Search Results (4,778)

Ventricular remodeling occurring in heart failure (HF) involves structural disarray of the sarcolemma T-tubule (TT)–sarcoplasmic reticulum (SR) dyad junctions, thereby disrupting the close apposition of L-type Ca²⁺ channels (Ca_V1.2) with ryanodine receptors (RyR2) that trigger SR Ca²⁺ release and myofilament contraction. In a rat ischemic heart failure model expressing low thyroid hormone (TH) function, we used 3D stochastic optical reconstruction microscopy (STORM) to image RyR2 clusters with Ca_V1.2 channels, and the associated protein junctophilin-2 (Jph2). We tested whether treatment with T3, the biologically active form of TH, throughout progression of the disease would preserve T-tubule structure and dyadic ion channel organization. Confocal microscopy of isolated cardiomyocytes (CMs) stained with ANEPPS membrane dye showed significantly decreased TT density in diseased CMs while T3 treatment attenuated TT disorganization. 3D STORM images of dyadic ion channels labeled with fluorescent-tagged antibodies to RyR-Dylight550, Jph-CF647 and Ca_V1.2/IgG-Dylight488 were captured. A density-based algorithm defined RyR2 clusters, and a 400 nm spherical 3D volume of interest around each RyR2 cluster’s centroid determined the number of Ca_V1.2 and Jph2 localizations associated with each RyR2 cluster. Analysis revealed significant reduction in RyR2 cluster size and number with reduced co-localized Jph2 in failing CMs. T3 treatment increased RyR2 cluster numbers and cluster volumes albeit non-significantly, with increased co-clustering of Jph2. The number of Ca_V1.2 co-localized with RyR2 clusters trended lower in the failing CMs. These results support maintaining TH homeostasis in optimizing the nanoscale organization of Ca²⁺ ion channels in triggering Ca²⁺ release and myofibrillar contraction in patients with heart disease. Full article

Background/Objectives: This review describes the advantages of new photonic-based approaches for assessing the activity of caries lesions. Many lesions have been arrested or are non-carious developmental defects, such as fluorosis, which do not require intervention. New methods are needed to assess lesion activity and avoid unnecessary removal of the tooth structure. Methods: At present, there are no reliable methods for assessing lesion activity in vivo. Nondestructive optical monitoring of lesion structure and the changes in light scattering that occur during drying offer the potential for lesion activity assessment during a single examination. Since optical diagnostic instruments exploit changes in the porosity and the permeability of the lesion, they have the potential to assess whether lesions are active and expanding or arrested and undergoing remineralization. Optical coherence tomography (OCT), Raman imaging and fluorescence loss, thermal and short-wavelength infrared (SWIR) reflectance measurements during lesion dehydration with forced air are presented. Results: Clinical studies have shown that optical coherence tomography is capable of showing distinct structural differences between active and arrested lesions on coronal and root surfaces. Differences in the kinetics of dehydration measured using reflectance measurements at SWIR wavelengths coincident with water absorption bands also show great potential. Conclusions: OCT and dehydration imaging at SWIR wavelengths have great potential for assessing lesion activity since they can also be used for caries screening, are safe for frequent monitoring and do not require the application of external agents. Full article

