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24 pages, 6660 KB  
Article
The Structural Proteins of Thermophilic Bacteriophage P23-77: Expression and Characterization
by Milad Kheirvari and Ebenezer Tumban
Int. J. Mol. Sci. 2025, 26(17), 8688; https://doi.org/10.3390/ijms26178688 (registering DOI) - 6 Sep 2025
Viewed by 158
Abstract
P23-77 is a thermophilic bacteriophage that infects Thermus thermophilus bacteria. The genome of the virus is enclosed in an icosahedral capsid. This capsid is made of the small major capsid protein (VP16), the large major capsid protein (VP17), and the minor capsid protein [...] Read more.
P23-77 is a thermophilic bacteriophage that infects Thermus thermophilus bacteria. The genome of the virus is enclosed in an icosahedral capsid. This capsid is made of the small major capsid protein (VP16), the large major capsid protein (VP17), and the minor capsid protein (VP11). In addition to these three structural proteins, membrane-associated proteins (VP15, VP19, VP20, VP22, and VP23) have been identified in the virus and may serve as scaffold proteins to help with viral assembly. Previous studies have expressed VP11, VP16, and VP17 in E. coli. A mixture of these proteins can lead to the formation of complexes. However, the potential to express membrane-associated proteins has never been explored. Here, we demonstrated, for the first time, the expression and co-expression of some membrane-associated proteins with capsid (coat) proteins, both in the natural host and in E. coli. Co-expression of these proteins did not result in the assembly of virus-like particles. We explored further strategies to express and purify some of the proteins for future studies. We observed that the insertion of a purification tag (Strep-II tag, but not a histidine tag) significantly reduced the expression levels of some of the proteins. Six of the eight structural proteins were successfully purified to homogeneity using different approaches. We showed that VP20 and VP22 migrated on SDS PAGE gel at sizes larger than their predicted molecular weights. Predicted 3D structures of the proteins show that most of them are helical in nature with disordered regions. The work presented here will help pave the way for the expression and purification of these proteins. This will help determine their 3D structures and may shed light on the requirements for viral assembly. Full article
(This article belongs to the Special Issue Molecular Biology of Extremophiles: Recent Advances)
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13 pages, 602 KB  
Article
Prophylactic Antibiotics in Vertebroplasty and Kyphoplasty: A Nationwide Analysis of Infection Rates and Antibiotic Use in South Korea
by Youngjin Kim, Young-Hoon Kim, Sukil Kim, Jun-Seok Lee, Sang-Il Kim, Joonghyun Ahn, So-Young Han and Hyung-Youl Park
Antibiotics 2025, 14(9), 901; https://doi.org/10.3390/antibiotics14090901 - 5 Sep 2025
Viewed by 155
Abstract
Background/Objectives: Vertebroplasty (VP) and kyphoplasty (KP) are widely performed minimally invasive procedures for osteoporotic vertebral compression fractures and vertebral metastases. Although generally safe, postoperative surgical site infections (SSIs) can lead to severe complications. The true incidence of SSIs and optimal prophylactic antibiotic [...] Read more.
Background/Objectives: Vertebroplasty (VP) and kyphoplasty (KP) are widely performed minimally invasive procedures for osteoporotic vertebral compression fractures and vertebral metastases. Although generally safe, postoperative surgical site infections (SSIs) can lead to severe complications. The true incidence of SSIs and optimal prophylactic antibiotic strategies remains unclear. This study evaluated SSI incidence and the impact of antibiotic timing and type using a nationwide quality assessment (QA) database in South Korea. Methods: We analyzed data from the 7th to 9th QA waves of the Health Insurance Review and Assessment (HIRA) Service, including 23,868 patients who underwent VP or KP. SSI incidence was compared across antibiotic timing groups (preoperative-only, postoperative-only, and combined) and antibiotic types. Multivariate logistic regression identified independent risk factors for SSIs. Results: SSI occurred in 47 patients (0.20% of 23,868 procedures). No infections were observed in the preoperative-only group, compared with 0.36% in the postoperative-only group and 0.19% in the pre- and postoperative group. The lowest incidence (0.16%) was seen with first- or second-generation cephalosporins. Multivariate analysis found no significant difference between the preoperative-only and the combined regimens, nor between first-/second-generation cephalosporins and broad-spectrum antibiotics. However, surgery at a tertiary hospital (aOR: 3.566) and malnutrition (aOR: 2.915) were independently associated with increased SSI risk. Conclusions: This nationwide study, the largest to date on VP and KP, demonstrated that SSIs are rare (0.2%). A single preoperative dose of first- or second-generation cephalosporins was as effective as combined or broader-spectrum regimens. Targeted preventive measures may be warranted for high-risk groups such as patients with malnutrition or those treated in tertiary hospitals. Full article
(This article belongs to the Special Issue Orthopedic Infections: Epidemiology and Antimicrobial Treatment)
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15 pages, 2419 KB  
Article
Development and 3D Printing of AESO-Based Composites Containing Olive Pit Powder
by Giovanna Colucci, Francesca Sacchi, Marta Checchi, Marianna Barbalinardo, Francesca Chiarini, Federica Bondioli, Carla Palumbo and Massimo Messori
J. Compos. Sci. 2025, 9(9), 479; https://doi.org/10.3390/jcs9090479 - 3 Sep 2025
Viewed by 317
Abstract
Bio-based polymeric composites were prepared by dispersing different amounts of olive pit (OP) powder within an acrylate epoxidized soybean oil (AESO) photocurable resin using tetrahydrofurfuryl acrylate (THFA) as diluent and (2,4,6-trimethylbenzoyl), phosphine oxide (BAPO) as photo-initiator, and they were photocured by Vat Photopolymerization [...] Read more.
