Search Results (2,191)

The barbel chub (Squaliobarbus curriculus) exhibits remarkable resistance to grass carp reovirus (GCRV), a devastating pathogen in aquaculture. To reveal the molecular basis of this resistance, we investigated complement factor D (DF)—a rate-limiting serine protease governing alternative complement pathway activation. Molecular cloning revealed that the barbel chub DF (ScDF) gene encodes a 1251-bp cDNA sequence translating into a 250-amino acid protein. Crucially, bioinformatic characterization identified a unique N-glycosylation site at Asn¹³⁹ in ScDF, representing a structural divergence absent in grass carp (Ctenopharyngodon idella) DF (CiDF). While retaining a conserved Tryp_SPc domain harboring the catalytic triad (His⁶¹, Asp¹⁰⁹, and Ser²⁰⁴) and substrate-binding residues (Asp¹⁹⁸, Ser²¹⁹, and Gly²²¹), sequence and phylogenetic analyses confirmed ScDF’s evolutionary conservation, displaying 94.4% amino acid identity with CiDF and clustering within the Cyprinidae. Expression profiling revealed constitutive ScDF dominance in the liver, and secondary prominence was observed in the heart. Upon GCRV challenge in S. curriculus kidney (SCK) cells, ScDF transcription surged to a 438-fold increase versus uninfected controls at 6 h post-infection (hpi; p < 0.001)—significantly preceding the 168-hpi response peak documented for CiDF in grass carp. Functional validation showed that ScDF overexpression suppressed key viral capsid genes (VP2, VP5, and VP7) and upregulated the interferon regulator IRF9. Moreover, recombinant ScDF protein incubation induced interferon pathway genes and complement C3 expression. Collectively, ScDF’s rapid early induction (peaking at 6 hpi) and multi-pathway coordination may contribute to barbel chub’s GCRV resistance. These findings may provide molecular insights into the barbel chub’s high GCRV resistance compared to grass carp and novel perspectives for anti-GCRV breeding strategies in fish. Full article

Museums may acquire archaeological artefacts discovered by non-specialists or amateur archaeologists, holding the potential to promote the safeguarding of cultural heritage by integrating the local community in their activities. However, this also creates an opportunity for the fraudulent sale of modern forgeries presented as archaeological artefacts, resulting in the need for a critical assessment of the artefact’s authenticity prior to acquisition by the museum. In 2019, the regional museum in Cáceres (Spain) was offered the opportunity to acquire a Phoenician-Punic head pendant, allegedly discovered in the vicinity of the city. The artefact’s authenticity was assessed by traditional approaches, including typological analysis and analysis of manufacture technique, which raised doubts about its purported age. VP-SEM-EDS analysis of the chemical composition of the different glass portions comprising the pendant was used for non-invasive determination of glassmaking recipes, enabling the identification of glass components incompatible with known Iron Age glassmaking recipes from the Mediterranean. Further comparison with historical and modern glassmaking recipes allowed for the identification of the artefact as a recent forgery made from glasses employing modern colouring and opacifying techniques. Full article

