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22 pages, 1656 KB  
Article
Investigation into the Multiphase Product Distribution and Evolution During Biomass Pyrolysis Using Wheat Straw and Pine Sawdust
by Jishuo Li, Kaili Xu, Xiwen Yao and Xingyu Luo
Energies 2025, 18(20), 5397; https://doi.org/10.3390/en18205397 (registering DOI) - 13 Oct 2025
Abstract
Understanding the formation mechanisms of three-phase products during biomass pyrolysis is essential for optimizing thermochemical conversion and enhancing the efficient utilization of renewable resources. In this study, wheat straw (WS) and pine sawdust (PS) were selected as representative feedstocks to investigate the thermal [...] Read more.
Understanding the formation mechanisms of three-phase products during biomass pyrolysis is essential for optimizing thermochemical conversion and enhancing the efficient utilization of renewable resources. In this study, wheat straw (WS) and pine sawdust (PS) were selected as representative feedstocks to investigate the thermal decomposition behavior and evolution characteristics of gas, liquid (tar), and solid (char) products during pyrolysis. Thermogravimetric analysis and kinetic modeling revealed that PS exhibited higher activation energy (75.44 kJ/mol) than WS (65.63 kJ/mol), indicating greater thermal resistance. Tar yield increased initially and then declined with temperature, peaking at 700 °C (37.79% for PS and 32.82% for WS), while the composition shifted from oxygenated compounds to polycyclic aromatic hydrocarbons as temperature rose. FTIR analysis demonstrated that most functional group transformations in char occurred below 400 °C, with aromatic structures forming above 300 °C and stabilizing beyond 700 °C. Gas product evolution showed that WS produced higher CO and H2 yields due to its composition, with CH4 generated in relatively lower amounts. These findings provide insights into biomass pyrolysis mechanisms and offer a theoretical basis for targeted regulation of product distributions in bioenergy applications. Full article
11 pages, 621 KB  
Article
Using Conversations, Listening and Leadership to Support Staff Wellness: The CALM Framework
by Usman Iqbal, Natalie Wilson, Robyn Taylor, Louise Smith and Friedbert Kohler
Int. J. Environ. Res. Public Health 2025, 22(10), 1558; https://doi.org/10.3390/ijerph22101558 (registering DOI) - 13 Oct 2025
Abstract
Healthcare workers’ (HCWs) wellness is a critical concern, particularly following the COVID-19 pandemic. Staff Wellness Rounding (SWR) has emerged as a leadership-driven strategy to support HCWs but research on its effectiveness remains limited. This study examines the impact of SWR within a large [...] Read more.
Healthcare workers’ (HCWs) wellness is a critical concern, particularly following the COVID-19 pandemic. Staff Wellness Rounding (SWR) has emerged as a leadership-driven strategy to support HCWs but research on its effectiveness remains limited. This study examines the impact of SWR within a large healthcare organisation in Australia and introduces the CALM (Conversation, Active Listening, Leadership Engagement, Mechanism for Feedback) Framework to enhance leadership-driven wellness initiatives. SWR was implemented across six acute hospitals and 14 community health centres in New South Wales, Australia (July to October 2021). A sequential mixed-methods design was used to evaluate SWR effectiveness, leadership engagement, and key components for a structured wellness approach. Phase One included a survey of 169 HCWs to capture their experiences, and Phase Two and Three comprised semi-structured interviews with SWR leaders, participants of SWR and analysis of 342 SWR records. Findings showed that informal conversations foster trust, active listening supports emotional well-being, and leadership engagement facilitates issue escalation. However, feedback mechanisms require improvement: 77.5% of HCWs felt able to escalate concerns but only 32.5% believed feedback was effectively addressed. These insights directly informed the development of the CALM Framework with implications for leadership training and digital wellness integration in healthcare settings. Full article
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56 pages, 661 KB  
Review
Analytical Methods for the Determination of Diamorphine (Heroin) in Biological Matrices: A Review
by Ahmed Ibrahim Al-Asmari
Toxics 2025, 13(10), 867; https://doi.org/10.3390/toxics13100867 (registering DOI) - 13 Oct 2025
Abstract
Diamorphine (DIM, heroin) is a semi-synthetic opioid that undergoes rapid conversion to 6-monoacetylmorphine and morphine, producing short-lived biomarkers that are difficult to capture during the process. This review critically explores the evolution of analytical techniques for quantitative DIM analysis in biological matrices from [...] Read more.
