Search Results (177)

Bacterial biofilms remain a major challenge in clinical infections due to their dense extracellular polymeric substance (EPS) matrix and strong resistance to conventional antibiotics. This study reports manganese dioxide (MnO₂) nanoparticles capable of autonomous navigation toward bacterial clusters, mechanical penetration of biofilm structures, redox-driven membrane disruption, and synergistic oxidative stress. The nanoparticles exhibit directional movement attributed to a combination of negatively charged surface potential, asymmetric topology, and catalytic reactivity toward bacterial metabolites. MnO₂ demonstrates potent antibiofilm activity against MRSA and MDR E. coli (>98% eradication) and partial activity against Pseudomonas aeruginosa. Time-lapse microscopy, EPR spectroscopy, XPS analysis, and SEM imaging reveal that MnO₂ disrupts both EPS and cell membranes while maintaining structural integrity throughout treatment. Cytotoxicity assays confirm ≥85% viability in human fibroblasts and keratinocytes at therapeutic concentrations. MnO₂ shows controlled biodegradation into Mn²⁺ ions, which participate in physiological pathways and undergo renal clearance. These findings support MnO₂ nanoparticles as promising biofilm-targeting agents for topical formulations, wound care, and implant coatings. Full article

Artificial intelligence (AI) is reshaping healthcare by supporting faster and more informed clinical decisions. However, the complexity of human health makes accurate predictive modeling challenging. In this study, we introduce a methodological framework for constructing intelligent digital twins of disease progression by combining patients’ sequential medical images with temporally aligned electronic health records (EHRs). EHRs in this context include structured clinical parameters such as laboratory test results, demographic characteristics, and medication information. The existing literature provides limited approaches that jointly forecast future medical images and clinical status using long-term historical data. Our framework integrates aligned temporal image sequences with these EHR features and employs either ConvLSTM or ViViT-based spatio-temporal encoders, optionally coupled with a generative module for future image synthesis. While awaiting access to patient datasets, we conducted an initial evaluation using a single-cell time-lapse microscopy dataset whose temporal dynamics resemble patient data. Both systems generate time-ordered image sequences that evolve under changing conditions, and the shifting nutrient environment in microfluidic channels parallels the temporal variations observed in patients’ EHR records. This preliminary study demonstrates the broader applicability of our model to datasets containing long-term sequential images and associated parameters, supporting its potential for future patient-specific digital twin development. Full article

Although combination therapies with mitogen activated protein kinase inhibitors remain among the most effective treatments for malignant melanoma, they are not universally applicable to all subtypes of this cancer, and their efficacy decreases in the presence of distant metastases. Drug resistance, often associated with elevated autophagy in tumor cells, and adverse effects of the treatment also reduce the survival time of melanoma patients. Therefore, the aim of our research was to assess the cytotoxicity of the combination of a late-stage autophagy blocker chloroquine with thymoquinone, a natural substance with anticancer potential and low toxicity towards healthy cells, in metastatic melanoma cell lines. Using the WST-1 assay, we examined the cytotoxicity of the combination of chloroquine and thymoquinone in melanoma WM9 and WM852 cell lines and assessed the type of their interactions. Additionally, using time-lapse bright field and fluorescent microscopy, we assessed changes in cell morphology during 48 h incubation with the tested compounds and their combinations, as well as their effect on autophagy. We identified an additive and sub-additive cytotoxic effect of thymoquinone/chloroquine combinations against WM9 and WM852 cells. Moreover, we found that thymoquinone combined with chloroquine caused an increase in autophagosome accumulation in WM9 cells, while attenuating the chloroquine’s anti-autophagic effect. A thorough understanding of the mechanism of drug interactions with natural substances is crucial for the development of new effective anticancer therapies. Full article

Heterotrophic nanoflagellates (HNANs) are central components of the microbial loop, transferring carbon from bacteria to higher trophic levels and facilitating nutrient recycling. While many HNANs are free-swimming, some exhibit enhanced feeding efficiency when attached to surfaces, including diatom frustules. Here, we describe the attachment behavior of a novel interception-feeding HNAN affiliated with the order Bicosoecida to centric diatoms common in North Carolina coastal waters. Using growth experiments, live observations, and time-lapse microscopy, we quantified attachment frequency and assessed its influence on diatom growth for three diatom species: Coscinodiscus sp., Odontella sp., and Rhizosolenia sp. HNAN attachment differed significantly among diatom taxa: Coscinodiscus sp. hosted the highest and most sustained numbers per frustule, whereas after normalizing for surface area, Rhizosolenia sp. exhibited the highest attachment efficiency. Diatom peak growth was 1.2 to 2.1-fold higher and occurred earlier in HNAN co-cultures than in controls, indicating microbial recycling by the HNAN stimulated growth. These findings highlight the nuanced ecological role attached HNANs might play as they exploit diatom-associated boundary layers to enhance bacterial encounter rates. The growth trajectories in our lab experiments suggests that attachment behavior in situ can play a role in driving diatom bloom dynamics and, therefore, play an important role for carbon cycling. Full article

