Search Results (1,143)

RNA interference (RNAi) is a powerful mechanism of post-transcriptional gene regulation in which small interfering RNA (siRNA) is utilized to target and degrade specific RNA sequences. In this study, experiments were conducted to evaluate the efficacy of combination siRNA therapy against enterovirus 71 (EV71) and the potential of this therapy to delay or prevent the emergence of resistance in vitro. siRNAs targeting multiple genes of EV71 were designed, and the effects of a cocktail of siRNAs on viral replication were assessed compared to those of single-siRNA treatment. Cotransfection of multiple siRNAs targeting different protein-coding genes of the EV71 genome effectively suppressed escape mutants resistant to RNAi. Combination therapy with siRNAs targeting multiple viral genes successfully prevented viral escape mutations over five passages. By contrast, serial passaging with a single siRNA led to the rapid emergence of resistance, with mutations identified in the siRNA target sites. The combination of siRNAs specifically targeting different regions demonstrated an additive effect and was more effective than individual siRNAs at inhibiting EV71 replication. This study supports the effectiveness of combination therapy using siRNAs targeting multiple genes of EV71 to inhibit viral replication and prevent the emergence of resistant escape mutants. Overall, the findings identify RNAi targeting multiple viral genes as a potential strategy for therapeutic development against viral diseases and for preventing the emergence of escape mutants resistant to antiviral RNAi. Full article

Extracellular vesicles (EVs) secreted by glioblastoma multiforme and other types of cancer cells are key factors contributing to the aggressiveness of the disease and its resistance to therapy. Chloroquine (CHQ), a lysosomal inhibitor, has shown potential as an enhancer of temozolomide (TMZ) cytotoxicity against glioblastoma cells. Since both CHQ and TMZ are known to modulate EV secretion, we sought to investigate their potential interplay in this process. Simultaneous treatment of TMZ-sensitive (U87-MG) and TMZ-resistant (U138-MG) glioblastoma cells with TMZ and CHQ led to a synergistic upregulation of EV secretion. Although CHQ did not enhance the TMZ cytotoxicity in U87-MG cells, it synergized with the latter to upregulate the release of extracellular nucleic acids implicating activation of unconventional secretory pathways. Synergistic upregulation of the autophagy markers LC3B-II and p62 by CHQ and TMZ in both cells and EVs indicates that secretory autophagy is likely involved in the observed unconventional secretion. Moreover, a significant enrichment of caveolin-1 in small EVs highlights their potential role in modulating tumor aggressiveness. The synergy in EV upregulation was not confined to the specific biological activity of TMZ and CHQ; similar effects were observed upon co-treatments with CHQ and etoposide (a topoisomerase inhibitor) and TMZ and Bafilomycin A1 (another lysosomal inhibitor). Heightened EV release was also observed in THP-1 monocytes and macrophages treated with Bafilomycin and TMZ, highlighting a broader, cell-type-independent mechanism. These findings indicate that combined DNA damage and lysosomal inhibition synergistically stimulate secretory autophagy and EV release, potentially impacting the tumor microenvironment and driving disease progression. Full article

This study evaluated the epidemiology and seasonal patterns of respiratory viruses in adults hospitalized with acute respiratory tract infections during two consecutive post-COVID-19 pandemic seasons. A retrospective study was conducted at the University Hospital “P. Giaccone”, Palermo, from September 2022 to September 2024. Multiplex molecular assays were used to detect the ten respiratory viruses most relevant from an epidemiological perspective in respiratory samples (n = 1110) of 1081 patients. A respiratory viral infection was identified in 29.6% of patients. The highest viral infection rate was observed in the 31–50 age group. Human rhinovirus/enterovirus (HRV/EV) was the most frequently detected (40%), followed by influenza A virus (IAV; 18.4%) and human coronaviruses (HuCoVs; 12.8%). Viral co-infections were identified in 10.9% of positive cases, with HRV/EV, adenovirus (ADV), and parainfluenza virus (PIV) being most frequently involved. Influenza and respiratory syncytial viruses (RSVs) showed a winter seasonality, while diverse circulation patterns were revealed for the other viruses. This study demonstrated a sustained circulation of respiratory viruses in adults hospitalized with severe respiratory symptoms, with HRV/EV accounting for most of them. Syndromic multiplex molecular testing, although limited to the detection of a small fraction of epidemiologically relevant known viruses, has proven to be a valuable tool, not only for diagnostic purposes but also for acquiring genotyping data and implementing epidemiological information from sentinel surveillance systems. Full article