Soil salinization poses a significant constraint to agricultural productivity. However, certain plant growth-promoting bacteria (PGPB) can mitigate salinity stress and enhance crop performance. In this study, a bacterial isolate, R-18, isolated from saline-alkali soil in Ningxia, China, was identified as Enterobacter bugandensis based on 16S rRNA gene sequencing. The isolate was characterized for its morphological, biochemical, and plant growth-promoting traits and was evaluated for its potential to alleviate NaCl-induced stress in maize (Zea mays L.) under hydroponic conditions. Isolate R-18 exhibited halotolerance, surviving at NaCl concentrations ranging from 2.0% to 10.0%, and alkaliphilic adaptation, growing at pH 8.0–11.0. Biochemical assays confirmed it as a Gram-negative bacterium, displaying positive reactions in the Voges–Proskauer (V–P) tests, catalase activity, citrate utilization, fluorescent pigment production, starch hydrolysis, gelatin liquefaction, and ammonia production, while testing negative for the methyl red and cellulose hydrolysis. Notably, isolate R-18 demonstrated multiple plant growth-promoting attributes, including nitrogen fixation, phosphate and potassium solubilization, ACC deaminase activity, and indole-3-acetic acid (IAA) biosynthesis. Under 100 mM NaCl stress, inoculation with isolate R-18 significantly enhanced maize growth, increasing plant height, stem dry weight, root fresh weight, and root dry weight by 20.64%, 47.06%, 34.52%, and 31.25%, respectively. Furthermore, isolate R-18 improved ion homeostasis by elevating the K⁺/Na⁺ ratio in maize tissues. Physiological analyses revealed increased chlorophyll and proline content, alongside reduced malondialdehyde (MDA) levels, indicating mitigated oxidative damage. Antioxidant enzyme activity was modulated, with decreased superoxide dismutase (SOD) and peroxidase (POD) activities but increased catalase (CAT) activity. These findings demonstrated that Enterobacter bugandensis R-18 effectively alleviated NaCl-induced growth inhibition in maize by enhancing osmotic adjustment, reducing oxidative stress, and improving ion balance. Full article

The Korteweg–de Vries (KdV) equation is a nonlinear evolution equation that reflects a wide variety of dispersive wave occurrences with limited amplitude. It has also been used to describe a range of major physical phenomena, such as shallow water waves that interact weakly and nonlinearly, acoustic waves on a crystal lattice, lengthy internal waves in density-graded oceans, and ion acoustic waves in plasma. The KdV equation is one of the most well-known soliton models, and it provides a good platform for further research into other equations. The KdV equation has several forms. The aim of this study is to introduce and investigate a (2+1)-dimensional generalized fifth-order KdV equation with power law nonlinearity (gFKdVp). The research methodology employed is the Lie group analysis. Using the point symmetries of the gFKdVp equation, we transform this equation into several nonlinear ordinary differential equations (ODEs), which we solve by employing different strategies that include Kudryashov’s method, the $(G^{\prime }/G)$ expansion method, and the power series expansion method. To demonstrate the physical behavior of the equation, 3D, density, and 2D graphs of the obtained solutions are presented. Finally, utilizing the multiplier technique and Ibragimov’s method, we derive conserved vectors of the gFKdVp equation. These include the conservation of energy and momentum. Thus, the major conclusion of the study is that analytic solutions and conservation laws of the gFKdVp equation are determined. Full article

Objectives: Most antiviral vaccines are created by inactivating the virus using chemical methods. The inactivation and production of viral vaccine preparations after the irradiation of viruses with accelerated electrons has a number of significant advantages. Determining the integrity of the genome of the resulting viral particles is necessary to assess the quality and degree of inactivation after irradiation. Methods: This work was performed on the Sabin 2 model polio virus. To determine the most sensitive and most radiation-resistant part, the polio virus genome was divided into 20 segments. After irradiation at temperatures of 25 °C, 2–8 °C, −20 °C, or −70 °C, the amplification intensity of these segments was measured in real time. Results: The best correlation between the amplification cycle and the irradiation dose at all temperatures was observed for segment 3D, left. Consequently, this section of the poliovirus genome is the least resistant to the action of accelerated electrons and is the most representative for determining genome integrity. The worst dependence was observed for the VP1 right section, which, therefore, cannot be used to determine genome integrity during inactivation. The electrochemical approach was also employed for a comparative assessment of viral RNA integrity before and after irradiation. An increase in the irradiation dose was accompanied by an increase in signals indicating the electrooxidation of RNA heterocyclic bases. The increase in peak current intensity of viral RNA electrochemical signals confirmed the breaking of viral RNA strands during irradiation. The shorter the RNA fragments, the greater the peak current intensities. In turn, this made the heterocyclic bases more accessible to electrooxidation on the electrode. Conclusions: These results are necessary for characterizing the integrity of the viral genome for the purpose of creating of antiviral vaccines. Full article