Thraustochytrium aureum ssp. strugatskii, a marine protist belonging to the class Labyrinthulea, exhibits a complex life cycle characterized by alternating motile and vegetative phases. Using an integrative multimodal microscopy approach, we reconstructed its full developmental cycle and analyzed the coordination between cellular morphology, subcellular architecture, and population-level behavior. Transmission and scanning electron microscopy, combined with fluorescence and time-lapse imaging, revealed the dynamics of nuclear division, organelle rearrangement, and zoospore formation. Morphometric analysis of serial ultrathin sections demonstrated distinct changes in mitochondrial distribution, Golgi apparatus, and lipid droplet abundance during transitions between stages. We have shown that vegetative cells undergo synchronized karyokinesis coupled with stable nuclear-to-cytoplasmic ratios, leading to the emergence of multinucleate stages prior to zoospore formation. The integration of ultrastructural and dynamic data enabled us to propose a systems-level model linking metabolic state, morphogenesis, and population structure. This model highlights feedback regulation between nutrient availability, biomass accumulation, and developmental synchronization. Our results establish that T. aureum ssp. strugatskii has good potential to serve as a tractable model organism for systems-level studies of protists and provide an initial framework for predictive modeling of its life cycle under controlled conditions. Full article

Background: The combination of manganese porphyrins (MnPs) and ascorbate (ASC) represents a promising redox-based therapeutic approach for selectively targeting cancer cells. We investigated the cytotoxic effects of two structurally distinct MnPs (MnTPPS and MnF₂BMet) with differing lipophilicity and potential membrane permeability in combination with ASC. Methods: Cancer cell lines (MCF-7, PANC-1, U87, T98G, AT-2) and normal human dermal fibroblasts (HDFs) were treated with MnTPPS and MnF₂BMet in the absence or presence of ASC. Viability, migration potential, and intracellular oxidative stress were assessed using single-cell methods. Results: MnPs alone exhibited no intrinsic cytostatic or cytotoxic activity, as confirmed by proliferation, viability, and motility assays. When combined with ASC, both MnTPPS and MnF₂BMet significantly enhanced ASC-induced oxidative stress, leading to lipid peroxidation, glutathione depletion, mitochondrial dysfunction, and cell membrane disruption. Time-lapse microscopy revealed rapid necrotic cell death under co-treatment. Catalase fully abolished cytotoxicity, indicating the essential role of hydrogen peroxide. In contrast, dehydroascorbate (DHA), which increases intracellular ASC levels, did not induce the same toxicity, suggesting that extracellular ROS generation contributes predominantly to the observed effects. Normal fibroblasts were minimally affected, supporting the MnPs–ASC system’s selectivity toward cancer cells. Conclusions: MnTPPS and MnF₂BMet enhance extracellular oxidation of ascorbate and subsequent ROS production, leading to selective oxidative-stress-mediated cancer cell death. This study supports the potential of MnPs–ASC redox catalysis as a complementary oxidative-stress-based anticancer strategy and highlights the need for further mechanistic and structure–activity investigations. Full article

The development of epidermis plays a central role in driving the morphogenesis of the Caenorhabditis elegans embryo. However, current research on epidermal morphogenesis focuses disproportionately on overt phenotypic abnormalities, potentially overlooking the crucial role of developmental timing. In this study, we developed a modular two-step deep learning-based image analysis pipeline. First, we used ResU-Net to extract completely developed embryos and suppress noise; second, ResNet was used to predict the corresponding embryonic stage. The predicted probabilities and their corresponding embryonic time points were subsequently utilized to construct a developmental timeline. Combining this pipeline with differential interference contrast time-lapse microscopy, we dynamically tracked the timeline of epidermal morphogenesis in RNAi-treated embryos (ajm-1, tes-1, leo-1) and mutant embryos (clk-1). By statistically comparing the duration of each embryonic stage, our approach enabled the detection of stage-specific developmental timing without relying on overt phenotypic abnormalities or fluorescent markers, successfully recapitulating and extending the known roles of these genes from a temporal perspective. Our work underscores the importance of incorporating developmental timing into morphogenetic analysis, offering a novel framework for revealing subtle developmental processes, deepening the understanding of morphogenetic dynamics, and bridging the methodological gap in C. elegans embryology. Full article

