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10 pages, 965 KB  
Article
Sandalwood Sesquiterpene (Z)-α-Santalol Exhibits In Vivo Efficacy Against Madurella mycetomatis in Galleria mellonella Larvae
by Shereen O. Abd Algaffar, Stephan Seegers, Shaoqin Zhou, Prabodh Satyal, William N. Setzer, Thomas J. Schmidt and Sami A. Khalid
Molecules 2025, 30(20), 4090; https://doi.org/10.3390/molecules30204090 (registering DOI) - 15 Oct 2025
Abstract
In continuation of our recent report on the in vitro activity of sandalwood essential oils and their major components, the sesquiterpenes (Z)-α- and -β-santalols, against the pathogenic fungus Madurella mycetomatis, causative agent of the neglected tropical disease eumycetoma, we have [...] Read more.
In continuation of our recent report on the in vitro activity of sandalwood essential oils and their major components, the sesquiterpenes (Z)-α- and -β-santalols, against the pathogenic fungus Madurella mycetomatis, causative agent of the neglected tropical disease eumycetoma, we have now tested these isolated constituents, as well as native Royal Hawaiian sandalwood essential oil obtained from Santalum paniculatum, for in vivo activity in a model system using infected Galleria mellonella larvae. Besides confirming the superior activity of (Z)-α-santalol over the (Z)-β-isomer and the crude essential oil in two further strains of M. mycetomatis in vitro, the former compound also turned out to extend the lifespan of the infected larvae significantly, in contrast to (Z)-β-santalol, the total oil, or the antifungal drug itraconazole. The present findings not only characterize (Z)-α-santalol as a natural compound with promising in vivo activity against eumycetoma but also inspire further studies as a potential for novel, more effective therapy and warranting further studies to understand its mechanism of action and potential clinical applications. Full article
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15 pages, 3635 KB  
Article
Development and Comparative Evaluation of Two Enzyme-Based Amperometric Biosensor Designs for Alanine Aminotransferase Determination in Biological Fluids
by Daryna Mruga, Yevhen Vakhovskyi, Veronika Bakhmat, Viktoriya Pyeshkova, Sergii Dzyadevych and Oleksandr Soldatkin
Micromachines 2025, 16(10), 1168; https://doi.org/10.3390/mi16101168 (registering DOI) - 15 Oct 2025
Abstract
Alanine aminotransferase (ALT) is a key biomarker of liver function. Compared with conventional assays for ALT detection—which are expensive, time-consuming, labor-intensive, and require experienced personnel—biosensors represent a promising alternative, but it remains unclear which biorecognitive enzymatic configuration offers the best analytical performance for [...] Read more.
Alanine aminotransferase (ALT) is a key biomarker of liver function. Compared with conventional assays for ALT detection—which are expensive, time-consuming, labor-intensive, and require experienced personnel—biosensors represent a promising alternative, but it remains unclear which biorecognitive enzymatic configuration offers the best analytical performance for ALT detection. This study presents the development and comparative evaluation of two amperometric biosensors based on oxidase biorecognition elements: pyruvate oxidase (POx) and glutamate oxidase (GlOx). Enzymes were immobilized onto platinum electrodes under optimized conditions using entrapment for POx (pH 7.4, enzyme loading 1.62 U/µL, PVA-SbQ concentration 13.2%) and covalent crosslinking for GlOx (pH 6.5, enzyme loading 2.67%, glutaraldehyde concentration 0.3%). Analytical parameters were systematically assessed, including linear range (1–500 U/L for POx vs. 5–500 U/L for GlOx), limit of detection (1 U/L for both), and sensitivity (0.75 vs. 0.49 nA/min at 100 U/L). The POx-based biosensor demonstrated higher sensitivity and lower detection limits, whereas the GlOx-based biosensor exhibited greater stability in complex solutions and reduced assay costs due to a simpler working solution. Moreover, while the POx-based system is uniquely suited for ALT determination, the GlOx-based sensor can be affected by AST activity in samples but may also be adapted for targeted AST detection. Overall, the study highlights a trade-off between sensitivity, robustness, and versatility in ALT biosensor design, providing guidance for the rational development of clinically relevant devices. Full article
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17 pages, 655 KB  
Article
Probable Depression Is Associated with Lower BMI Among Women on ART in Kinshasa, the Democratic Republic of Congo: A Cross-Sectional Study
by Annie Kavira Viranga, Ignace Balaw’a Kalonji Kamuna, Paola Mwanamoke Mbokoso, Celestin Nzanzu Mudogo and Pierre Akilimali Zalagile
Nutrients 2025, 17(20), 3230; https://doi.org/10.3390/nu17203230 (registering DOI) - 15 Oct 2025
Abstract
Background: Women living with HIV (WLHIV) in low-income urban settings face multiple intersecting nutritional risks from food insecurity, poor dietary quality, and mental health problems. We evaluated the prevalence of household food insecurity and inadequate dietary diversity, examining their associations with depressive [...] Read more.
Background: Women living with HIV (WLHIV) in low-income urban settings face multiple intersecting nutritional risks from food insecurity, poor dietary quality, and mental health problems. We evaluated the prevalence of household food insecurity and inadequate dietary diversity, examining their associations with depressive symptoms, antiretroviral therapy (ART)-related factors, and body mass index (BMI) among WLHIV attending routine ART clinics in Kinshasa, The Democratic Republic of Congo. This study addresses critical gaps in understanding the interplay between mental health and nutrition in the context of HIV care, with significant implications for improving health outcomes among vulnerable populations. Methods: In this clinic-based cross-sectional study (February–April 2024), we enrolled 571 women on ART in Masina 2, Kinshasa. Household food insecurity was measured using the Household Food Insecurity Access Scale (HFIAS), dietary diversity was assessed using the Minimum Dietary Diversity for Women (MDD_W; inadequate ≤ 5 food groups in 24 h), and probable depression was assessed using the Hopkins Symptom Checklist-10 (HSCL-10), which is a validated screening tool. We obtained baseline BMIs from clinic records at ART induction, which we measured again upon survey completion. We used analysis of covariance (ANCOVA) to model follow-up BMI, adjusting for baseline values, age, ART duration, self-reported adherence, household food insecurity, dietary diversity, and probable depression. Sensitivity analyses included change-score and mixed-effects models. Results: The prevalence of any household food insecurity was high (75%; 95% CI:71.5–78.6), with 57.6% (95% CI:53.5–61.6) of the participants experiencing inadequate dietary diversity (MDD_W < 5). Furthermore, forty-two per cent (95% CI:38.4–46.5) experienced depressive symptoms and sixty-eight percent (95% CI: 64.4–72.0) adhered to antiretroviral therapy (ART). The mean MDD_W was 4.3, with a low consumption rate of animal-source foods. Baseline BMI was associated with follow-up values (adjusted βunstandardized, 0.48 kg/m2 per 1 kg/m2 baseline, 95% CI 0.38–0.59; p < 0.001). Probable depression was independently associated with a lower follow-up BMI (adjusted βunstandardized, −0.99 kg/m2; 95% CI −1.72 to −0.26; p = 0.008). Time since ART initiation showed a slight positive association with BMI (adjusted βunstandardized, 0.10 kg/m2 per year). Self-reported ART adherence, household food insecurity, and dietary diversity were not independently associated with follow-up BMI in fully adjusted models. The interaction between age and probable depression did not suggest heterogeneity between age groups (p = 0.503). Conclusions: In our cohort, food insecurity and poor dietary diversity were widespread but did not significantly correlate with BMI, while probable depression, a potentially modifiable factor, was independently associated with lower BMI after accounting for baseline nutritional status. These findings highlight the need for HIV care programs integrating mental health screening and services with nutrition-sensitive interventions to support recovery and long-term health among WLHIV. Full article
(This article belongs to the Section Nutrition and Public Health)
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21 pages, 13748 KB  
Article
Integrated Assessment of Anthropogenic Carbon, Nitrogen, and Phosphorus Inputs: A Panjin City Case Study
by Tianxiang Wang, Simiao Wang, Li Ye, Guangyu Su, Tianzi Wang, Rongyue Ma and Zipeng Zhang
Water 2025, 17(20), 2962; https://doi.org/10.3390/w17202962 (registering DOI) - 15 Oct 2025
Abstract
Energy consumption and environmental pollution pose significant challenges to sustainable development. This study develops a comprehensive coupled framework model that advances the quantitative integration of carbon (C), nitrogen (N), and phosphorus (P) cycles driven by multiple anthropogenic pollution sources. This paper used Panjin [...] Read more.
