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18 pages, 2763 KiB  
Article
Enhancing Students’ Interest in Physics Concepts with a Low-Cost STEM Tool Focused on Motivation in Rural Areas of Developing Countries
by René Flores-Godínez, Antonio Alarcón-Paredes, Iris Paola Guzmán-Guzmán, Yanik Ixchel Maldonado-Astudillo and Gustavo Adolfo Alonso-Silverio
Educ. Sci. 2025, 15(8), 994; https://doi.org/10.3390/educsci15080994 (registering DOI) - 5 Aug 2025
Abstract
Physics concepts are considered an essential component of STEM (science, technology, engineering, and mathematics) education and fundamental for economic and technological development in the world. However, there can be student academic underperformance, such as the school environment, learning media and infrastructure, student interest [...] Read more.
Physics concepts are considered an essential component of STEM (science, technology, engineering, and mathematics) education and fundamental for economic and technological development in the world. However, there can be student academic underperformance, such as the school environment, learning media and infrastructure, student interest and emotions, as well as social and economic development factors in communities. These problems are even more acute in rural areas of developing countries, where poverty is high and teachers often lack the necessary technological skills. The aim of this study was to evaluate the impact of a low-cost STEM tool focused on motivation in learning, in terms of five variables of interest in physics in rural areas, as well as the durability of the tools used to learn 12 physics concepts. A quasi-experimental study was conducted with the participation of 78 high school students, with an average age of 15.82 years, in a rural area of Guerrero, Mexico. The results showed that using the STEM tool significantly increased students’ interest in learning methodology, active participation, and attitude towards physics, facilitating the teacher’s work. In addition, the 3D construction kit used in the experimentation, besides being low-cost, proved to be affordable and durable, making it ideal for use in rural areas. Full article
(This article belongs to the Special Issue Interdisciplinary Approaches to STEM Education)
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17 pages, 3099 KiB  
Article
Assessment of Fish Community Structure and Invasion Risk in Xinglin Bay, China
by Shilong Feng, Xu Wang, Liangmin Huang, Jiaqiao Wang, Lin Lin, Jun Li, Guangjie Dai, Qianwen Cai, Haoqi Xu, Yapeng Hui and Fenfen Ji
Biology 2025, 14(8), 988; https://doi.org/10.3390/biology14080988 (registering DOI) - 4 Aug 2025
Viewed by 164
Abstract
A total of 32 fish species were detected in Xinglin Bay using a combination of environmental DNA metabarcoding (eDNA) and traditional morphological survey methods (TSM), covering eight orders, fifteen families, and twenty-six genera. The dominant order was Perciformes, accounting for 43.75% of the [...] Read more.
A total of 32 fish species were detected in Xinglin Bay using a combination of environmental DNA metabarcoding (eDNA) and traditional morphological survey methods (TSM), covering eight orders, fifteen families, and twenty-six genera. The dominant order was Perciformes, accounting for 43.75% of the total species. Among the identified species, there were ten non-native fish species. Compared with the TSM, the eDNA detected 13 additional fish species, including two additional non-native fish species—Gambusia affinis (Baird and Girard, 1853) and Micropterus salmoides (Lacepède, 1802). In addition, the relative abundance of fish from both methods revealed that tilapia was overwhelmingly dominant, accounting for 80.75% and 75.68%, respectively. Furthermore, the AS-ISK assessment revealed that all non-native fish species were classified as medium or high-risk, with five identified as high-risk species, four of which belong to tilapia. These findings demonstrated that tilapia are the dominant and high-risk invasive species in Xinglin Bay and should be prioritized for management. Population reduction through targeted harvesting of tilapia is recommended as the primary control strategy. Additionally, the study highlights the effectiveness of eDNA in monitoring fish community structure in brackish ecosystems. Full article
(This article belongs to the Special Issue Advances in Aquatic Ecological Disasters and Toxicology)
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19 pages, 10865 KiB  
Article
Evaluation of Immunoprotective Activities of White Button Mushroom (Agaricus bisporus) Water Extract Against Major Pathogenic Bacteria (Aeromonas hydrophila or Vibrio fluvialis) in Goldfish (Carassius auratus)
by Shujun Sun, Jing Chen, Pan Cui, Xiaoxiao Yang, Yuhan Zheng, Zijian Ma, Yong Liu and Xiang Liu
Animals 2025, 15(15), 2257; https://doi.org/10.3390/ani15152257 - 1 Aug 2025
Viewed by 168
Abstract
The white button mushroom (Agaricus bisporus) is a widely cultivated edible and medicinal mushroom, which contains various active substances, and has application value against pathogenic bacteria in aquaculture. Firstly, A. bisporus water extract (AB-WE) was prepared. Through the detection kits, it [...] Read more.
