Search Results (110)

Transmitochondrial cybrid technology is a key approach for elucidating the effects of mitochondrial DNA (mtDNA) mutations in defined nuclear genetic backgrounds and for studying nuclear–mitochondrial interactions. However, its application is limited by the availability of suitable recipient cell lines and by technically demanding enucleation procedures. We report three advances in cybrid technology: (1) enucleation using mitomycin C, a widely used agent for generating feeder layers in stem cell culture, which does not depend on cell attachment and provides a gentler alternative to actinomycin D; (2) selection of cybrids using mitochondrial uncouplers, which can reduce background survival of non-cybrid cells; and (3) cryopreservation of enucleated donor cells in liquid nitrogen, preserving fusion competence and increasing experimental flexibility. Additionally, we validate newly developed mtDNA-free (ρ⁰) derivatives of HCT116, HT1080, and U2OS cell lines as recipients for cybrid generation. These advances facilitated donor cell preparation, improved cybrid selection, and enhanced experimental flexibility, including the demonstration of preserved fusion competence of enucleated HeLa cells after 10 years of cryostorage. The ρ⁰ derivatives of HCT116, HT1080, and U2OS cells were confirmed as effective recipients. Together, these improvements enhance the efficiency and accessibility of transmitochondrial cybrid technology and are expected to facilitate its broader application. Full article

This paper reviews recent work on dynamic virtual power plants (DVPPs) using an Energy–Information–Market framework. It addresses the important problem of how DVPPs can support low-inertia power system operation and feeder-level stability under high renewable penetration. First, system-level studies on low-inertia operation and frequency control are used to frame quantitative requirements on rate of change of frequency, nadir, and quasi-steady-state limits. Second, energy-layer models are surveyed, including participation-factor-based DVPP controllers, grid-forming architectures, model-free frequency regulation, and robust frequency-constrained scheduling for allocating virtual inertia and fast frequency response (FFR) across distributed energy resource fleets. Third, information-layer and market-layer models are reviewed, covering stochastic and robust bidding, distribution locational marginal price-based clearing, peer-to-peer and community markets, privacy-preserving coordination, and emerging governance and cybersecurity schemes for DVPP participation. Across these strands, much of the literature remains centred on steady-state active and reactive power dispatch, with dynamic security enforced as constraints rather than formulated as verifiable and tradable services. This review identifies gaps in dynamic metrics and benchmarks, forecasting of available inertia and FFR capacity, market-physics co-design, multi-aggregator interaction, and experimentally validated DVPP implementations. These findings suggest that DVPPs can “sell stability” at the feeder level only through co-designed control, information, and market mechanisms and outline a research roadmap for this purpose. Full article

The conversion of primed pluripotent stem cells to a naive-like state has emerged as a critical strategy for enhancing developmental potential and broadening applications in regenerative medicine. Conditioned media (CM)-based approaches provide a supportive microenvironment enriched with secreted factors that may facilitate this state transition without extensive genetic or chemical manipulation. In this study, we investigated the potential of human Wharton’s Jelly-derived mesenchymal stem cell-conditioned media (hWJ-MSCs-CM) and mouse embryonic fibroblasts CM (MEFs-CM) to support the conversion of primed rhesus monkey embryonic stem cells (rhESCs) into a naive-like state. The rhESCs were cultured under feeder-free and feeder conditions using both hWJ-MSCs-CM and MEFs-CM, exhibiting distinct morphological changes during conversion. Immunofluorescence analysis demonstrated the expression of pluripotency and naive markers under both conditions. Gene expression analysis further confirmed the upregulation of naive-specific genes and downregulation of primed markers, with statistically significant differences between groups. Additionally, epigenetic reprogramming was assessed, revealing differential effects of the CM sources on the reversion to a naive state. These findings highlight the potential of hWJ-MSCs-CM as a supportive system for naive-like state induction in primate ESCs. Full article