Energy consumption and environmental pollution pose significant challenges to sustainable development. This study develops a comprehensive coupled framework model that advances the quantitative integration of carbon (C), nitrogen (N), and phosphorus (P) cycles driven by multiple anthropogenic pollution sources. This paper used Panjin city as a case study to analyze the dynamic changes and interconnections among C, N, and P. Results indicated that net anthropogenic carbon inputs (NAIC) increased by 33% from 2016–2020, while net anthropogenic nitrogen inputs (NAIN) and net anthropogenic phosphorus inputs (NAIP) decreased by 14% and 28%, respectively. The primary driver of NAIC was energy consumption, while wetlands were the dominant carbon sequestration sink. Agricultural production was identified as the primary source of NAIN and NAIP, and approximately 4.5% of NAIN and 2.9% of NAIP were discharged into receiving water bodies. We demonstrate that human activities and natural processes exhibit dual attributes, producing positive and negative environmental effects. The increase in carbon emissions drives economic growth and industrial restructuring; however, the enhanced economic capacity also strengthens the ability to mitigate pollution through environmental protection measures. Similarly, natural ecosystems, including forests and grasslands, contribute to carbon sequestration and the release of non-point source pollution. The comprehensive environmental impact assessment of C, N, and P revealed that the comprehensive environmental index for Panjin city exhibited an improved trend. The factors of energy structure, energy efficiency, and economic scale promoted NAIC growth, with the economic scale factor alone accounting for 93% of the total increment. Environmental efficiency factor and population size factor were the primary drivers in reducing NAIN and NAIP discharges into the receiving water bodies. We propose a novel management model, ecological restoration, clean energy utilization, resource recycling, and pollution source reduction to achieve systemic governance of C, N, and P inputs. Full article
(This article belongs to the Special Issue Science and Technology for Water Purification, 2nd Edition)
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23 pages, 3750 KB  
Article
CaP Nanoparticles Improve the Effect of dsRNA on Gene Expression, Growth, and Mycotoxin Production of Toxigenic Fusarium graminearum
by Alexander A. Stakheev, Polina Bagdasarova, Eugene A. Rogozhin, Victoria Tikhomirova, Ekaterina Popova, Assel Akhmetova, Olga Kost, Natalia O. Kalinina, Michael Taliansky and Sergey K. Zavriev
Int. J. Mol. Sci. 2025, 26(20), 10021; https://doi.org/10.3390/ijms262010021 (registering DOI) - 15 Oct 2025
Abstract
Fusarium species and the mycotoxins produced by them represent a significant problem for agriculture and human health. Thus, the development of novel management strategies and tools is of high importance. Spray-induced gene silencing (SIGS), based on the natural mechanism of RNA interference (RNAi), [...] Read more.
Fusarium species and the mycotoxins produced by them represent a significant problem for agriculture and human health. Thus, the development of novel management strategies and tools is of high importance. Spray-induced gene silencing (SIGS), based on the natural mechanism of RNA interference (RNAi), has been considered as a highly specific and ecologically safe alternative to chemical fungicides, the use of which is restricted by the emergence of resistant strains and environmental concerns. At the same time, massive application of SIGS is challenged by the degradability of RNA molecules in the environment. Nanoparticles have been widely applied to protect RNA from degradation and improve its action. The aims of this study were to evaluate whether RNAi-mediated silencing of the regulatory FgVe1 gene leads to inhibition of growth, mycotoxin production, and pathogenicity of Fusarium graminearum and whether the use of CaP nanoparticles (CaPs) as double-stranded RNA (dsRNA) carriers enhances and prolongs the silencing effect. It was shown that dsRNA treatment of fungal liquid cultures resulted in 19.78-fold silencing of FgVe1 expression as well as inhibition of expression of genes related to secondary metabolism, including those involved in trichothecene and aurofusarin biosynthesis, thus leading to a reduction in DON accumulation and changes in culture color. The results also demonstrated that naked dsRNA and CaPs:dsRNA nanocomplexes differed in their abilities to induce a high silencing effect at different time points. Naked dsRNA proved more effective in inducing silencing in the early stages of fungal growth, whereas application of nanocomplexes provided a prolonged effect up to 10 days in liquid cultures and up to 14 days on detached leaves. The obtained data can be considered as a basis for the further development of new efficient SIGS-based plant protection strategies. Full article
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20 pages, 2132 KB  
Article
Research on the Influencing Factors of the Cropland Abandonment Behavior of Different Typical Types of Farming Households: Based on a Survey in Mountainous Areas
by Yingbin Feng, Jingjing Li and Dedong Feng
Land 2025, 14(10), 2057; https://doi.org/10.3390/land14102057 (registering DOI) - 15 Oct 2025
Abstract
Cropland abandonment (CA) is a critical environmental issue globally, with balancing food security and ecological protection vital for sustainable development. This study explores CA behavior differences and drivers between out-of-poverty farming households (OPFHs) and non-poverty farming households (NPFHs) in China′s mountainous areas, using [...] Read more.
Cropland abandonment (CA) is a critical environmental issue globally, with balancing food security and ecological protection vital for sustainable development. This study explores CA behavior differences and drivers between out-of-poverty farming households (OPFHs) and non-poverty farming households (NPFHs) in China′s mountainous areas, using stepwise regression on survey data from 321 households in Liping County, Guizhou. The results show that: (1) The differences in CA behaviors between the two types of farming households are mainly reflected at the farmer level and the plot level. Plot integrity is a common influencing factor of CA areas for both types of farming households. (2) The driving factors affecting the area of CA by OPFHs also include the average age of the labor force, the proportion of the resident population in the total household registration population, and plot type, while the drivers affecting the area of CA by NPFHs include per capita income, non-agricultural income, per capita cropland area, and commuting time. (3) The differences in CA behavior and its driving factors between OPFHs and NPFHs in mountainous areas are characterized by diversity and interaction. Based on the results of the study, we propose the management of farming households and cropland, which can contribute to rural revitalization in China and the world, to a certain extent. Full article
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17 pages, 1447 KB  
Article
Optimization of Loop-Mediated Isothermal Amplification for Avian Influenza Detection
by Anastasia Glazunova, Timofey Sevskikh, Dmitry Kudryashov, Irina Sindryakova, Olga Kolbasova, Maria Erokhina, Andrey Mukhin, Denis Kolbasov and Ilya Titov
Animals 2025, 15(20), 2983; https://doi.org/10.3390/ani15202983 (registering DOI) - 15 Oct 2025
Abstract
Avian influenza viruses (AIV) cause severe economic losses in poultry production and pose zoonotic threats, necessitating rapid, field-deployable diagnostics. While real-time PCR is the gold standard, its use is limited in resource-constrained settings. This study aimed to develop and validate optimized loop-mediated isothermal [...] Read more.
