Search Results (196)

For many therapeutic agents to be effective against intracellular targets, they must first be able to penetrate the cell membrane. Current methodologies for assessing internalization, such as confocal microscopy and conventional flow cytometry, are limited by low throughput or an inability to provide precise spatial information on signal localization. Here, we present a comprehensive, semi-automated analytical pipeline for investigating compound internalization based on imaging flow cytometry, which is designed to address these limitations. Our workflow details the procedure from sample preparation and data acquisition on an Amnis FlowSight cytometer to analysis using IDEAS 6.2 software with a custom-designed template. Key features of our approach include the automated discrimination of signal between the plasma membrane and cytoplasmic compartments, the calculation of an internalization coefficient, and the introduction of a novel parameter—signal distribution entropy—to quantify the uniformity of the compound distribution within cells. For the statistical analysis, we developed FluoSta v1.0, a software tool that automates descriptive statistics and analysis of variance (ANOVA with Tukey’s post hoc test) and facilitates data visualization. The pipeline’s utility was demonstrated in a series of model experiments, including a comparative assessment of the internalization efficiency of PS- versus PS/LNA-modified compounds in MT-4 cell cultures. Full article

Decarbonising aquaculture support vessels is pivotal to reducing greenhouse gas (GHG) emissions across both the aquaculture and maritime sectors. This study evaluates the technical and economic feasibility of deploying hydrogen as a marine fuel for a 14.95 m net cleaning vessel (NCV) operating in Tasmania, Australia. The analysis retains the vessel’s original layout and subdivision to enable a like-for-like comparison between conventional diesel and hydrogen-based systems. Two options are evaluated: (i) replacing both the main propulsion engines and auxiliary generator sets with hydrogen-based systems—either proton exchange membrane fuel cells (PEMFCs) or internal combustion engines (ICEs); and (ii) replacing only the diesel generator sets with hydrogen power systems. The assessment covers system sizing, onboard hydrogen storage integration, operational constraints, lifecycle cost, and GHG abatement. Option (i) is constrained by the sizes and weights of PEMFC systems and hydrogen-fuelled ICEs, rendering full conversion unfeasible within current spatial and technological limits. Option (ii) is technically feasible: sixteen 700 bar cylinders (131.2 kg H₂ total) meet one day of onboard power demand for net-cleaning operations, with bunkering via swap-and-go skids at the berth. The annualised total cost of ownership for the PEMFC systems is 1.98 times that of diesel generator sets, while enabling annual CO₂ reductions of 433 t. The findings provide a practical decarbonisation pathway for small- to medium-sized service vessels in niche maritime sectors such as aquaculture, while clarifying near-term trade-offs between cost and emissions. Full article

Selenium nanoparticles (Se NPs) have received increasing attention as a new alternative source to other forms of selenium in nutritional dietary supplements; however, the limited stability and pronounced tendency of selenium nanoparticles (Se NPs) to aggregate in aqueous environments have significantly constrained their practical applications. In this study, Poria cocos polysaccharide-modified Se NPs (PCP-Se NPs) were synthesized by the selenite/ascorbic acid chemical reduction method. PCP-Se NPs exhibited a uniformly dispersed spherical morphology with an average particle size of 66.64 ± 0.30 nm, and displayed an amorphous crystal structure. Compared to unmodified Se NPs, the PCP-Se NPs exhibited low Se release (8.83 ± 0.73%) after simulated gastrointestinal digestion, and they had excellent storage stability and salt ion stability. PCP-Se NPs exhibited potent antioxidant activity manifested by the effective scavenging of DDPH and ABTS radicals. PCP-Se NPs were efficiently internalized by RAW264.7 cells and released into the cytoplasm by a lysosomal escape mechanism, thereby effectively reducing intracellular inflammatory factor levels (the levels of MPO, NO, iNOS, TNF-α, IL-1β, and IL-10 in the PCP-Se NPs treatment group were 0.38 ± 0.013-fold, 0.26 ± 0.02-fold, 0.36 ± 0.02-fold, 0.57 ± 0.03-fold, 0.35 ± 0.02-fold, and 2.07 ± 0.16-fold that of the LPS group, respectively), alleviating oxidative stress (the levels of CAT, SOD, GSH, and MDA in the PCP-Se NP-treated group were 2.48 ± 0.02-fold, 1.91 ± 0.11-fold, 3.16 ± 0.28-fold, and 0.46 ± 0.03-fold that of the LPS group, respectively), and maintaining mitochondrial membrane potential stability. This study provides a basis and reference for improving the stability of Se NPs and developing novel selenium-enriched dietary supplements. Full article