The white button mushroom (Agaricus bisporus) is a widely cultivated edible and medicinal mushroom, which contains various active substances, and has application value against pathogenic bacteria in aquaculture. Firstly, A. bisporus water extract (AB-WE) was prepared. Through the detection kits, it was found that the polysaccharide, protein, and polyphenol components of AB-WE were 9.11%, 3.3%, and 1.5%, respectively. The 246 compounds were identified in AB-WE, and the major small-molecule components included L-Isoleucine, L-Tyrosine, L-Valine, and Linoleic acid by HPLC-Q Exactive-Orbitrap-MS. Secondly, the AB-WE was evaluated for its immunological activities through dietary administration and pathogen challenge (Aeromonas hydrophila and Vibrio fluvialis) in goldfish (Carassius auratus). The results showed that the levels of immune factors of acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM) increased (p < 0.05) in goldfish, and the relative percentage survival of AB-WE against A. hydrophila and V. fluvialis were 80.00% (p < 0.05) and 81.82% (p < 0.05), respectively. The AB-WE reduced the bacterial content in renal tissue, enhanced the phagocytic activity of leukocytes, and exhibited antioxidant and anti-inflammatory effects by reducing the expression of antioxidant-related factors and inflammatory factors. Through histopathological and immunofluorescence techniques, it was found that AB-WE maintained the integrity of visceral tissues and reduced renal tissue apoptosis and DNA damage. Therefore, AB-WE exhibits immunoprotective activity against A. hydrophila and V. fluvialis infections in fish, and holds promise as an immunotherapeutic agent against major pathogenic bacteria in aquaculture. Full article
(This article belongs to the Section Aquatic Animals)
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13 pages, 373 KiB  
Article
Impact Assessment of Rural Electrification Through Photovoltaic Kits on Household Expenditures and Income: The Case of Morocco
by Abdellah Oulakhmis, Rachid Hasnaoui and Youness Boudrik
Economies 2025, 13(8), 224; https://doi.org/10.3390/economies13080224 - 31 Jul 2025
Viewed by 177
Abstract
This study evaluates the socio-economic impact of rural electrification through photovoltaic (PV) systems in Morocco. As part of the country’s broader energy transition strategy, decentralized renewable energy solutions like PV kits have been deployed to improve energy access in isolated rural areas. Using [...] Read more.
This study evaluates the socio-economic impact of rural electrification through photovoltaic (PV) systems in Morocco. As part of the country’s broader energy transition strategy, decentralized renewable energy solutions like PV kits have been deployed to improve energy access in isolated rural areas. Using quasi-experimental econometric techniques, specifically propensity score matching (PSM) and estimation of the Average Treatment Effect on the Treated (ATT), the study measures changes in household income, expenditures, and economic activities resulting from PV electrification. The results indicate significant positive effects on household income, electricity spending, and productivity in agriculture and livestock. These findings highlight the critical role of decentralized renewable energy in advancing rural development and poverty reduction. Policy recommendations include expanding PV access with complementary support measures such as microfinance and technical training. Full article
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21 pages, 4846 KiB  
Article
Bioactive Chalcone-Loaded Mesoporous Silica KIT-6 Nanocarrier: A Promising Strategy for Inflammation and Pain Management in Zebrafish
by Maria Kueirislene Amâncio Ferreira, Francisco Rogenio Silva Mendes, Emmanuel Silva Marinho, Roberto Lima de Albuquerque, Jesyka Macedo Guedes, Izabell Maria Martins Teixeira, Ramon Róseo Paula Pessoa Bezerra de Menezes, Vinicius Patricio Santos Caldeira, Anne Gabriella Dias Santos, Marisa Jádna Silva Frederico, Antônio César Honorato Barreto, Inês Domingues, Tigressa Helena Soares Rodrigues, Jane Eire Silva Alencar de Menezes and Hélcio Silva dos Santos
Pharmaceutics 2025, 17(8), 981; https://doi.org/10.3390/pharmaceutics17080981 - 30 Jul 2025
Viewed by 526
Abstract
Background/Objectives: The incorporation of bioactive molecules into mesoporous carriers is a promising strategy to improve stability, solubility, and therapeutic efficacy. In this study, we report for the first time the encapsulation of the synthetic chalcone 4-Cl into KIT-6 mesoporous silica and evaluate [...] Read more.