Bio-based polymeric composites were prepared by dispersing different amounts of olive pit (OP) powder within an acrylate epoxidized soybean oil (AESO) photocurable resin using tetrahydrofurfuryl acrylate (THFA) as diluent and (2,4,6-trimethylbenzoyl), phosphine oxide (BAPO) as photo-initiator, and they were photocured by Vat Photopolymerization (VP) using a Liquid Crystal Display (LCD) 3D printer. Formulation viscosity was studied because of its important role in a VP process able to influence the printability of the final parts. Different 3D printed architectures were successfully realized with good resolution and accuracy, high level of detail, and flexibility. The effect of OP addition was investigated by thermal (TGA and DSC), morphological (SEM and PSD), viscoelastic (DMA), and mechanical (tensile testing) characterization. The filler led to an increase in the Tg, storage modulus, and tensile properties, underlining the stiffening effect induced by the OP particles onto the polymeric starting resin. This underlines the possibility to apply these bio-based composites in many application fields by valorizing agro-wastes, developing more sustainable materials, and taking advantages of VP 3D printing, such as low costs, minimal wastage, and customized geometry. Biocompatibility tests were also successfully carried out. The results clearly indicate that the AESO-based composites promote cell adhesion and viability. Full article
(This article belongs to the Special Issue Sustainable Polymer Composites: Waste Reutilization and Valorization)
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13 pages, 2179 KB  
Article
Direct Intercellular Transport Mode of Filovirus Nucleocapsids
by Catarina Harumi Oda Ibrahim and Yuki Takamatsu
Int. J. Mol. Sci. 2025, 26(17), 8485; https://doi.org/10.3390/ijms26178485 - 1 Sep 2025
Viewed by 298
Abstract
Intercellular pathways of viral infection in host cells offer advantages, such as efficiency of viral spread and immune surveillance evasion, compared to cell-free viral infection. Therefore, some enveloped viruses present both cell-to-cell and cell-free forms of infection in the host organisms. In this [...] Read more.
Intercellular pathways of viral infection in host cells offer advantages, such as efficiency of viral spread and immune surveillance evasion, compared to cell-free viral infection. Therefore, some enveloped viruses present both cell-to-cell and cell-free forms of infection in the host organisms. In this study, we investigated the occurrence of Ebola virus (EBOV) and Marburg virus (MARV) nucleocapsid exchange in vitro between interconnected Huh7 cells using live-cell imaging methods. Moreover, through plasmid transfection methods, we demonstrated that nucleocapsid-like structures (NCLSs) formed with EBOV NP, VP35, VP24, and VP30 proteins can also be transported intercellularly to non-transfected cells through cell-to-cell contact regions in a process involving interaction with the host cell actin cytoskeleton. Our results provide further evidence of cell-to-cell transport as a mechanism of filovirus spread and support the need for further research in this field to develop new intervention methods targeting this transmission pathway. Full article
(This article belongs to the Section Molecular Biology)
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21 pages, 2002 KB  
Article
Grey Wolf Optimizer Based on Variable Population and Strategy for Moving Target Search Using UAVs
by Ziyang Li, Zhenzu Bai and Bowen Hou
Drones 2025, 9(9), 613; https://doi.org/10.3390/drones9090613 - 31 Aug 2025
Viewed by 254
Abstract
Unmanned aerial vehicles (UAVs) are increasingly favored for emergency search and rescue operations due to their high adaptability to harsh environments and low operational costs. However, the dynamic nature of search path endpoints, influenced by target movement, limits the applicability of shortest path [...] Read more.