Background: Healthcare-associated infections (HAIs) remain a critical patient safety challenge. Hand hygiene is considered the most effective preventive measure, yet traditional monitoring captures only compliance, not technique quality. This prospective before–after study evaluated whether real-time visual feedback via the Semmelweis UV-fluorescence system is associated with improved hand hygiene quality, measured by ATP bioluminescence and microbiological culture. Methods: Three clinical departments (the Intensive Care Unit, Hematology, and Gynecology) at a Romanian tertiary hospital were purposively selected. Seventy-one healthcare workers (HCWs) were enrolled. The 12-week study comprised Phase 1 (baseline, weeks 1–4), Phase 2 (active intervention with Semmelweis feedback, weeks 5–8), a one-week washout (week 9), and Phase 3 (sustainability assessment, weeks 10–12). Paired ATP-CFU samples were collected weekly. Within-group comparisons used Kruskal–Wallis H tests with post hoc Dunn’s tests and Bonferroni correction. Secondary outcomes included Semmelweis global and zone-specific coverage and the correlation between subject-level Semmelweis coverage and ATP bioluminescence (Spearman’s rho). Results: A total of 781 paired ATP-CFU samples and 497 Semmelweis evaluations were analyzed. Mean ATP declined from 195.9 RLU at baseline to 148.2 RLU in Phase 2 (−24.4%) and 154.8 RLU in Phase 3 (−21.0%; Kruskal–Wallis H = 102.73, p < 0.001). CFU/mL declined from 84.8 to 66.2 (−21.9%) and 70.7 (−16.6%; H = 22.48, p < 0.001). Post hoc comparisons confirmed significant Phase 1 versus Phase 2 and Phase 1 versus Phase 3 differences for both markers (all p < 0.01), while Phase 2 versus Phase 3 was non-significant, indicating stabilization at an improved level. Subject-level Semmelweis coverage correlated negatively with ATP (rho = −0.665, 95% CI −0.778 to −0.510, p < 0.001), supporting construct validity at the operator level. Semmelweis global coverage was 93.1% (Phase 2) and 90.6% (Phase 3); interdigital spaces showed the highest inadequacy rate (73.9% protocol-based, 92.5% targeted). Conclusions: Real-time visual feedback via UV-fluorescence imaging was associated with significant and sustained improvements in hand hygiene quality beyond baseline. ATP, CFU, and Semmelweis assessments captured complementary, non-redundant dimensions, supporting multimodal evaluation. Interdigital spaces and fingertips remained persistent failure points requiring targeted educational reinforcement. Full article

Background: Glucocorticoid-induced osteoporosis (GIOP) is marked by impaired osteogenesis and reduced bone formation. Isobavachin (IBA), a flavonoid from Psoralea corylifolia, shows multiple potentials in anti-inflammatory and bone metabolism regulations, but its effects against GIOP remain unclear. This study investigated the osteoprotective effects and potential mechanism of IBA using zebrafish GIOP model. Methods: osteoprotective effects of IBA was assessed by fluorescence imaging in a prednisolone-induced zebrafish model, following with osteogenic gene expressions measured by RT-qPCR. Potential targets and pathways of IBA was filtered and predicted by network pharmacology, molecular docking, and molecular dynamics (MD) simulations, and finally validated with a pharmacological rescue experiment using a PI3K-specific inhibitor. Results: IBA improved bone mineralization and upregulated osteogenesis-related genes. Network pharmacology identified the PI3K-Akt pathway as a key pathway, with ESR1, GSK3B, MTOR, and CCND1 as core targets. PI3K inhibition attenuated the osteoprotective effects of IBA and suppressed downstream osteogenic gene expression. Conclusions: IBA alleviates GIOP by modulating the ESR1-associated PI3K-Akt signaling pathway and may serve as a multi-target therapeutic candidate for osteoporosis. Full article

Regenerative medicine integrates interdisciplinary approaches towards restoring the function of diseased organs. This study examined alterations that occurred in the liver under a high-fat diet (HFD) with the development of obesity and fatty liver, and changes in metabolic homeostasis and glucose levels, in mice. HFD nutrition causes hyperglycemia, leading to the formation and accumulation of advanced glycation end-products (AGEs) promoting protein post-translational modifications (PTMs) and introducing crosslinking in the extracellular matrix (ECM). Using histological and gene expression analyses, we detected an increase in adiposity, as well as in ECM protein deposition in the liver. Further, decellularization of the liver yielded the isolated ECM organ scaffold, allowing us to analyze the chemical modification in proteins by various imaging methods combined with spectroscopy. The measurements of intrinsic protein fluorescence are consistent with increased AGE-associated levels. SEM allows for the visualization of ECM fiber thickening as a result of protein crosslinking. Using cathodoluminescence, a label-free imaging method, we confirmed the protein modifications. The combination of innovative technologies highlights the ECM structural alterations associated with impaired glucose regulation and liver adiposity. These findings provide novel views on liver-scaffold ECM structure under metabolic diseases that will play a significant role in accelerating the understanding of effective regenerative therapies. Full article