Background/Objectives: Cesarean section (CS) is among the most common surgical procedures worldwide and is associated with a markedly increased risk of postpartum infection, including surgical site infection (SSI). International guidelines recommend routine prophylaxis for all CSs, but in Slovenia, it has traditionally been reserved for high-risk procedures, with limited SSI surveillance data. The aim of this study was to determine the incidence of SSI within 30 days after CS and to evaluate the impact of universal prophylaxis implemented in a regional secondary care teaching hospital. Methods: We conducted a retrospective observational cohort study including all CS performed during 2023 (risk-based-only prophylaxis) and 2024 (universal prophylaxis) at Izola General Hospital, Slovenia. SSI was defined according to ECDC criteria and identified from inpatient and outpatient records up to 30 days postoperatively. Logistic regression was used to assess associations between prophylaxis, clinical variables, and SSI. Results: Among 1055 deliveries (208 CS; 99 in 2023, 109 in 2024), the rate of antimicrobial prophylaxis increased from 58.6% to 89.0% (p < 0.001). The overall 30-day SSI incidence was 7.2%, with no significant difference between the pre- and post-implementation periods (8.1% vs. 6.4%, p = 0.644). Most infections (86.7%) were diagnosed after discharge and were superficial incisional SSI (60%). In multivariable analysis, prophylaxis was independently protective (adjusted OR 0.11; 95% CI 0.02–0.58; p = 0.009), while prelabor rupture of membranes (PROM) and higher maternal weight significantly increased SSI risk. Conclusions: Antibiotic prophylaxis was independently associated with a reduced risk of SSI following SC; however, the absolute infection rate did not decline significantly and remained moderate after implementation. PROM and higher maternal weight were additional independent risk factors. These findings support universal prophylaxis with optimization for high-risk women and ongoing hospital and national surveillance to improve CS safety. Full article

Total internal reflection fluorescence (TIRF) microscopy excites fluorophores within a few hundred nanometers of the sample–substrate interface, enabling high-contrast imaging near the cell membrane. When cultured cells differentiate, the membrane in contact with the coverslip generally acquires basal characteristics, while the opposite membrane develops apical features. Consequently, conventional TIRF microscopy is limited to imaging the basal surface. We developed an immersed-prism TIRF (IP-TIRF) microscope, in which a prism immersed in the culture medium generates TIR at the cell/medium–prism interface, illuminating the apical membrane and reducing cytosolic background. In proof-of-principle experiments, we imaged fluorescent beads and 3xmNeonGreen-tagged intraflagellar transport (IFT) particles in cilia, and compared the performance with confocal microscopy. In cellular regions where both methods can be applied (such as the IFT base pool), on average, IP-TIRF achieved approximately 1.8 times the contrast-to-noise ratio (CNR~31) compared to confocal microscopy. Furthermore, IFT-particle motion was detected in IP-TIRF image sequences and Kymographs of cilia, with adequate spatial resolution. Kymograph analysis revealed an average anterograde IFT velocity of 0.156 ± 0.071 µm/s and an average retrograde velocity of 0.020 ± 0.007 µm/s, approximately one-quarter and one-twentieth, respectively, of the values reported for mammalian primary cilia, which we attribute to acquisition at room temperature rather than physiological conditions. Therefore, these velocity measurements should be regarded as proof-of-principle demonstrations obtained at room temperature, not as validated physiological transport rates. Our IP-TIRF method provides a high-resolution, cost-effective, and broadly accessible approach for imaging the apical membrane in live cells. Full article

Shale gas wastewater from hydraulic fracturing poses significant environmental risks due to its high salinity and complex inorganic composition. This study investigates the behavior of major and trace inorganic constituents across a full-scale treatment train in the Sichuan Basin, China. Despite multi-stage processes including equalization, flocculation, flotation, biological reactors, membrane filtration, and clarification, key inorganic species such as Cl, Na, Br, Sr, Li, and B remained largely persistent in the final effluent with values of 13,760, 8811, 70, 95.9, 26.6, and 60.2 mg/L, respectively. Geochemical tracers including Br/Cl (average: 0.0022 mM/mM), Na/Br (average: 125 mg/mg), and Sr/Ca (average: 0.15 mM/mM) ratios, combined with halide endmember mixing models, revealed that salinity primarily originated from highly evaporated formation brines, with limited evidence for halite dissolution or external contamination. Elevated Sr (average: 89.3 mg/L) and Ca (average: 274 mg/L) levels relative to Mg (average: 32 mg/L) suggest significant water–rock interaction. Environmental risk assessments showed that concentrations of several elements in treated effluent greatly exceeded national and international discharge or reuse standards. These findings underscore the limitations of conventional treatment technologies and highlight the urgent need for advanced processes and regulatory frameworks that address the unique challenges of high-TDS (total dissolved solids) unconventional wastewater. Full article