Vibrio parahaemolyticus is a major foodborne pathogen worldwide, responsible for seafood-associated poisoning. Among its toxin genes, tdh2 is the most critical. To investigate the role of tdh2 in V. parahaemolyticus under gastrointestinal conditions, we constructed tdh2 deletion and complementation strains and compared their survival under acid (pH 3 and 4) and bile stress (2%). The results showed that tdh2 expression was significantly upregulated under cold (4 °C) and bile stress (0.9%). Survival assays and PI staining revealed that the tdh2 mutant strain (VP: △tdh2) was more sensitive to acid and bile stress than the wild-type (WT), and this sensitivity was rescued by tdh2 complementation. These findings suggest that tdh2 plays a protective role in enhancing V. parahaemolyticus tolerance to acid and bile stress. In the VP: △tdh2 strain, seven genes were significantly upregulated and six were downregulated as a result of tdh2 deletion. These genes included VPA1332 (vtrA), VPA1348 (vtrB), VP2467 (ompU), VP0301 and VP1995 (ABC transporters), VP0527 (nhaR), and VP2553 (rpoS), among others. Additionally, LC-MS/MS analysis identified 12 differential metabolites between the WT and VP: △tdh2 strains, including phosphatidylserine (PS) (17:2 (9Z,12Z) /0:0 and 20:1 (11Z) /0:0), phosphatidylglycerol (PG) (17:0/0:0), flavin mononucleotide (FMN), and various nucleotides. The protective mechanism of tdh2 may involve preserving cell membrane permeability through regulation of ompU and ABC transporters and enhancing electron transfer efficiency via regulation of nhaR. The resulting reduction in ATP, DNA, and RNA synthesis—along with changes in membrane permeability and electron transfer due to decreased FMN—likely contributed to the reduced survival of the VP: △tdh2 strain. Meanwhile, the cells actively synthesized phospholipids to repair membrane damage, leading to increased levels of PS and PG. This study provides important insights into strategies for preventing and controlling food poisoning caused by tdh⁺ V. parahaemolyticus. Full article

Poly(N-vinyl-2-pyrrolidone), or PVP, nanogels loaded with gadolinium nitrate (Gd(NO₃)₃·6H₂O) were synthesized by ionizing irradiation, aiming for potential applications in magnetic resonance imaging (MRI). A comprehensive characterization of PVP and Gd aqueous solutions with different VP-monomer-to-Gd ratios was conducted before and after irradiation. The results indicate a complexation between PVP and Gd ions before irradiation. The size of the nanogels exhibited a strong dependence on several factors, including PVP molecular weight, concentration, temperature, and the precise timing of Gd introduction relative to the irradiation process. A quantification study was conducted to investigate the impact of molecular weight, the VP/Gd ratio, and Gd addition before or after the irradiation process on the concentration of free Gd ions. These findings offer valuable insights into optimizing the synthesis of Gd-loaded PVP nanogels for potential applications, highlighting the critical factors that influence their size and stability. Full article

Porcine Rotavirus (PoRV), a predominant causative agent of neonatal diarrhea in piglets, shares substantial genetic homology with human rotavirus and represents a considerable threat to both public health and the global swine industry in the absence of specific antiviral interventions. The VP6 protein, an internal capsid component, is characterized by exceptional sequence conservation and robust immunogenicity, rendering it an ideal candidate for viral genotyping and vaccine development. In the present study, the recombinant plasmid pET28a(+)-VP6 was engineered to facilitate the high-yield expression and purification of the VP6 antigen. BALB/c mice were immunized to generate monoclonal antibodies (mAbs) through hybridoma technology, and the antigenic specificity of the resulting mAbs was stringently validated. Subsequently, a panel of truncated protein constructs was designed to precisely map linear B-cell epitopes, followed by comparative conservation analysis across diverse PoRV strains. Functional validation demonstrated that all three mAbs exhibited high-affinity binding to VP6, with a peak detection titer of 1:3,000,000 and exclusive specificity toward PoRVA. These antibodies effectively recognized representative genotypes such as G3 and X1, while exhibiting no cross-reactivity with unrelated viral pathogens; however, their reactivity against other PoRV serogroups (e.g., types B and C) remains to be further elucidated. Epitope mapping identified two novel linear B-cell epitopes, ¹²⁸YIKNWNLQNR¹³⁷ and ¹³⁸RQRTGFVFHK¹⁴⁷, both displaying strong sequence conservation among circulating PoRV strains. Collectively, these findings provide a rigorous experimental framework for the functional dissection of VP6 and reinforce its potential as a valuable diagnostic and immunoprophylactic target in PoRV control strategies. Full article