Avian influenza viruses (AIV) cause severe economic losses in poultry production and pose zoonotic threats, necessitating rapid, field-deployable diagnostics. While real-time PCR is the gold standard, its use is limited in resource-constrained settings. This study aimed to develop and validate optimized loop-mediated isothermal amplification (LAMP) protocols for AIV detection directly at sample collection sites. We optimized Real-Time RT-LAMP and colorimetric LAMP assays targeting the conserved M gene, using primers described in the literature. Analytical sensitivity was assessed with a plasmid standard (106–100 copies/μL); specificity was evaluated against 27 AIV strains (H1–H12) and heterologous avian viruses (Newcastle disease, infectious bronchitis, Gumboro, and laryngotracheitis viruses). Reverse transcription was integrated into the LAMP reaction. Real-Time LAMP with SYBR Green achieved 100% analytical sensitivity (95% CI: 80–100; detection limit: Ct 38), while colorimetric LAMP (cresol red, malachite green, calcein) detected 102 plasmid copies (Ct 32) with 91.67% sensitivity (95% CI: 76.1–100). No cross-reactivity occurred. These optimized LAMP protocols offer sensitivity and specificity comparable to PCR, require minimal equipment, and enable rapid AIV screening, significantly enhancing early detection and epidemiological surveillance in field conditions. Full article
(This article belongs to the Section Poultry)
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16 pages, 2875 KB  
Article
Clarification of Copper Sulfide Precipitates by Polymeric Microfiltration Membranes
by Michelle Quilaqueo, Nicolás Barraza, Lorena Barros, Karla Pérez, René Ruby-Figueroa, Elizabeth Troncoso and Humberto Estay
Processes 2025, 13(10), 3292; https://doi.org/10.3390/pr13103292 (registering DOI) - 15 Oct 2025
Abstract
The recovery of copper from metallurgical effluents is critical for advancing sustainable mining and circular economy practices. This study evaluated a hybrid process combining copper sulfide precipitation with clarification using polymeric polyvinylidene fluoride (PVDF) microfiltration membranes. Laboratory-scale experiments were performed under controlled cyanide [...] Read more.
The recovery of copper from metallurgical effluents is critical for advancing sustainable mining and circular economy practices. This study evaluated a hybrid process combining copper sulfide precipitation with clarification using polymeric polyvinylidene fluoride (PVDF) microfiltration membranes. Laboratory-scale experiments were performed under controlled cyanide conditions (100 mg/L free CN, 1800 mg/L Cu2+), focusing on permeate flux behavior, fouling mechanisms, and cleaning strategies. Optimal performance was achieved at moderate transmembrane pressures (<2.0 bar) and higher flow rates, which provided a balance between productivity and fouling control. Flux decline was attributed to a combination of pore blocking and cake layer formation, confirming the multifactorial nature of fouling dynamics. Cleaning tests revealed that oxidizing solutions (HCl + H2O2) restored up to 96% of the initial permeability, while combined treatments with NaCN achieved complete recovery (>100%), albeit with potential risks of membrane aging under prolonged exposure. A techno-economic assessment comparing polymeric and ceramic membranes revealed similar capital and operational costs, with polymeric membranes offering slight reductions in CAPEX (10%) and OPEX (2.3%). Overall, the findings demonstrate the technical feasibility and economic competitiveness of polymeric membranes for copper sulfide clarification, while emphasizing the need to improve long-term chemical resistance to ensure reliable industrial-scale implementation. Full article
(This article belongs to the Section Separation Processes)
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26 pages, 1100 KB  
Review
Assessment of Dental Fear and Anxiety Tools for Children: A Review
by Mohammed Barry, Mustafa Alnami, Yazeed Thamer Alshobaili, Osama M. Felemban and Heba Jafar Sabbagh
Healthcare 2025, 13(20), 2597; https://doi.org/10.3390/healthcare13202597 (registering DOI) - 15 Oct 2025
Abstract
Background and Objectives: Accurate assessment of Dental fear and anxiety (DFA) is crucial for effective management, yet the vast array of measurement tools presents a dilemma for clinicians and researchers in selecting appropriate methods. This literature review aims to provide a comprehensive review [...] Read more.
Background and Objectives: Accurate assessment of Dental fear and anxiety (DFA) is crucial for effective management, yet the vast array of measurement tools presents a dilemma for clinicians and researchers in selecting appropriate methods. This literature review aims to provide a comprehensive review and comparison of the commonly used DFA measurement tools in the pediatric dentistry literature. Methods: A Comprehensive literature search was conducted in December 2024 using the electronic databases PubMed, MEDLINE, ScienceDirect, and Google Scholar. The search was limited to studies focused on identifying research relevant to DFA in children. For each DFA tool, information on its structure, validity, reliability, strengths, limitations, and target population was recruited and tabulated. A comparison between the DFA tools was then conducted. Quality assessment was performed using Patient-Reported Outcome Measures (PRO). Results: The search identified 15 subjective and 5 objective tools. Subjective tools included self-reported scales and pictorial analogs, while objective measures involved physiological monitoring and behavioral observation. While subjective tools offer valuable insights into a child’s self-perceived anxiety, their applicability is influenced by age and cognitive development. Objective measures provide quantifiable data, but require specialized equipment and trained observers. Tools combining simplicity, visual aids, and robust validation were found to be most practical for clinical use. Conclusions: A wide range of valid and reliable tools are available to assess DFA in children. Selection should be tailored to the child’s age, cognitive abilities, clinical setting, and clinician training. Combining subjective and objective assessments may enhance diagnostic accuracy. Full article
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16 pages, 9307 KB  
Article
Projected Heat-Stress in Sheep and Cattle in Greece Under Future Climate Change Scenarios
by Dimitris K. Papanastasiou, Athanasios I. Gelasakis, Georgios Papadopoulos, Dimitrios Melas, Kostas Douvis, Ioannis Faraslis, Stavros Keppas, Ioannis Stergiou, Anastasia Poupkou, Dimitris Voloudakis, Athena Progiou, John Kapsomenakis and Nikolaos Katsoulas
Agriculture 2025, 15(20), 2141; https://doi.org/10.3390/agriculture15202141 (registering DOI) - 15 Oct 2025
Abstract
It is well established that exposure to heat-stress conditions significantly impacts the physiology, health, welfare, and productivity of both sheep and cattle. The aim of this study was to apply the Temperature Humidity Index (THI) in order to assess the impact of future [...] Read more.
