Inherited retinal degenerative diseases, such as retinitis pigmentosa, cause progressive photoreceptor loss and irreversible vision decline, yet effective treatments remain unavailable. Our previous studies demonstrated that Lycium barbarum glycopeptide delays photoreceptor degeneration in a chemically induced retinitis pigmentosa model, primarily through antiinflammatory mechanisms. In this study, we extended these findings to an inherited retinitis pigmentosa model to further elucidate the neuroprotective actions of Lycium barbarum glycopeptide. Lycium barbarum glycopeptide was orally administered daily to rd10 mice beginning at postnatal day 8, prior to photoreceptor degeneration, and retinal function and morphology were evaluated at postnatal day 25, the peak of rod apoptosis. Behavioral assays, electroretinography, immunofluorescence staining, proteomic profiling, and western blotting were performed to assess the therapeutic effects and molecular mechanisms of Lycium barbarum glycopeptide. Lycium barbarum glycopeptide treatment significantly improved visual performance in rd10 mice, as shown by enhanced optomotor responses and black.white transition behavior. Electroretinography analysis revealed increased scotopic a-wave amplitudes, indicating improved photoreceptor function. Histological evaluation showed preservation of outer nuclear layer thickness and maintenance of rod and cone opsin expression. Lycium barbarum glycopeptide also reduced microglial and Muller glial activation in a region-dependent manner. Proteomic and biochemical analyses revealed that Lycium barbarum glycopeptide upregulated key phototransduction proteins while concurrently downregulating pro-inflammatory mediators such as interleukin-6, nuclear factor kappa B, cyclooxygenase-2, and tumor necrosis factor-α. Collectively, these results demonstrate that Lycium barbarum glycopeptide protects against inherited photoreceptor degeneration by improving retinal function and structure, alleviating neuroinflammation, and supporting phototransduction recovery. This work extends our previous findings and highlights Lycium barbarum glycopeptide as a promising therapeutic candidate for inherited retinal degenerative diseases.

With the expanding deployment of nanoscale zero-valent iron (nZVI) in groundwater remediation, the frequency and duration of microbial exposure in aquatic niches are steadily increasing. Despite extensive research on nZVI-induced toxicity, microbial survival strategies and adaptive resistance mechanisms under repeated or prolonged nZVI exposure remain largely uncharacterized, particularly in anaerobic subsurface environments. Herein, using Escherichia coli as a model Gram-negative facultative anaerobe, we systematically explored bacterial responses to repeated nZVI exposure under aerobic and anaerobic conditions. After 20 consecutive nZVI exposure-cultivation cycles, bacterial strains developed stronger resistance under anaerobic conditions than those under aerobic ones. Mechanistically, the resistant strain exhibited increased biofilm biomass and thickness (1.55-2.00-fold those of the original strain, p < 0.001), reduced intracellular Fe2+ accumulation, elevated catalase activity, and enhanced maintenance of membrane potential and lipid homeostasis. Transcriptomic profiling further revealed that nZVI repressed ribosomal and energy metabolic functions, while activating stress-responsive pathways (e.g., two-component systems, transport, global metabolic regulation), thereby mediating resistance via coordinated multimodal mechanisms. Notably, nZVI-resistant strains exhibited cross-resistance to Ag and Cu nanoparticles, together with stable phenotypic inheritance after subculturing and potential intraspecific transferability in conjugation assays. This study reveals the microbial resistance-associated mechanisms under repeated nZVI exposure in a model system, providing a mechanistic basis for assessing the ecological safety of nZVI and optimizing its environmental applications.