Background/Objectives: The incorporation of bioactive molecules into mesoporous carriers is a promising strategy to improve stability, solubility, and therapeutic efficacy. In this study, we report for the first time the encapsulation of the synthetic chalcone 4-Cl into KIT-6 mesoporous silica and evaluate its cytotoxicity, toxicological profile, and pharmacological activities (antinociceptive, anti-inflammatory, and anxiolytic) using an in vivo zebrafish (Danio rerio) model. Methods: Zebrafish were orally dosed with 4-Cl, 4-Cl/KIT-6, or KIT-6 (4, 20, 40 mg/kg) and mortality was recorded for 96 h. For analgesia, zebrafish pretreated with 4-Cl, 4-Cl/KIT-6, KIT-6, or morphine received a tail stimulus (0.1% formalin). Locomotor activity (quadrant crossings) was monitored for 30 min to assess analgesia (neurogenic: 0–5 min; inflammatory: 15–30 min). For inflammation, abdominal edema and weight gain were assessed 4 h after intraperitoneal carrageenan (1.5%). Zebrafish (n = 6/group) received 4-Cl, 4-Cl/KIT-6, or KIT-6 (4, 20, 40 mg/kg, p.o.). Controls received ibuprofen (100 mg/kg, p.o.) or 3% DMSO. Weight was measured hourly for 4 h post-carrageenan (difference between baseline and hourly weights). Results: Physicochemical characterizations confirmed successful encapsulation without compromising the ordered structure of KIT-6, as evidenced by a significant reduction in surface area and pore volume, indicating efficient drug incorporation. In vivo assays demonstrated that the 4-Cl/KIT-6 formulation maintained the pharmacological activities of the free chalcone, reduced toxicity, and, notably, revealed a significant anxiolytic effect for the first time. Conclusions: These findings highlight KIT-6 as a promising platform for chalcone delivery systems and provide a solid basis for future preclinical investigations. Full article
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28 pages, 8824 KiB  
Article
Platform Approaches in the AEC Industry: Stakeholder Perspectives and Case Study
by Layla Mujahed, Gang Feng and Jianghua Wang
Buildings 2025, 15(15), 2684; https://doi.org/10.3390/buildings15152684 - 30 Jul 2025
Viewed by 232
Abstract
The architecture, engineering, and construction (AEC) industry faces challenges related to inefficiencies and fragmentation that highlight the need for advanced construction technologies and drive interest in innovative solutions such as the platform approach to design. This study assessed platform-based building design through (1) [...] Read more.
The architecture, engineering, and construction (AEC) industry faces challenges related to inefficiencies and fragmentation that highlight the need for advanced construction technologies and drive interest in innovative solutions such as the platform approach to design. This study assessed platform-based building design through (1) interviews with practitioners from China, Jordan, and the UK, which helped to define the platform approach in the AEC industry and the challenges involved, and (2) a residential building design simulation conducted to evaluate the potential of the platform approach. The simulated design’s materials costs, energy efficiency, and construction time were compared with those of the traditional building design. The results of the comparison corroborate the interview findings concerning practitioners’ perspectives on platform definition, benefits, challenges, and implementation. The findings also demonstrate the potential of the platform approach to enhance productivity and scalability through modularization, kit-of-parts configuration, and standardization. This research bridges the gap between theory and practice by supporting shareholder perspectives on building design and construction with the results of a simulated platform approach to a real-world design project. This research addresses the urgent need to better understand and test the platform approach to achieve material, energy, and construction time savings through collaborative and practice-informed design. Full article
(This article belongs to the Section Architectural Design, Urban Science, and Real Estate)
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18 pages, 2111 KiB  
Article
Modelling Renewable Energy and Resource Interactions Using CLEWs to Support Thailand’s 2050 Carbon Neutrality Goal
by Nat Nakkorn, Surasak Janchai, Suparatchai Vorarat and Prayuth Rittidatch
Sustainability 2025, 17(15), 6909; https://doi.org/10.3390/su17156909 - 30 Jul 2025
Viewed by 338
Abstract
This study utilises the Open Source Energy Modelling System (OSeMOSYS) in conjunction with the Climate, Land, Energy, and Water systems (CLEWs) framework to investigate Thailand’s energy transition, which is designed to achieve carbon neutrality by 2050. Two scenarios have been devised to evaluate [...] Read more.