Diamorphine (DIM, heroin) is a semi-synthetic opioid that undergoes rapid conversion to 6-monoacetylmorphine and morphine, producing short-lived biomarkers that are difficult to capture during the process. This review critically explores the evolution of analytical techniques for quantitative DIM analysis in biological matrices from 1980 to 2025. It synthesizes findings across blood, plasma, urine, hair, sweat, and postmortem samples, emphasizing matrix-specific challenges and forensic applicability. Unlike previous opioid reviews that primarily focused on metabolites, this work highlights analytical methods capable of successfully detecting diamorphine itself alongside its key metabolites. This review examines 32 studies spanning three decades and compares three core analytical methods: gas chromatography–mass spectrometry (GC–MS), high-performance liquid chromatography (HPLC) with optical detection and liquid chromatography–mass spectrometry (LC–MS). Key performance metrics include sensitivity, sample preparation workflow, hydrolysis control, metabolite coverage, matrix compatibility, automation potential and throughput. GC–MS remains the workhorse for hair and sweat ultra-trace screening after derivatization. HPLC with UV, fluorescence or diode-array detection enables robust quantification of morphine and its glucuronides in pharmacokinetic and clinical settings. LC–MS facilitates the multiplexed analysis of DIM, its ester metabolites and its conjugates in a single, rapid run under gentle conditions to prevent ex vivo degradation. Recent advances such as high-resolution mass spectrometry and microsampling techniques offer new opportunities for sensitive and matrix-adapted analysis. By integrating validation parameters, forensic applicability, and evolving instrumentation, this review provides a practical roadmap for toxicologists and analysts navigating complex biological evidence. Full article
(This article belongs to the Special Issue Current Issues and Research Perspectives in Forensic Toxicology)
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13 pages, 276 KB  
Article
Sperm Quality and Welfare of Sexually Mature Boars Supplemented with Partially Fermentable Insoluble Fiber
by Daniela Ferreira de Brito Mandu, Vivian Schwaab Sobral, Juliana Cristina Rego Ribas, Maria Fernanda de Castro Burbarelli, Cristiny Santos Braga, Rodrigo Garófallo Garcia, Ibiara Correia de Lima Almeida Paz, Claudia Marie Komiyama and Fabiana Ribeiro Caldara
Life 2025, 15(10), 1597; https://doi.org/10.3390/life15101597 - 13 Oct 2025
Abstract
Dietary fiber plays an important role in animal nutrition by influencing gut health, feed intake, and metabolism. In swine production, studies suggest that fibers may also affect reproductive traits, but findings remain inconsistent, especially in adult boars. This study evaluated the effects of [...] Read more.
Dietary fiber plays an important role in animal nutrition by influencing gut health, feed intake, and metabolism. In swine production, studies suggest that fibers may also affect reproductive traits, but findings remain inconsistent, especially in adult boars. This study evaluated the effects of partially fermentable insoluble fiber (PFIF) on semen quality, behavior, and general health of adult boars. Thirty animals were assigned to a completely randomized design with two treatments: (1) CON: no fiber supplementation, and (2) PFIF: fiber supplementation (35 g/animal/day). Fiber was provided once daily for 120 consecutive days. During the period, semen was collected weekly and analyzed macroscopically and microscopically using the Computer-Assisted Sperm Analysis (CASA) system. Behavior was recorded weekly, one and three hours after feeding, based on a pre-established ethogram. Feed intake, perineal, and fecal scores were also evaluated. Fiber supplementation did not affect total motility, progressive motility, sperm concentration, fecal or perineal scores, or behavior. However, improvements were observed in sperm kinematics, with higher straight-line distance (DSL), linearity (LIN), and straightness (STR), as well as a tendency for increased straight-line velocity (VSL) and wobble (WOB). Conversely, a higher incidence of proximal cytoplasmic droplets was recorded in the fiber group, indicating more sperm maturation defects. Supplemented animals also showed reduced feed intake compared with controls, suggesting a satiety effect of the fiber. In conclusion, PFIF supplementation (35 g/animal/day offered once daily) in adult boars produced mixed outcomes, with improved sperm kinematics but increased maturation defects and only minor changes in feeding behavior, indicating a limited and inconsistent physiological response. Full article
(This article belongs to the Special Issue Animal Reproduction and Health)
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13 pages, 3481 KB  
Article
Small Extracellular Vesicles Derived from NF2-Associated Schwannoma Cells Modulate Tumor Progression and Immunity via HSP90
by Ying Wang, Yuan Ren, Qi Zhang, Chao Zhang, Minjun Yan, Xin Ma, Bo Wang, Peng Li and Pinan Liu
Curr. Oncol. 2025, 32(10), 569; https://doi.org/10.3390/curroncol32100569 (registering DOI) - 13 Oct 2025
Abstract
In-depth exploration of tumor immune suppression mechanisms may provide new therapeutic options for NF2-associated tumors. In this study, we found that sEVs secreted by NF2-associated schwannomas (NF2-EVs) facilitate the conversion of CD14+ monocytes into an MDSC-like phenotype, showcasing MDSC-like inhibitory functions. Moreover, [...] Read more.