Anticancer strategies targeting the DNA damage response are largely centered on a number of false hypotheses. For example, engaging apoptosis in solid tumors is universally assumed to represent a tumor suppression response. But what is “apoptosis”, really? Time-lapse microscopy and other single-cell assays have revealed that engaging apoptosis in solid tumor cells is accompanied by anastasis, the homeostatic process of cell recovery from late stages of apoptosis, even after the formation of apoptotic bodies. Furthermore, apoptotic cells secrete a variety of prosurvival factors that contribute to overall tumor repopulation. Not surprisingly, numerous clinical studies reported since the 1990s have demonstrated that increased apoptosis in solid tumors is associated with cancer aggressiveness rather than representing a favorable clinical outcome. Another major false hypothesis pertains to the role of wild-type p53 in regulating apoptosis. Several recent articles addressing the challenges that have been encountered in implementing p53-based cancer therapies assume that p53 is pro-apoptotic. This assumption, which has become an almost indisputable fact, is shocking given that by mid-2000s it was already well established that p53 serves to inhibit apoptosis through upregulating ~40 anti-apoptotic proteins. The complexity of cancer cell response to therapeutic agents is discussed herein with a focus on the significance of p53-p21^WAF1 signaling in suppressing the apoptosis–anastasis tumor repopulation pathway. Full article

Intravital two-photon microscopy enables deep-tissue imaging of subcellular structures in live animals, but its original spatial resolution and image quality are limited by scattering, motion, and low signal-to-noise ratios. To address these challenges, we used a combination of tissue stabilization, denoising methods, and motion correction, together with resolution enhancement algorithms, including enhanced Super-Resolution Radial Fluctuations (eSRRF) and deconvolution, to acquire high-fidelity time-lapse images of internal organs. We applied this imaging pipeline to image genetically labeled mitochondria in vivo, in Dendra2 mice. Our results demonstrate that the eSRRF-combined method, compared to other evaluated algorithms, significantly shows improved spatial resolution and mitochondrial structure visualization, while each method exhibiting distinct strengths in terms of noise tolerance, edge preservation, and computational efficiency. These findings provide a practical framework for selecting enhancement strategies in intravital imaging studies targeting dynamic subcellular processes. Full article

Encapsulation of fish oil within oleogels can potentially prevent oxidation and enable its use in food with programmable release within the gastrointestinal tract. Here, we report on the formation of oleogels from two different fish oils—salmon oil (SO) and cod liver oil (CLO)—using different concentrations of either rice bran wax (RBW) or myristic acid (MA) as gelators. The gels were assessed with respect to their structural, thermal, viscosity, digestive, and oxidative properties. Polarized light microscopy (POM) revealed that RBW consistently produced dense, interconnected crystalline networks across both oils, while MA formed larger, spherulitic crystals that were more sensitive to the oil type. This was further supported by time-lapse imaging, showing faster crystal growth of MA in cod liver oil. Viscosity studies indicate that the molecular weight and concentration of gelator, as well as the type of fish oil (SO vs. CLO), significantly impact the shear stability of the oleogels. Thermal and viscosity analyses confirmed that RBW-based oleogels exhibited higher crystallization temperatures and stronger viscoelastic behaviour. Based on oxidative stability measurements—as measured by peroxide value (PV) analysis—encapsulation within oleogels does not lead to significant oxidation of the fish oils and also attenuates further oxidation upon storage. The fish oil oleogels were stable when exposed to either simulated gastric or intestinal fluids (SGF and SIF, respectively), but decomposed after sequential exposure first to SGF and then to SIF. These findings could broaden the range of food products which can be fortified with fish oils. Full article

Alzheimer’s disease (AD) is characterized by amyloid-beta (Aβ) plaque accumulation and neurodegeneration. This study identified gyrophoric acid, a lichen-derived phenolic metabolite, as a dual-action Aβ42 inhibitor preventing aggregation and disassembling of mature Aβ42 fibrils. Integrated in silico studies revealed that gyrophoric acid was a strong thermodynamic stabilizer of Aβ42 (MM–GBSA: −27.3 kcal/mol) via entropically driven hydrophobic interactions and disruption of aggregation-prone conformations (100 ns MD simulations). Through biochemical analysis of the fluorescent dye thioflavin T (ThT), gyrophoric acid induced rapid Aβ42 fibril disassembly within 5 h, with time-lapse confocal microscopy quantitatively confirming the near-complete dissolution of large aggregates by 24 h. ADMET profiling revealed favorable pharmacokinetics (moderate oral absorption: 48.5–57.3%; low toxicity) and Lipinski’s rule compliance. These results establish gyrophoric acid as a promising natural bioactive compound for anti-AD therapeutics with a unique hydrophobic-stabilization mechanism. Full article