The present study investigated the decline in human fertility by analyzing the multielemental profile of seminal plasma and its relationship with seminal parameters and sperm biomarkers. Twenty-nine donor seminal plasma samples were examined using inductively coupled plasma–tandem mass spectrometry (ICP-MS/MS). Method optimization demonstrated that robust plasma conditions, including internal standardization and helium (He) collision gas, were essential to achieve reliable quantification. These conditions mitigated matrix effects and spectroscopic interferences, despite lower sensitivity. Elements such as copper (Cu), iron (Fe), manganese (Mn), strontium (Sr), titanium (Ti), vanadium (V), and chromium (Cr) were quantified, and several significant correlations were identified. Specifically, Cu was negatively correlated with seminal volume and positively correlated with sperm concentration and spontaneous acrosome reacted sperm, but negatively correlated with medium mitochondrial membrane potential (MMP); Mn showed negative associations with sperm vitality and medium MMP; Fe showed a negative correlation with motile sperm concentration (4 h); V was positively correlated with acrosome reacted sperm after acrosome reaction induction and with very low/medium MMP, whereas it was negatively associated with tyrosine phosphorylation; and Cr also showed a negative correlation with tyrosine phosphorylation. As, Mo, and Pb were detected in a few samples, limiting correlation analysis. From a functional perspective, elements such as As and Pb, as well as excess Cu or Fe, may contribute to oxidative stress by enhancing reactive oxygen species (ROS) generation and impairing antioxidant defenses. Conversely, essential metals, including Mn and Cu, at physiological concentrations act as cofactors of antioxidant enzymes and play a protective role against oxidative damage. Full article

Hydrogen proton exchange membrane fuel cells (PEMFCs) are a promising clean energy technology for automotive applications owing to their high efficiency and environmentally friendly characteristics. Efficient hydrogen recirculation is critical for sustaining the PEMFC performance, and ejector-based systems offer a passive, energy-efficient solution. However, traditional ejectors suffer from performance degradation across varying fuel-cell loads owing to their limited adaptability. To address this limitation, this study investigated the internal flow behavior and recirculation performance of single- and coaxial-nozzle ejectors, focusing on the influence of the diameter ratio between the mixing chamber and nozzle throat. Numerical simulations were performed to evaluate the flow structures and recirculation ratios under various operating conditions. The diameter ratio between the mixing chamber and the nozzle throat played a crucial role in determining the flow uniformity and recirculation efficiency. Specifically, lower diameter ratios reduce the recirculation ratio across all operating conditions, whereas higher diameter ratios exhibit diminished performance only under very low power outputs (≤4 bar) but show enhanced performance at medium-to-high outputs. These findings suggest that tailoring the geometric parameters of coaxial-nozzle ejectors can significantly improve hydrogen recirculation adaptability in PEMFC systems, thereby supporting more stable and efficient operation across a wide range of vehicle load conditions. Full article

This review provides a critical and technically grounded assessment of continuous electrocoagulation processes (CEPs) for the treatment of industrial inorganic pollutants, emphasizing recent innovations, methodological developments, and practical outcomes. A comprehensive literature survey indicates that 53 studies published over the past 25 years have investigated CEPs for inorganic contaminant removal, with 36 focusing on standalone electrocoagulation systems and 17 exploring integrated CEPs approaches. Recent advancements in reactor design, such as enhanced internal mixing, optimized electrode geometry, and modular configurations, have significantly improved treatment efficiency, scalability, and operational stability. Evidence indicates that CEPs can achieve high removal efficiencies for a wide range of inorganic contaminants, including fluoride, arsenic, heavy metals (e.g., chromium, lead, nickel, iron), nitrates, and phosphates, particularly under optimized operating conditions. Compared to conventional treatment methods, CEPs offer several advantages, such as simplified operation, reduced chemical consumption, lower sludge generation, and compatibility with renewable energy sources and complementary processes like membrane filtration, flotation, and advanced oxidation. Despite these promising outcomes, industrial-scale implementation remains constrained by non-standardized reactor designs, variable operational parameters, electrode passivation, high energy requirements, and limited long-term field data. Furthermore, few studies have addressed the modeling and optimization of integrated CEPs systems, highlighting critical research gaps for process enhancement and reliable scale-up. In conclusion, CEPs emerge as a novel, adaptable, and potentially sustainable approach to industrial inorganic wastewater treatment. Its future deployment will rely on continued technological refinement, standardization, validation under real-world conditions, and alignment with regulatory and economic frameworks. Full article