Predicting the acoustic response of coated elastic systems remains a challenging problem in nondestructive evaluation, particularly when interfacial mechanical contrast and layer thickness jointly govern complex dispersive behavior. In this work, generalized Rayleigh-wave propagation in dental restorative bilayer systems is theoretically investigated within a scanning acoustic microscopy framework. Four coating materials composite resin, amalgam, cobalt chromium alloy, and gold alloy are examined on glass-ionomer, silicate, and zinc-phosphate cement substrates. The Rayleigh critical angle and surface wave velocity are analyzed as functions of the normalized thickness h/λ T over the range 0-2. The results reveal two distinct propagation regimes separated at h/λ T ≈ 1. In the interaction-dominated regime, anomalous and non-monotonic dispersion emerges from strong layer-substrate mechanical coupling and is governed by a single dimensionless mismatch parameter ξ=(E L /E S )/(ρ L /ρ S ). In the asymptotic regime, dispersion vanishes as propagation becomes layer controlled. A four-type acoustic classification (Types A, B, C, D) is established from the joint analysis of ξ, the Anomaly Severity Index (ASI), and the Velocity Sensitivity Index (VSI), with a near unity linear ξ-VSI correlation (R 2 =0.98). The natural enamel-dentin bilayer (ξ=3.31, VSI=+79.8 %) is identified as the acoustic reference target for restorative design. These findings provide a unified and clinically actionable framework for evaluating acoustic compatibility in dental restorative assemblies.

Systolic heart failure is increasingly recognized as a disease of impaired cardiac energy metabolism. Here, the authors show that cardiac myosin super-relaxation (an energy-conserving myosin state) is destabilized in failing human myocardium. Importantly, this loss of energy conservation is associated with elevated post-translational modifications (phosphorylation and acetylation) rather than gross structural changes in thick filament organization. Hence, targeting post-translational modifications signaling pathways opens new therapeutic opportunities.

Accurate evaluation of corrosion in metal plates is essential across many industries, particularly in shipbuilding. Among available techniques, non-destructive testing (NDT) is especially valuable, as it enables the assessment of material degradation without compromising structural integrity. Within this domain, guided ultrasonic waves have attracted increasing attention in recent years due to their potential for estimating statistical corrosion parameters, such as mean thickness loss and its standard deviation. However, practical application of these methods remains challenging because of the complex interaction between wave propagation and spatially varying thickness profiles. This study presents an advanced methodology for precise estimation of corrosion parameters using deep learning-based inverse models. The corrosion morphology is modeled as a stochastic field, while guided wave responses, obtained through numerical simulations and recorded by a sensor network, are postprocessed to extract statistical descriptors. In particular, the mean vector and covariance matrix of sensor signals, including wave packet location, width, and amplitude, are computed. To ensure robustness, feature scaling and elliptic envelope-based covariance estimation are employed, effectively mitigating the influence of outliers, which are especially pronounced at higher levels of thickness loss and surface roughness. The inverse model establishes a mapping between extracted feature vectors and the corresponding corrosion parameters, enabling their direct prediction. This framework addresses the intrinsic non-uniqueness of the problem, where identical statistical descriptors may arise from infinitely many realizations of the stochastic corrosion field. Extensive numerical investigations demonstrate that reliable feature extraction, particularly robust handling of outliers, combined with the ability of deep neural networks to capture nonlinear relationships, is critical for accurate parameter estimation. Prediction performance is further improved by aggregating responses from multiple independent realizations of the stochastic corrosion field. Overall, the results demonstrate that the proposed framework, combining robust statistical feature extraction, covariance-based feature representation and nonlinear inverse modeling, provides improved prediction accuracy compared with the previously developed feature-based linear regression model and the conventional time-of-flight approach under the considered numerical simulation conditions.