Background: Cultivated oral mucosal epithelial transplantation (COMET/CAOMECS) is an autologous, immunosuppression-sparing option for ocular surface reconstruction in limbal stem cell deficiency (LSCD). After two decades, indications, platforms and outcome definitions vary, and COMET’s position relative to limbal-derived epithelium remains uncertain. Methods: We conducted a PRISMA-ScR scoping review of human clinical studies (PubMed, 2000–30 December 2025) with hand-searching and regulatory sources. Eligible reports included COMET/CAOMECS series and comparative cohorts (CLET/ACLET, SLET, KLAL/CLAL). The primary outcome was anatomical success (stable epithelialised cornea without recurrent persistent epithelial defect, progressive conjunctivalisation or uncontrolled neovascularisation at last assessment). Given heterogeneity in definitions and analytic frames (fixed-time vs. Kaplan–Meier [KM]), results were synthesised narratively by indication and platform. Results: Twenty-five reports (893 eyes; 821 patients) were included. Aetiologies were predominantly burns and SJS/TEN. Across amniotic membrane-based mixed-aetiology series, 12-month anatomical success clustered around 55–70%. Aggregated descriptively across COMET eyes, 211/467 (45%) had a stable surface at last follow-up. Epithelialisation was generally rapid in quiet AM-based reconstructions and slower with severe adnexal disease or carrier-free platforms. Mean BCVA improved from 1.8 ± 0.7 to 1.4 ± 0.7 logMAR (471 eyes); ≥2-line gains occurred in 308/471 (65.4%). A matched comparison suggested better 12-month survival, less neovascularisation and better BCVA with substrate-free versus AM-carried COMET; a biomaterial-/feeder-free platform reconstructed most eyes but failed more often with four-quadrant symblepharon. Observational comparative cohorts suggested higher surface survival and average visual gain with limbal-derived epithelium, at the cost of systemic immunosuppression. Conclusions: In appropriately selected bilateral LSCD, COMET offers immunosuppression-sparing reconstruction with moderate, durable surface stability and clinically meaningful visual gains when performed on a quiet, optimised surface. Platform refinements—particularly substrate-free constructs—and prospective, indication-defined comparative studies with harmonised outcomes are needed to define COMET’s role relative to limbal-derived epithelium. Full article

The increasing complexity of modern electrical substations—driven by renewable integration, advanced automation, and asset aging—necessitates a transition from reactive maintenance toward intelligent, data-driven strategies. Predictive maintenance (PdM), supported by artificial intelligence, enables early fault detection and remaining useful life (RUL) estimation, while Digital Twin (DT) technology provides synchronized cyber–physical representations for situational awareness and risk-free validation of maintenance decisions. This study proposes a five-layer DT-enabled PdM architecture integrating standards-based data acquisition, semantic interoperability (IEC 61850, CIM, and OPC UA Part 17), hybrid AI analytics, and cyber-secure decision support aligned with IEC 62443. The framework is validated using utility-grade operational data from the SS1 substation of the Badra Oil Field, comprising approximately one million multivariate time-stamped measurements and 139 confirmed fault events across transformer, feeder, and environmental monitoring systems. Fault detection is formulated as a binary classification task using event-window alignment to the 1 min SCADA timeline, preserving realistic operational class imbalance. Five supervised learning models—a Random Forest, Gradient Boosting, a Support Vector Machine, a Deep Neural Network, and a stacked ensemble—were benchmarked, with the ensemble embedded within the DT core representing the operational predictive model. Experimental results demonstrate strong performance, achieving an F1-score of 0.98 and an AUC of 0.995. The results confirm that the proposed DT–AI framework provides a scalable, interoperable, and cyber-resilient foundation for deployment-ready predictive maintenance in modern substation automation systems. Full article