Glottic insufficiency, often caused by laryngeal nerve injury, impairs voice quality and breathing. Current treatments, such as hyaluronic acid injection, require frequent reapplication every 3–6 months. This study aimed to investigate the therapeutic potential of small extracellular vesicles (sEVs) derived from Wharton’s Jelly mesenchymal stem cells (WJMSCs) incorporated into genipin-crosslinked gelatin hydrogels (GCGHs) for promoting vocal fold fibroblast (VFFs) regeneration in vitro. WJMSCs were isolated from umbilical cords, expanded to passage 4, and used for sEV isolation via tangential flow filtration (TFF). The sEVs (585.89 ± 298.93 µg/mL) were characterized using bicinchoninic acid assay (BCA), nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and Western blot. Seven concentrations of sEVs were tested on VFFs to evaluate cytotoxicity and proliferation, identifying 75 µg/mL as the optimal dose. GCGHs were then combined with WJMSCs and sEVs and evaluated for physicochemical properties, degradation, biocompatibility, and immune response. The hydrogels were injectable within 20 min and degraded in approximately 42 ± 0.72 days. The optimal sEV concentration significantly enhanced VFFs proliferation (166.59% ± 28.11) and cell viability (86.16% ± 8.55, p < 0.05). GCGH-MSCs showed the highest VFFs viability (82.04% ± 10.51) and matrix contraction (85.98% ± 1.25) compared to other groups. All hydrogel variants demonstrated minimal immune response when co-cultured with peripheral blood mononuclear cells (PBMCs). GCGH is a promising scaffold for delivering WJMSCs and sEVs to support VFF regeneration, with demonstrated biocompatibility and regenerative potential. Further in vivo studies are warranted to validate these findings. Full article

The need for prostate cancer (PCa)-specific biomarkers that enable more accurate detection of the disease and better prediction of tumor aggressiveness remains ongoing due to the low cancer specificity of PSA screening. Several potential mRNA markers for diagnosing PCa, in tissue and urine, have been reported in the literature. In this study, we aim to explore the potential of selected prostate-specific molecules and transcripts contained in small extracellular vesicles (sEVs) in semen to predict PCa risk reclassification for patients with moderately elevated PSA levels—a clinical scenario where identifying truly non-invasive biomarkers is especially critical. RT-qPCR analysis in semen sEVs successfully showed differential expression of KLK3 and PCA3 genes between PCa and healthy controls, whereas CREB3L4, CCNQ and DUSP23 levels were related to the severity or degree of PCa affectation. Our findings also present strong evidence that classifiers based on combined long transcript levels in semen sEVs serve as effective biomarkers. They can be used alone or in combination with blood PSA and/or semen citric acid levels to improve the diagnosis of PCa and assess its severity and disease progression with high accuracy. This strategy would allow a more comprehensive assessment, increase prognostic accuracy, and facilitate accurate clinical decision-making in the management of PCa. Full article

Extracellular vesicles (EVs) are rapidly gaining recognition as critical mediators of inter-cellular communication during viral infections. To contribute to fill the gap in knowledge regarding the role of EVs in adenovirus infection, we used human adenovirus type 4 of species Mastadenovirus exoticum (HAdV-E4), a prevalent respiratory and ocular pathogen, and characterized the cargo and biological properties of EVs released by HAdV-E4-infected A549 lung epithelial cells at a pre-lytic stage of infection. Using immunocapture-based isolation and multi-omics approaches, we found that infection profoundly alters the EV uploaded proteome and small non-coding RNA repertoire. Mass spectrometry identified 268 proteins unique to EVs purified from infected cells (AdV-EVs), with enrichment in pathways supporting vesicle trafficking and viral protein translation, and importantly also a few virus-encoded proteins. A small RNA transcriptome analysis showed differential uploading in AdV-EVs of various small non-coding RNAs, including snoRNAs, as well as the presence of virus associated RNAs I and II. Notably, AdV-EVs contained viral genomic DNA and could initiate productive infection upon delivery to naïve cells in the absence of detectable viral particles. Our data suggest that EVs released during the HAdV-E4 infection may serve as vehicles for non-lytic viral dissemination and highlight their possible role in intra-host dissemination Full article