This study utilises the Open Source Energy Modelling System (OSeMOSYS) in conjunction with the Climate, Land, Energy, and Water systems (CLEWs) framework to investigate Thailand’s energy transition, which is designed to achieve carbon neutrality by 2050. Two scenarios have been devised to evaluate the long-term trade-offs among energy, water, and land systems. Data were sourced from esteemed international organisations (e.g., the IEA, FAO, and OECD) and national agencies and organised into a tailored OSeMOSYS Starter Data Kit for Thailand, comprising a baseline and a carbon neutral trajectory. The baseline scenario, primarily reliant on fossil fuels, is projected to generate annual CO2 emissions exceeding 400 million tons and water consumption surpassing 85 billion cubic meters by 2025. By the mid-century, the carbon neutral scenario will have approximately 40% lower water use and a 90% reduction in power sector emissions. Under the carbon neutral path, renewable energy takes the front stage; the share of renewable electricity goes from under 20% in the baseline scenario to almost 80% by 2050. This transition and large reforestation initiatives call for consistent investment in solar energy (solar energy expenditures exceeding 20 billion USD annually by 2025). Still, it provides notable co-benefits, including greater resource sustainability and better alignment with international climate targets. The results provide strategic insights aligned with Thailand’s National Energy Plan (NEP) and offer modelling evidence toward achieving international climate goals under COP29. Full article
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17 pages, 2885 KiB  
Article
Silanization-Modified Lignin Nanoparticles for Paper Coating with Enhanced Liquid and Vapor Barriers, Frication Resistance, and Self-Cleaning Properties
by Wen Chen, Ren’ai Li, Yunfeng Cao, Chunjie Ye, Zhulan Liu and Huining Xiao
Polymers 2025, 17(15), 2066; https://doi.org/10.3390/polym17152066 - 29 Jul 2025
Viewed by 270
Abstract
Paper’s inherent hydrophilicity and porosity cause inadequate barrier properties, failing under high humidity/temperature. This study successfully developed a hydrophobic nanocoating agent (xLNPs-OTS) through silanization modification using D276 (lignin nanoparticles with a diameter of 276 nm) as the substrate and OTS (octadecyltrichlorosilane) as the [...] Read more.
Paper’s inherent hydrophilicity and porosity cause inadequate barrier properties, failing under high humidity/temperature. This study successfully developed a hydrophobic nanocoating agent (xLNPs-OTS) through silanization modification using D276 (lignin nanoparticles with a diameter of 276 nm) as the substrate and OTS (octadecyltrichlorosilane) as the functionalizing agent. By applying the coating to paper surfaces followed by a hot-pressing process, the paper achieved comprehensive performance enhancements, including superior water, oil, and vapor barrier properties, thermal stability, mechanical strength, frictional resistance, and self-cleaning capabilities. The Cobb 60 value of LOTSC3.5T120t30 (the coating made from the OTS silanized lignin with the coating amount of 3.5 g/m2 and a hot-pressing at 120 °C for 30 min) coated paper is as low as 3.75 g/m2, and can withstand hot water at 100 °C for 60 min. The Cobb 60 value of the LOTSC20T120t30 (the coating made from the OTS silanized lignin with the coating amount of 20 g/m2 and a hot-pressing at 120 °C for 30 min) coated paper is reduced to 0.9 g/m2, the Kit grade is 6, and all coated papers are endowed with self-cleaning features. This study advances lignin’s high-value utilization, driving sustainable packaging and supporting eco-friendly paper material development. Full article
(This article belongs to the Special Issue Advances in Lignocellulose Research and Applications)
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18 pages, 3968 KiB  
Article
Design, Development, and Clinical Validation of a Novel Kit for Cell-Free DNA Extraction
by Ekin Çelik, Hande Güner, Gizem Kayalı, Haktan Bagis Erdem, Taha Bahsi and Hasan Huseyin Kazan
Diagnostics 2025, 15(15), 1897; https://doi.org/10.3390/diagnostics15151897 - 29 Jul 2025
Viewed by 305
Abstract
Background: Cell-free DNA (cfDNA) has become a cornerstone of liquid biopsy applications, offering promise for early disease detection and monitoring. However, its widespread clinical adoption is limited by variability in pre-analytical processing, especially during isolation. Current extraction methods face challenges in yield, purity, [...] Read more.
