Membrane blebbing is a dynamic cellular phenomenon that plays a key role in numerous biological processes such as migration, apoptosis, cytokinesis, and extracellular vesicle (EV) formation. This phenomenon is traditionally observed via high-resolution imaging techniques on fixed samples, since it is difficult to quantify and monitor on live cells. This aspect limits our understanding of their temporal evolution and mechanical behavior. In this work, we present and validate a novel approach based on Atomic Force Microscopy (AFM) force spectroscopy combined with a custom data analysis pipeline that is capable to detect and map bleb- and EV-like structures directly on live cells. The method leverages the mechanical signatures embedded in force-distance (FD) curves-specifically, the presence of breakthrough points, which indicate the rupture or collapse of soft, vesicular structures. A custom MATLAB algorithm was developed to automatically identify and spatially map FD curves exhibiting EV-like mechanical behavior, based on signal smoothing, inflection point analysis, and user-defined force thresholds. This algorithm was at first validated against isolated EVs in liquid and then applied to live cultures of MDA-MB-231 breast cancer cells, monitored over 3, 7, 10, and 14 days to follow the temporal changes in membrane activity. This method enables the quantitative mapping of vesicle- and bleb-associated curves on the cell surface, highlighting their spatial distribution. A progressive increase in the number of EV-like events was observed up to Day 10, followed by a decline at Day 14-suggesting a dynamic remodeling of the membrane during cell culture maturation or stress. Moreover, they exhibit a preferential localization at the cell periphery. To validate the correspondence between EV-like force signatures and blebs, standard AFM high-resolution images were acquired on the same cells fixed and then analyzed using a Circular Hough Transform-based algorithm to detect and count blebs based on their circular geometry. The time trends obtained from both techniques showed good agreement, confirming the robustness of the force spectroscopy-based detection on live samples. This approach represents a significant advancement in the use of AFM for live-cell analysis. By extracting meaningful biophysical information from FD curves beyond standard elasticity metrics, we demonstrate that AFM spectroscopy can serve as a high-content, non-invasive tool for the detection of nanoscale membrane phenomena.

The DES gene encodes the IF (intermediate filament) protein desmin, which connects different multiprotein complexes, such as the cardiac desmosomes, and is highly important for the structural integrity of cardiomyocytes. Pathogenic DES mutations cause filament assembly defects leading to cardiomyopathies. However, most DES variants listed in genetic disease databases are currently classified as variants of unknown significance. Here, we characterized 21 different DES variants of unknown significance and 18 additional proline variants, localized in a highly conserved stretch at the C terminus of the desmin coil-2 subdomain. We inserted desmin variants via site-directed mutagenesis and investigated the filament assembly in transfected cell lines and cardiomyocytes derived from induced pluripotent stem cells by confocal microscopy. In addition, we purified recombinant wild-type and mutant desmin and analyzed the filament formation by atomic force microscopy. Coexpression with wild-type desmin delivered by adeno-associated virus was used to model the heterozygous status of cardiomyopathy patients. Twelve DES variants of unknown significance formed cytoplasmic aggregates, which were likewise verified by atomic force microscopy. Of note, these 12 variants disturb the filament assembly even when coexpressed with wild-type desmin. Using a proline screen, we showed that proline residues localized at nearly each of the positions in this stretch cause filament assembly defects. By modeling the tetrameric structure of desmin, we demonstrated that specific heptad positions, as well as positions of intramolecular and intermolecular ion bridge sites, are particularly susceptible to mutations that promote desmin aggregation. In summary, our study demonstrated that the highly conserved stretch at the C terminus of the coil-2 subdomain is a hotspot region, where several pathogenic DES mutations cause an aberrant desmin aggregation. Based on our molecular data, we suggest reclassifying the aggregate-forming variants as likely pathogenic mutations rather than variants of unknown significance. Our study may have relevance for the genetic counseling of cardiomyopathy patients with similar DES variants.

Finite element modelling for elucidating surface topography influence on cell-substrate interaction on fatty acid-modified PCL substrate.

Evaluation of bonding performance and enhancement mechanism of tannin isocyanate modified asphalt

Multiscale mechanical heterogeneity and structural gradients in the annulus fibrosus-endplate interface in the spine characterized using AFM nanomechanical testing.

Empagliflozin-mediated protection of heparan sulfate-rich endothelial glycocalyx during vascular inflammation.