Nucleocytoplasmic trafficking is a highly regulated process that allows the cell to control the partitioning of proteins and nucleic acids between the cytosolic and nuclear compartments. The Ebola virus minor matrix protein VP24 (eVP24) hijacks this process by binding to a region on the NPI-1 subfamily of karyopherin alpha (KPNA) nuclear importers. This region overlaps with the activated transcription factor STAT1 binding site on KPNAs, preventing STAT1 nuclear localization and activation of antiviral gene transcription. However, the molecular interactions of eVP24-KPNA5 binding that lead to the nuclear localization of eVP24 remain poorly characterized. Here, we show that trafficking of eVP24 into the nucleus by KPNA5 requires simultaneous binding of cargo. We also describe the conformational dynamics of KPNA5 and interactions with eVP24 and cargo nuclear localization sequences (NLS) using biophysical approaches. Our results reveal that eVP24 binding to KPNA5 does not impact cargo NLS binding to KPNA5, indicating that simultaneous binding of both cellular cargo and eVP24 to KPNA5 is likely required for nuclear trafficking. Together, these results provide a biophysical basis for how Ebola virus VP24 protein gains access to the nucleus during Ebola virus infection. Full article

Stable oxygen isotopes in tree rings (δ¹⁸O) serve as important proxies for climate change and offer unique advantages for climate reconstruction in arid and semi-arid regions. We established an annual δ¹⁸O chronology spanning 1964–2023 using Juniperus excelsa tree-ring samples collected from the Alborz Mountains in Iran. We analyzed relationships between δ¹⁸O and key climate variables: precipitation, temperature, Palmer Drought Severity Index (PDSI), vapor pressure (VP), and potential evapotranspiration (PET). Correlation analysis reveals that tree-ring δ¹⁸O is highly sensitive to hydroclimatic variations. Tree-ring cellulose δ¹⁸O shows significant negative correlations with annual total precipitation and spring PDSI, and significant positive correlations with spring temperature (particularly maximum temperature), April VP, and spring PET. The strongest correlation occurs with spring PET. These results indicate that δ¹⁸O responds strongly to the balance between springtime moisture supply (precipitation and soil moisture) and atmospheric evaporative demand (temperature, VP, and PET), reflecting an integrated signal of both regional moisture availability and energy input. The pronounced response of δ¹⁸O to spring evaporative conditions highlights its potential for capturing high-resolution changes in spring climatic conditions. Our δ¹⁸O series remained stable from the 1960s to the 1990s, but showed greater interannual variability after 2000, likely linked to regional warming and climate instability. A comparison with the δ¹⁸O variations from the eastern Alborz Mountains indicates that, despite some differences in magnitude, δ¹⁸O records from the western and eastern Alborz Mountains show broadly similar variability patterns. On a larger climatic scale, δ¹⁸O correlates significantly and positively with the Niño 3.4 index but shows no significant correlation with the Arctic Oscillation (AO) or the North Atlantic Oscillation (NAO). This suggests that ENSO-driven interannual variability in the tropical Pacific plays a key role in regulating regional hydroclimatic processes. This study confirms the strong potential of tree-ring oxygen isotopes from the Alborz Mountains for reconstructing hydroclimatic conditions and high-frequency climate variability. Full article