In-depth exploration of tumor immune suppression mechanisms may provide new therapeutic options for NF2-associated tumors. In this study, we found that sEVs secreted by NF2-associated schwannomas (NF2-EVs) facilitate the conversion of CD14+ monocytes into an MDSC-like phenotype, showcasing MDSC-like inhibitory functions. Moreover, these NF2-EVs are capable of enhancing tumor cell proliferation. Through proteomic analysis and subsequent validation of the NF2-EVs, we identified elevated levels of HSP90. When we knocked down HSP90 expression in tumor cells, the sEVs secreted showed diminished capacity to convert monocytes into MDSCs and a reduced ability to promote tumor cell proliferation. Conversely, sEVs secreted by tumor cells that overexpress HSP90 displayed the opposite effects. Further mechanistic studies revealed that HSP90 could influence the expression of AKT/p-AKT and ERK/p-ERK. Our results suggest that NF2 tumor cells could regulate the AKT/p-AKT and ERK/p-ERK pathways to promote tumor cell proliferation and the formation of an immunosuppressive microenvironment by secreting sEVs’ HSP90, offering valuable insights into the involvement of HSP90 in exosome-mediated communication within the context of NF2-related schwannomatosis (NF2-SWN). This information has the potential to inform the design of effective immunotherapeutic protocols and offer new treatment options for NF2-SWN patients. Full article
(This article belongs to the Section Neuro-Oncology)
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22 pages, 10459 KB  
Article
Effect of Extracellular Matrix Derived from Porcine Tissue on Stemness of Porcine Spermatogonial Stem Cells
by Donghyeon Kim, Min-Gi Han, Yoseop Jeon, Hyoyoung Maeng, Youngseok Choi, Kwonho Hong, Jeong Tae Do and Hyuk Song
Int. J. Mol. Sci. 2025, 26(20), 9937; https://doi.org/10.3390/ijms26209937 (registering DOI) - 13 Oct 2025
Abstract
The extracellular matrix (ECM) supports spermatogonial stem cell (SSC) function by mimicking biochemical and structural features of the native niche. However, optimal feeder systems and ECM materials remain key limitations in porcine SSC (pSSC) cultures. We developed a porcine-derived ECM (pECM) from porcine [...] Read more.
