate links are modeled as elastic 3D Timoshenko beam elements. The model efficiency is additionally reinforced by using surrogate model approach, with corresponding material parameter identification carried out by multi-objective Bayesian optimization framework to reproduce experimental results. The performance of the proposed model is illustrated by reproducing experimental results obtained from concrete cube compression test and three-point bending test under low-cycle fatigue loading, where the errors between experimental and numerical results are reduced by 82% (stress) and 88% (energy) for the cube test and by 86% (force) and 93% (energy) for the bending test, relative to the initial dataset error. Full article

Background/Objectives: Alzheimer’s disease (AD) is a progressive neurodegenerative disorder for which no disease-modifying therapy that halts or substantially slows disease progression is currently available. Although antibody therapies targeting amyloid β (Aβ) have recently received FDA approval, their high cost, limited efficacy, and potential adverse effects highlight the need for alternative solutions. Therefore, the development of low-molecular-weight compounds capable of reducing toxic Aβ aggregates is of considerable interest. In this study, we investigated the effects of 2,3,4-trihydroxybenzophenone (THB) on the inhibition and disassembly of Aβ_1–42 aggregates through in vitro and in vivo experiments. Methods: In vitro assays were performed to evaluate the effects of THB on Aβ_1–42 aggregation and fibril disassembly. Cell viability assays and hippocampal slice electrophysiology were conducted to assess neurotoxicity and synaptic function. In vivo effects were examined in Aβ_1–42 aggregate-injected mice and in 5 Familial AD mutations (5XFAD) mice using behavioral, histological, and electrophysiological analyses. Results: THB inhibited Aβ_1–42 aggregation in a concentration-dependent manner and promoted the disassembly of preformed fibrils. THB attenuated Aβ_1–42-induced Neuro2a cell death and restored Aβ_1–42 aggregate-associated long-term potentiation (LTP) deficits in hippocampal slices. In Aβ_1–42 aggregate-injected and 5XFAD mice, THB reduced amyloid pathology and neuroinflammatory markers and improved synaptic function and memory performance. Conclusions: These findings suggest that THB modulates pathogenic Aβ_1–42 assemblies and provides a structural basis for the development of small-molecule modulators of Aβ_1–42 aggregation with potential preventive or disease-modifying applications in AD. Full article

Accurate prediction of tuna distribution is essential for sustainable fisheries management. This study develops a two-stage hybrid model combining Convolutional Neural Networks (CNN), Recurrent Neural Networks (RNN), and Random Forest (RF) to predict tuna distribution around drifting fish aggregating devices (DFAD) in the Western and Central Pacific Ocean (WCPO). Echo-sounder buoy data from DFAD were aggregated into 2° × 2° grid cells and matched with oceanographic variables from the Copernicus Marine Service. Random Forest-based variable importance analysis identified primary productivity (27%), chlorophyll-a (22%), and dissolved oxygen (18%) as the three dominant environmental drivers. The CNN-RNN component extracts spatiotemporal features from multi-layer ocean data, while the RF classifier performs binary classification of tuna aggregation zones (high-yield vs. low-yield). All five models (Decision Tree, RF, CNN, Transformer, and CNN-RNN-RF) were evaluated on 557 samples using 5-fold stratified cross-validation, with each fold further split 80:20 for training and validation. The proposed CNN-RNN-RF model achieved the highest performance with an AUC of 0.830, accuracy of 82.6%, and F1-scores of 86.3% (high-yield) and 76.2% (low-yield), outperforming the best baseline model (RF: AUC 0.761, accuracy 75.4%). Predicted high-yield zones showed strong consistency with fishing log records, demonstrating the potential of integrating echo-sounder data with hybrid deep learning for data-driven tuna fisheries management. Full article

In bulk heterojunction (BHJ) organic solar cells (OSCs) employing perylene diimide (PDI)-based non-fullerene acceptors, excessive intermolecular interactions among PDI units lead to severe aggregation and pronounced donor–acceptor phase separation, both of which critically limit device performance. To address these issues, numerous structurally engineered PDI derivatives have been developed. In particular, twisted multi-PDI architectures designed to suppress intermolecular aggregation have shown improved morphological control; however, such twisted structures are often highly amorphous, which reduces electron-transport efficiency and constrains OSC performance. In this work, we introduce a mixed-acceptor strategy combining a twisted PDI dimer (SF-PDI₂) with a planar monomeric PDI (m-PDI) to balance aggregation and morphological uniformity. Ternary blend OSCs consisting of PTB7-Th as the donor and these two PDI acceptors exhibit systematic performance variations depending on their relative ratios. At the optimized composition (SF-PDI₂:m-PDI = 90:10 by weight), the device outperforms single-acceptor systems, which is attributed to controlled aggregation arising from the complementary structural features of the two PDI acceptors. This study demonstrates that combining mixed PDI acceptors with similar molecular moieties enables precise control of aggregation, improving both morphology and photovoltaic performance. Full article