Background: Cell-free DNA (cfDNA) has become a cornerstone of liquid biopsy applications, offering promise for early disease detection and monitoring. However, its widespread clinical adoption is limited by variability in pre-analytical processing, especially during isolation. Current extraction methods face challenges in yield, purity, and reproducibility. Methods: We developed and optimized SafeCAP 2.0, a novel magnetic bead-based cfDNA extraction kit, focusing on efficient recovery, minimal genomic DNA contamination, and PCR compatibility. Optimization involved systematic evaluation of magnetic bead chemistry, buffer composition, and reagent volumes. Performance was benchmarked against a commercial reference kit (Apostle MiniMax) using spiked oligonucleotides and plasma from patients with stage IV NSCLC. Results: The optimized protocol demonstrated superior recovery with a limit of detection (LoD) as low as 0.3 pg/µL and a limit of quantification (LoQ) of 1 pg/μL with no detectable PCR inhibition. In comparative studies, SafeCAP 2.0 showed equivalent or improved performance over the commercial kit. Clinical validation using 47 patient plasma samples confirmed robust cfDNA recovery and fragment integrity. Conclusions: SafeCAP 2.0 offers a cost-effective, high-performance solution for cfDNA extraction in both research and clinical workflows. Its design and validation address key pre-analytical barriers, supporting integration into routine diagnostics and precision medicine platforms. Full article
(This article belongs to the Section Pathology and Molecular Diagnostics)
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16 pages, 1423 KiB  
Article
Measurement of Oxidative Stress Index in 102 Patients with Peyronie’s Disease
by Gianni Paulis, Andrea Paulis, Giovanni De Giorgio and Salvatore Quattrocchi
Metabolites 2025, 15(8), 503; https://doi.org/10.3390/metabo15080503 - 29 Jul 2025
Viewed by 247
Abstract
Background: Peyronie’s disease (PD) is a chronic inflammatory condition that affects the penile albuginea. Oxidative stress (OS) plays a crucial role in the development of the disease, prompting us to investigate OS levels at the site of the disease and in peripheral [...] Read more.
Background: Peyronie’s disease (PD) is a chronic inflammatory condition that affects the penile albuginea. Oxidative stress (OS) plays a crucial role in the development of the disease, prompting us to investigate OS levels at the site of the disease and in peripheral blood. This article presents our second study in which the OS was evaluated by calculating the OS index (OSI) in blood samples taken directly from the penile corpora cavernosa of patients with PD. Our innovative diagnostic method, which focuses on the analysis of oxidative stress (OS) in the corpora cavernosa of the penis, allows us to accurately identify the “chemical” signals (OS levels) of the pathology in the area where it is present. Methods: Our study included 102 PD patients from our Peyronie’s care center and 100 control cases. To conduct a comprehensive OS analysis, we measured both the total oxidant status (TOS) and total antioxidant status (TAS) and calculated the oxidative stress index (OSI) as OSI = TOS/TAS × 100. Blood samples were collected from the penis and a vein in the upper extremity, and OS was measured using d-ROMs and PATs (FRAS kit). Results: Pearson’s analyses revealed a significant statistical correlation between penile OSI values and PD plaque volumes (p = 0.003), while no correlation was found between systemic OSI values and plaque volumes (p = 0.356). Penile OSI values decreased significantly after PD plaque removal (p < 0.0001). A comparison of penile OSI values in PD patients (post plaque removal) and the control group showed no significant differences (p = 0.418). Conclusions: The lack of correlation between systemic OSI values and Peyronie’s plaque volume suggests that direct sampling from the site of the disease is preferable for OS studies. Conducting a penile OSI study could provide a precise oxidative marker dependent on plaque volume. In addition, the penile OSI study can biochemically monitor the therapeutic result, alongside penile ultrasound imaging. Full article
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17 pages, 1525 KiB  
Article
Clonidine Protects Endothelial Cells from Angiotensin II-Induced Injury via Anti-Inflammatory and Antioxidant Mechanisms
by Bekir Sıtkı Said Ulusoy, Mehmet Cudi Tuncer and İlhan Özdemir
Life 2025, 15(8), 1193; https://doi.org/10.3390/life15081193 - 27 Jul 2025
Viewed by 405
Abstract
Background: Cerebral aneurysm (CA) is a focal or diffuse pathological dilation of the cerebral arterial wall that arises due to various etiological factors. It represents a serious vascular condition, particularly affecting the elderly, and carries a high risk of rupture and neurological morbidity. [...] Read more.