Unmanned aerial vehicles (UAVs) are increasingly favored for emergency search and rescue operations due to their high adaptability to harsh environments and low operational costs. However, the dynamic nature of search path endpoints, influenced by target movement, limits the applicability of shortest path models between fixed points in moving target search problems. Consequently, the moving target search problem using UAVs in complex environments presents considerable challenges, constituting an NP-hard problem. The Grey Wolf Optimizer (GWO) is known for addressing such problems. However, it suffers from limitations, including premature convergence and instability. To resolve these constraints, a Grey Wolf Optimizer with variable population and strategy (GWO-VPS) is developed in this work. GWO-VPS implements a variable encoding scheme for UAV movement patterns, combining motion-based encoding with path-based encoding. The algorithm iteratively alternates between global optimization and local smoothing phases. The global optimization phase incorporates: (1) a Q-learning-based strategy selection; (2) position updates with obstacle avoidance and energy consumption reduction; and (3) adaptive exploration factor. The local smoothing phase employs four path smoothing operators and Q-learning-based strategy selection. Experimental results demonstrate that GWO-VPS outperforms both enhanced GWO variants and standard algorithms, confirming the algorithm’s effectiveness in UAV-based moving target search simulations. Full article
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13 pages, 2048 KB  
Article
Dual Energy CT-Derived Liver Extracellular Volume Fraction for Assessing Liver Functional Reserve in Patients with Liver Cirrhosis
by Seok Jin Hong, Ji Eun Kim, Jae Min Cho, Ho Cheol Choi, Mi Jung Park, Hye Young Choi, Hwa Seon Shin, Jung Ho Won, Wonjeong Yang and Hyun Ok Kim
Medicina 2025, 61(9), 1561; https://doi.org/10.3390/medicina61091561 - 30 Aug 2025
Viewed by 300
Abstract
Background and Objectives: The extracellular volume fraction (fECV) of the liver, as measured by contrast-enhanced computed tomography (CT), has been shown to correlate closely with the histological stages of hepatic fibrosis. This study aimed to investigate the diagnostic performance of a liver [...] Read more.
Background and Objectives: The extracellular volume fraction (fECV) of the liver, as measured by contrast-enhanced computed tomography (CT), has been shown to correlate closely with the histological stages of hepatic fibrosis. This study aimed to investigate the diagnostic performance of a liver extracellular volume fraction derived from dual-energy CT (DECT) for evaluating liver functional reserve based on the Child–Pugh class in cirrhotic patients, compared with other noninvasive markers. Materials and Methods: This retrospective study included 258 patients with liver cirrhosis who underwent contrast-enhanced DECT. The fECV was measured using iodine maps derived from equilibrium phase images obtained 3 min after contrast injection at 100/140 Sn kVp. Statistical analyses included Welch’s ANOVA with post hoc tests, Spearman’s correlation, and ROC analysis. The area under the curve (AUC) was compared among fECV and other noninvasive markers (aspartate transaminase to platelet ratio index [APRI], Fibrosis-4 [FIB-4], and model for end-stage liver disease [MELD]) using DeLong’s test. Intra- and interobserver reliability of fECV was assessed with the intraclass correlation coefficient (ICC). The area under the receiver operating characteristic curve (AUC) for differentiating Child–Pugh classes was compared among the fECV and other noninvasive markers (aspartate transaminase to platelet ratio index [APRI], Fibrosis-4 [FIB-4], and model for end-stage liver disease [MELD]). Results: The fECV increased significantly with advancing Child–Pugh classes (p < 0.001), showing a moderate correlation with Child–Pugh class (r = 0.53). The mean differences in fECV among the Child–Pugh classes were 8.88 between A and B (95% confidence interval [CI], 5.85–11.92; p < 0.001) and 7.42 between B and C (95% CI, 1.92–12.91: p < 0.001). The AUC for differentiating Child–Pugh classes A and B demonstrated no significant differences among the fECV (0.84), APRI (0.83, p > 0.99) and FIB-4 (0.83, p > 0.99), except for MELD, which had a significantly higher AUC (0.94, p = 0.047). For differentiating Child-Pugh classes B and C, the fECV demonstrated a significantly higher AUC (0.78), compared with FIB-4 (0.50, p = 0.038) and APRI (0.49, p = 0.037), whereas no significant difference was observed between fECV and MELD (0.92, p = 0.12). The intra- and interobserver reliabilities of the fECV measurements were excellent (ICC, 0.93; 95% CI, 0.91–0.95 and 0.91; 95% CI, 0.88–0.92, respectively). Conclusions: DECT derived fECV is a useful noninvasive marker for assessing liver functional reserve based on the Child–Pugh classification. Full article
(This article belongs to the Section Gastroenterology & Hepatology)
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13 pages, 1635 KB  
Article
Molecular Characterization of Emerging and Uncommon Enteroviruses C104, C105, and C109 in Respiratory Samples from Maryland, USA, 2018–2024
by Amary Fall, Ting X. Zhuang, Alaina Dodge, Omar Abdullah, Julie M. Norton, David Villafuerte, Andrew Pekosz, Eili Klein and Heba H. Mostafa
Viruses 2025, 17(9), 1183; https://doi.org/10.3390/v17091183 - 29 Aug 2025
Viewed by 475
Abstract
Background: While enteroviruses (EVs) are recognized causes of diverse illnesses, little is known about the epidemiology and molecular characteristics of uncommon enterovirus C (EV-C) types, including EV-C104, EV-C105, and EV-C109. Methods: We conducted genomic surveillance of EVs at the Johns Hopkins [...] Read more.