The extracellular matrix (ECM) supports spermatogonial stem cell (SSC) function by mimicking biochemical and structural features of the native niche. However, optimal feeder systems and ECM materials remain key limitations in porcine SSC (pSSC) cultures. We developed a porcine-derived ECM (pECM) from porcine feet and evaluated its effectiveness in supporting pSSC maintenance and proliferation under feeder-dependent conditions. We examined protein molecular weight distribution and pECM extract composition. Surface characterization was performed using scanning electron microscopy and atomic force microscopy. We compared pECM with conventional coatings, including gelatin and non-coated controls, using morphological analysis, WST-1 assay, cell cycle analysis, and gene/protein expression of SSC markers. pECM promoted larger, well-defined pSSC colonies and enhanced stemness-related marker expression, including PGP9.5, Thy-1, PLZF, GFRA1, NANOG, and VASA. Additionally, pECM facilitated active pSSC proliferation while suppressing feeder overgrowth, contributing to a stable and functional co-culture environment. Conversely, gelatin supported early feeder proliferation but led to growth saturation, whereas N/C showed delayed attachment and reduced viability. These findings suggest that pECM mimics the native SSC niche and improves pSSC culture. The dual function of pECM in regulating feeder behavior and enhancing pSSC maintenance highlights its potential as a biomaterial for species lacking established feeder-free protocols. Full article
(This article belongs to the Special Issue Molecular Research in Animal Reproduction)
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23 pages, 2242 KB  
Article
Using Large Language Models to Analyze Interviews for Driver Psychological Assessment: A Performance Comparison of ChatGPT and Google-Gemini
by Ruifen Sun, Xinni Hu, Yang Shao, Zhongbin Luo, Bin Liu and Yuzhu Cheng
Symmetry 2025, 17(10), 1713; https://doi.org/10.3390/sym17101713 - 13 Oct 2025
Abstract
This study examines the application of large language models (LLMs) in analyzing subjective driver perceptions during tunnel driving simulations, comparing the effectiveness of questionnaires and interviews. Building on previous research involving driver simulations, we recruited 29 new participants, collected their perceptions via questionnaires, [...] Read more.
This study examines the application of large language models (LLMs) in analyzing subjective driver perceptions during tunnel driving simulations, comparing the effectiveness of questionnaires and interviews. Building on previous research involving driver simulations, we recruited 29 new participants, collected their perceptions via questionnaires, and conducted follow-up interviews. The interview data were analyzed using three LLMs: GPT-3.5, GPT-4, and Google-Gemini. The results revealed that while GPT-4 provides more in-depth and accurate analysis, it is significantly slower than GPT-3.5. Conversely, Google-Gemini demonstrated a balance between analysis quality and speed, outperforming the other models overall. Despite the challenge of occasional misunderstandings, LLMs still have the potential to enhance the efficiency and accuracy of subjective data analysis in transportation research. Full article
(This article belongs to the Section Computer)
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14 pages, 390 KB  
Article
Deviant Behavior in Young People After COVID-19: The Role of Sensation Seeking and Empathy in Determining Deviant Behavior
by Marta Floridi, Allison Uvelli, Benedetta Tonini, Simon Ghinassi, Silvia Casale, Gabriele Prati, Giacomo Gualtieri, Alessandra Masti and Fabio Ferretti
COVID 2025, 5(10), 173; https://doi.org/10.3390/covid5100173 - 12 Oct 2025
Abstract
Background: The COVID-19 pandemic significantly impacted adolescent development, increasing behavioral problems and emotional distress. This study aimed to examine the impact of sensation seeking, empathy, and COVID-19-related stressors on deviant behavior in adolescents. Methods: A cross-sectional study was conducted with 638 Italian adolescents [...] Read more.
Background: The COVID-19 pandemic significantly impacted adolescent development, increasing behavioral problems and emotional distress. This study aimed to examine the impact of sensation seeking, empathy, and COVID-19-related stressors on deviant behavior in adolescents. Methods: A cross-sectional study was conducted with 638 Italian adolescents and young adults (M = 18.8 years, SD = 3.51) recruited from schools, universities, and the general population in Tuscany and Emilia-Romagna. Participants completed validated measures assessing sensation seeking, empathy, COVID-19-related stress, and deviant behaviors. Multiple regression analyses examined predictors of deviant behavior, while mediation analyses tested whether empathy mediated the relationship between sensation seeking and deviant behavior. Results: Correlation analyses show a positive association between sensation seeking and deviant behavior and a weaker positive association with COVID-19 isolation. Conversely, affective empathy demonstrated negative correlations with both deviant behavior and sensation seeking. COVID-19 stress demonstrated differentiated effects: social isolation increased deviance, whereas fear of contagion was protective. Mediation analysis revealed that affective empathy partially mediated the relationship between sensation seeking and deviance. Conclusions: This study demonstrates that sensation seeking is a primary risk factor for deviant behavior in adolescents and young adults, while affective empathy acts as a protective mechanism that partially mediates this relationship. Furthermore, COVID-19-related stressors have shown complex effects, with social isolation amplifying the risk of deviance, while fear of contagion promotes more inhibited behavior. These findings underscore the importance of considering both stable personality traits and situational stressors when seeking to understand the pathways leading to adolescent behavioral problems during periods of social crisis. Full article
(This article belongs to the Section COVID Public Health and Epidemiology)
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31 pages, 532 KB  
Article
AI in the Classroom: GPT Usage and Learner Typologies in Programming
by Di Wang, Xingbo Dong, Zheng Fang, Lillian Yee Kiaw Wang and Zhe Jin
Educ. Sci. 2025, 15(10), 1353; https://doi.org/10.3390/educsci15101353 - 12 Oct 2025
Abstract
This study examines GPT usage in university-level programming education, focusing on patterns and correlations in students’ learning behaviors. A survey of 438 students from four universities was conducted to analyze their adoption of AI, learning dispositions, and behavioral patterns. The research aimed to [...] Read more.