Arrhythmogenic cardiomyopathy (ACM) is an inherited cardiac desmosome disease, as more than 50% of affected patients carry pathogenic variants in desmosome protein-coding genes. In this study, we focused on the role and mechanisms of pathogenic and non-pathogenic autoantibodies against intercalated disc (ICD) proteins such as desmoglein2 (DSG2) in ACM patients, healthy relatives (HR), and murine ACM models. IgG fractions from ACM patients, HR, healthy controls, and murine ACM models were isolated. Besides ELISA and cleavage assay, dissociation assay, immunostaining, Triton-X-100 assay, Western blots, and atomic force microscopy were performed in murine cardiac slices, HL-1 cells, or induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs). IgG fractions from ACM patients and HR, but not murine ACM model-derived or grouped healthy controls IgG (G-HC), revealed positive ICD staining. Three out of six ACM patients derived IgGs that reduced cardiomyocyte cohesion. Pathogenic autoantibodies, bound to DSG2 in healthy and ACM hiPSC-CMs, cleaved and reduced DSG2 interaction at the molecular level. We investigated GSK-3β contribution to the cardiomyocyte cohesion loss and observed GSK-3β reduced baseline cohesion in cultured cardiomyocytes and cardiac slices. Among five ACM-IgGs, three HR-IgGs tested, three pathogenic ACM-IgGs activated GSK-3β upstream of p38MAPK, leading to phosphorylation and junctional loss of β-catenin. GSK-3β inhibition rescued the loss of cell cohesion in ACM hiPSC-CMs. Pathogenic autoantibodies targeting DSG2 are present in ACM patients and impair cardiomyocyte cohesion in a GSK-3β-dependent manner. In contrast, autoantibodies are absent in murine ACM models and are non-pathogenic in some patients and HR.

The structural basis of texture in cooked taro: Ion-mediated pectin cross-linking in mother corms versus pectin disassembly in baby corms.

Cell mechanics, elasticity and viscoelasticity, are key markers of biological states like cancer. Atomic force microscopy (AFM) is ideal for such studies, but its low throughput limits large-scale use. Two solutions exist: automation for higher throughput, or high-density measurements for richer data. The latter enables machine learning (ML)-based classification, with viscoelastic parameters offering unique insights beyond static measures like Young's modulus. This study used dynamic mechanical analysis (DMA) to classify cells, focusing on viscoelastic descriptors (storage/loss moduli) across frequencies. Normal (RWPE-1) and grade IV cancerous (PC3-GFP) prostate cells were probed at 1-200Hz, generating 304 features per cell. The fuzzy logic-based LAMDA algorithm, trained on 19 selected features, classified cells using 40 samples per line. PC3-GFP cells showed higher deformability and heterogeneity, behaving more like viscous fluids at low frequencies. The model achieved 79% classification accuracy. Adding features improved performance, suggesting fewer training samples may suffice with rich datasets. A sensitivity-optimized threshold reduced false negatives in cancer detection. Combining viscoelastic analysis with ML effectively discriminates normal and malignant cells. Future work could refine training and integrate new features, though acquisition time remains a challenge. This approach offers a promising framework for mechanome-based diagnostics, with applications in cancer and stem cell research.

Physico-chemical behavior of magnesium-doped hydroxyapatite/chitosan composite layers in simulated physiological conditions.

Investigation of the impact of catechol on the fluidity of lipid monolayers.

Mechanical stiffening and delayed cell death of endothelial cells exposed to impulsive pressure.

Glycosylation pattern controls solubility, micellization, and aggregation of structurally defined ivy-derived triterpenoid saponins

Background: Copper oxide nanoparticles (CuO NPs) have attracted increasing attention for their versatile applications in catalysis, biosensing, anticancer therapy, and other biomedical fields. Green synthesis using microbial extracellular components offers a sustainable and eco-friendly alternative to conventional chemical methods. Objective: This study aimed to elucidate the biosynthetic process of copper oxide nanoparticles (Cu(NO₃)₂ NPs) utilizing extracellular components from the environmental isolate Lactobacillus plantarum as a natural reducing and stabilizing agent. Methodology: For nanoparticle synthesis, 1 g of copper nitrate was added to 10 mL of extracellular filtrate. The obtained CuO NPs were characterized using Atomic Force Microscopy (AFM), Fourier Transform Infrared Spectroscopy (FTIR), and Field Emission Scanning Electron Microscopy (FE-SEM). Antibiotic resistance profiles of the bacterial isolates and the antibacterial activity of the CuO NPs were assessed. Results: FE-SEM analysis revealed spherical CuO nanoparticles forming nano-cluster aggregates. All isolates exhibited 100 % resistance to cefotaxime and 91 % resistance to ceftriaxone and lincomycin, while resistance to other antibiotics ranged from 18 % to 73 %. Multidrug resistance was observed in all isolates, with at least four antibiotics resisted. The biosynthesized CuO NPs showed strong antibacterial activity, producing inhibition zones of 33 mm against Staphylococcus saprophyticus (150 % concentration) and 19 mm against Burkholderia cepacia (100 % concentration). Conclusion: Lactobacillus plantarum proved effective for the eco-friendly biosynthesis of CuO NPs. Given the high prevalence of antibiotic resistance, these nanoparticles demonstrate promising potential as alternative antimicrobial agents against multidrug-resistant pathogens.