Background: While enteroviruses (EVs) are recognized causes of diverse illnesses, little is known about the epidemiology and molecular characteristics of uncommon enterovirus C (EV-C) types, including EV-C104, EV-C105, and EV-C109. Methods: We conducted genomic surveillance of EVs at the Johns Hopkins Health System between 2018 and 2024 (a total of 3715 samples), identifying EV-C104, EV-C105, and EV-C109 in respiratory samples. VP4-VP2 and whole-genome sequencing were used to assess genetic diversity and intra-host evolution. Results: Five EV-C105 infections were identified primarily in pediatric patients, presenting with a range of clinical features including fever, gastrointestinal symptoms, and cerebellitis. Prolonged EV-C104 and EV-C109 infections were identified in two immunocompromised adults. EV-C104 persisted for over five months and showed evidence of viral genomic changes (intra-host evolution). EV-C109 was detected over a four-month period. Phylogenetic analysis revealed a novel EV-C105 clade (C3) closely related to recent USA strains. EV-C104 genomes aligned with genotype B sequences from the USA and Europe, while EV-C109 sequences were similar to 2014–2015 strains from the Netherlands. Conclusions: These findings highlight the emergence, persistence, and genetic evolution of uncommon EV-C types in Maryland, especially among immunocompromised hosts, emphasizing the importance of continued genomic surveillance and clinical correlations. Full article
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28 pages, 2367 KB  
Article
A Polyomavirus-Positive Merkel Cell Carcinoma Mouse Model Supports a Unified Origin for Somatic and Germ Cell Cancers
by Wendy Yang, Sara Contente and Sarah Rahman
Cancers 2025, 17(17), 2800; https://doi.org/10.3390/cancers17172800 - 27 Aug 2025
Viewed by 572
Abstract
Background/Objectives: The Germ Cell Theory of cancer posits that human primordial germ cells (hPGCs) are the cells of origin for malignancies. While this theory is well established for germ cell cancers, a germ cell origin for somatic cancers has been largely overlooked despite [...] Read more.