This study examines GPT usage in university-level programming education, focusing on patterns and correlations in students’ learning behaviors. A survey of 438 students from four universities was conducted to analyze their adoption of AI, learning dispositions, and behavioral patterns. The research aimed to understand the current state of GPT adoption and the connection between student learning approaches and their use of these technologies. Findings show that while a majority of students use GPT applications, the frequency and depth of this engagement vary significantly. Students who favor self-directed learning tend to leverage this technology more for personalized learning and self-assessment. Conversely, students more accustomed to traditional teaching methods use it more conservatively. The study identified four distinct learner groups through cluster analysis, each with unique interaction styles. Furthermore, a correlation analysis indicated that learning orientations, such as Technology-Driven Learning and intrinsic motivation, are positively associated with more frequent and effective GPT use. Full article
(This article belongs to the Section Technology Enhanced Education)
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16 pages, 612 KB  
Article
Determinants of Quality of Life Among Saudi Adults with Sciatica: A Cross-Sectional Study
by Mohammad A. Jareebi, Abdullah J. Almalki, Abdulkarim Zain Suwaydi, Ehab F. Hakami, Mnar H. Moafa, Saud N. Wadani, Fatimah N. Hakami, Shatha K. Alharbi, Malik I. Hakami, Abdulmajid E. Abuhadi, Majed A. Ryani, Ahmed A. Bahri, Yahya H. Khormi, Ibrahim A. Hakami and Abdulwahab A. Aqeeli
Medicina 2025, 61(10), 1824; https://doi.org/10.3390/medicina61101824 (registering DOI) - 11 Oct 2025
Abstract
Background and Objectives: Sciatica is a common condition associated with significant pain and reduced quality of life (QoL). However, its impact and associated determinants remain underexplored in Saudi Arabia. Therefore, this study aimed to examine determinants of quality of life among Saudi adults [...] Read more.
Background and Objectives: Sciatica is a common condition associated with significant pain and reduced quality of life (QoL). However, its impact and associated determinants remain underexplored in Saudi Arabia. Therefore, this study aimed to examine determinants of quality of life among Saudi adults with sciatica. Materials and Methods: A cross-sectional study was conducted in Saudi Arabia between February and March 2024 using an online Arabic questionnaire disseminated via social media. Participants (n = 927) aged ≥18 years completed a 50-item questionnaire covering sociodemographic characteristics, clinical history, lifestyle factors, and the Short Form-12 (SF-12) QoL scale. Results: Of the 927 adults (mean age 30 ± 11 years), 76% (n = 706) were female and 24% (n = 221) male; 10% (n = 92) reported sciatica. Overall, 57% (n = 531) reported good QoL. Multivariate analysis revealed that increasing age (Odds ratio (OR) = 1.04; 95% CI: 1.01–1.07), urban residence (OR = 1.49; 95% CI: 1.11–2.00), and higher income (>15,000 Saudi Arabia Riyal (SAR); OR = 1.53; 95% CI: 1.03–2.27) were associated with better QoL. Conversely, diabetes (OR = 0.42; 95% CI: 0.22–0.79), arthritis (OR = 0.52; 95% CI: 0.32–0.83), and sciatica duration >1 year (OR = 0.29; 95% CI: 0.12–0.69) were associated with poorer QoL. Gender, body mass index (BMI), smoking, and physical activity showed no significant associations. Conclusions: Sciatica, prolonged symptom duration, and comorbidities such as diabetes and arthritis negatively affect QoL in Saudi adults, whereas socioeconomic and demographic factors contribute positively. The results underscore the importance of early intervention and tailored support for sciatica patients with long-standing symptoms or specific comorbidities to improve health outcomes. Longitudinal studies are warranted to assess causality and the impact of interventions. Full article
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58 pages, 1997 KB  
Review
Immunomodulatory Activities of Emerging Rare Ginsenosides F1, Rg5, Rk1, Rh1, and Rg2: From Molecular Mechanisms to Therapeutic Applications
by Chang-Eui Hong and Su-Yun Lyu
Pharmaceuticals 2025, 18(10), 1529; https://doi.org/10.3390/ph18101529 - 11 Oct 2025
Abstract
Ginsenosides, the primary bioactive components of Panax ginseng, have demonstrated significant immunomodulatory potential. While major ginsenosides have been extensively studied, rare ginsenosides produced through deglycosylation, heating, and steaming show enhanced biological activities with improved bioavailability. This review aimed to comprehensively analyze the [...] Read more.