Background/Objectives: Norovirus is a major cause of acute gastroenteritis worldwide. Considering its highly infectious and transmissible nature, rapid and accurate diagnostic tools are of utmost importance for the effective control of outbreaks in the context of point-of-care testing (POCT). In this study, we developed a genogroup-specific multiplex reverse transcriptase loop-mediated isothermal amplification assay to detect the human norovirus genogroups I and II (GI and GII, respectively). Methods: For the comprehensive detection of clinically relevant genotypes, two sets of primers were incorporated into the assays targeting the RdRp-VP1 junction: one against GI.1 and GI.3, and the other for GII.2 and GII.4. Following optimization of the reaction variables, we standardized the reaction conditions at 65 °C with 6 mM MgSO₄, 1.4 mM dNTPs, 7.5 U WarmStart RTx Reverse Transcriptase, and Bst DNA polymerase at 8 U and 10 U for GI and GII, respectively. Amplification was monitored in real-time using a thermocycler platform to ensure precise quantification and detection. Finally, the assay was evaluated through portable isothermal detection device to test its feasibility in on-site settings. Results: Both assays detected the template down to 10²–10³ copies per reaction and showed high target selectivity, yielding no non-specific amplification across 39 enteric pathogens. These assays enabled prompt detection of GI within 10–12 min and of GII within 12–17 min after the reaction was initiated. Onsite validation reveals all template detection below 15 min, demonstrating its potential feasibility in point-of-care applications. Including the sample preparation time, test results were obtained in less than 1 h. Conclusions: This method is a rapid, reliable, and scalable solution for detecting human norovirus in POCT settings for both clinical diagnosis and public health surveillance. Full article

The structural analysis of shelterbelts forms the foundation of their planning and management, yet the scientific and effective quantification of shelterbelt structures requires further investigation. This study developed an innovative heterogeneous analytical framework, integrating three key methodologies: the LeWoS algorithm for wood–leaf separation, TreeQSM for structural reconstruction, and 3D alpha-shape spatial quantification, using terrestrial laser scanning (TLS) technology. This framework was applied to three typical farmland shelterbelts in the Ulan Buh Desert oasis, enabling the first precise quantitative characterization of structural components during the leaf-on stage. The results showed the following to be true: (1) The combined three-algorithm method achieved ≥90.774% relative accuracy in extracting structural parameters for all measured traits except leaf surface area. (2) Branch length, diameter, surface area, and volume decreased progressively from first- to fourth-order branches, while branch angles increased with ascending branch order. (3) The trunk, branch, and leaf components exhibited distinct vertical stratification. Trunk volume and surface area decreased linearly with height, while branch and leaf volumes and surface areas followed an inverted U-shaped distribution. (4) Horizontally, both surface area density (Scd) and volume density (Vcd) in each cube unit exhibited pronounced edge effects. Specifically, the Scd and Vcd were greatest between 0.33 and 0.60 times the shelterbelt’s height (H, i.e., mid-canopy). In contrast, the optical porosity (Op) was at a minimum of 0.43 H to 0.67 H, while the volumetric porosity (Vp) was at a minimum at 0.25 H to 0.50 H. (5) The proposed volumetric stratified porosity (Vsp) metric provides a scientific basis for regional farmland shelterbelt management strategies. This three-dimensional structural analytical framework enables precision silviculture, with particular relevance to strengthening ecological barrier efficacy in arid regions. Full article

Amyotrophic lateral sclerosis (ALS) remains a progressive neurodegenerative disease, lacking effective causal therapies. The Wobbler mouse model harboring a spontaneous autosomal recessive mutation in the vacuolar protein sorting associated protein (Vps54), has emerged as a valuable model for investigating ALS pathophysiology and potential treatments. This model exhibits cellular and phenotypic parallels to human ALS, including protein aggregation, microglia and astrocyte activation, as well as characteristic disease progression at distinct stages. Exploring the underlying pathomechanisms and identifying therapeutic targets requires a comprehensive analysis of gene and protein expression. In this study, we examined the expression of three well-established housekeeping genes and proteins—calnexin, ß-actin, and ßIII-tubulin—in the cervical spinal cord of the Wobbler model. These candidates were selected based on their demonstrated stability across various systems like animal models or cell culture. Calnexin, an integral protein of the endoplasmic reticulum, ß-actin, a structural component of the cytoskeleton, and ß-tubulin III, a component of microtubules, were quantitatively assessed using quantitative reverse transcription-polymerase chain reaction (RT-PCR) for gene expression and Western blotting for protein expression. Our results revealed no significant differences in the expression of CANX, ACTB, and TUBB3 between spinal cords of wild-type and Wobbler mice at the symptomatic stage (p40) at both the gene and protein levels. These findings suggest that the pathophysiological alterations induced by the Wobbler mutation do not significantly affect the expression of these crucial housekeeping genes and proteins at p40. Overall, this study provides a basis for further investigations using the Wobbler mouse model, while highlighting the potential use of calnexin, ß-actin, and ßIII-tubulin as reliable reference genes and proteins in future research to aid in the discovery for effective therapeutic interventions. Full article