Platelet shortage poses a significant barrier to research and transfusion therapies because native megakaryocytes (MKs) are scarce in blood. To overcome this limitation, pluripotent stem cell–derived MKs (PSC-MKs) offer a standardized, donor-independent platform for research and therapeutic development, including disease modeling and ex vivo platelet production. Here, we report a chemically defined, feeder-free protocol to generate MKs from human pluripotent stem cells (hPSCs). The protocol combines the small molecule MPL agonist Butyzamide, macrophage colony-stimulating factor (M-CSF), and three-dimensional (3D) suspension culture, achieving high efficiency and reproducibility. Butyzamide replaced recombinant thrombopoietin (TPO), yielding comparable CD41⁺/CD42b⁺ populations and enhanced polyploidization. M-CSF accelerated nuclear lobulation and induced 4N MKs, while 3D culture increased yield, cell size, and substrate detachment. Multiple independent assays confirmed mature MK hallmarks, multi-nuclei, demarcation membranes, granules, and elevated mitochondrial respiration. Single-cell RNA sequencing outlined a continuous trajectory from early progenitors to functionally specialized MK subsets. This platform enables reliable MK supply for mechanistic studies and in vitro platelet production, advancing both basic research and therapeutic development. Full article

The extracellular matrix (ECM) supports spermatogonial stem cell (SSC) function by mimicking biochemical and structural features of the native niche. However, optimal feeder systems and ECM materials remain key limitations in porcine SSC (pSSC) cultures. We developed a porcine-derived ECM (pECM) from porcine feet and evaluated its effectiveness in supporting pSSC maintenance and proliferation under feeder-dependent conditions. We examined protein molecular weight distribution and pECM extract composition. Surface characterization was performed using scanning electron microscopy and atomic force microscopy. We compared pECM with conventional coatings, including gelatin and non-coated controls, using morphological analysis, WST-1 assay, cell cycle analysis, and gene/protein expression of SSC markers. pECM promoted larger, well-defined pSSC colonies and enhanced stemness-related marker expression, including PGP9.5, Thy-1, PLZF, GFRA1, NANOG, and VASA. Additionally, pECM facilitated active pSSC proliferation while suppressing feeder overgrowth, contributing to a stable and functional co-culture environment. Conversely, gelatin supported early feeder proliferation but led to growth saturation, whereas N/C showed delayed attachment and reduced viability. These findings suggest that pECM mimics the native SSC niche and improves pSSC culture. The dual function of pECM in regulating feeder behavior and enhancing pSSC maintenance highlights its potential as a biomaterial for species lacking established feeder-free protocols. Full article

Green ammonia has emerged as a promising alternative fuel for maritime decarbonization, owing to its carbon-free combustion, favorable volumetric energy density, and well-established logistics infrastructure compared to other alternatives. However, critical gaps persist in the development of an integrated fuel supply framework, which hinders the large-scale adoption of ammonia-fueled vessels. Therefore, this paper proposes an onshore wind-powered green ammonia plant located along the Gaolan–Yangpu feeder route. The plant comprises PEM electrolysis, nitrogen separation, Haber–Bosch synthesis, and storage facilities. An optimal plant configuration is subsequently derived through hourly simulations based on wind power generation and a priority-based capacity expansion algorithm. Key findings indicate that a stable ammonia supply—synchronized with monsoon wind patterns and capable of fueling vessels with 10 MW propulsion systems consuming around 680 tons per fortnight—requires a 72 MW onshore wind farm, a 63 MW PEM electrolyzer, 3.6 MW of synthesis facility, and 3205 tons of storage. This configuration yields a levelized cost of ammonia (LCOA) of approximately USD 700/ton, with wind turbines and electrolyzers (including replacement costs) accounting for over 70% of the total cost. Sensitivity analysis further shows that wind turbine and electrolyzer prices are the primary factors affecting ammonia costs. Although variations in operational parameters may significantly alter final configuration, they cause only minor (±1%) fluctuations in the levelized cost without significantly altering its overall trend. Full article