Time-lapse microscopy of mesenchymal stem cell (MSC) cultures allows for the quantitative observation of their self-renewal, proliferation, and differentiation. However, the rigorous comparison of two conditions, baseline (A) versus perturbation (B) (the addition of molecular factors, environmental shifts, genetic modification, etc.), remains difficult because morphology, division timing, and migratory behavior are highly heterogeneous at the single-cell scale. MSCs can be used as an in vitro model to study cell morphology and kinetics in order to assess the effect of, for example, gene therapy and prime editing in the near future. By combining static, frame-wise morphology with dynamic descriptors, we can obtain weight profiles that highlight which morphological and behavioral dimensions drive divergence. In this study, we present A/B Cells Policy: a modular, open-source Python package implementing a robust cell tracking pipeline. It integrates a YOLO-based architecture as a two-stage assignment framework with fallback and recovery passes, re-identification of lost tracks, and lineage reconstruction. The framework links descriptive statistics to a transferable system, opening up avenues for regenerative medicine, pharmacology, and early translational pipelines. It does this by providing an interpretable, measurement-based bridge between in vitro imaging and in silico intervention strategy planning. Full article

This study presents a novel method for high-precision detection and quantitative evaluation of the spatial relationship between cells and amyloid-$\beta$ (A$\beta$) in time-lapse brightfield microscopy images. Achieving accurate detection of non-fluorescent cells and A$\beta$ deposits requires high-quality video images free from noise, distortion, and frame-to-frame luminance flicker. To this end, we employ a robust preprocessing pipeline that combines multi-frame integration with vignetting correction to enhance image quality and reduce luminance variability across frames. Key preprocessing steps include background correction via two-dimensional polynomial fitting, temporal smoothing of luminance fluctuations, histogram matching for luminance normalization, and dust artifact removal based on intensity thresholds. This enhanced imaging approach enables accurate identification of A$\beta$ aggregates, which typically appear as jelly-like structures and are difficult to detect under standard brightfield conditions. Furthermore, we introduce a quantitative index to assess the spatial relationship between cells and A$\beta$ concentrations, facilitating detailed analysis under varying A$\beta$ levels. Full article

Similarly to the short-lived messenger nitric oxide (NO), the more stable carbon monoxide (CO) molecule can also activate soluble guanylyl cyclase (sGC) to increase cGMP levels. However, CO-induced cGMP production is much less efficient. Using an accessible invertebrate model, we dissect a potential interaction between the canonical NO/sGC/cGMP and CO signalling pathways during development. The embryonic midgut of locusts is innervated by neurons that migrate in four discrete chains on its outer surface. Transcellular diffusing NO stimulates enteric neuron migration via cGMP signalling. The application of an NO donor results in virtually all enteric neurons being cGMP-immunoreactive while CO increases cGMP production only in approximately 33% of the migrating neurons. Cellular CO release appears to act as a slow down signal for motility. We quantify how CO specifically increases the interneuronal distance during chain migration. Moreover, time-lapse microscopy shows that CO reduces the directionality of the migrating neurons. These findings support the function of NO and CO as antagonistic signals for the coordination of collective cell migration during the development of the enteric nervous system. These experiments and the resulting insights into basic scientific questions prove once more that locust embryos are not only preparations for basic research, but also relevant models for screening of drugs targeting NO and CO signalling pathways as well as for isolating compounds affecting neuronal motility in general. Full article

Full-field optical coherence tomography (FF-OCT) is a high-resolution interferometric imaging technique that enables label-free visualization of cellular structural changes. In this study, we employed a custom-built time-domain FF-OCT system to monitor morphological alterations in HeLa cells undergoing doxorubicin-induced apoptosis and ethanol-induced necrosis at the single-cell level. Apoptotic cells showed characteristic features such as echinoid spine formation, cell contraction, membrane blebbing, and filopodia reorganization. In contrast, necrotic cells exhibited rapid membrane rupture, intracellular content leakage, and abrupt loss of adhesion structure. These dynamic events were visualized using high-resolution tomography and three-dimensional surface topography mapping. Furthermore, FF-OCT-based interference reflection microscopy (IRM)-like imaging effectively highlighted changes in cell–substrate adhesion and cell boundary integrity during the cell death process. Our findings suggest that FF-OCT is a powerful imaging platform for distinguishing cell death pathways and assessing dynamic cellular states, with potential applications in drug toxicity testing, anticancer therapy evaluation, and regenerative medicine. Full article