Background/Objectives: Systematic lymphadenectomy is recommended in apparent early-stage epithelial ovarian cancer (EOC) to assess nodal status, but it is associated with significant morbidity and lacks survival benefit. Sentinel lymph node (SLN) mapping may offer a less invasive alternative, although evidence remains limited due to the complexity of ovarian lymphatic drainage and methodological heterogeneity across studies. This prospective study evaluates the feasibility and diagnostic accuracy of a true hybrid ^99mTc–indocyanine green (ICG) tracer for SLN mapping in apparent early-stage EOC. Methods: A prospective observational study was conducted at a tertiary oncology center between 2021 and 2026. Patients presenting with a suspicious ovarian mass (Group A) or requiring restaging after adnexectomy for confirmed EOC (Group B) underwent SLN mapping using a hybrid ^99mTc–ICG tracer injected into the infundibulopelvic (IPL) and/or utero-ovarian ligament (UOL). SLNs were identified using gamma detection and near-infrared fluorescence imaging. All malignant cases underwent complete surgical staging including systematic pelvic and para-aortic lymphadenectomy. SLNs were ultrastaged and compared with the final nodal status. Results: Forty patients were included; 20 (50%) had malignant tumors. The overall SLN detection rate was 92.5% (37/40), with 100% in malignant cases. Among malignant tumors, 3/20 (15%) had metastatic SLNs, all accurately detected (false-negative rate 0%). Sensitivity and negative predictive value were 100%. Combined pelvic and para-aortic drainage was the most frequent pattern (75%). Conclusions: SLN mapping may represent a feasible and potentially accurate staging strategy in apparent early-stage EOC. In the present study, a hybrid ^99mTc–ICG tracer was associated with high detection rates and complete concordance with final nodal status. These findings support further multicenter validation to define its potential role as an alternative to systematic lymphadenectomy. Full article

Tobramycin is a highly hydrophilic aminoglycoside antibiotic with limited cellular permeability and negligible oral bioavailability, necessitating parenteral administration. This study aimed to develop drug delivery systems based on nano-sized colloidal assemblies (NCAs) incorporating tobramycin ion pairs to enhance its lipophilicity, potential for transition to the oral route, and antimicrobial activities. Tobramycin was ionically paired with oleic acid, lauric acid, and fluorescein and formulated into NCA preconcentrates (F1–F5) using combinations of Tween 80, DMSO, and propylene glycol. The resulting formulations formed stable nanodroplets upon dilution (9.50–16.30 nm) with narrow size distributions (polydispersity index; PDI < 0.3) and moderate negative zeta potentials (−4.99 to −11.13 mV). In vitro release studies indicated sustained drug release for ion-paired systems compared to the rapid release of free tobramycin. Cytotoxicity evaluation in Caco-2 cells demonstrated high biocompatibility at 1:10,000 and 2:10,000 dilutions, while concentration-dependent toxicity at higher doses suggested enhanced intracellular delivery. Cellular uptake studies revealed significantly higher tobramycin internalization (p < 0.001) from formulations F1–F3, with uptake values in the range of 81.76–96.14% compared to free drug, which showed zero or negligible uptake. Fluorescein-labeled formulations (F4 and F5) further confirmed enhanced uptake, demonstrating strong intracellular fluorescence. This was supported by visual observation, UV–Vis absorbance (70.5–84.8% relative to positive control), and confocal microscopy imaging. Antimicrobial activities against P. aeruginosa and S. aureus were comparable between formulations F1–F5 and free tobramycin (inhibition zones of 16–18 mm), utilizing the same tobramycin concentration in the diluting medium. These findings validate the effectiveness of the formulated NCAs in facilitating intracellular delivery of tobramycin while preserving biocompatibility and similar antimicrobial activities. Moreover, the uptake of fluorescein provides indirect evidence supporting the enhanced internalization of tobramycin in analogous ion-paired formulations. This strategy holds promise for overcoming intestinal barriers and improving oral bioavailability, potentially enabling the transition of tobramycin from parenteral to oral administration. Full article