It is well established that exposure to heat-stress conditions significantly impacts the physiology, health, welfare, and productivity of both sheep and cattle. The aim of this study was to apply the Temperature Humidity Index (THI) in order to assess the impact of future climate conditions on the thermal stress exposure of sheep and cattle in Greece. The Weather Research and Forecasting (WRF) model was used as a high-resolution regional climate model to simulate climate conditions for two decades in Greece at a 10 Km spatial resolution and a 1 h temporal resolution. The WRF model was applied to two emission scenarios, namely SSP2-4.5 (intermediate) and SSP5-8.5 (worst-case). Projections were made for the near-future decade (2046–2055), with the decade (2005–2014) serving as the reference period for comparative analysis. The data analysis indicated that under the SSP2-4.5 emission scenario, the mean temperature is projected to increase by 1.2–1.4 °C and 1.4–1.6 °C across 38% and 58% of the country’s territory, respectively. Increases higher than 1.6 °C are projected across 32% of the Greek territory under the SSP5-8.5 emission scenario. The mean THI (sheep) and mean THI (adj) (cattle) are projected to increase by 5–10% and by 4% across 74% and 82% of the Greek territory, respectively, when considering the SSP2-4.5 emission scenario. Slightly more severe mean heat-stress conditions were projected when considering the SSP5-8.5 emission scenario. The analysis of the hourly THI values showed that sheep and cattle are expected to experience heat-stress conditions during extended periods in the future, in which hot weather will prevail. Specifically, the number of severe/danger heat-stress hours is projected to double in the greater part of the country. To mitigate the adverse effects of climate-change-induced thermal stress on animal productivity, health, and welfare, the implementation of adaptation measures and best management practices is strongly recommended for sheep and cattle farmers. These measures encompass improvements in breeding strategies, livestock housing and microclimate management, nutritional interventions, and the adoption of precision livestock farming technologies. Given the outstanding economic, social, and environmental importance of sheep and cattle farming in Greece, effective adaptation to and mitigation of climate change impacts represent urgent priorities to ensure the long-term sustainability and resilience of the livestock sector. Full article
(This article belongs to the Special Issue The Threats Posed by Environmental Factors to Farm Animals)
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21 pages, 1489 KB  
Article
Effects of Waterlogging at Different Developmental Stages on Growth, Yield and Physiological Responses of Forage Maize
by Chang-Woo Min, Il-Kyu Yoon, Min-Jun Kim, Jeong-Sung Jung, Md Atikur Rahman and Byung-Hyun Lee
Agronomy 2025, 15(10), 2389; https://doi.org/10.3390/agronomy15102389 (registering DOI) - 15 Oct 2025
Abstract
Waterlogging (WL) is an abiotic stress that severely limits crop yield. However, limited research has addressed the effects of long-term WL stress at different developmental stages on the yield and physiological responses of forage maize. In this study, forage maize plants were subjected [...] Read more.
Waterlogging (WL) is an abiotic stress that severely limits crop yield. However, limited research has addressed the effects of long-term WL stress at different developmental stages on the yield and physiological responses of forage maize. In this study, forage maize plants were subjected to 14-day WL stress at the emergence (E), four-leaf (V4), eleven-leaf (V11), and tasseling (VT) stages. Plant height significantly decreased by 60% at the E stage and 48% at the V4 stage when exposed to 14-day WL. Leaf area decreased by 79% at the E stage, and the number of green leaves decreased most significantly at the VT stage. Chlorophyll fluorescence (Fv/Fm) and the relative chlorophyll content index (RCI) decreased most significantly at the V4 stage. The lysigenous aerenchyma formation rate of the roots increased significantly after 14-day WL at the V4 stage, whereas the number of adventitious roots increased most significantly at the V11 stage. The hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents, which are indicative of the root oxidation state, exhibited the highest increase at the E stage. In addition, at the E and V4 stages, the expression of genes related to energy metabolism and lysigenous aerenchyma formation in the roots was upregulated after 14-day WL. The total dry matter (DM) of maize after harvest decreased most significantly when exposed to 14-day WL at the V4 stage, while acid detergent fiber (ADF) and neutral detergent fiber (NDF) increased with the developmental stages. Consequently, total digestible nutrients (TDNs) and the relative feed value (RFV) decreased with advancing developmental stages, with the highest decrease at the VT stage. These results demonstrate that effective drainage management during the early developmental stage (V4) is more important to prevent forage maize yield loss due to prolonged WL stress, which is expected to increase in frequency due to climate change, and management during the later developmental stage (VT) is critical to prevent decreases in feed values. These findings provide valuable insights into the physiological responses of forage maize to WL stress. Full article
(This article belongs to the Section Crop Breeding and Genetics)
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19 pages, 7895 KB  
Article
SpiKon-E: Hybrid Soft Artificial Muscle Control Using Hardware Spiking Neural Network
by Florian-Alexandru Brașoveanu, Mircea Hulea and Adrian Burlacu
Biomimetics 2025, 10(10), 697; https://doi.org/10.3390/biomimetics10100697 (registering DOI) - 15 Oct 2025
Abstract
Artificial muscles play a key role in the future of humanoid robotics and medical devices, with research on wire-driven joints leading the field. While electric servo motors were once at the forefront, the focus has shifted toward materials that react to changes in [...] Read more.
Artificial muscles play a key role in the future of humanoid robotics and medical devices, with research on wire-driven joints leading the field. While electric servo motors were once at the forefront, the focus has shifted toward materials that react to changes in the environment (smart materials), including pneumatic silicone actuators and temperature-reactive metallic alloys, aiming to replicate human muscle actuation for improved performance. Initially designed for rigid actuators, control strategies were adapted to address the unique dynamics of artificial muscles. Although current controllers offer satisfactory performance, further optimization is necessary to mimic natural muscle control more rigorously. This study details the design and implementation of a novel system that mimics biological muscle. This system is designed to replicate the full range of motion and control functionalities, which can be utilized in various applications. This research has three significant contributions in the field of sustainable soft robotics. First, a novel shape memory alloy-based linear actuator is introduced, which achieves significantly higher displacements compared to traditional SMA wire-driven systems through a guiding mechanism. Second, this linear actuator is integrated into a hybrid soft actuation structure, which features a silicone PneuNet as the end effector and a force sensor for real-time pressure feedback. Lastly, a hardware Spiking Neural Network (HW-SNN) is utilized to control the exhibited force at the actuator’s endpoint. Experimental results showed that the displacement with the control system is significantly higher than that of the traditional control-based shape memory alloy systems. The system evaluation demonstrates good performance, thus advancing actuation and control in humanoid robotics. Full article
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37 pages, 8931 KB  
Article
Predicting the Properties of Polypropylene Fiber Recycled Aggregate Concrete Using Response Surface Methodology and Machine Learning
by Hany A. Dahish and Mohammed K. Alkharisi
Buildings 2025, 15(20), 3709; https://doi.org/10.3390/buildings15203709 (registering DOI) - 15 Oct 2025
Abstract
The use of recycled coarse aggregate (RCA) concrete and polypropylene fibers (PPFs) presents a sustainable alternative in concrete production. However, the non-linear and interactive effects of RCA and PPF on both fresh and hardened properties are not yet fully quantified. This study employs [...] Read more.