Ginsenosides, the primary bioactive components of Panax ginseng, have demonstrated significant immunomodulatory potential. While major ginsenosides have been extensively studied, rare ginsenosides produced through deglycosylation, heating, and steaming show enhanced biological activities with improved bioavailability. This review aimed to comprehensively analyze the immunomodulatory mechanisms, structure-activity relationships (SARs), therapeutic applications, and clinical translation strategies of five emerging rare ginsenosides: F1, Rg5, Rk1, Rh1, and Rg2. We conducted a comprehensive literature review examining the production methods, immunological effects, molecular mechanisms, pharmacokinetics, safety profiles, and clinical applications of these five compounds. Analysis focused on chemical structures, immune cell modulation, signaling pathways, disease model efficacy, and bioavailability enhancement strategies. Ginsenoside F1 uniquely demonstrated immunostimulatory effects, enhancing natural killer (NK) cell cytotoxicity and macrophage phagocytosis through mitogen-activated protein kinase (MAPK)/nuclear factor-κB (NF-κB) activation. Conversely, Rg5, Rk1, Rh1, and Rg2 exhibited anti-inflammatory properties via distinct mechanisms: Rg5 through Toll-like receptor 4 (TLR4)/NF-κB inhibition, Rk1 via triple pathway modulation (NF-κB, p38 MAPK, signal transducer and activator of transcription (STAT)), Rh1 by selective p38 MAPK and STAT1 inhibition, and Rg2 through modulation of both central nervous system (neuroinflammation) and peripheral organ systems. Structure-activity analysis revealed that sugar moiety positions critically determine immunological outcomes. Crucially, advanced delivery systems including nanostructured lipid carriers, self-microemulsifying systems, and specialized liposomes have overcome the major translational barrier of poor bioavailability, achieving up to 2.6-fold improvements and enabling clinical development. Safety assessments demonstrated favorable tolerability profiles across preclinical and clinical studies. These five rare ginsenosides represent promising immunomodulatory agents with distinct therapeutic applications. F1’s unique immunostimulatory properties position it for cancer immunotherapy, while the complementary anti-inflammatory mechanisms of Rg5, Rk1, Rh1, and Rg2 offer opportunities for precision medicine in inflammatory diseases. Advanced formulation technologies and optimized production methods now enable their significant clinical translation potential, providing promising therapeutic options for immune-related disorders pending further development. Full article
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20 pages, 1372 KB  
Article
α-Linolenic Acid Production in Aspergillus oryzae via the Overexpression of an Endogenous Omega-3 Desaturase Gene
by Hiroki Kikuta, Hirotoshi Sushida, Tsuyoshi Tanaka, Eiichi Kotake, Wakako Tsuzuki, Ryota Hattori, Satoshi Suzuki, Ken-Ichi Kusumoto and Junichi Mano
Fermentation 2025, 11(10), 585; https://doi.org/10.3390/fermentation11100585 (registering DOI) - 11 Oct 2025
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Abstract
α-Linolenic acid (ALA) is an important essential omega-3 (ω-3) polyunsaturated fatty acid for the maintenance of human health. Although ALA has traditionally been obtained from plant sources, microbial fermentation has emerged as a promising alternative for its sustainable and cost-effective production. However, most [...] Read more.