Background: Parvovirus B19 (B19V) can cause severe relapsing episodes of pure red cell aplasia in immunocompromised individuals, which are commonly treated with intravenous immunoglobulins (IVIGs). Few data are available on B19V intra-host evolution and the role of humoral immune selection. Here, we report the dynamics of genomic mutations and subsequent protein changes during relapsing infection. Methods: Longitudinal plasma samples from immunocompromised patients with relapsing B19V infection in the period 2011–2019 were analyzed using whole-genome sequencing to evaluate intra-host evolution. The impact of mutations on the 3D viral protein structure was predicted by deep neural network modeling. Results: Of the three immunocompromised patients with relapsing infections for 3 to 9 months, one patient developed two consecutive nonsynonymous mutations in the VP1/2 region: T372S/T145S and Q422L/Q195L. The first mutation was detected in multiple B19V IgG-seropositive follow-up samples and resolved after IgG seroreversion. Computational prediction of the VP1 3D structure of this mutant showed a conformational change in the proximity of the antibody binding domain. No conformational changes were predicted for the other mutations detected. Discussion: Analysis of relapsing B19V infections showed mutational changes occurring over time. Resulting amino acid changes were predicted to lead to a conformational capsid protein change in an IgG-seropositive patient. The impact of humoral response and IVIG treatment on B19V infections should be further investigated to understand viral evolution and potential immune escape. Full article

Background: Migration of the peritoneal end of the ventriculoperitoneal shunt (VPS) into the scrotum is a rare but recognized complication. Inguinoscrotal migration typically occurs as a result of increased intra-abdominal pressure combined with a patent processus vaginalis. A 14-month-old pediatric patient presented to the emergency department with abdominal pain, vomiting, and swelling of the right scrotum that had persisted for two days. The patient had a history of a head injury that had resulted in a large secondary arachnoid cyst for which a VPS had been placed at eight months of age. Examination of the inguinoscrotal region revealed a swollen and painful right side of the scrotum with a hydrocele and a palpable distal portion of the ventriculoperitoneal catheter in the right groin extending to the scrotum. After a brief preoperative preparation, the patient underwent laparoscopic abdominal emergency exploration, during which shunt repositioning and laparoscopic closure of the patent processus vaginalis were performed. Methods: A systematic review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Results: A total of 30 case reports and six case series were included, analyzing 52 pediatric patients with scrotal migration of the VPS. The median age at presentation was 24 months (range: 1–169 months). The indication for VPS placement was hydrocephalus. Migration of the VPS catheter occurred on the right side in 34 cases. The median interval from VPS placement to the onset of symptoms was 9.0 months (range: 1 day–72 months). The most frequently reported symptoms were scrotal/inguinoscrotal swelling (n = 50), vomiting (n = 7), and fever (n = 3). Diagnostic methods included abdominal X-ray (n = 43), ultrasound (n = 5), scrotal transillumination test (n = 5), and computed tomography (n = 1). Regarding treatment, surgical repositioning of the VPS catheter into the peritoneal cavity was performed in 47 patients (90.4%), with no intraoperative or postoperative complications reported. Conclusions: Laparoscopic repositioning of the VPS into the peritoneal cavity, combined with closure of the processus vaginalis, appears to be a safe and effective treatment option for scrotal migration of the VPS. However, further well-designed studies are warranted to provide more comprehensive, generalizable, and unbiased evidence regarding this complication in the pediatric population. Full article