Background: Cerebral aneurysm (CA) is a focal or diffuse pathological dilation of the cerebral arterial wall that arises due to various etiological factors. It represents a serious vascular condition, particularly affecting the elderly, and carries a high risk of rupture and neurological morbidity. Clonidine (CL), an α2-adrenergic receptor agonist, has been reported to suppress aneurysm progression; however, its underlying molecular mechanisms, especially in relation to cerebral endothelial dysfunction, remain unclear. This study aimed to investigate the potential of CL to mitigate CA development by modulating apoptosis, inflammation, and oxidative stress in an Angiotensin II (Ang II)-induced endothelial injury model. Methods: Human brain microvascular endothelial cells (HBMECs) were used to establish an in vitro model of endothelial dysfunction by treating cells with 1 µM Ang II for 48 h. CL was administered 2 h prior to Ang II exposure at concentrations of 0.1, 1, and 10 µM. Cell viability was assessed using the MTT assay. Oxidative stress markers, including reactive oxygen species (ROS) and Nitric Oxide (NO), were measured using 2′,7′–dichlorofluorescin diacetate (DCFDA). Gene expression levels of vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP-2 and MMP-9), high mobility group box 1 (HMGB1), and nuclear factor kappa B (NF-κB) were quantified using RT-qPCR. Levels of proinflammatory cytokines; tumor necrosis factor-alpha (TNF-α), Interleukin-6 (IL-6), and interferon-gamma (IFN-γ); were measured using commercial ELISA kits. Results: Ang II significantly increased ROS production and reduced NO levels, accompanied by heightened proinflammatory cytokine release and endothelial dysfunction. MTT assay revealed a marked decrease in cell viability following Ang II treatment (34.18%), whereas CL preserved cell viability in a concentration-dependent manner: 44.24% at 0.1 µM, 66.56% at 1 µM, and 81.74% at 10 µM. CL treatment also significantly attenuated ROS generation and inflammatory cytokine levels (p < 0.05). Furthermore, the expression of VEGF, HMGB1, NF-κB, MMP-2, and MMP-9 was significantly downregulated in response to CL. Conclusions: CL exerts a protective effect on endothelial cells by reducing oxidative stress and suppressing proinflammatory signaling pathways in Ang II-induced injury. These results support the potential of CL to mitigate endothelial injury in vitro, though further in vivo studies are required to confirm its translational relevance. Full article
(This article belongs to the Section Pharmaceutical Science)
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14 pages, 384 KiB  
Article
Outbreak Caused by VIM-1- and VIM-4-Positive Proteus mirabilis in a Hospital in Zagreb
by Branka Bedenić, Gernot Zarfel, Josefa Luxner, Andrea Grisold, Marina Nađ, Maja Anušić, Vladimira Tičić, Verena Dobretzberger, Ivan Barišić and Jasmina Vraneš
Pathogens 2025, 14(8), 737; https://doi.org/10.3390/pathogens14080737 - 26 Jul 2025
Viewed by 287
Abstract
Background/objectives: Proteus mirabilis is a frequent causative agent of urinary and wound infections in both community and hospital settings. It develops resistance to expanded-spectrum cephalosporins (ESCs) due to the production of extended-spectrum β-lactamases (ESBLs) or plasmid-mediated AmpC β-lactamases (p-AmpCs). Recently, carbapenem-resistant isolates of [...] Read more.
Background/objectives: Proteus mirabilis is a frequent causative agent of urinary and wound infections in both community and hospital settings. It develops resistance to expanded-spectrum cephalosporins (ESCs) due to the production of extended-spectrum β-lactamases (ESBLs) or plasmid-mediated AmpC β-lactamases (p-AmpCs). Recently, carbapenem-resistant isolates of P. mirabilis emerged due to the production of carbapenemases, mostly belonging to Ambler classes B and D. Here, we report an outbreak of infections due to carbapenem-resistant P. mirabilis that were observed in a psychiatric hospital in Zagreb, Croatia. The characteristics of ESBL and carbapenemase-producing P. mirabilis isolates, associated with an outbreak, were analyzed. Materials and methods: The antibiotic susceptibility testing was performed by the disk-diffusion and broth dilution methods. The double-disk synergy test (DDST) and inhibitor-based test with clavulanic and phenylboronic acid were applied to screen for ESBLs and p-AmpCs, respectively. Carbapenemases were screened by the modified Hodge test (MHT), while carbapenem hydrolysis was investigated by the carbapenem inactivation method (CIM) and EDTA-carbapenem-inactivation method (eCIM). The nature of the ESBLs, carbapenemases, and fluoroquinolone-resistance determinants was investigated by PCR. Plasmids were characterized by PCR-based replicon typing (PBRT). Selected isolates were subjected to molecular characterization of the resistome by an Inter-Array Genotyping Kit CarbaResisit and whole-genome sequencing (WGS). Results: In total, 20 isolates were collected and analyzed. All isolates exhibited resistance to amoxicillin alone and when combined with clavulanic