Understanding anterior segment crowding is essential for identifying preoperative risk factors for malignant glaucoma (MG) following phacoemulsification, intraocular lens implantation, and goniosynechialysis (PEI‒GSL) in primary angle-closure glaucoma (PACG). This retrospective case-control study compared 24 PACG patients who developed postoperative MG (MG group) to 24 randomly selected controls (control group). Preoperative ultrasound biomicroscopy (UBM) metrics included anterior chamber depth (ACD), lens vault (LV), ciliary process distance (CCD), ciliary body thickness, anterior ciliary body placement, trabecular‒ciliary process angle (TCPA), and simulated cilio-lenticular distance (sCLD). To eliminate observer bias, all exported raw UBM images were completely anonymized, stripped of all identifiable metadata, randomized, and assigned unique computer-generated hashes before undergoing single-blinded evaluation. The MG group exhibited significantly shorter sCLD, greater LV, shallower ACD, and narrower TCPA preoperatively, with standardized mean differences exceeding 0.8. After False Discovery Rate correction, these differences remained significant (adjusted P < 0.05). Crucially, multivariable logistic regression adjusting for axial length (AL) demonstrated that a shortened sCLD (P < 0.001), an increased LV (P < 0.001), and a narrowed TCPA (adjusted P = 0.0099) maintained highly robust independent associations with MG development. The MG group showed a trend of weakened parameter coordination, particularly between AL and ACD (r = 0.334 vs. 0.718). Exploratory principal component analysis identified an architectural "anterior segment crowding" axis (explaining 62.2% of the variance), with the MG group clustering negatively. An exploratory, unweighted composite ciliary block score showed a large separation effect size (Cohen's d = 1.995, P < 0.001). In conclusion, preoperative shortened sCLD, increased LV, and narrowed TCPA constitute an anatomical risk cluster independently associated with postoperative MG. While anterior segment crowding characterizes this cohort, these multivariate models function strictly as exploratory, sample-specific constructs and explicitly lack external clinical validation.

Volumetric muscle loss (VML) is a devastating injury in which extensive skeletal muscle loss overwhelms endogenous healing. Despite their therapeutic efficacy, acellular constructs fail to provide a comprehensive set of bioactive cues required for muscle development. Notably, very few constructs provide bioelectrical cues necessary for muscle development. To address that need, this study aimed to develop a myogenic platform that provides a comprehensive set of bioactive cues, including bioelectrical cues that can be delivered wirelessly, to promote skeletal muscle development. To achieve this, we engineered a microchanneled, piezoelectric, 3D-printed platform based on a unique interpenetrating hydrogel network of gelatin methacrylate and poly(acrylic acid) that incorporates barium titanate nanoparticles. Rheological studies showed stable gel-like viscoelasticity and shear-thinning behavior of the hydrogels, indicating their suitability for extrusion-3D printing. Physicochemical characterization demonstrated the platform's structural fidelity, precise microchannel definition, pronounced flexibility, and self-adhesiveness. While the platform's stiffness was comparable to that of skeletal muscle, it also generated a surface potential of ∼90 mV when deformed by ultrasound (US), highlighting its piezoelectric nature and ability to harness electromechanically triggered bioelectrical cues. FE-SEM and immunofluorescence analyses revealed that microchannels and US-induced piezoelectricity from the platform can synergistically help form dense, compact, and aligned myotubes. Notably, we demonstrate that the US-induced piezoelectric platforms can promote early-stage myotube development, enhance myogenic progression, and help form mature, elongated, thick myotubes over time. Finally, we observe that the piezoelectric platforms exhibit increased FAK and MAPK activation in adherent cells over time, suggesting that these pathways can contribute to transducing electromechanical cues that promote myotube maturation. Overall, this is the first-of-its-kind comprehensive study to engineer a platform that delivers a comprehensive set of bioactive cues, specifically wirelessly delivered bioelectrical cues, resulting in a spatiotemporally bioinstructive microenvironment for skeletal muscle development. We believe this unique platform could be a promising regenerative therapy for VML.

Multiple studies have demonstrated that the number, length, and gap width (thickness) of ER-mitochondria contacts, or mitochondria-associated membranes (MAMs), influence their biological roles. Our previous work showed that the stabilization of tight MAMs, characterized by a gap width of approximately 7 nm, leads to an increase in amyloid β (Aβ) levels, whereas the presence of loose MAMs, with a gap width of around 40 nm, reduces Aβ production in a three-dimensional (3D) neural model of Alzheimer's disease (AD). To investigate the effects of MAMs with different gap widths, ER- and mitochondria-targeted FRET (Förster Resonance Energy Transfer) biosensors-ER-CFP (cyan fluorescent protein) and Mito-YFP (yellow fluorescent protein), respectively-were developed to quantify tight MAMs (<10 nm gap width) in contrast to loose or non-MAMs. FRET occurs when the donor fluorophore (CFP) and the acceptor fluorophore (YFP) are within 10 nm of each other, making this system suitable for assessing MAM proximity. This protocol outlines the use of spectral ratiometric FRET to measure the extent of tight MAM formation.