Background/Objectives: The Germ Cell Theory of cancer posits that human primordial germ cells (hPGCs) are the cells of origin for malignancies. While this theory is well established for germ cell cancers, a germ cell origin for somatic cancers has been largely overlooked despite clinical observations of malignant somatic transformation (MST), wherein germ cell cancers give rise to diverse somatic cancer phenotypes, often without additional mutations. Methods: To test the Germ Cell Theory experimentally in somatic cancer, we established a virus-driven MST model linking hPGC-like cells (hPGCLCs) to Merkel cell polyomavirus (MCPyV)-positive Merkel cell carcinoma (MCC), a highly aggressive somatic cancer with a germ cell cancer-like, low-mutation epigenetic profile. The MCPyV genome was transduced into human induced pluripotent stem cells (hiPSCs) or hPGC-like cells by lentiviral transfection, followed by xenotransplantation. Results: Virus-positive MCC (VP-MCC)-like tumors were consistently induced without additional oncogenic mutations. These tumors recapitulated VP-MCC’s high-grade neuroendocrine carcinoma histology and molecular profiles. DNA methylation analysis revealed near-complete global hypomethylation in VP-MCC-like tumors, matching the unique epigenetic state of late-stage hPGCs. Notably, pluripotent intermediates were neither necessary nor sufficient for MST; transformation required acquisition of a late-hPGC-like epigenetic state. Conclusions: This is the first MST model of a somatic cancer arising through an aberrant germline-to-soma transition. Our findings unify VP-MCC and germ cell cancer biology, challenge mutation- and soma-centric paradigms, and provide a tractable platform to investigate developmental and epigenetic mechanisms of oncogenesis. This MST model supports a unifying germ cell origin for both germ cell and non-germ cell somatic malignancies. Full article
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20 pages, 6738 KB  
Article
Evaluation of the Acceptability and Feasibility of the Social Attention and Communication Surveillance-Revised (SACS-R) Tool for Early Identification of Autism in Preterm Infants: The Identify and Act Study
by Gayatri Athalye-Jape, Sarah Pillar, Sudharshana Saminathan, Kexian Wu, Stephanie Sherrard, Emma Dudman and Mary Sharp
Children 2025, 12(9), 1130; https://doi.org/10.3390/children12091130 - 27 Aug 2025
Viewed by 424
Abstract
Introduction: Preterm birth is associated with a 3.3-fold increased likelihood of autism diagnosis, with lower gestational age conferring higher likelihood. In Australia, autism is typically diagnosed at around age four, potentially missing the optimal neuroplasticity window before age two. The Social Attention and [...] Read more.
Introduction: Preterm birth is associated with a 3.3-fold increased likelihood of autism diagnosis, with lower gestational age conferring higher likelihood. In Australia, autism is typically diagnosed at around age four, potentially missing the optimal neuroplasticity window before age two. The Social Attention and Communication Surveillance—Revised (SACS-R) tool identifies early autism signs in children aged 11–30 months, enabling pre-emptive intervention. Aims: This quality improvement (QI) study assessed the acceptability, and feasibility of SACS-R for early detection of autism traits in 12-month-old infants born very preterm/VP (gestation < 32 weeks), from both caregiver and clinician perspectives. Methods: From September 2024 to February 2025, 47 VP infants attending the 12-month Neonatal Follow-up Clinic (NNFU) at King Edward Memorial Hospital (KEMH), Western Australia, were assessed using SACS-R. Caregivers completed acceptability and feasibility questionnaires; clinicians completed similar surveys. Forty-seven infants met inclusion criteria; 12 clinicians provided responses. Results: Of 47 infants, 4 (8.5%) were identified as having a high likelihood of autism and referred for early intervention. Among caregivers, 29 (61%) provided complete acceptability responses and 28 (59%) feasibility responses, both predominantly positive. Clinicians reported high satisfaction (83%) and ease of use (91%), with 74% supporting routine implementation. Concerns included parental understanding and overlap with other assessments. Conclusions: Our QI study indicates that the SACS-R is highly acceptable and feasible in neonatal follow-up for preterm infants. Larger-scale evaluation of diagnostic accuracy and practical refinements based on feedback are warranted to support routine integration in early surveillance programs. Full article
(This article belongs to the Special Issue Long-Term Child and Family Outcomes After High-Risk Birth)
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15 pages, 2702 KB  
Article
Evaluation of Hypovirus Infection on the Vesicular Protein Expression Pattern of Cryphonectria parasitica by TMT-Based Proteomics Analysis
by Zishan Zeng, Nanxin Lin, Tao Lu, Jian Xu, Zheng Zhang, Fang Wang and Jinzi Wang
Biology 2025, 14(9), 1123; https://doi.org/10.3390/biology14091123 - 25 Aug 2025
Viewed by 387
Abstract
Hypovirus infection is known to reduce the pathogenicity of Cryphonectria parasitica, the causative agent of chestnut blight. Isoforms derived from a viral protein p48 have been discovered in host mitochondria and vesicles, which may contribute to virulence attenuation, as reported in earlier [...] Read more.