The widespread production and poor management of plastic waste have led to the pervasive presence of microplastics (MPs) in environmental and biological systems. Among various polymers, polyethylene (PE) is the most widely produced plastic globally, primarily due to its use in single-use packaging. Its persistence in ecosystems and resistance to degradation processes result in the continuous formation of PE-derived MPs. These particles have been detected in human biological matrices, including blood, lungs, placenta, and even the brain, raising increasing concerns about their bioavailability and potential health effects. Once internalized, PE MPs can interact with cellular membranes, induce oxidative stress, inflammation, and apoptosis, and interfere with epigenetic regulatory pathways. In vitro studies on epithelial, immune, and neuronal cells reveal concentration-dependent cytotoxicity, mitochondrial dysfunction, membrane disruption, and activation of pro-inflammatory cytokines. Moreover, recent findings suggest that PE MPs can induce epithelial-to-mesenchymal transition (EMT), senescence, and epigenetic dysregulation, including altered expression of miRNAs and DNA methyltransferases. These cellular changes highlight the potential role of MPs in disease development, especially in cardiovascular, metabolic, and possibly cancer-related conditions. Despite growing evidence, no standardized method currently exists for quantifying MPs in human samples, complicating comparisons across studies. Further, MPs can carry harmful additives and environmental contaminants such as bisphenols, phthalates, dioxins, and heavy metals, which enhance their toxicity. Global estimates indicate that humans ingest and inhale tens of thousands of MPs particles each year, yet long-term human research remains limited. Given these findings, it is crucial to expand research on PE MP toxicodynamics and to establish regulatory policies to reduce their release. Promoting alternative biodegradable materials and improved waste management practices will be vital in decreasing human exposure to MPs and minimizing potential health risks. Full article

Determining the active substance content in the tested product is an essential part of research for overall assessment of the quality of a medicinal substance. This role can be successfully performed by membrane electrodes that are selective for a specific drug. The novelty of the presented research is the development of the first ion-selective electrode with a polymer membrane phase with the octenidine (OCT) function. Classical ion-selective electrodes (ISE), polymer electrodes with an internal Ag/AgCl electrode, and electrode bodies with glassy carbon were used for the research. The membranes were prepared based on cation exchangers from the borate group and neutral cyclodextrin. All sensors have good parameters, e.g., the polymer electrode with KtpClPB is characterised by a wide linear range of −logc 6−3, a low limit of detection 5 × 10⁻⁷ M, and a near-Nernstian, reproducible slope of characteristics of 31.41 ± 1.14 mV/decade. It can be seen that a stable, reversible potential and a short response time were achieved for this sensor. The obtained favourable selectivity coefficients of the electrode determined in relation to excipients allowed direct determination of octenidine, e.g., in lozenges. The results obtained with the calibration curve method show a recovery of 97% and a precision of SD 2.3 mg/L, which indicates that the data are consistent with the pharmacopoeia requirements. Full article

The synergistic effect of using D2EHPA and TOPO together to enhance the extraction of samarium(III) from aqueous media via emulsion liquid membrane (ELM) technology was explored. D2EHPA in binary mixtures with TBP and in ternary mixtures with TOPO and TBP was also tested. Among the tested extractants, a binary mixture of 0.1% (w/w) D2EHPA and 0.025% (w/w) TOPO achieved 100% samarium(III) extraction at a low loading. This mixture outperformed D2EHPA-TBP and other systems because D2EHPA strongly binds to Sm(III) ions, while TOPO increases the solubility and transport efficiency of metal complexes. Additionally, process factors that optimize performance and minimize emulsion breakage were examined. Key insights for successfully implementing the process include the following: 5 min emulsification with 0.75% Span 80 in kerosene at pH 6.7 (natural), 250 rpm stirring, a 1:1 internal/membrane phase volume ratio, a 20:200 treatment ratio, and a 0.2 N HNO₃ stripping agent. These insights produced stable, fine droplets, enabling complete recovery and rapid carrier regeneration without emulsion breakdown. Extraction kinetics accelerate with temperature up to 35 °C but declined above this limit due to emulsion rupture. The activation energy was calculated to be 33.13 kJ/mol using pseudo-first-order rate constants. This suggests that the process is diffusion-controlled rather than chemically controlled. Performance decreases with Sm(III) feed concentrations greater than 200 mg/L and in high-salt matrices (Na₂SO₄ > NaCl > KNO₃). Integrating these parameters yields a scalable, low-loading ELM framework capable of achieving complete Sm(III) separation with minimal breakage. Full article