Vesicles formed from amphiphilic copolymers, alone or blended with phospholipids, offer superior mechanical and chemical stability compared to conventional lipid vesicles. This makes them an attractive chassis, one that can be further developed and expanded to enable specific applications, such as drug delivery, diagnostic biosensing, construction of artificial cells, and bioinspired micro- and nanoreactors. In particular, functionalization through membrane protein incorporation is essential for many of these applications. Here, we present a protocol for preparing membrane protein-functionalized large unilamellar vesicles (LUVs) from the graft copolymer PDMS-g-PEO via detergent-mediated reconstitution. Furthermore, we describe how these large vesicles can be converted to giant unilamellar vesicles (GUVs) using a fusion-electroformation approach. The protocol covers fluorescence labeling of membrane proteins, preparation of polymer and hybrid LUVs, membrane protein reconstitution, size distribution analysis via dynamic light scattering (DLS), assessment of protein activity via oxygen consumption measurements, preparation of protein-functionalized GUVs, and analysis of protein insertion and proton pumping activity in GUVs via confocal microscopy. Representative results demonstrate the formation of monodisperse proteo-LUVs with a polydispersity index (PDI) below 0.2, and successful generation of proteo-GUVs ranging from 5-35 µm in diameter. Protein activity is confirmed by oxygen consumption measurements in both polymer LUVs (18.2 nmol/min/mL) and hybrid LUVs (26.9 nmol/min/mL). Protein insertion into GUVs is quantified via fluorescence intensity, yielding 20.6 ± 3.7 a.u. for polymer GUVs and 26.2 ± 5.0 a.u. for hybrid GUVs. Proton pumping activity in GUVs, monitored via an encapsulated pH-sensitive dye, is consistent with protein functionality, with inward proton pumping being predominant. The copolymer's mechanical softness and lipid-like bilayer thickness (~5.3 nm) support efficient protein insertion and preservation of functionality.

Conventional preclinical models used in dental research, including animal models and static cell cultures, present important limitations in reproducing the physiological conditions of the dentin-pulp interface and predicting human biological responses to dental biomaterials. Organ-on-a-chip (OoC) technology provides an alternative approach by enabling the recreation of tissue-specific microenvironments within controlled microfluidic systems. This manuscript describes a step-by-step protocol for the fabrication and assembly of a dentin-pulp complex OoC device incorporating a human dentin disc within a three-layer polydimethylsiloxane (PDMS) microfluidic platform. The protocol includes dentin disc preparation, fabrication of microstructured molds using photolithography, PDMS casting and curing, plasma-assisted layer bonding, device assembly, leak testing, and dental pulp stem cell seeding under static culture conditions. Critical fabrication parameters influencing device performance are discussed, including control of PDMS layer thickness, plasma surface treatment, and integration of the dentin disc within the device to minimize leakage. Cell adhesion and viability within the device were evaluated using scanning electron microscopy and confocal live/dead staining. Representative results demonstrate successful integration of the dentin disc within the microfluidic assembly, maintenance of leak-free conditions, and adhesion of viable dental pulp stem cells to the dentin surface. This protocol provides a reproducible approach for the fabrication of a dentin-pulp complex OoC platform that may support future studies investigating cellular responses to dental biomaterials and microenvironmental stimuli.