Hypovirus infection is known to reduce the pathogenicity of Cryphonectria parasitica, the causative agent of chestnut blight. Isoforms derived from a viral protein p48 have been discovered in host mitochondria and vesicles, which may contribute to virulence attenuation, as reported in earlier work using two-dimensional electrophoresis (2-DE). In this study, a total of 1739 fungal proteins were identified in fungal vesicles through Tandem Mass Tag (TMT)-based quantitative proteomics. The infection of CHV1-EP713 was associated with 75 up-regulated and 201 down-regulated proteins, predominantly involved in vesicular transport process and related cellular functions, including protein folding, membrane fusion, retrograde transport, autophagy, and ER stress responses. The down-regulation of calnexin, COPI, ArfGAP, importin-β, and Atg8 is consistent with impairments in protein folding, retrograde transport, and autophagy. Meanwhile, the up-regulation of clathrin, dynamin, Vps10p, HSP70, and t-SNAREs indicated enhanced trafficking to vacuoles and increased stress response activity. Overall, our findings indicate that hypoviral infection is associated with extensive alterations in the vesicular transport system of C. parasitica, likely mediated through changes in the abundance of multiple key protein regulators. These alterations may underlie attenuation of virulence by impacting crucial cellular processes. Full article
(This article belongs to the Section Infection Biology)
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15 pages, 763 KB  
Article
Immune Checkpoint Dysregulation in Aneurysmal Subarachnoid Hemorrhage: A Prospective Study of sCTLA-4 and sPD-L1 as Biomarkers of Symptomatic Vasospasm
by Reka Varnai, Gábor J. Szebeni, Nikolett Gémes, Attila Schwarcz, Tihamer Molnar, Csaba Olah and Peter Csecsei
Int. J. Mol. Sci. 2025, 26(17), 8228; https://doi.org/10.3390/ijms26178228 - 25 Aug 2025
Viewed by 494
Abstract
Aneurysmal subarachnoid hemorrhage (aSAH) is a severe stroke subtype often complicated by symptomatic cerebral vasospasm (sVP), contributing to delayed cerebral ischemia and poor outcomes. Immune dysregulation, particularly T-cell imbalances and pro-inflammatory cytokines, is implicated in vasospasm development. Soluble immune checkpoint proteins—CTLA-4 (sCTLA-4) and [...] Read more.
Aneurysmal subarachnoid hemorrhage (aSAH) is a severe stroke subtype often complicated by symptomatic cerebral vasospasm (sVP), contributing to delayed cerebral ischemia and poor outcomes. Immune dysregulation, particularly T-cell imbalances and pro-inflammatory cytokines, is implicated in vasospasm development. Soluble immune checkpoint proteins—CTLA-4 (sCTLA-4) and PD-L1 (sPD-L1)—regulate immune homeostasis and may serve as biomarkers or modulators of inflammation in aSAH. This prospective cohort study included 179 aSAH patients, divided into sVP+ (n = 48) and sVP− (n = 131), plus 50 healthy controls. Serum sCTLA-4 and sPD-L1 levels were measured on days 1, 5, and 9 post-ictus using Luminex xMAP. Associations with clinical outcomes were analyzed using non-parametric statistics and hierarchical clustering. Both sCTLA-4 and sPD-L1 were significantly elevated in sVP+ patients versus sVP− and controls, increasing over time. sCTLA-4 was significantly higher in sVP+ on days 5 (p = 0.001) and 9 (p < 0.001), and sPD-L1 on days 5 and 9 (both p < 0.001). Clustering revealed distinct expression patterns between sVP+ and sVP− groups. Elevated sCTLA-4 and sPD-L1 levels are associated with sVP after aSAH and may serve as biomarkers for early immune dysfunction, offering insights into potential therapeutic targets. Full article
(This article belongs to the Section Molecular Neurobiology)
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17 pages, 6170 KB  
Article
Immunogenicity of Virus-like Particles Based on VP1 Protein of Bovine Norovirus
by Zhigang Ma, Xuelian Ma, Xinyu Tao, Yong Huang, Qian Jiang, Xiaojun Ding, Fang Min, Yichen Chu, Ru Li, Xinying Zhang, Lu Liu, Caiyun Zhang, Qi Zhong and Gang Yao
Vet. Sci. 2025, 12(9), 802; https://doi.org/10.3390/vetsci12090802 - 24 Aug 2025
Viewed by 528
Abstract
Bovine Norovirus (BNoV) is a member of the enterovirus family that can cause gastroenteritis in calves. This virus poses a significant risk to calf growth and development as well as to the long-term sustainability of the cattle industry in China and elsewhere. No [...] Read more.