α-Linolenic acid (ALA) is an important essential omega-3 (ω-3) polyunsaturated fatty acid for the maintenance of human health. Although ALA has traditionally been obtained from plant sources, microbial fermentation has emerged as a promising alternative for its sustainable and cost-effective production. However, most of the present approaches rely on genetically modified organisms, which present regulatory and consumer-acceptance concerns. In this study, we aimed to develop a high-ALA-producing strain of Aspergillus oryzae, a Generally Recognized As Safe (GRAS) microorganism widely used in food production in Japan, through self-cloning, a form of genetic engineering that utilizes only the host’s own DNA. To achieve this, an endogenous ω-3 desaturase gene (fad3), which catalyzes the conversion of linoleic acid to ALA, was identified via BLASTP analysis. Subsequently, a multicopy A. oryzae strain (Aofad3-MC) overexpressing fad3 was constructed. This strain increased ALA production, with ALA comprising 30.7% of the total lipids. Furthermore, k-mer analysis confirmed the absence of foreign vector sequences, verifying that Aofad3-MC was constructed through self-cloning. In addition to the identification of the A. oryzae ω-3 desaturase gene, this study provides a microbial platform for the sustainable production of ALA, with potential applications across the food, feed, and related industries. Full article
(This article belongs to the Special Issue Metabolic Engineering, Strain Modification and Industrial Application)
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22 pages, 4622 KB  
Article
Elucidating the Mechanistic Role of Exogenous Melatonin in Salt Stress Tolerance of Maize (Zea mays L.) Seedlings: An Integrated Physiological, Metabolomic, and Proteomic Profiling Analysis
by Zhichao Wang, Linhao Zong, Qiqi Cai, Yinjie Fu, Zhiping Gao and Guoxiang Chen
Plants 2025, 14(20), 3129; https://doi.org/10.3390/plants14203129 (registering DOI) - 10 Oct 2025
Viewed by 112
Abstract
Maize (Zea mays L.), as a globally significant cereal crop, exhibits high sensitivity to salt stress during early seedling stages. Although melatonin (MT) has demonstrated potential in mitigating abiotic stresses, the specific mechanisms underlying MT-mediated alleviation of salt stress in maize seedlings [...] Read more.
Maize (Zea mays L.), as a globally significant cereal crop, exhibits high sensitivity to salt stress during early seedling stages. Although melatonin (MT) has demonstrated potential in mitigating abiotic stresses, the specific mechanisms underlying MT-mediated alleviation of salt stress in maize seedlings remain unclear. In this study, we established four treatment groups: control (CK), melatonin treatment (MT), salt stress (NaCl), and combined treatment (NaCl_MT). Metabolomic and proteomic analyses were performed, supplemented by photosynthesis-related experiments as well as antioxidant-related experiments. Metabolomic analysis identified key metabolites in MT-mediated salt stress mitigation. Both metabolomic and proteomic analyses underscored the critical roles of photosynthetic and antioxidant pathways. Salt stress significantly decreased the net photosynthetic rate (Pn) by 67.7%, disrupted chloroplast ultrastructure, and reduced chlorophyll content by 41.6%. Conversely, MT treatment notably mitigated these detrimental effects. Moreover, MT enhanced the activities of antioxidant enzymes by approximately 10–20% and reduced the accumulation of oxidative stress markers by around 10–25% in maize seedlings under salt stress. In conclusion, this study conducted a systematic and multidimensional investigation into the mitigation of salt stress in maize seedlings by MT. Our results revealed that MT enhances antioxidant systems, increases chlorophyll content, and alleviates damage to chloroplast ultrastructure, thereby improving photosystem II performance and strengthening photosynthesis. This ultimately manifests as improved seedling phenotypes under salt stress. These findings provide a meaningful entry point for breeding salt-tolerant maize varieties and mitigating the adverse effects of salinized soil on maize growth and yield. Full article
(This article belongs to the Special Issue Abiotic Stress Responses in Plants—Second Edition)
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29 pages, 19561 KB  
Article
Empirical Analysis of the Impact of Two Key Parameters of the Harmony Search Algorithm on Performance
by Geonhee Lee and Zong Woo Geem
Mathematics 2025, 13(20), 3248; https://doi.org/10.3390/math13203248 - 10 Oct 2025
Viewed by 76
Abstract
Metaheuristic algorithms are widely utilized as effective tools for solving complex optimization problems. Among them, the Harmony Search (HS) algorithm has garnered significant attention for its simple structure and excellent performance. The efficacy of the HS algorithm is heavily dependent on the configuration [...] Read more.