Background/Objectives: Sentinel lymph node (SLN) biopsy is a minimally invasive alternative to systematic lymphadenectomy for endometrial cancer staging. However, optimization of SLN identification during robot-assisted surgery remains an important clinical issue. This prospective exploratory study evaluated the feasibility and mapping characteristics of dual-tracer SLN mapping combining radioisotope (RI) and indocyanine green (ICG) in robot-assisted surgery for clinical stage IA endometrial cancer. Methods: Ten patients with clinical stage IA endometrioid carcinoma (grade 1 or 2) who underwent robot-assisted surgery at our institution between June 2025 and March 2026 were prospectively enrolled. Technetium-99m phytate was injected cervically the day before surgery, followed by SPECT-CT imaging. ICG was administered intraoperatively. SLNs were identified using both RI mapping and near-infrared fluorescence imaging. All patients subsequently underwent pelvic lymphadenectomy. SLN detection rates, concordance between tracers, intraoperative findings, and perioperative outcomes were evaluated. Results: The SLN detection rate with the dual-tracer method was 100% (10/10). RI mapping alone achieved bilateral detection in 30% (3/10), whereas ICG fluorescence imaging identified SLNs in all patients (100%). All RI-positive SLNs were also identified by ICG fluorescence imaging. In several cases, multiple fluorescent lymph nodes were observed, and gamma-probe assessment aided intraoperative SLN selection. No lymph node metastases were identified. One Clavien–Dindo grade IIIa complication (pelvic hematoma requiring intervention) occurred. No adverse events related to ICG were observed. Conclusions: Dual-tracer SLN mapping using RI and ICG was feasible in robot-assisted surgery for clinical stage IA endometrial cancer. ICG fluorescence imaging provided sensitive lymphatic visualization, whereas RI mapping offered complementary information for intraoperative SLN identification in selected cases. Further prospective studies with larger cohorts are required to clarify the clinical utility of the dual-tracer approach. Full article

Research on the cataloging of microstructures and chemical compound localization in peanut hulls in relation to fungal tolerance remains limited. The hull (pericarp) is the first physical interface with the soil environment and may contribute to defense against fungal invasion. Here, hull microstructure and histochemical localization of alkaloid-like compounds, cellulose, lignin, starch, and total proteins were characterized across reproductive developmental stages R3–R6 in three commercially grown cultivars (Georgia-06G, Georgia-12Y, and Georgia-18RU). Stained sections were examined by light and fluorescence microscopy, and images were quantified in Fiji-ImageJ as stained area percentage. Among the compounds studied, the highest area percentages were observed at later stages (R5 and R6). Alkaloid-like compounds, cellulose, and starch were higher at the R5 stages of G-18 (9.61 ± 0.75), G-12Y (22.96 ± 5.84), and G-06 (6.31 ± 1.13) respectively, while lignin and total proteins were highest at the R6 stage of G-18 (respectively, 14.49 ± 1.43 and 13.90 ± 1.45). The lowest histochemical presence for most metabolites occurred in the early stages (R3–R4). This indicates that hull maturation is accompanied by increased physical (sclerenchyma and lignified cells) and biochemical (alkaloid-like compounds, proteins) features consistent with protective roles. As the analysis was based on representative sections and regions of interest (ROI)-level quantification, the results are intended to guide future studies on hull-mediated defense and breeding for Aspergillus tolerance. Full article

In this study, probe 1, a novel mitochondria-targeted fluorescent probe for the colorimetric and ratiometric detection of Hg²⁺, was developed. Upon addition of Hg²⁺ to the solution of 1, distinct spectral changes were observed. The absorption spectra underwent a blue shift from 510 nm to 450 nm (Δλ = 60 nm), and the solution color changed from red to pale yellow under daylight. Concurrently, a significant blue shift occurred from 645 nm to 540 nm (Δλ = 105 nm) in the fluorescence spectra. There were remarkable variations in the fluorescence intensity ratio of F_{540 nm}/F_{645 nm} with the R/R₀ value reaching up to 824-fold, and the fluorescence color changed from red to green under a 365 nm UV lamp. Probe 1 featured a large Stokes shift of 135 nm, high sensitivity with an LOD of 25.5 nm, and excellent selectivity for Hg²⁺ even in the presence of other analytes. Furthermore, 1 was successfully applied for the ratiometric imaging of intracellular Hg²⁺ and was confirmed to localize specifically within mitochondria. Full article