acid, cefuroxime, cefotaxime, ceftriaxone, cefepime, imipenem, ceftazidime–avibactam, ceftolozane–tazobactam, gentamicin, amikacin, and ciprofloxacin. There was uniform susceptibility to ertapenem, meropenem, and cefiderocol. The DDST and combined disk test with clavulanic acid were positive, indicating the production of an ESBL. The MHT was negative in all except one isolate, while the CIM showed moderate sensitivity, but only with imipenem as the indicator disk. Furthermore, eCIM tested positive in all of the CIM-positive isolates, consistent with a metallo-β-lactamase (MBL). PCR and sequencing of the selected amplicons identified VIM-1 and VIM-4. The Inter-Array Genotyping Kit CarbaResist and WGS identified β-lactam resistance genes blaVIM, blaCTX-M-15, and blaTEM genes; aminoglycoside resistance genes aac(3)-IId, aph(6)-Id, aph(3″)-Ib, aadA1, armA, and aac(6′)-IIc; as well as resistance genes for sulphonamides sul1 and sul2, trimethoprim dfr1, chloramphenicol cat, and tetracycline tet(J). Conclusions: This study revealed an epidemic spread of carbapenemase-producing P. mirabilis in two wards in a psychiatric hospital. Due to the extensively resistant phenotype (XDR), therapeutic options were limited. This is the first report of carbapenemase-producing P. mirabilis in Croatia. Full article
(This article belongs to the Special Issue Emerging and Neglected Pathogens in the Balkans)
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18 pages, 529 KiB  
Article
Perspectives on Mail-Based Fecal Testing for Colorectal Cancer Screening in Bulgaria: A Survey of Gastroenterologists
by Kostadin Yordanov Dimitrov, Vladislav Velchev, Nely Danailova, Elena Staneva, Teodor Koparanov, Trifon Diankov, Teodora Gencheva, Bozhidar Valkov, Eleonora Hristova-Atanasova, Georgi Iskrov and Rumen Stefanov
Gastroenterol. Insights 2025, 16(3), 25; https://doi.org/10.3390/gastroent16030025 - 26 Jul 2025
Viewed by 300
Abstract
Background: Bulgaria carries a high burden of colorectal cancer (CRC) but, at the start of this study, lacked a nationwide organized screening program. Understanding specialist views (particularly on mail-based fecal testing) is essential for effective policy development. Objective: The objective is to assess [...] Read more.
Background: Bulgaria carries a high burden of colorectal cancer (CRC) but, at the start of this study, lacked a nationwide organized screening program. Understanding specialist views (particularly on mail-based fecal testing) is essential for effective policy development. Objective: The objective is to assess the attitudes towards, practices of, and perceived barriers to CRC screening among Bulgarian gastroenterologists, with a focus on the feasibility of mail-based fecal occult blood testing (FOBT). Methods: A cross-sectional survey of 38 gastroenterologists examined clinical use of FOBT, screening method preferences, and perceived systemic and patient-level barriers to CRC screening. Results: Among respondents, 57.89% reported using FOBT in clinical practice, and 71.05% indicated they would undergo the test themselves and recommend it to relatives. Colonoscopy was the preferred diagnostic tool for 84.21% of participants; however, the existing literature raises concerns about its feasibility for large-scale population screening. Key systemic barriers, rated on a 5-point Likert scale, included financial constraints (mean = 3.08), inadequate infrastructure (2.89), and healthcare workforce shortages (2.71). Patient-level barriers were led by low health literacy (4.13), lack of motivation (3.95), and procedural fears (3.26). A majority (84.38%) believed that mail-based FOBT would increase screening uptake, and 57.89% supported annual distribution of test kits. Nearly all respondents (97.37%) favored initiating screening at age 50. Conclusions: This study highlights strong support among Bulgarian gastroenterologists for a national CRC screening program, with particular endorsement of mail-based FOBT. Despite acknowledged systemic and population-level barriers, the findings suggest that such an approach could increase screening coverage, promote early detection, and support the strategic rollout of Bulgaria’s emerging cancer control initiatives. Full article
(This article belongs to the Section Gastrointestinal Disease)
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13 pages, 8639 KiB  
Article
In-Depth Characterization of L1CAM+ Extracellular Vesicles as Potential Biomarkers for Anti-CD20 Therapy Response in Relapsing–Remitting Multiple Sclerosis
by Shamundeeswari Anandan, Karina Maciak, Regina Breinbauer, Laura Otero-Ortega, Giancarlo Feliciello, Nataša Stojanović Gužvić, Oivind Torkildsen and Kjell-Morten Myhr
Int. J. Mol. Sci. 2025, 26(15), 7213; https://doi.org/10.3390/ijms26157213 - 25 Jul 2025
Viewed by 760
Abstract
The effective suppression of inflammation using disease-modifying therapies is essential in the treatment of multiple sclerosis (MS). Anti-CD20 monoclonal antibodies are commonly used long-term as maintenance therapies, largely due to the lack of reliable biomarkers to guide dosing and evaluate treatment response. However, [...] Read more.