Metaheuristic algorithms are widely utilized as effective tools for solving complex optimization problems. Among them, the Harmony Search (HS) algorithm has garnered significant attention for its simple structure and excellent performance. The efficacy of the HS algorithm is heavily dependent on the configuration of its internal parameters, with the Harmony Memory Considering Rate (HMCR) and Pitch Adjusting Rate (PAR) playing pivotal roles. These parameters determine the probabilities of using the Random Generation (RG), Harmony Memory Consideration (HMC), and Pitch Adjustment (PA) operators, thereby controlling the balance between exploration and exploitation. However, a systematic empirical analysis of the interaction between these parameters and the characteristics of the problem at hand remains insufficient. Thus, this study conducts a comprehensive empirical analysis of the performance sensitivity of the HS algorithm to variations in HMCR and PAR values. The analysis is performed on a suite of 23 benchmark functions, encompassing diverse characteristics such as unimodality/multimodality and separability/non-separability, along with 5 real-world optimization problems. Through extensive experimentation, the performance for each parameter combination was evaluated on a rank-based system and visualized using heatmaps. The results experimentally demonstrate that the algorithm’s performance is most sensitive to the HMCR value across all function types, establishing that setting a high HMCR value (≥0.9) is a prerequisite for securing stable performance. Conversely, the optimal PAR value showed a direct correlation with the topographical features of the problem landscape. For unimodal problems, a low PAR value between 0.1 and 0.3 was more effective, whereas for complex multimodal problems with numerous local optima, a relatively higher PAR value between 0.3 and 0.5 proved more efficient in searching for the global optimum. This research provides a guideline into the parameter settings of the HS algorithm and contributes to enhancing its practical applicability by proposing a systematic parameter tuning strategy based on problem characteristics. Full article
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15 pages, 852 KB  
Article
Effect of Influenza Vaccination on the Disease Severity and Viral Load Among Adult Outpatients and Inpatients
by Alexander Domnich, Vincenzo Paolozzi, Giada Garzillo, Andrea Orsi and Giancarlo Icardi
Vaccines 2025, 13(10), 1046; https://doi.org/10.3390/vaccines13101046 - 10 Oct 2025
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Abstract
Background: Some studies suggest that, thanks to the mechanisms of immune-mediated attenuation, influenza vaccination reduces severity of influenza illness in breakthrough infections. This study aimed to assess whether influenza vaccination attenuates severity of laboratory-confirmed influenza among Italian adults. Methods: This secondary [...] Read more.
Background: Some studies suggest that, thanks to the mechanisms of immune-mediated attenuation, influenza vaccination reduces severity of influenza illness in breakthrough infections. This study aimed to assess whether influenza vaccination attenuates severity of laboratory-confirmed influenza among Italian adults. Methods: This secondary analysis included all influenza cases detected during respiratory surveillance studies conducted in outpatient and inpatient settings in Genoa (Italy), throughout the 2023/2024 and 2024/2025 seasons. Here, we compared viral load and the count of influenza-related symptoms in outpatients, alongside all-cause in-hospital mortality and radiologically confirmed pneumonia in inpatients, between vaccinated and unvaccinated adults. Results: The study included 188 influenza cases diagnosed in primary care and 281 influenza cases identified among inpatients. Of these, 37.2% and 31.7%, respectively, were vaccinated, constituting breakthrough infections. Compared to unvaccinated adults, vaccinated outpatients had a slightly lower viral load (difference in cycle threshold values of 1.36 corresponding to about 0.51 log10 reduction in the number of copies/mL; p = 0.077), primarily driven by influenza A(H1N1)pdm09. Vaccinated outpatients also reported 9% fewer influenza-related symptoms than unvaccinated counterparts [prevalence ratio 0.91; 95% confidence interval (CI): 0.84, 0.99]. Among hospitalized older adults, influenza vaccination was associated with 64% reduced odds of in-hospital death (odds ratio 0.36; 95% CI: 0.12, 0.94). Conversely, no association between vaccination and development of pneumonia was found. Conclusions: This study corroborates the idea that influenza vaccination attenuates disease severity in breakthrough infections. These effects are, however, dependent on the measure of severity used. Full article
(This article belongs to the Special Issue The Effectiveness of Influenza Vaccine)
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