Bovine Norovirus (BNoV) is a member of the enterovirus family that can cause gastroenteritis in calves. This virus poses a significant risk to calf growth and development as well as to the long-term sustainability of the cattle industry in China and elsewhere. No specific treatment or vaccine is currently available; thus, the development of a safe and effective vaccine is paramount. Here, we describe a strategy to assemble BNoV virus-like particles (VLPs) using the insect baculovirus expression system (BEV) to express the major structural protein, VP1, and demonstrate their potentiality as vaccines. The results showed that the BNoV-VLP self-assembled into complete spherical particles with a diameter of approximately 40 nm. When it was immunized in mice, the levels of specific IgG and IgA antibodies peaked at weeks 6 and 7 post-immunization, respectively, with maximum titers of 1:25,600 and 1:200. Moreover, we observed a significant increase in the CD4+/CD8+ T-cell ratio in splenic lymphocytes of immunized mice (p < 0.05), accompanied by a significant increase in TNF-α+CD4+ T-cells and TNF-α+CD8+ T-cells (p < 0.05). These results demonstrate that BNoV-VLPs are promising vaccine candidates for providing immunoprotection in the future. These studies support the significant practical implications of using a scientific basis for the development of a BNoV-VLP vaccine. Full article
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19 pages, 4449 KB  
Article
Characterization of the NFAT Gene Family in Grass Carp (Ctenopharyngodon idellus) and Functional Analysis of NFAT1 During GCRV Infection
by Yao Shen, Yitong Zhang, Chen Chen, Shitao Hu, Jia Liu, Yiling Zhang, Tiaoyi Xiao, Baohong Xu and Qiaolin Liu
Fishes 2025, 10(9), 422; https://doi.org/10.3390/fishes10090422 - 22 Aug 2025
Viewed by 361
Abstract
Nuclear factors of activated T cells (NFATs) are pivotal regulatory factors of immune responses, primarily by modulating T cell activity and regulating inflammatory cytokine gene transcription. The grass carp reovirus (GCRV) triggers a serious hemorrhagic condition, posing a significant threat to sustainable grass [...] Read more.
Nuclear factors of activated T cells (NFATs) are pivotal regulatory factors of immune responses, primarily by modulating T cell activity and regulating inflammatory cytokine gene transcription. The grass carp reovirus (GCRV) triggers a serious hemorrhagic condition, posing a significant threat to sustainable grass carp (Ctenopharyngodon idella) aquaculture. However, the precise function of NFAT in the host’s defense against GCRV infection is mostly undefined. This study comprehensively identified and characterized the NFAT genetic family in grass carp, cloned grass carp NFAT1 (CiNFAT1), and investigated its expression and function during GCRV infection. Eight NFAT genes encoding seventeen isoforms have been detected within the grass carp’s genomic sequence, distributed across six different chromosomes. Comparative analysis revealed homology with zebrafish NFATs. CiNFAT1 possesses a 2697 bp open reading frame, encoding 898 amino acids, and contains conserved Rel homology domain (RHD) and NFAT-homology (IPT) domains. Quantitative PCR (qPCR) revealed ubiquitous CiNFAT1 expression in healthy grass carp tissues, with the highest expression in gills and skin and the lowest in liver. Following GCRV challenge in vivo, CiNFAT1 expression in immune tissues (liver, spleen, kidney, gill, intestine) showed dynamic changes over time. In vitro experiments in CIK cells demonstrated that CiNFAT1 expression peaked at 12 h post-GCRV infection. Further functional studies revealed that overexpression of CiNFAT1 significantly reduced GCRV replication at 36 h post-infection. This reduction was accompanied by elevated expression of type I interferon (IFN-I) and interferon regulatory factor 7 (IRF7) at 24 and 36 h, respectively, as well as modulated IL-2, IL-8, and IL-10. Conversely, RNA interference-mediated knockdown of CiNFAT1 enhanced GCRV VP5 and VP7 mRNA levels and suppressed IL-2 and IL-8 expression. These results suggest that CiNFAT1 contributes to anti-GCRV immunity by promoting antiviral and inflammatory cytokine responses, thereby inhibiting viral replication. This study provides a foundational understanding of the NFAT genetic family in grass carp and highlights an important role of CiNFAT1 in mediating the body’s inherent defense mechanism against GCRV infection, offering insights for disease control strategies in aquaculture. Full article
(This article belongs to the Special Issue Molecular Design Breeding in Aquaculture)
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19 pages, 3042 KB  
Article
Characterization of GmABI3VP1 Associated with Resistance to Soybean Cyst Nematode in Glycine max
by Shuo Qu, Miaoli Zhang, Gengchen Song, Shihao Hu, Weili Teng, Yongguang Li, Xue Zhao, Rongxia Guan and Haiyan Li
Agronomy 2025, 15(8), 2005; https://doi.org/10.3390/agronomy15082005 - 21 Aug 2025
Viewed by 429
Abstract
The ABI3 transcription factor is a key regulator in plant growth and development. Through transcriptome analysis of the resistant soybean cultivar ‘Dongnong L10′ and the susceptible cultivar ‘Heinong 37′ exposed to soybean cyst nematode race 3 (SCN 3) stress, the differentially expressed gene [...] Read more.