The use of recycled coarse aggregate (RCA) concrete and polypropylene fibers (PPFs) presents a sustainable alternative in concrete production. However, the non-linear and interactive effects of RCA and PPF on both fresh and hardened properties are not yet fully quantified. This study employs Response Surface Methodology (RSM) and the Random Forest (RF) algorithm with K-fold cross-validation to predict the combined effect of using recycled coarse aggregate (RCA) as a partial replacement for natural coarse aggregate and polypropylene fiber (PPF) on the engineering properties of RCA-PPF concrete, addressing the critical need for a robust, data-driven modeling framework. A dataset of 144 tested samples obtained from literature was utilized to develop and validate the prediction models. Three input variables were considered in developing the proposed prediction models, namely, RCA, PPF, and curing age (Age). The examined responses were compressive strength (CS), tensile strength (TS), ultrasonic pulse velocity (UPV), and water absorption (WA). To assess the developed models, statistical metrics were calculated, and analysis of variance (ANOVA) was employed. Afterwards, the responses were optimized using optimization in RSM. The optimal results of responses by maximizing TS, CS, and UPV and minimizing WA were achieved at a PPF of 3% by volume of concrete and an RCA of approximately 100% replacing natural coarse aggregate, highlighting optimal reuse of recycled aggregate, with an AGE of 83.6 days. The RF model demonstrated superior performance, significantly outperforming the RSM model. Feature importance analysis via SHAP values was employed to identify the most effective parameters on the predictions. The results confirm that ML techniques provide a powerful and accurate tool for optimizing sustainable concrete mixes. Full article
(This article belongs to the Section Building Structures)
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10 pages, 1892 KB  
Article
The Complete Chloroplast Genome of Camellia tianeensis (Camellia L.) and Phylogenetic Relationships with Other Plants of the Genus Camellia
by Juyan Chen, He Li and Lunxiu Deng
Genes 2025, 16(10), 1217; https://doi.org/10.3390/genes16101217 (registering DOI) - 15 Oct 2025
Abstract
Background/Objectives: Species within section Chrysantha represent the only camellias known to produce golden-yellow petals. The primary objectives of this study were to characterize the chloroplast genome structure of Camellia tianeensis and to elucidate its phylogenetic position with sect. Chrysantha. Methods: The complete [...] Read more.
Background/Objectives: Species within section Chrysantha represent the only camellias known to produce golden-yellow petals. The primary objectives of this study were to characterize the chloroplast genome structure of Camellia tianeensis and to elucidate its phylogenetic position with sect. Chrysantha. Methods: The complete chloroplast genome of C. tianeensis was sequenced, assembled, and annotated. Phylogenetic inference was conducted using maximum likelihood and Bayesian methods based on complete chloroplast genomic sequences. Results: The chloroplast genome of C. tianeensis is 156,865 bp in length and exhibits a typical quadripartite structure consisting of a large single-copy (LSC) region (86,579 bp), a small single-copy (SSC) region (18,236 bp), and two inverted repeat (IR) regions (26,025 bp each). The genome encodes 164 genes, including 111 protein-coding genes, 45 tRNAs, and 8 rRNA genes. The overall GC content was 37.32%, with regional values of 35.33% (LSC), 30.59% (SSC), and 42.99% (IRs). Sixty-nine simple sequence repeats (SSRs) were identified, predominantly mononucleotide repeats, Thirty-eight dispersed repeats were categorized into three types (forward, reverse, and palindromic), with no complement repeats detected. Phylogenetic analysis strongly supported that C. tianeensis is a member within sect. Chrysantha. Conclusions: C. tianeensis is phylogenetically closely related to C. huana, forming a well-supported clade. This study enhances the molecular research available for sect. Chrysantha and provides a genomic foundation for future phylogenetic and taxonomic studies in this group. Full article
(This article belongs to the Section Plant Genetics and Genomics)
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13 pages, 3455 KB  
Article
Genome-Wide Identification and Analysis of the Fatty Acid Export Family Revealed the Role of GmFAX8 in Improving Soybean Oil Accumulation
by Yan Zhang, Yina Zhu, Xiuli Rui, Yuan Li, Jie Wang, Yuhang Zhan, Yongguang Li, Xue Zhao, Yingpeng Han and Xunchao Zhao
Plants 2025, 14(20), 3166; https://doi.org/10.3390/plants14203166 (registering DOI) - 15 Oct 2025
Abstract
Fatty acid transporters (FAXs) play an important role in fatty acid synthesis by facilitating transport fatty acids from the plastid to the endoplasmic reticulum. This process is essential for providing precursor substances necessary for triglycerides (TAGs). Although FAX genes have been identified in [...] Read more.
Fatty acid transporters (FAXs) play an important role in fatty acid synthesis by facilitating transport fatty acids from the plastid to the endoplasmic reticulum. This process is essential for providing precursor substances necessary for triglycerides (TAGs). Although FAX genes have been identified in variety of plant species, the identification and molecular functions of the GmFAX gene members in soybean are still unclear. In this study, soybean FAX genes were identified through the utilization of the Phytozome (v13) and NCBI online websites. Subsequently, phylogenetic trees, expression patterns, gene structures, and qRT-PCR were analyzed. A total of eight GmFAX members were identified at the whole genome level, and further phylogenetic analysis revealed that these members can be categorized into four subfamilies. In addition, all members of GmFAX contain a highly conserved domain Tmemb_14. Through qRT-PCR analysis, it was found that the expression level of the GmFAX8 gene is relatively high in leaves and stems. Further investigation revealed that the total fatty acid content in hairy roots overexpressing the GmFAX8 gene was significantly greater than that observed in the control strain. The results presented above suggest that the GmFAX8 gene may play an important role in the accumulation of oil within soybeans. Full article
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26 pages, 7247 KB  
Article
DyslexiaNet: Examining the Viability and Efficacy of Eye Movement-Based Deep Learning for Dyslexia Detection
by Ramis İleri, Çiğdem Gülüzar Altıntop, Fatma Latifoğlu and Esra Demirci
J. Eye Mov. Res. 2025, 18(5), 56; https://doi.org/10.3390/jemr18050056 (registering DOI) - 15 Oct 2025
Abstract
Dyslexia is a neurodevelopmental disorder that impairs reading, affecting 5–17.5% of children and representing the most common learning disability. Individuals with dyslexia experience decoding, reading fluency, and comprehension difficulties, hindering vocabulary development and learning. Early and accurate identification is essential for targeted interventions. [...] Read more.
Dyslexia is a neurodevelopmental disorder that impairs reading, affecting 5–17.5% of children and representing the most common learning disability. Individuals with dyslexia experience decoding, reading fluency, and comprehension difficulties, hindering vocabulary development and learning. Early and accurate identification is essential for targeted interventions. Traditional diagnostic methods rely on behavioral assessments and neuropsychological tests, which can be time-consuming and subjective. Recent studies suggest that physiological signals, such as electrooculography (EOG), can provide objective insights into reading-related cognitive and visual processes. Despite this potential, there is limited research on how typeface and font characteristics influence reading performance in dyslexic children using EOG measurements. To address this gap, we investigated the most suitable typefaces for Turkish-speaking children with dyslexia by analyzing EOG signals recorded during reading tasks. We developed a novel deep learning framework, DyslexiaNet, using scalogram images from horizontal and vertical EOG channels, and compared it with AlexNet, MobileNet, and ResNet. Reading performance indicators, including reading time, blink rate, regression rate, and EOG signal energy, were evaluated across multiple typefaces and font sizes. Results showed that typeface significantly affects reading efficiency in dyslexic children. The BonvenoCF font was associated with shorter reading times, fewer regressions, and lower cognitive load. DyslexiaNet achieved the highest classification accuracy (99.96% for horizontal channels) while requiring lower computational load than other networks. These findings demonstrate that EOG-based physiological measurements combined with deep learning offer a non-invasive, objective approach for dyslexia detection and personalized typeface selection. This method can provide practical guidance for designing educational materials and support clinicians in early diagnosis and individualized intervention strategies for children with dyslexia. Full article
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24 pages, 3583 KB  
Article
Explainable Model of Hybrid Ensemble Learning for Prostate Cancer RNA-Seq Classification via Targeted Feature Selection
by Ahmet Demiröz and Nesrin Aydın Atasoy
Electronics 2025, 14(20), 4050; https://doi.org/10.3390/electronics14204050 (registering DOI) - 15 Oct 2025
Abstract
High dimensional, small sample ribonucleic acid sequencing (RNA-seq) data pose a major challenge for reliable classification due to the curse of dimensionality and the risk of overfitting. This study addresses that challenge for prostate cancer by coupling machine learning (ML) based feature selection [...] Read more.