Vibrating feeders are used to empty bulk materials from storage bins, to feed and dispense materials into weighing bins or dispensers, or to feed materials evenly and smoothly into downstream equipment. The harmonic oscillation of the trough can be provided by an electromagnetic oscillator, which consists of an electromagnet consisting of a core and a coil with a given number of coil turns and armature. The aim of this paper has been to verify whether the working condition of the vibrating feeder, i.e., its fault-free operation and the ability to transport the required mass amount of material, can be described on a basis of the measured vibration values using acceleration sensors. This paper describes three experimental methods that allow us with the use of force sensors to measure the adhesion force of the electromagnet and the deformation force of the bent leaf springs, and the use of acceleration sensors to measure the vibration on the trough and on the steel frame of the vibrating feeder. The highest average value of the effective vibration velocity (56.7 mm·s⁻¹) in the horizontal plane was measured on a steel frame of a vibrating feeder using FR4 Epoxy leaf springs with a stiffness of 47.8 N·mm⁻¹ and a weight of 2.57 kg of conveyed material per trough. The lowest average value of the effective vibration velocity (24.6 mm·s⁻¹) has been measured at a weight of 5.099 kg of material conveyed on the trough. We can state that from the analysis of the measured vibration velocities transmitted to the steel frame of the vibrating feeders, it is possible to monitor the partial phases of their operation and diagnose any faults that may occur. It is also possible to determine whether the optimal amount of bulk material is being loaded onto the trough. Full article

The dipteran family Chironomidae is the most widely distributed and, frequently, the most abundant group of insects in freshwater environments. Ecologically, the group plays an important role in trophic cycles and detritus processing in aquatic ecosystems and can be the most productive group of secondary producers in running waters. The annual secondary production of Chironomidae was estimated using emergence trap data from riffles and pools from two stations on the middle reaches of the Shinano River: Tokida Bridge Area (slope 1/180) and Iwano Bridge Area (1/1000). Emergence traps were set up for 24 h on the river’s water surface at three points each in the riffles and pools monthly from April 2015 to March 2019. Five subfamilies of Chironomidae were recorded during the investigation period: Chironominae, Orthocladiinae, Tanypodinae, Diamesinae, and Prodiamesinae. In the Shinano River (Tokida + Iwano area), Orthocladiinae and Diamesinae were the dominant subfamilies in terms of both density (63.2% and 19.0%, respectively) and biomass (62.2% and 25.2%, respectively). Each year’s annual secondary production measured as ash free dry weight (AFDW) was 11.33–55.04 g/m²/year in Tokida and 5.83–38.43 g/m²/year in Iwano. The average annual secondary production of detritus feeder Chironomidae (all except for Tanypodinae) was 24.46 ± 20.38 (ranging from 11.13 in 2015 to 54.67 in 2018) in the Tokida area and 19.61 ± 16.38 (ranging from 5.41 in 2016 to 37.53 in 2017) in the Iwano area. On the other hand, that of carnivorous Chironomidae (Tanypodinae) was 0.22 ± 0.10 (ranging from 0.13 in 2016 to 0.37 in 2018) in the Tokida area and 0.66 ± 0.24 (ranging from 0.42 in 2016 to 0.90 in 2017) in the Iwano area, and overall secondary production averaged 22.48 g/m²/year in the middle reaches of the Shinano River. These values were higher than in previous reports. Full article

Zebrafish (Danio rerio) has become a pivotal vertebrate model in biomedical research, renowned for its genetic similarity to humans, optical transparency, rapid embryonic development, and amenability to experimental manipulation. In recent years, the derivation of cell lines from zebrafish embryos has unlocked new possibilities for in vitro studies across developmental biology, toxicology, disease modeling, and genetic engineering. These embryo-derived cultures offer scalable, reproducible, and ethically favorable alternatives to in vivo approaches, enabling high-throughput screening and mechanistic exploration under defined conditions. This review provides a comprehensive overview of protocols for establishing and maintaining zebrafish embryonic cell lines, emphasizing culture conditions, pluripotency features, transfection strategies, and recent innovations such as genotype-defined mutant lines generated via CRISPR/Cas9 and feeder-free systems. We also highlight emerging applications in oncology, regenerative medicine, and functional genomics, positioning zebrafish cell lines as versatile platforms bridging animal models and next-generation in vitro systems. Its continued optimization holds promise for improved reproducibility, reduced animal use, and expanded translational impact in biomedical research. Full article