Liquid biopsy, which analyzes tumor secretomes in biological fluids, allows us to not only diagnose cancer, but also evaluate the effectiveness of antitumor therapy, predict the prognosis of the disease, and select targeted therapy. One of the promising sources for identifying tumor markers using liquid biopsy is exosomes—small extracellular vesicles (sEVs) (30–150 nm in size) that are secreted by all types of cells, including tumor cells, to exchange information. It is known that during the maturation process, mainly biologically active proteins and non-coding RNA are packaged into exosomes, and tumor cells secrete significantly more exosomes than normal cells. Taking into account the involvement of microRNAs in the mechanisms of carcinogenesis, their high stability in EVs, and ease of detection, exosomal microRNAs are the most promising tumor markers for creating panels that can serve as a guide both for clarifying diagnostics and for making therapeutic decisions on effective cancer treatment, including breast cancer (BC). The purpose of this review is to summarize information on the shortcomings of modern methods for diagnosing early BC, the involvement of exosomal microRNAs in BC dissemination (impact on the immune system, epithelial–mesenchymal transition, proliferation, invasion, migration, angiogenesis, and metastasis), and the high diagnostic potential of exosomal microRNAs for detecting early BC. Full article

The rapid growth of Electric Vehicles (EVs) risks causing grid congestion. Smart charging strategies can help to prevent overload while ensuring timely charging, thereby reducing the need for costly infrastructure upgrades. We study EV charging from a scheduling perspective, assuming an aggregator manages charging while respecting network cable capacities. In our model, vehicles depart only after charging is complete, so delays are possible. Our aim is to minimize these delays. We consider a network of parking lots, some of which are equipped with solar panels, where the demand that can be served is limited by the cables connecting them to the grid. We propose novel scheduling strategies that combine an online variant of well-known schedule generation schemes with a destroy-and-repair heuristic. We evaluate their effectiveness in a case study with data from the city of Utrecht. Without control, network cables would be overloaded 60–70% of the time. Our strategies completely eliminate overload, introducing an average delay of just over 1.5 min per EV in high-occupancy scenarios. This demonstrates that scheduling can significantly increase the number of EVs charged without compromising grid safety at the cost of a rather small delay. We also highlight the importance of accounting for grid topology and show the benefits of using flexible charging rates. Full article

Historical chemical production sites often harbor irregularly distributed solid waste landfills, posing significant environmental risks. Traditional drilling methods, while accurate, are inefficient for comprehensive characterization due to high costs and spatial limitations. This study aims to develop an integrated geophysical drilling approach to accurately delineate the spatial distribution and volume of landfilled solid waste (predominantly organic pollutants) at two decommissioned chemical plant sites (total area: 8954 m²). Methods: We combined (1) geophysical surveys (transient electromagnetic (TEM, 50 profiles, 2936 points), high-density resistivity (HDR, 2 profiles, 192 points), and ground-penetrating radar (GPR, 22 profiles, 1072.1 m)) and (2) systematic drilling verification (136 boreholes, ≤10 m × 10 m density). Anomalies were interpreted through integrating geophysical responses, historical records, and borehole validation. Spatial modeling was conducted using Kriging interpolation in EVS software. The results show that (1) the anomalies exhibited a “sparse multi-point distribution” across zones A2 (primary waste concentration), A4, and A6, which were differentiated into solid waste, foundations, contaminated soil, voids, and cracks; (2) drilling confirmed solid waste at nine locations (A2: “multi-point, small-quantity” residues; A6: contaminated clay layers with garbage) with irregular thicknesses (0.2–1.3 m); (3) TEM identified diagnostic medium–high-resistivity anomalies (e.g., 28–37 m in A4L3), while GPR detected 17 shallow anomalies (only one validated as waste); and (4) the total waste volume was quantified as 266.9 m³. The methodology reduced the field effort by ∼35% versus drilling-only approaches, resolved geophysical limitations (e.g., HDR’s volume effect overestimating the thickness), and provided a validated framework for efficient characterization of complex historical landfills. Full article