The effective suppression of inflammation using disease-modifying therapies is essential in the treatment of multiple sclerosis (MS). Anti-CD20 monoclonal antibodies are commonly used long-term as maintenance therapies, largely due to the lack of reliable biomarkers to guide dosing and evaluate treatment response. However, prolonged use increases the risk of infections and other immune-mediated side effects. The unique ability of brain-derived blood extracellular vesicles (EVs) to cross the blood–brain barrier and reflect the central nervous system (CNS) immune status has sparked interest in their potential as biomarkers. This study aimed to assess whether blood-derived L1CAM+ EVs could serve as biomarkers of treatment response to rituximab (RTX) in patients with relapsing-remitting MS (RRMS). Serum samples (n = 25) from the baseline (month 0) and after 6 months were analyzed from the RTX arm of the ongoing randomized clinical trial OVERLORD-MS (comparing anti-CD20 therapies in RRMS patients) and were compared with serum samples from healthy controls (n = 15). Baseline cerebrospinal fluid (CSF) samples from the same study cohort were also included. EVs from both serum and CSF samples were characterized, considering morphology, size, and concentration, using transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The immunophenotyping of EV surface receptors was performed using flow cytometry with the MACSPlex exosome kit, while label-free quantitative proteomics of EV protein cargo was conducted using a proximity extension assay (PEA). TEM confirmed the presence of EVs with the expected round morphology with a diameter of 50–150 nm. NTA showed significantly higher concentrations of L1CAM+ EVs (p < 0.0001) in serum total EVs and EBNA1+ EVs (p < 0.01) in serum L1CAM+ EVs at baseline (untreated) compared to in healthy controls. After six months of RTX therapy, there was a significant reduction in L1CAM+ EV concentration (p < 0.0001) and the downregulation of TNFRSF13B (p = 0.0004; FC = −0.49) in serum total EVs. Additionally, non-significant changes were observed in CD79B and CCL2 levels in serum L1CAM+ EVs at baseline compared to in controls and after six months of RTX therapy. In conclusion, L1CAM+ EVs in serum showed distinct immunological profiles before and after rituximab treatment, underscoring their potential as dynamic biomarkers for individualized anti-CD20 therapy in MS. Full article
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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14 pages, 1694 KiB  
Article
The Role of MLPA in Detecting Syndromic Submicroscopic Copy Number Variations in Normal QF-PCR Miscarriage Specimens
by Gabriela Popescu-Hobeanu, Mihai-Gabriel Cucu, Alexandru Calotă-Dobrescu, Luminița Dragotă, Anca-Lelia Riza, Ioana Streață, Răzvan Mihail Pleșea, Ciprian Laurențiu Pătru, Cristina Maria Comănescu, Ștefania Tudorache, Dominic Iliescu and Florin Burada
Genes 2025, 16(8), 867; https://doi.org/10.3390/genes16080867 - 24 Jul 2025
Viewed by 330
Abstract
Background/Objectives: Miscarriage is an increasingly common event worldwide arising from various factors, and identifying its etiology is important for planning and managing any future pregnancies. It is estimated that about half of early pregnancy loss cases are caused by genetic abnormalities, while [...] Read more.
Background/Objectives: Miscarriage is an increasingly common event worldwide arising from various factors, and identifying its etiology is important for planning and managing any future pregnancies. It is estimated that about half of early pregnancy loss cases are caused by genetic abnormalities, while a significantly lower rate is found in late pregnancy loss. Multiplex ligation-dependent probe amplification (MLPA) can detect small changes within a gene with precise breakpoints at the level of a single exon. The aim of our study was to identify the rate of copy number variations (CNVs) in spontaneous pregnancy loss samples after having previously tested them via quantitative fluorescence PCR (QF-PCR), with no abnormal findings. Methods: DNA was extracted from product-of-conception tissue samples, followed by the use of an MLPA kit for the detection of 31 microdeletion/microduplication syndromes (SALSA® MLPA® Probemix P245 Microdeletion Syndromes-1A, MRC-Holland, Amsterdam, The Netherlands). Results: A total of 11 (13.1%) out of the 84 successfully tested samples showed CNVs. Duplications accounted for 9.5% of the analyzed samples (eight cases), while heterozygous or hemizygous deletions were present in three cases (3.6%). Among all the detected CNVs, only three were certainly pathogenic (3.6%), with two deletions associated with DiGeorge-2 syndrome and Rett syndrome, respectively, and a 2q23.1 microduplication syndrome, all detected in early pregnancy loss samples. For the remaining cases, additional genetic tests (e.g., aCGH/SNP microarray) are required to establish CNV size and gene content and therefore their pathogenicity. Conclusions: MLPA assays seem to have limited value in detecting supplementary chromosomal abnormalities in miscarriages. Full article
(This article belongs to the Section Human Genomics and Genetic Diseases)
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