Background/Objectives: Our objective is to develop hormone-producing pituitary cells that can function in the same manner as the human body and provide more effective treatments than current hormone replacement therapy. We have already established a technique for generating hypothalamic–pituitary organoids using feeder-free human pluripotent stem cells (hPSCs) and demonstrated their effectiveness in vivo through transplantation into hypopituitary mouse models. To prospectively determine the upper limit of transplanting adenohypophyseal cells into humans, we investigated the human maximum secretion capacity of adrenocorticotropic hormone (ACTH) and growth hormone (GH). Methods: We analyzed data from 28 patients with pituitary adenomas, among whom 16 evinced no abnormality of ACTH secretion and 12 showed no GH secretion on corticotropin-releasing hormone (CRH) and growth hormone-releasing hormone-2 (GHRP-2) stimulation testing. Results: The average ACTH peak value after CRH stimulation tests was 97.2 pg/mL, and the average GH peak value after GHRP-2 stimulation tests was 25.1 ng/mL. Conclusions: These data will likely serve as benchmarks of ACTH and GH secretion when transplanting cultured cells into humans. Full article

This article reviews the scientific literature discussing the microbial interactions between water microbiota, live food microbiota, fish larvae immune system and gut microbiota, and biofilm microbial communities in rearing systems for marine fish larvae. Fish gut microbiota is the first line of defense against opportunistic pathogens, and marine fish larvae are vulnerable to high mortalities during the first weeks after hatching. The bacterial colonization of fish larvae is a dynamic process influenced by environmental and host-related factors. The bacteria transferred to larvae from the eggs can influence the composition of the gut microbiota in the early stages of fish. Fish larvae ingest free-living microorganisms present in the water, as marine fish larvae drink water for osmoregulation. In marine aquaculture systems, the conventional feeding–rearing protocol consists of zooplankton (rotifers, Artemia, and copepods). These live food organisms are filter-feeders. Once transferred to a new environment, they quickly adopt the microflora of the surrounding water. So, the water microbiota is similar to the microbiota of the live food at the time of ingestion of live food by the larvae. In aquaculture rearing systems, bacterial biofilms may harbor opportunistic pathogenic bacteria and serve as a reservoir for those microbes, which may colonize the water column. The methods applied for the study of fish larvae microbiota were reviewed. Full article

The successful application of primordial germ cells (PGCs) is an ideal method for generating gene-edited birds. However, barriers to efficient DNA transfection in PGCs lead to low transfection efficiency, limiting the generation of genetically modified chickens. The current study utilized chemical transfection and electroporation methods to determine the optimal transfection conditions for the PGC line under feeder- and serum-free medium. Among the tested methods, the Lonza electroporation system exhibited the highest transduction efficiency, with a previously unreported rate of 71.13 ± 1.26%. Optimal transfection conditions were achieved using 4 µg of DNA and 100 µL of Entranster^TM-E in 1 × 10⁶ PGCs. Furthermore, the optimal electroporation conditions resulted in low cell death and normal expression of pluripotency-related genes, highlighting the low cytotoxicity. The resulting electroporation models were then used to deliver the enhanced green fluorescent protein (EGFP) gene to the Z chromosome with a Cas9-gRNA plasmid, achieving a 7-day insertion efficiency of 14.63 ± 1.07%. Our study highlights the vast potential of electroporation technology for the transfection of PGCs. Full article

Feed conversion ratio (FCR) is a key indicator of pig productivity, but its measurement is labor-intensive and time-consuming. This study aimed to construct a predictive model for cumulative feeding intake (CFI), which could help estimate FCR more efficiently and reduce the time and effort needed for measurements. This study included a total of 987 Yorkshire boars raised in specific pathogen-free environments, with feeding and growth data collected using automatic feeders. The segmented R package and Bayesian ridge regression (BRR) were used to build a predictive model for CFI. The results showed that the optimal body weight range for predicting FCR was 80–110 kg. The BRR model achieved 80% accuracy for CFI prediction, and FCR calculated from predicted CFI showed 81.4% similarity to the corrected FCR. The results clearly demonstrate that even with a limited training dataset, the BRR model has good predictive potential for FCR. The findings of this study could reduce the selection pressure on FCR traits, decrease production costs, and shorten measurement periods, ultimately benefiting the swine industry significantly. Full article