Brushless DC (BLDC) motors are commonly used in electric vehicles (EVs) because of their efficiency, small size and great torque-speed performance. These motors have a few benefits such as low maintenance, increased reliability and power density. Nevertheless, BLDC motors are highly nonlinear and their dynamics are very complicated, in particular, under changing load and supply conditions. The above features require the design of strong and adaptable control methods that can ensure performance over a broad spectrum of disturbances and uncertainties. In order to overcome these issues, this paper uses a Fractional-Order Proportional-Integral-Derivative (FOPID) controller that offers better control precision, better frequency response, and an extra degree of freedom in tuning by using non-integer order terms. Although it has the benefits, there are three primary drawbacks: (i) it is not real-time adaptable, (ii) it is hard to choose appropriate initial gain values, and (iii) it is sensitive to big disturbances and parameter changes. A new control framework is suggested to address these problems. First, a Reinforcement Learning (RL) approach based on Deep Deterministic Policy Gradient (DDPG) is presented to optimize the FOPID gains online so that the controller can adjust itself continuously to the variations in the system. Second, Snake Optimization (SO) algorithm is used in fine-tuning of the FOPID parameters at the initial stages to guarantee stable convergence. Lastly, cascade control structure is adopted, where FOPID controllers are used in the inner (current) and outer (speed) loops. This construction adds robustness to the system as a whole and minimizes the effect of disturbances on the performance. In addition, the cascade design also allows more coordinated and smooth control actions thus reducing stress on the power electronic switches, which reduces switching losses and the overall efficiency of the drive system. The suggested RL-enhanced cascade FOPID controller is verified by Hardware-in-the-Loop (HIL) testing, which shows better performance in the aspects of speed regulation, robustness, and adaptability to realistic conditions of operation in EV applications. Full article

The understanding of medulloblastoma (MB) progression is limited by the lack of minimally invasive monitoring methods. Extracellular vesicles (EVs) carrying disease-specific signatures are promising for liquid biopsies, but clinical translation is hindered by inconsistent isolation techniques. This study compares small EVs (sEVs) and their proteomes from blood plasma (BP) and cerebrospinal fluid (CSF) in MB. Using ultrafiltration and size exclusion chromatography (UF-SEC), we isolated sEVs from pediatric patient samples. sEV proteins from matched CSF-BP samples from MB patients (MBCSF/MBBP), healthy BP controls (HCBP), and MB cell lines (MBCL) were analyzed by liquid chromatography-tandem mass spectrometry, subjected to Gene Ontology and Cytoscape analyses, and compared to published MB, CSF, and EV datasets. By optimizing UF-SEC for small volumes, we found that CSF-sEVs are smaller and elute in later SEC fractions. Proteins linked to the extracellular matrix (ECM) were enriched in MBCSF and MBCL, while integrin binding showed inconsistent patterns between MBCSF and MBBP. MBBP and HCBP showed no significant differences. Fourteen proteins from MB datasets were identified in our analysis and primarily enriched in CSF. These findings support CSF-sEVs as more informative than BP-sEVs for MB diagnosis and monitoring, emphasize the need for fluid-specific sEV isolation, and suggest that ECM components and integrins may mediate MB progression. Full article