High dimensional, small sample ribonucleic acid sequencing (RNA-seq) data pose a major challenge for reliable classification due to the curse of dimensionality and the risk of overfitting. This study addresses that challenge for prostate cancer by coupling machine learning (ML) based feature selection with a hybrid ensemble classifier. RNA-seq datasets retrieved from the University of California Santa Cruz (UCSC) Xena platform have been pre-processed, and the number of features has been reduced to 30 through systematic feature selection. Three complementary learners have been combined into a majority voting ensemble to improve robustness and generalization. Performance has been assessed using evaluation metric criteria and the area under the curve. The results have been corroborated on liver, lung, and thyroid cancer datasets from the literature. The proposed hybrid ensemble method has been achieved 97.82% accuracy with Lasso feature selection. Compared with conventional single model approaches, the proposed hybrid model has yielded accurate and reliable predictions. Additionally, biologically meaningful information has been presented using explainable artificial intelligence techniques regarding the importance of genes. These findings suggest that the joint use of feature selection and hybrid ensembles provides a practical and interpretable framework for classifying high dimensional genomic profiles under limited sample sizes. Full article
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10 pages, 210 KB  
Article
Use of Videos as Disability Educational Tools for Medical Students
by Ovuokerie Addoh, Alyssa Axelrod, Caroline Gormley, Sarah Mahasin, Brionn Tonkin, Alexander Senk and Erica Seidel
Int. Med. Educ. 2025, 4(4), 41; https://doi.org/10.3390/ime4040041 (registering DOI) - 15 Oct 2025
Abstract
People with disabilities (PWDs) face unique challenges in accessing equitable medical care, due in part to physicians’ limited training in disability-related competencies. This study explored a novel approach to bridging this gap by integrating selected videos and reflective essays into medical education. Third- [...] Read more.
People with disabilities (PWDs) face unique challenges in accessing equitable medical care, due in part to physicians’ limited training in disability-related competencies. This study explored a novel approach to bridging this gap by integrating selected videos and reflective essays into medical education. Third- and fourth-year medical students watched four videos depicting diverse experiences of PWDs and submitted reflective essays, resulting in the analysis of 90 essays by six investigators. Through thematic analysis, 50 concept codes were distilled into six key themes: recovery process, student insight, adjustment to disability, patient-doctor relationship, social model of disability, and support systems. The reflections revealed a growing awareness among students of the need to individualize care and address systemic barriers, though some medicalized views of disability were noted. These findings underscore the potential of video-based education to enhance disability competence in medical training by fostering critical reflection and empathy. However, to maximize impact, educators should prioritize diverse portrayals of disability and challenge sensationalized or “overcoming” narratives common in such media. Overall, this study demonstrates that widely available media can be a valuable, scalable tool for advancing disability education in medical curricula. Full article
18 pages, 1382 KB  
Review
Microbiome–Aging–Wrinkles Axis of Skin: Molecular Insights and Microbial Interventions
by Varun Challa, Santosh Kumar Prajapati, Surabhi Gangani, Dhananjay Yadav, Lalitha Lekkala, Shalini Jain and Hariom Yadav
Int. J. Mol. Sci. 2025, 26(20), 10022; https://doi.org/10.3390/ijms262010022 (registering DOI) - 15 Oct 2025
Abstract
Skin aging is a complex biological process influenced by both intrinsic factors such as hormonal changes, genetic programming, and immunosenescence and extrinsic stressors including ultraviolet (UV) radiation (particularly UV-A and UV-B), pollution, and lifestyle habits. One of the most prominent manifestations of skin [...] Read more.
Skin aging is a complex biological process influenced by both intrinsic factors such as hormonal changes, genetic programming, and immunosenescence and extrinsic stressors including ultraviolet (UV) radiation (particularly UV-A and UV-B), pollution, and lifestyle habits. One of the most prominent manifestations of skin aging is wrinkle formation, which arises from the progressive degradation of key extracellular matrix (ECM) components like collagen and elastin. Emerging evidence highlights the skin microbiome as a critical, yet underappreciated, modulator of these structural changes. This review summarizes current understanding of how aging alters skin structure and microbial composition, and how these changes contribute to wrinkle development. Age-associated skin is characterized by reduced hydration, sebum production, and barrier integrity, accompanied by a shift in microbial communities. These microbial shifts promote local inflammation, matrix metalloproteinase (MMP) activation, and oxidative stress, all of which accelerate ECM degradation. We further discuss how commensal microbes and their bioactive products such as probiotics and postbiotics can counteract wrinkle formation. Clinical studies support the efficacy of strains such as Lactobacillus plantarum HY7714 and Bifidobacterium breve in improving skin elasticity and reducing wrinkle depth. Additionally, this review highlights the emerging role of microbiome-based interventions in skincare, including oral supplements, topical formulations, and postbiotic-enriched products. Overall, we emphasized the therapeutic potential of microbiome modulation as a novel strategy for maintaining skin health and preventing wrinkle formation during aging. Full article
(This article belongs to the Special Issue Microbiome Connections in Age-Related Diseases)
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5 pages, 178 KB  
Editorial
A Needle in a Haystack: Looking for Gaps in Treatment and Education in Emergency Medicine—Editorial
by Filip Jaskiewicz and Klaudiusz Nadolny
Healthcare 2025, 13(20), 2598; https://doi.org/10.3390/healthcare13202598 (registering DOI) - 15 Oct 2025
Abstract
When this Special Issue, “A Needle in a Haystack: Looking for Gaps in Treatment and Education in Emergency Medicine,” was first announced, its aim was to highlight the unique nature of emergency medicine as a discipline [...] Full article
22 pages, 2455 KB  
Article
Bioactive Potential of Protein Extracts Derived from Dried Wolffia globosa on In Vitro Antioxidant Activities and Pro-Inflammatory Cytokine Production
by Ruttiros Khonkarn, Krai Daowtak, Oranit Kraseasintra, Thitiya Luetragoon, Kanchana Usuwanthim, Kraisorn Taynawa and Kanokphon Chanphong
Molecules 2025, 30(20), 4092; https://doi.org/10.3390/molecules30204092 (registering DOI) - 15 Oct 2025
Abstract
This study investigated the contamination, composition, and functional properties of Wolffia globosa from northern Thailand. The results showed that the heavy metal content of dried W. globosa complied with Thai regulations, ensuring its safety. Its proximate analysis revealed high protein levels with lysine, [...] Read more.