Nanostructured electrochemical biosensor modified with a plant-derived protein for serine protease detection in prostate cancer.

Enhancing higher-order modal responses in atomic force microscopy with a segmented constraint methodology.

3D polyurethane scaffolds for exploring osteogenic differentiation and mechanical stimulation of mesenchymal stromal cells.

This study investigated the influence of mass ratios on the properties of soybean peptide aggregate (SPA)-soluble soybean polysaccharide (SSPS) composite particles (CPs) fabricated under ultrasonic assistance. The physicochemical and emulsifying properties of CPs were systematically evaluated. Results indicated that the mass ratio played a critical role in determining CPs characteristics. Increasing SSPS content initially led to an enlargement in particle size, followed by a subsequent reduction. Both ζ-potential and three-phase contact angle (θ) decreased with higher SSPS content, with a θ of 87.26° observed at a 1:1 ratio. CPs with SPA-SSPS ratios of 3:1 and 2:1 exhibited lower emulsifying activity and stability compared to SPA alone, whereas ratios between 1:1 and 1:3 demonstrated significantly enhanced emulsifying performance. Mechanistic studies revealed that nanoparticle formation was primarily driven by electrostatic interactions and hydrogen bonding, as confirmed by ζ-potential and Fourier-transform infrared spectroscopy analyses. Atomic force microscopy further indicated the structural encapsulation of SPA by SSPS. Among the tested ratios, the emulsions stabilized by CPs with a 1:1 ratio displayed the smallest droplet size, highest viscosity, and maximal free fatty acid release rate, alongside improved viscoelastic properties (G' and G″) and phase stability. However, higher SSPS ratios were associated with increased free radical content. PRACTICAL APPLICATIONS: These findings provide new insights into the role of mass ratios in modulating the functional properties of SPA-SSPS CPs and offer a promising strategy for developing high-performance Pickering emulsifiers in food and pharmaceutical applications.

Unveiling mechanistic insights of inactivated Aspergillus niger spore by fishery-waste-derived chitosan via synchrotron radiation tomography.

• Intentional oxidation of the chalcogenide active layer for OTS selectors. • Co-optimization of Se/Te composition and intentional oxidation. • Streamlined synthesis of multi-component systems via oxygen incorporation without additional sputtering targets. • Demonstrating the feasibility of high-density integration using a two-terminal via-hole structure. In the push for next-generation memory technologies, crossbar selector devices play a critical role in suppressing sneak-path currents and enabling three-dimensional integration. This study investigates the operation of ovonic threshold switch devices based on Se-alloyed TeO x , focusing on how interfacial oxidation and electrode materials influence the switching behavior. A small amount of oxygen was intentionally incorporated during the deposition step, enabling stable selector operation without the need for additional dopants. Devices fabricated with Pt, W and Cr electrodes exhibited threshold switching with sub-50 ns responses and selectivity exceeding 10 3 , with Pt delivering the best endurance (>10 3 cycles). In contrast, Ti, Ni and Al electrodes formed interfacial oxide layers, as confirmed by cross-sectional transmission electron microscopy, energy-dispersive X-ray spectroscopy, and electron energy loss spectroscopy, leading to resistive random-access memory-like behavior or switching failures. Optimal performance was consistently observed at a specific Se sputtering power (10 W), corresponding to low areal density and improved film uniformity, as verified by X-ray photoelectron spectroscopy, Rutherford backscattering spectrometry, atomic force microscopy, and Raman spectroscopy. These findings demonstrate that selector performance capabilities are governed not only by the active-layer composition but also by electrode-induced interfacial defects. This work offers concrete design guidelines for reliable, low-voltage selector integration in future high-density memory architectures.