This paper presents a DC microgrid architecture with autonomous decentralized control that exhibits high resilience against increasingly common threats, such as natural disasters and cyber-physical attacks, as well as its operational characteristics under normal circumstances. The proposed system achieves autonomous decentralized cooperative control by combining a battery-integrated DC baseline, in which multiple distributed small-scale batteries are directly connected to the grid baseline, with a weakly coupled grid architecture in which each power device is loosely coupled via the grid baseline. Unlike conventional approaches that assign grid formation, inertial support, and power balancing functions to DC/DC converters, the proposed approach delegates these fundamental grid roles to the distributed batteries. This configuration simplifies the control logic of the DC/DC converters, limiting their role to power exchange only. To evaluate system performance, a four-family DC microgrid model incorporating a typical Japanese home environment, including an EV charger, was constructed in MATLAB/Simulink R2025a and subjected to one-year simulations. The results showed that with approximately 5 kW of PV panels and a 20 kWh battery capacity per household, a stable power supply could be maintained throughout the year, with more than 50% of the total power consumption covered by solar energy. Furthermore, the predicted battery life was over 20 years, confirming the practicality and economic viability of the proposed residential microgrid design. Full article

AA1050 aluminum was hydrostatically extruded at room temperature to true strains of 0.9 and 3.2, and at cryogenic temperature to a true strain of 0.9. As a result of the extrusion process, the yield strength (YS) increased by 130–160% to 120–130 MPa, and the ultimate tensile strength (UTS) rose by 64–81% to 125–140 MPa. The hardness reached 46–49 HV. YS and UTS values correspond to mechanical properties typical of the H6 or H8 temper designations, with unusually high elongation at break ranging from 15% to 16.4%. Differences in lattice parameters, crystallite size, and lattice strain between samples deformed under various conditions—as well as those annealed after deformation—were within the margin of measurement uncertainty. This indicated that differences in defect density between the samples were relatively small, due to dynamic recovery occurring during extrusion. However, positron annihilation spectroscopy demonstrated that the cryo-cooled material extruded at a true strain of 0.9, as well as the one extruded at RT at a true strain of 3.2, exhibited significantly higher mean lattice defect concentrations compared to the sample extruded at RT at a true strain of 0.9. The predominant defects detected were vacancies associated with dislocations. The extrusion parameters also significantly affected the crystallographic texture. In particular, they altered the relative proportions of the <111> and <100> components in the axial texture, with the <100> component becoming dominant in cryogenically extruded samples. This trend was further intensified during recrystallization, which enhanced the <100> component even more. Recrystallization of the deformed materials occurred in the temperature range of 520–570 K. The activation energy for grain boundary migration during recrystallization was estimated to be approximately 1.5 eV. Full article

Small extracellular vesicles (sEVs), typically <200 nm in diameter, have emerged as key mediators of intercellular communication, transferring bioactive molecules such as proteins, lipids, and nucleic acids between cells. This review focuses on the growing significance of sEVs in the context of sepsis, a life-threatening syndrome caused by a dysregulated immune response to infection. Sepsis remains a major global health challenge due to its complex pathophysiology, rapid progression, and the limitations of current diagnostic tools, which often fail to detect the condition early or accurately assess the host’s immune status. As interest grows in precision diagnostics, sEVs have gained attention for their potential as biomarkers in liquid biopsy—a minimally invasive approach that analyzes circulating vesicles to monitor disease. Small EVs reflect the physiological state of their cells of origin and can provide real-time insights into immune activation, inflammation, and pathogen presence. This review explores the mechanisms by which sEVs contribute to immune modulation in sepsis, recent advances in understanding their biogenesis and uptake, and their diagnostic and prognostic potential. By highlighting the role of sEVs in sepsis, we aim to underscore their promise in improving early detection, guiding therapeutic decisions, and advancing personalized medicine. Full article

Breast cancer has now surpassed lung cancer as the leading cause of cancer-related deaths among women worldwide. Given the urgent need for more effective treatment, extracellular vesicles (EVs) have gained attention as versatile and promising drug delivery systems. Derived from a variety of cell types, EVs can be loaded with therapeutic cargo or engineered to present specific surface ligands and receptors. These EV modifications enable them to overcome many limitations associated with conventional therapies. In this review, we highlight current methodologies for loading small molecule drugs, RNA-based therapeutics, and proteins into EVs through both pre-isolation (endogenous) and post-isolation (exogenous) methods. We further discuss recent advances in EV surface engineering strategies aimed at improving tumor-specific targeting and immunotherapeutic efficacy in breast cancer. Full article