This study investigated the contamination, composition, and functional properties of Wolffia globosa from northern Thailand. The results showed that the heavy metal content of dried W. globosa complied with Thai regulations, ensuring its safety. Its proximate analysis revealed high protein levels with lysine, leucine, and phenylalanine as the principal essential amino acids. The protein was effectively extracted using the alkaline extraction method, followed by precipitation induced by acid or heat. The precipitates and supernatants resulting from various acid- or heat-induced protein precipitation were obtained. The highest protein content was found in the pH 3 precipitate (51.15 ± 6.71%). In contrast, the pH 5 supernatant exhibited the most potent antioxidant activities (2.22 ± 0.05 mmol Trolox/mg and 4.55 ± 0.18 mmol Fe2+/mg), as determined by ABTS and FRAP assays, respectively. Additionally, a strong correlation was observed between phenolic content and antioxidant activity. Both supernatant and precipitate protein extracts from W. globosa exhibited no cytotoxicity in THP-1 cells and displayed anti-inflammatory effects by decreasing the production of IL-1β and IL-6. They also downregulated phospho-NF-κB, phospho-IκB-α, and COX-2, consistent with reduced NF-κB pathway activation. These findings position W. globosa as a promising, sustainable plant-based protein with bioactive and functional properties, making it a viable candidate for functional food formulations that enhance dietary health and add value to local agricultural resources. Full article
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15 pages, 3340 KB  
Article
Antifungal Drug Efficacy Profiles Against Vaginal Candida albicans: A Multi-Drug Comparative Analysis
by Mohammad Zubair and Yazeed Albalawi
J. Clin. Med. 2025, 14(20), 7266; https://doi.org/10.3390/jcm14207266 (registering DOI) - 15 Oct 2025
Abstract
Background: Candida albicans infects most reproductive-aged women, causing a prevalent infection known as Vulvovaginal Candida. As there has been an increase in resistance to widely used antifungal agents, particularly fluconazole, used in infections, local susceptibility profiles are needed to inform treatment [...] Read more.
Background: Candida albicans infects most reproductive-aged women, causing a prevalent infection known as Vulvovaginal Candida. As there has been an increase in resistance to widely used antifungal agents, particularly fluconazole, used in infections, local susceptibility profiles are needed to inform treatment options. Methods: This comparative observational study was carried out to determine the in vitro susceptibility of six antifungal compounds [fluconazole, voriconazole, itraconazole, ketoconazole, nystatin, and amphotericin B] to 163 vaginal Candida albicans isolates obtained in three hospitals in Tabuk, Saudi Arabia. MIC50, MIC90, and MFC values were calculated in Broth microdilution tests according to the standards of CLSI M27-A3. Friedman and Wilcoxon signed-rank tests were used to carry out statistical analysis. Results: It was observed that Amphotericin B and itraconazole recorded the lowest MIC and MFC, revealing better antifungal action. The worst performer was fluconazole with MIC50 (13.79 μg/mL), MIC90 (27.59 μg/mL), and MFC (37.93 μg/mL), and 85% resistance. It was found that there are significant differences between antifungal agents (p < 0.001), and amphotericin B and itraconazole always performed best compared to fluconazole and voriconazole. Conclusions: The results shows antifungal effectiveness as Amphotericin B and itraconazole are the most effective against vaginal Candida albicans isolates. There is a high rate of resistance to fluconazole, suggesting it should no longer be the first choice of treatment in this area. These findings highlight the need for local monitoring of drug resistance to guide treatment choices and emphasize the importance of using antifungals properly to prevent increased resistance. Full article
(This article belongs to the Section Pharmacology)
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11 pages, 1157 KB  
Systematic Review
Impact of Vaccinating Adult Women Who Are HPV-Positive or with Confirmed Cervical SIL with the 9-Valent Vaccine—A Systematic Review
by Dominik Pruski, Sonja Millert-Kalińska, Robert Jach, Jakub Żurawski and Marcin Przybylski
Viruses 2025, 17(10), 1377; https://doi.org/10.3390/v17101377 (registering DOI) - 15 Oct 2025
Abstract
Infection with oncogenic human papillomavirus (HPV) remains a leading cause of cervical cancer and other HPV-related diseases. This situation persists despite the availability of effective prophylactic vaccines. While global vaccination programs have significantly reduced the incidence of HPV in adolescents and young adults, [...] Read more.
Infection with oncogenic human papillomavirus (HPV) remains a leading cause of cervical cancer and other HPV-related diseases. This situation persists despite the availability of effective prophylactic vaccines. While global vaccination programs have significantly reduced the incidence of HPV in adolescents and young adults, many women presenting with HPV infection or squamous intraepithelial lesions (SIL) were not covered by primary prevention. This review was performed with the aim of evaluating the impact of administering the 9-valent HPV vaccine in adult women who are HPV-positive or have histologically confirmed cervical precancerous lesions. Following the PRISMA 2020 guidelines, a search was performed in the MEDLINE, Scopus, and Cochrane Library databases. A total of 653 studies were retrieved, of which 7 studies, including 19,414 women, met the inclusion criteria. According to the literature, vaccination was linked to significant reductions in persistent HPV infection, progression of SIL, and recurrence of high-grade lesions after surgical removal. Complete HPV remission was achieved in up to 72.4% of vaccinated women, compared to 45.7% among unvaccinated controls. Vaccination after conization lowered the recurrence risk of CIN2+ lesions by 87%, with benefits seen regardless of timing. The most significant effect was observed when vaccine administration was performed before the surgical procedure. Furthermore, HPV vaccination notably enhanced viral clearance and decreased the likelihood of repeated surgical interventions. Despite differences in study design and follow-up definitions, the overall evidence supports additional vaccination in HPV-positive adult women as an effective measure to reduce recurrence and promote viral remission. These findings emphasize the need for clear guidelines and wider access to HPV vaccination for adult populations. Full article
(This article belongs to the Special Issue Viral Infections in Gynecological Diseases)
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21 pages, 4695 KB  
Article
Comparative Proteomics Analysis Reveals Differential Immune Responses of Paralichthys olivaceus to Edwardsiella tarda Infection Under High and Low Temperature
by Xiaojuan Chen, Lejia Luo, Beibei Zhang, Xiaowei Zhou, Kaipeng Zhang, Panpan Zhang and Bin Sun
Biology 2025, 14(10), 1417; https://doi.org/10.3390/biology14101417 (registering DOI) - 15 Oct 2025
Abstract
Fluctuating water temperatures and bacterial pathogens such as Edwardsiella tarda pose a serious threat to mariculture, resulting in significant economic losses within the flounder industry. A previous study revealed that elevated temperature enhanced E. tarda dissemination in flounder tissues. However, the underlying mechanism [...] Read more.
Fluctuating water temperatures and bacterial pathogens such as Edwardsiella tarda pose a serious threat to mariculture, resulting in significant economic losses within the flounder industry. A previous study revealed that elevated temperature enhanced E. tarda dissemination in flounder tissues. However, the underlying mechanism has not been fully explained, especially the changes in protein level. In this study, label-free proteomics was utilized to investigate the impact of high temperature (23 °C) and low temperature (15 °C) on flounder immune response to E. tarda infection. Our results identified 317 differentially abundant proteins (DAPs) in the low-temperature group (LI-LC) and 302 DAPs in the high-temperature group (HI-HC). GO and KEGG analyses of DAPs revealed numerous immune-related proteins and pathways. Twenty-six key DAPs in the LI-LC group and twenty-seven key DAPs in the HI-HC group were further identified and formed extensive interaction networks, respectively. Through the analysis of key immune-related DAPs that were specifically identified in both groups via Venn diagram analysis, we demonstrated that the endocytosis capacity and complement activity were enhanced in the HI-HC group, while histone abundance and RNA transport function were, respectively, increased and severely interfered with in the LI-LC group. These findings highlight a clear divergence in the immune response of flounder to E. tarda infection between 15 °C and 23 °C, providing valuable insights into how temperature variation influences antibacterial immunity in fish. Full article
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13 pages, 280 KB  
Article
Entomopathogenic Nematodes and Bioactive Compounds of Their Bacterial Endosymbionts Act Synergistically in Combination with Spinosad to Kill Phthorimaea operculella (Zeller, 1873) (Lepidoptera: Gelechiidae), a Serious Threat to Food Security
by Ebubekir Yüksel, Rachid Lahlali, Aydemir Barış, Muhammad Sameeullah, Furkan Ulaş, Abdurrahman Sami Koca, Essaid Ait Barka, Mustafa İmren and Abdelfattah Dababat
Microorganisms 2025, 13(10), 2368; https://doi.org/10.3390/microorganisms13102368 (registering DOI) - 15 Oct 2025
Abstract
As a staple food, potato (Solanum tuberosum L.) (Solanaceae) is one of the most produced food crops to ensure food security. The potato tuber moth (PTM), Phthorimaea operculella (Zeller, 1873) (Lepidoptera: Gelechiidae), is a major pest of potato, damaging both the growing [...] Read more.