The optimal surgical margin for cutaneous melanoma remains controversial, particularly in anatomically constrained areas such as the acral regions. We conducted a retrospective cohort study of 79 patients with invasive cutaneous melanoma treated at National Cheng Kung University Hospital between 2012 and 2024, including 51 cases of acral lentiginous melanoma (ALM). Patients were categorized as having adequate surgical margins (per NCCN guidelines) or inadequate but histologically tumor-free margins. Survival outcomes, including overall survival (OS), progression-free survival (PFS), local recurrence-free survival (LRFS), and melanoma-specific survival (MSS), were analyzed using Kaplan-Meier estimates and Cox proportional hazards models after a median follow-up of 60 months. No significant differences were observed in OS, LRFS, or MSS between the two groups. Although univariate analysis suggested a difference in PFS, the association did not persist in multivariate analysis, where margin inadequacy was not independently associated with poorer outcomes (OS: HR 1.81, 95% CI 0.84-3.90, p = 0.13; PFS: HR 1.87, 95% CI 0.90-3.87, p = 0.093). Subgroup analyses of ALM and high-risk tumors (Breslow thickness > 2 mm) yielded consistent results, showing no significant differences in oncologic outcomes. In conclusion, our study showed that inadequate but histologically tumor-free excision margins may not independently compromise survival outcomes in cutaneous melanoma. A narrower margin may be acceptable in selected patients when wider excision is precluded by anatomic or functional constraints.

Concentrated surfactant pastes often form lamellar liquid-crystalline phases whose processing depends on alignment, confinement, temperature, and defect topology. Here, we investigate temperature-driven lamellar contraction and parabolic focal conic defect (pFCD) formation in 70 wt% sodium lauryl ether sulfate (SLEnS) lamellar pastes using small-angle X-ray scattering (SAXS), confined cross-polarized microscopy, and confined oscillatory rheometry. SAXS measurements show that the lamellar repeat spacing decreases with increasing temperature in both SLE1S and SLE3S, corresponding to one-dimensional, bilayer-normal negative thermal expansion. This contraction was observed in both bulk-thickness and confined SAXS geometries, indicating that the nanoscale thermal response is not specific to sample thickness, chemistry, or instrument configuration. Under confinement, squeeze-flow alignment appears to produce coherent lamellar stacks that cannot freely accommodate the temperature-dependent spacing decrease. Heating these aligned lamellae produces thermal undulations and optically resolved pFCD fleurettes. SLE3S forms pFCDs at a lower onset temperature and completes defect development over a lower temperature range, whereas SLE1S forms brighter and larger final pFCD textures. Confined rheometry shows non-monotonic changes in storage and loss moduli over the same temperature range as lamellar contraction and pFCD development, suggesting coupling between nanoscale contraction, mesoscale defect formation, and bulk viscoelasticity. Together, these results establish a confinement-mediated mechanism by which lamellar negative thermal expansion generates compressive strain, undulatory defect formation, and altered mechanics in industrially relevant concentrated surfactant pastes.

Chiral molecular organization at semiconductor interfaces is vital for advancing quantum and chirality-induced spin-selective (CISS) technologies yet achieving atomic-level control remains challenging. Here, we show that the adsorption and nanoscale assembly of enantiopure [7]-helicene on Zinc Oxide (ZnO) is dictated primarily by the identity of the topmost surface atom rather than by macroscopic surface polarity. Using atomic force microscopy (AFM), circular dichroism (CD), force field molecular dynamics (FFMD), and density functional tight-binding (DFTB), we examine four ZnO terminations and find that polar O-terminated (0001) and both nonpolar surfaces reproducibly host uniform helicene nanoislands with a thickness of about 4 nm, while the polar Zn-terminated (0001) facet largely suppresses adsorption experimentally. FFMD and DFTB analyses provide a picture of the underlying interaction landscape, offering valuable theoretical support for the experimentally observed facet selectivity. The robust formation of oxygen-driven nanoislands identifies surface oxygen as an atomic-scale selector for chiral self-assembly, enabling deterministic patterning of ultrathin 2D chiral domains on oxide semiconductors. These insights establish a design rule for integrating helicene-based chiral architecture into spintronic and quantum devices.