The ABI3 transcription factor is a key regulator in plant growth and development. Through transcriptome analysis of the resistant soybean cultivar ‘Dongnong L10′ and the susceptible cultivar ‘Heinong 37′ exposed to soybean cyst nematode race 3 (SCN 3) stress, the differentially expressed gene GmABI3VP1 was identified. The GmABI3VP1 gene was then cloned and analyzed through bioinformatics, subcellular localization, and qRT-PCR analysis of resistant and susceptible soybean germplasms, as well as overexpression and gene editing of soybean hairy roots followed by SCN 3 identification analysis. It was found that the protein encoded by GmABI3VP1 is an acidic and hydrophilic protein with transmembrane domains. It has a collinear relationship with Arabidopsis and is widely distributed in plants. Through the analysis of promoter elements, it was shown that this gene contains multiple hormone-responsive promoter elements like ABRE/ABRE3a/ABRE/4a/as-1 and stress-responsive elements such as Myb/MYC/MYc. Transient expression in tobacco indicated that the GmABI3VP1 gene is located in the nucleus. The transcription of GmABI3VP1 responds to the stress of SCN, and its transcriptional level is relatively high in the roots of resistant materials. Genetic transformation mediated by Agrobacterium rhizogenes was used to obtain GmABI3VP1 gene overexpressed and CRISPR-Cas9 gene-edited soybean hairy roots. In comparison to the wild type (WT), the density of nematodes per area was notably lower in hairy roots overexpressing (OX) the gene, whereas the density of SCN per unit area (per cm of lateral root length) significantly increased in gene-edited (KO) soybean hairy roots. Through SCN phenotyping, GmABI3VP1 was identified as a contributor to SCN 3 resistance. This study provides initial insights into the role of the GmABI3VP1 gene in SCN resistance, establishing a robust basis for future research on the mechanisms underlying SCN disease resistance and offering valuable genetic reservoirs for SCN 3 resistance. Full article
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Article
Elemental Composition Analysis of Major Refined Petroleum Fuel Products in Ghana
by Robert Wilson and Calvin Kwesi Gafrey
Fuels 2025, 6(3), 62; https://doi.org/10.3390/fuels6030062 - 19 Aug 2025
Viewed by 341
Abstract
Samples of refined petroleum fuels from the three major oil-marketing companies (GOIL Company Limited, Total Energies Ghana Limited and Shell Vivo Ghana Limited) in Ghana have been analysed for elemental concentrations using an X-ray fluorescence facility at the National Nuclear Research Institute, Ghana [...] Read more.
Samples of refined petroleum fuels from the three major oil-marketing companies (GOIL Company Limited, Total Energies Ghana Limited and Shell Vivo Ghana Limited) in Ghana have been analysed for elemental concentrations using an X-ray fluorescence facility at the National Nuclear Research Institute, Ghana Atomic Energy Commission. The samples were acquired from seven different fuel service stations where customers directly purchase refined petroleum fuels such as diesel, petrol and kerosene. The X-ray fluorescence method was considered for the study because sample preparation does not require the addition of reagents, and the fluorescence measurements involve a direct electron transition effect. The fluorescence study was carried out to estimate the concentrations of sulphur and other contaminants in the major refined petroleum fuel products patronised in Ghana. The average sulphur concentration in the samples of diesel products were 17.543, 25.805 and 26.813 ppm in DFS, DE and DXP samples compared to 22.258, 22.623 and 15.748 ppm in petrol samples of PE, PXP and VP. Also, the sulphur concentration of sample KE, kerosene products, is 33.250 ppm. Among the diesel samples, DE and DXP recorded the highest but most comparable average concentration of elemental contaminants, and DFS the least, while PXP recorded the least among the petrol samples. The study estimated the concentrations of four heavy metal elements that are toxic to biological life (Hg, Pb, Cr and Mn) to be less than 10.0 ppm, except Cr. The study concluded that most of the elemental contaminants of heavy metals in the samples were relatively less than ultra-low levels. Therefore, exhaust emissions may have little impact on the environment. Also, the content of the ash-producing metal elements in each sample of the seven refined fuel products is between 10.0 and 50.0 ppm. Since the concentration of sulphur and a few other elemental contaminants could not meet the internationally accepted standard (<10.0 ppm), the imported refined fuel products used in Ghana may be considered relatively good but not environmentally safe. Full article
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