As a staple food, potato (Solanum tuberosum L.) (Solanaceae) is one of the most produced food crops to ensure food security. The potato tuber moth (PTM), Phthorimaea operculella (Zeller, 1873) (Lepidoptera: Gelechiidae), is a major pest of potato, damaging both the growing and storage processes. In recent years, green pest control strategies have been gaining importance to reduce the adverse effects of chemicals and protect the environment. Entomopathogenic nematodes (EPNs) and their bacterial endosymbionts (Xenorhabdus and Photorhabdus spp.) have been one of the top topics studied in sustainable pest control approaches. In the present study, the two most common EPN species, Steinernema feltiae and Heterorhabditis bacteriophora, and their bacterial associates, Xenorhabdus bovienii and Photorhabdus luminescens subsp. kayaii were evaluated against PTM larvae separately and in combination with spinosad. The survival rates of infective juveniles (IJs) of EPNs were over 92% after 72 h of direct exposure to spinosad. Co-application of EPNs and bioactive compounds (BACs) of endosymbiotic bacteria with spinosad induced synergistic interactions and achieved the maximum mortality (100%) in PTM larvae 48 h post-treatment. Spinosad and BAC combinations were highly efficient in controlling the PTM larvae and provided LT50 values below 23.0 h. Gas chromatography mass spectrometry (GC-MS) analysis identified 29 compounds in total, 20 of which belonged to P. luminescens subsp. kayaii. The results indicate that the integration of EPNs and BACs of endosymbiotic bacteria with spinosad presents a synergistic interaction and enhances pest control efficacy. Full article
14 pages, 1707 KB  
Article
Vaginal Microbiome and Functional Pathway Alterations in Preterm Premature Rupture of Membranes Revealed by 16S rRNA Sequencing
by Sangho Nam, Subeen Hong, In Yang Park and Sun Shin
Life 2025, 15(10), 1604; https://doi.org/10.3390/life15101604 (registering DOI) - 15 Oct 2025
Abstract
Preterm prelabor rupture of membranes (PPROM) is a leading cause of preterm birth and significant neonatal morbidity. The vaginal microbiome is implicated in its pathogenesis, but its detailed characteristics and functional consequences remain to be fully elucidated. This study aimed to provide a [...] Read more.
Preterm prelabor rupture of membranes (PPROM) is a leading cause of preterm birth and significant neonatal morbidity. The vaginal microbiome is implicated in its pathogenesis, but its detailed characteristics and functional consequences remain to be fully elucidated. This study aimed to provide a comprehensive, multi-faceted analysis of the vaginal microbiome and its functional potential in pregnant women with PPROM compared to healthy term controls. We collected vaginal fluid samples from eight PPROM and seven healthy control (HC) pregnant women. The vaginal microbiome was analyzed using 16S rRNA gene sequencing. We assessed community composition and state types (CSTs), alpha and beta diversity, co-occurrence networks, and predicted functional pathways using PICRUSt2. A molecular bacterial vaginosis (molBV) score was also calculated to determine the clinical relevance of the dysbiosis. The PPROM microbiome was characterized by a significant depletion of Lactobacillus crispatus–dominated communities (CST I) and a shift towards L. iners–dominated (CST III) or polymicrobial (CST IV) communities, which was consistent with a BV-positive molBV score. Alpha diversity was significantly higher in the PPROM group, and beta diversity analysis confirmed a distinct microbial structure between the two groups. Co-occurrence network analysis revealed a collapse of the protective, Lactobacillus-centered network in the PPROM group, which was replaced by a densely interconnected network of anaerobic bacteria with Gardnerella vaginalis as a key hub. Functionally, the PPROM microbiome was enriched for amino acid biosynthesis pathways, in contrast to the HC group, which was enriched for nucleotide and peptidoglycan biosynthesis. PPROM appears to be linked with a complex vaginal dysbiosis that encompasses significant alterations in microbial composition, diversity, interactions, and functional potential. These findings highlight the vaginal microbiome as a critical factor in the pathogenesis of PPROM and suggest its potential for risk stratification and as a therapeutic target to improve pregnancy outcomes. Full article
(This article belongs to the Special Issue The Emerging Role of Microbiota in Health and Diseases)
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15 pages, 421 KB  
Article
Occurrence and Molecular Characterization of Extended-Spectrum Beta-Lactamase (ESBL)-Producing Escherichia coli in Broilers in Indonesia
by Nur Hidayatullah, Imron Suandy, Montira Intanon, Thomas Alter, Oli Susanti, Ajeng Herpianti, Sani Susanty, Riska Desitania and Nattakarn Awaiwanont
Antibiotics 2025, 14(10), 1030; https://doi.org/10.3390/antibiotics14101030 (registering DOI) - 15 Oct 2025
Abstract
Extended-spectrum β-lactamase-producing Escherichia coli (ESBL-E. coli) are widespread in the food chain, but nationwide surveillance in Indonesian broiler production is limited. This study investigated the occurrence, antimicrobial resistance, phylogenetic diversity, and molecular characteristics of ESBL-E. coli from broilers in Indonesia. [...] Read more.
Extended-spectrum β-lactamase-producing Escherichia coli (ESBL-E. coli) are widespread in the food chain, but nationwide surveillance in Indonesian broiler production is limited. This study investigated the occurrence, antimicrobial resistance, phylogenetic diversity, and molecular characteristics of ESBL-E. coli from broilers in Indonesia. A total of 2182 E. coli isolates from broiler cecal samples across three regions during the period 2018–2020 were analyzed. Antimicrobial susceptibility testing and ESBL phenotyping were performed following the CLSI guidelines. ESBL resistance genes and phylogenetic groups were detected using multiplex/quadruplex PCR. ESBL-E. coli (9.9%) was most frequently observed in the western (15.2%) region, followed by the central (8.0%) and eastern (7.2%) regions. A total of 85 resistance patterns were identified, with 98.5% exhibiting multidrug resistance. The blaCTX-M gene was detected in 97.5% of isolates, predominantly blaCTX-M-1 (97.5%), while blaCTX-M-9 was found in 2.5%. The blaTEM gene was present in 33.0% of ESBL isolates; however, blaSHV and blaOXA-1 were absent. Phylogenetic group A predominated (42.0%), followed by E (22.5%), B1 (20.5%), F (10.5%), C (2.5%), and D (2.0%). This study demonstrates a significant occurrence of ESBL-E. coli in Indonesian broilers with regional variation and blaCTX-M predominance. The high rate of multidrug resistance poses a serious public health concern, emphasizing the urgent need for antimicrobial stewardship and enhanced surveillance programs. Full article
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