This study aimed to systematically compare the functional properties of the four major components (albumin, globulin, prolamin, and glutelin) of protein from Yunnan deep-veined walnuts to screen for protein-based carrier materials with good processing adaptability and the ability to efficiently encapsulate the active ingredient rutin. In addition, the binding and molecular interactions between the preferred protein and rutin were analyzed. The results indicated that albumin exhibited superior performance compared to the other three components in solubility, emulsifying properties, foaming properties, and gel properties, and demonstrated the strongest processing applicability. Further analysis revealed that albumin possessed an excellent amino acid composition (essential amino acid content accounting for 42.30%) and antioxidant activity (with the highest ABTS scavenging rate reaching 85.71 ± 0.26%), which indicated its considerable potential as a functional carrier. Loading rutin onto albumin yielded a walnut albumin–rutin complex (WA@Rut), which significantly enhanced the thermal stability of albumin (with the thermal denaturation temperature elevated to 108.72 °C) and the storage stability of rutin (66.16 ± 5.05% retention after 22 days of storage). Combined analyses of FT-IR spectroscopy, intrinsic fluorescence spectroscopy, molecular docking, and molecular dynamics simulations confirmed that rutin primarily bound to albumin via hydrogen bonding and electrostatic interactions, and formed a stable complex structure. SEM images revealed that the composite surface was smooth and exhibited a flake-like morphology. In conclusion, walnut albumin is a protein resource with significant functional potential in Yunnan deep-veined walnuts, and it exhibits strong processing applicability and enables efficient encapsulation and protection of active ingredients. This study provides novel strategies and theoretical foundations for the high-value utilization of walnut protein. Full article

Background/Objectives: The toxic side effects and resistance-associated limitations of conventional chemotherapeutic agents necessitate the development of more effective and selective combination strategies incorporating naturally derived compounds. In this study, the cytotoxic, apoptotic, oxidative stress-associated, and immunomodulatory effects of thymoquinone (TQ), a bioactive compound derived from Nigella sativa, and docetaxel (Dos), a taxane-based chemotherapeutic agent, were investigated alone and in combination in OVCAR3 ovarian cancer cells using integrated two-dimensional (2D) and three-dimensional (3D) experimental models. Materials and Methods: Cell viability was evaluated following treatment with TQ (10–500 µM), Dos (1–500 nM), and the TQ + Dos combination, and synergistic interactions were assessed by IC₅₀⁻ and combination index-based analyses. Apoptosis and cell cycle distribution were analyzed by flow cytometry. Cytokine levels were determined using ELISA, whereas apoptosis- and cell cycle-associated gene expression profiles were evaluated by RT-qPCR. Active caspase-3 expression was assessed by immunocytochemistry. Intracellular reactive oxygen species (ROS) accumulation was examined using DCFH-DA-based fluorescence imaging and antioxidant rescue experiments using N-acetyl-L-cysteine (NAC). In addition, the antitumor activity of the combination was further evaluated in OVCAR3-derived 3D tumor spheroid models using spheroid morphology, ATP-based viability, and live/dead fluorescence imaging analyses. Results: The TQ + Dos combination demonstrated enhanced cytotoxic and apoptotic activity in OVCAR3 cells compared with single-agent treatments and induced marked G2/M cell cycle arrest. Combination treatment increased pro-apoptotic gene expression and was associated with reduced expression of anti-apoptotic markers and modulated inflammatory cytokine profiles. Fluorescence-based analyses demonstrated marked intracellular ROS accumulation following TQ + Dos treatment, whereas NAC pretreatment partially attenuated oxidative stress and restored viability, suggesting partial involvement of ROS-associated mechanisms in treatment-induced cytotoxicity. Importantly, the combination maintained stronger cytotoxic and growth-inhibitory effects than either monotherapy in 3D ovarian cancer spheroids, where combination treatment induced pronounced spheroid shrinkage, viability loss, and structural disruption. Relatively lower toxicity observed in HaCaT cells suggested partial selectivity toward cancer cells. Conclusions: Collectively, these in vitro findings suggest that the TQ + Dos combination produces greater cytotoxic, apoptotic, and growth-inhibitory effects than either agent alone in ovarian cancer models and is associated with alterations in apoptosis-, cell cycle-, and oxidative stress-related responses. The observation of these effects in 3D spheroid models supports further investigation of this combination in more advanced preclinical systems. Full article