Red blood cells represent a widely used cellular model in cytotoxicity studies, particularly in hemocompatibility assessments. As enucleated cells, which are abundant and easily accessible in both humans and animals, red blood cells allow for rapid, reproducible, and low-cost evaluation of the toxicity of bioactive compounds, whether natural, synthetic, or nanoparticulate. From a functional perspective, the red blood cell membrane is highly sensitive to physical and chemical environmental changes (osmolarity, temperature, pH, and the presence of oxidizing agents). This sensitivity makes red blood cells an effective biosensor for detecting membrane damage, hemolysis, oxidative stress, methemoglobin formation, and aggregation processes. Therefore, in vitro tests using red blood cells allow for the preliminary evaluation in preclinical development, particularly for the early screening of cytotoxicity, membrane-disruptive effects, and hemocompatibility of small molecules, nanomaterials, and blood-contacting biomaterials. These techniques include hemocompatibility tests, evaluation of oxidative and osmotic damage, and evaluation of erythrocyte aggregation and function. However, the use of red blood cells as a cytotoxicity model also has significant limitations. As anucleate cells, erythrocytes lack organelles such as nuclei, mitochondria, or lysosomes, which prevents the evaluation of their effects on key intracellular processes such as protein synthesis, cell signaling, apoptosis, or endoplasmic reticulum stress. This lack of cellular complexity limits their usefulness as a sole model in studies of systemic toxicity or tissue-specific cytotoxicity. These tools offer an effective preliminary approach to anticipating risks in biomedical and pharmacological research. Full article

The therapeutic potential of prodigiosin as a hydrophobic anticancer agent can be enhanced by various approaches, one of which is the loading of PG into extracellular vesicles. Drug distribution and stability in aqueous media play a crucial role in targeting and accumulation, thereby enabling the attainment of therapeutically effective drug concentrations. Extracellular vesicles are nano-sized, cell-derived vesicles with a lipid bilayer membrane. Extracellular vesicles can be utilized as drug carriers for both water-soluble and non-water-soluble therapeutic agents. We hypothesized that microvesicles could effectively address the current challenges of prodigiosin delivery. Several different techniques have been developed for fabricating extracellular vesicles. These include microvesicles induction by cytochalasin B treatment as well as cell cultivation in serum depleted media. In our study, prodigiosin, like cytochalasin B, demonstrated efficacy in microvesicles formation based on protein quantification and Nanoparticle Tracking Analysis. In addition, Nanoparticle Tracking Analysis showed that vesicles from mesenchymal stem cells are more stable under ultrasound exposure. Microvesicles encapsulating prodigiosin, compared to unmodified naïve ones, demonstrated slightly increased zeta potentials and hydrodynamic diameters, which probably contributed to better stability. We demonstrated that ultrasonic treatment for the loading of prodigiosin does not significantly increase the proportion of prodigiosin-positive microvesicles in comparison with microvesicles induced with prodigiosin; moreover, this method cannot be considered as optimal due to its disadvantages, such as particle aggregation. Prodigiosin-induced and prodigiosin-loaded microvesicles from mesenchymal stem cells were significantly smaller and less polydisperse in size. Overall, prodigiosin encapsulated in extracellular vesicles might be more suitable for medical and clinical applications compared to pure forms of PG due to their cell membrane compatibility. Full article

Automated analysis of liquid-based cervical cytology is increasingly supported by digital microscopy and deep learning. However, model generalization remains challenging due to scanner- and laboratory-induced domain shifts affecting color, texture, and morphology. In this study, we present a robust cell-level classification framework for liquid-based Pap smear cytology based on deep transfer learning, designed to operate under heterogeneous acquisition conditions. We construct a multi-source dataset by integrating three widely used public reference repositories (SIPaKMeD, Herlev, CRIC Cervix) with a proprietary cohort comprising 416 Whole Slide Images (WSIs) collected from two medical centers and digitized using different scanning systems. All labels are harmonized into four Bethesda categories (NILM, ASC-US, LSIL, HSIL), and cell-centered 224 × 224 patches are used as standardized inputs for model development and benchmarking. We evaluate state-of-the-art CNN backbones (ResNet50, EfficientNetB0, VGG16) and perform systematic ablation across data-source combinations to quantify robustness under acquisition variability. Among the evaluated models, ResNet50 yields the best overall performance on the independent test set (accuracy = 0.91; macro-F1 = 0.91), consistently outperforming EfficientNetB0 and VGG16. Importantly, incorporating proprietary multi-center WSI-derived data improves robustness to scanner-induced variation compared to training on public data alone. These findings demonstrate that combining diverse data sources can mitigate domain shift in cell-level cervical cytology classification. While clinically actionable screening requires slide-level aggregation (e.g., MIL-based WSI inference), the proposed classifier provides a robust component that can be integrated into end-to-end WSI screening pipelines in future work. Full article

Thrombosis remains a leading cause of cardiovascular morbidity and mortality. During thrombosis, activated platelets and endothelial cells expose phosphatidylserine (PS) on their outer membranes, creating a surface that accelerates clot formation. Current antithrombotic therapies, such as heparin and warfarin, carry significant bleeding risks, highlighting the need for safer alternatives. In response, we developed a PS-targeting liposomal formulation composed of Zn-dipicolylamine (DPA)-cyanine-3[22,22] and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (molar ratio 3:97). This DPA-harboring liposome (DPAL) binds selectively to PS-rich surfaces such as activated platelets and has demonstrated efficacy in reducing thrombosis in mouse models, with minimal bleeding. In the present study, we examined the interaction of DPAL with albumin, the most abundant plasma protein and a key transporter in the bloodstream, to assess the potential for harmful protein aggregation or structural disruption. Using dynamic light scattering and intrinsic protein fluorescence, we found that, unlike warfarin and heparin, DPAL does not induce any large protein aggregates or cause significant conformational changes near the tryptophan residue when mixed with human serum albumin, suggesting a favorable interaction profile. In addition, we used transwell permeability assays and CyQUANT cell proliferation assays to assess the cytotoxicity of DPAL in cultured human umbilical vein endothelial cells (HUVECs). Our results showed that DPAL does not compromise endothelial barrier integrity in HUVEC monolayers nor the cells’ viability. Our current and previous findings together suggest that DPAL could offer a promising approach to modulate harmful coagulation pathways and provide a new targeted therapeutic strategy for managing thrombotic disorders. Full article