Background: The proportion of people suffering from neurodegenerative conditions, such as Alzheimer’s disease (AD), is increasing in the population year on year. Despite the constant effort of researchers, these conditions remain incurable and can only be managed by alleviation or delaying of symptoms. The lack of suitable treatment is caused by constricted access to the brain, limited by the brain-blood barrier. The aim of this work was to investigate two pegylated gold nanoparticles as potential carriers of therapeutic siRNA and their impact on the cellular functions of Human Brain Endothelial Cells. Methods and Results: Nanoparticles AuNP14a and AuNP14b complexed with siRNA were internalized by HBEC-5i cells and located in the cytoplasm. The genotoxicity assay proved that the nucleus was not affected and complexed nanoparticles did not cause DNA damage. The reactive oxygen species formation and mitochondrial membrane potential changes were measured and showed an adaptive response of cells after compound administration. Results obtained in a cytotoxicity assay conducted on astrocytes and pericytes, which are components of the blood–brain barrier, confirmed the biosafety of tested nanoparticles. Conclusions: In summary, it was shown that AuNP14a and AuNP14b are promising candidates as nanocarriers for therapeutic nucleic acids through biological barriers. Full article

Biomaterials modified by antibacterial substances, including nanoparticles, open new opportunities for the effective treatment of infected wounds. Unfortunately, most publications focused only on experiments in vitro, with limited understanding of their potential for the clinic. This study evaluates the effectiveness in vivo of electrospun chitosan/polylactic acid (Ch/PLA) membranes enriched with silver nanoparticles (AgNPs) for purulent wound treatment. The composite biomaterial integrates chitosan’s biocompatibility and antimicrobial activity with PLA’s structural integrity, while AgNPs enhance antibacterial efficacy against major wound pathogens, including Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia aureus. A full-thickness purulent wound model was established in a rat model, and the animals were divided into three treatment groups: (i) Ch/PLA, (ii) Ch/PLA-AgNPs, and (iii) PLA-chlorhexidine (control). Wound healing was monitored over 21 days through macroscopic evaluation, histology, immunohistochemistry, and microbiological analysis. The Ch/PLA-AgNPs membranes significantly reduced bacterial colonization within 4–6 days, promoted granulation tissue formation, and accelerated epithelialization compared to the non-modified Ch/PLA scaffold. By day 15, complete wound closure was observed in the Ch/PLA-AgNPs group, comparable to PLA-chlorhexidine-treated wounds. Immunohistochemical analysis revealed a controlled inflammatory response with a balanced macrophage M1/M2 transition, supporting efficient tissue regeneration. Furthermore, systemic toxicity assessments indicated no significant adverse effects on internal organs. These findings demonstrate that electrospun Ch/PLA-AgNPs membranes effectively accelerate purulent wound healing by combining antimicrobial protection with biocompatible tissue support. This innovative approach presents a promising alternative to conventional wound dressings and paves the way for clinical applications in managing infected wounds. Full article

The growing scarcity of freshwater worldwide has increased interest in forward osmosis (FO) membranes as a promising solution for water desalination and wastewater treatment. This study investigates the enhancement of thin-film composite (TFC) FO membranes via the incorporation of carboxyl-functionalized multiwalled carbon nanotubes (COOH-MWCNTs) into the polyethersulfone (PES) support layer. The membranes were fabricated using a combination of phase inversion and interfacial polymerization techniques, with COOH-MWCNTs incorporated into the membrane support layers at different concentrations (0–0.75 wt.%). Comprehensive characterization was carried out using various analytical methods and mechanical testing to evaluate the physicochemical and structural properties of the membranes. The modified membranes demonstrated improved hydrophilicity, enhanced mechanical and thermal stability, and improved surface charge properties. Performance tests using a 1 M NaCl draw solution showed that the optimized membrane (0.5 wt.% COOH-MWCNTs) attained a 161% enhancement in water flux (7.48 LMH) compared to the unmodified membrane (2.86 LMH), while also reducing internal concentration polarization (ICP). The antifouling properties were also significantly improved, with a flux recovery rate of 91.92%, attributed to enhanced electrostatic repulsion as well as surface and microstructural modifications. Despite a moderate rise in reverse solute flux, the specific reverse solute flux (J_s/J_w) remained within acceptable limits. These findings highlight the potential of COOH-MWCNT-modified membranes in enhancing FO desalination performance, offering a promising option for next-generation water purification technologies. Full article