Background/Objectives: Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with epithelial remodeling, impaired mucociliary clearance, and altered nitric oxide (NO) metabolism. However, cell type-specific mechanisms underlying epithelial NO signaling remain poorly understood. This study investigated NO-related signaling in differentiated human sinonasal epithelial cells. Methods: Human sinonasal tissues were obtained from patients with CRSwNP (n = 20) and control subjects (n = 20). Air–liquid interface (ALI) cultures were established from donor-derived epithelial cells. Ciliated and non-ciliated cells were identified by immunostaining for acetylated α-tubulin and BCAM. Expression of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) was analyzed by quantitative RT-PCR. Intracellular NO-related fluorescence signals were evaluated using DAF-FM fluorescence imaging. Results: CRSwNP tissues exhibited significantly increased iNOS expression and elevated iNOS/eNOS ratios, whereas eNOS expression did not differ significantly from that in controls. ALI cultures reproduced differentiated sinonasal epithelium containing both ciliated and non-ciliated cell populations. DAF-FM fluorescence signals were significantly higher in ciliated cells than in non-ciliated cells (80.3 ± 25.3 vs. 49.3 ± 21.1). Non-selective NOS inhibition markedly reduced fluorescence signals in both cell types, whereas selective iNOS inhibition reduced but did not abolish signals in ciliated cells. Conclusions: NO-related signaling appears to differ among epithelial cell subtypes. Persistence of fluorescence signals after selective iNOS inhibition suggests a contribution of constitutive NOS activity in ciliated cells, whereas non-ciliated cells appear to rely more heavily on iNOS-dependent pathways. These findings support the hypothesis that altered epithelial NO signaling contributes to epithelial dysfunction and impaired mucociliary homeostasis in CRSwNP. Full article

In the Iberian Pyrite Belt (IPB), long-term persistence of mine waste piles poses environmental challenges. The present work studies the Trimpancho Mining Complex in northern IPB with exposed mine waste and acidic waters in the proximity to the Chança River, a tributary of the Guadiana international river. A multidisciplinary approach is proposed, using hyperspectral reflectance spectroscopy, portable X-ray fluorescence (pXRF), multispectral Unmanned Aerial Vehicle (UAV) and Sentinel-2 images. Spectroscopic, geochemical and remote sensing methods were applied to characterise the mining area. Comparison of hyperspectral data with spectral libraries were used to validate mineralogy. Multispectral UAV data is used for custom band-ratios and adapted to Sentinel-2 images. Results grouped the samples into four groups. Spectroscopy is indicative of clays (white mica and smectite group), hematite/goethite, jarosite, and arsenopyrite and pyrite (exclusive to the Group 2); iron-rich samples reach maximum reflectance earlier than iron-poor samples. Geochemical studies show an increase in content of heavy metal such as As, Cu, Fe, Pb, and Zn from Group 1 < Group 3 ≈ Group 4 < Group 2, but Group 4 showed elevated Pb and Zn. Custom false colour composition highlighted the groups in UAV and satellite, thus constituting cost-effective tools for finding contamination sources. Full article

As thoracic surgeons more frequently address smaller lung lesions and perform lung-sparing resections, their objective is to resect an adequate specimen and margin without removing excess healthy lung tissue. Although perioperative lung nodule localization has been in practice for decades, the existing and emerging techniques used for the identification of targeted and occult lesions are more widely utilized today than they were in the past. In this review, we detail the logic behind this increase in use, classify the techniques into preoperative and intraoperative categories, and define the specific modalities available. Where applicable, we review the published data comparing techniques, detailing efficacy and safety. In the preoperative space, we describe standard computed tomography (CT)-guided localization, virtual-assisted lung mapping, electromagnetic navigation bronchoscopy, robotic-assisted bronchoscopy, and novel fiducial markers. In the intraoperative space, we describe classical localization techniques, novel applications of intraoperative cone-beam CT, and fluorescence-guided surgery and intraoperative molecular imaging (IMI). Lastly, we review emerging approaches for intraoperative molecular imaging including a report on agents in early-stage clinical trials and a brief survey of promising preclinical models. With each approach mentioned, we analyze the potential benefits and hazards, and appraise the evidence for (or against) the use of any specific modality. Full article