Search Results (785)

Background/Objectives: Porcine circovirus type 2 (PCV2) infection causes porcine circovirus disease (PCVD), a global immunosuppressive disease in pigs. Its clinical manifestations include post-weaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS), which cause significant economic losses to the swine industry. The Cap protein, which is the major protective antigen of PCV2, can self-assemble to form virus-like particles (VLPs) in the insect baculovirus expression system. Few studies have compared the expression of Cap proteins in different baculovirus expression systems. Methods: In this study, we compared two commonly commercialized baculovirus construction systems with the Cap protein expression in various insect cells. Results: The results demonstrate that the flashBAC system expressed the Cap protein at higher levels than the Bac-to-Bac system. Notably, when expressing four copies of the Cap protein, the flashBAC system achieved the highest protein yield in High Five cells, where it reached 432 μg/mL at 5 days post-infection (dpi) with 27 °C cultivation. Animal experiments confirmed that the purified Cap protein effectively induced specific antibody production in mice and swine. Conclusions: This study provides critical data for optimizing the production of the PCV2 Cap protein, which is of great significance for reducing the production cost of PCV2 vaccines and improving the industrial production efficiency. Full article

A circular tri-trophic system integrating aquaponics, i.e., combined cultivation of crops and fish, with insect rearing is presented for lettuce cultivation. The nutrition cycle among crops, insects and fish turns waste into resource, thereby increasing the sustainability of this food production system. A comprehensive evaluation of the system’s efficiency was performed, including the growth, functional and resource use efficiency traits of lettuce, the dynamics of which were followed in a pilot-scale aquaponics greenhouse, under three treatments: conventional hydroponics (HP) as the control, coupled aquaponics (CAP) with crops irrigated with fish-derived water, and decoupled aquaponics (DCAP), where fish-derived water was amended with fertilizers to reach the HP target. The main findings indicate comparable physiological performance between DCAP and HP, despite the slightly lower yield observed in the former. The CAP treatment exhibited a significant decrease in biomass accumulation and functional impairments, which were attributed to reduced nutrient levels in lettuce leaves. The DCAP treatment exhibited a 180% increase in fertilizer use efficiency compared to the HP treatment. We conclude that the tri-trophic cropping system with the implementation of DCAP variant is an effective system that enables the combined production of crops and fish, the latter being fed with sustainably derived insect protein. The tri-trophic system improves the environmental impact and sustainability of lettuce production, while making circularity feasible. Full article

Circular RNA (CircRNA) is a single-stranded RNA arising from back splicing. CircRNAs interact with mRNA, miRNA, and proteins. These interactions regulate various life processes, including transcription, translation, cancer progression, anticancer drug resistance, and metabolism. Due to a lack of cap and poly(A) tails, circRNAs show exceptional stability and resistance to RNase degradation. CircRNAs exhibit dysregulated expression patterns in various cancers and influence cancer progression. Stability and regulatory roles in cancer progression make circRNAs reliable biomarkers and targets for the development of anticancer therapeutics. The dysregulated expression of circRNAs is associated with resistance to anticancer drugs. Enhanced glycolysis by circRNAs leads to resistance to anticancer drugs. CircRNAs have been known to regulate the response to chemotherapy drugs and immune checkpoint inhibitors. Exogenous circRNAs can encode antigens that can induce both innate and adaptive immunity. CircRNA vaccines on lipid nanoparticles have been shown to enhance the sensitivity of cancer patients to immune checkpoint inhibitors. In this review, we summarize the roles and mechanisms of circRNAs in anticancer drug resistance and glycolysis. This review discusses clinical applications of circRNA vaccines to overcome anticancer drug resistance and enhance the efficacy of immune checkpoint inhibitors. The advantages and disadvantages of circRNA vaccines are also discussed. Overall, this review stresses the potential value of circRNAs as new therapeutic targets and diagnostic/prognostic biomarkers for cancer Full article

(1) Background: The highly pathogenic avian influenza virus H5N1 clade 2.3.4.4b is an emerging threat that poses a great risk to the poultry industry. A few human cases have been linked to the infection with this clade in many parts of the world, including the USA. Unfortunately, there are no specific vaccines or antiviral drugs that could help prevent and treat the infection caused by this virus in birds. Our major objective is to identify/repurpose some (novel/known) antiviral compounds that may inhibit viral replication by targeting some key viral proteins. (2) Methods: We used state-of-the-art machine learning tools such as molecular docking and MD-simulation methods from Biovia Discovery Studio (v24.1.0.321712). The key target proteins such as hemagglutinin (HA), neuraminidase (NA), Matrix-2 protein (M2), and the cap-binding domain of PB2 (PB2/CBD) homology models were validated through structural assessment via DOPE scores, Ramachandran plots, and Verify-3D metrics, ensuring reliable structural representations, confirming their reliability for subsequent in silico approaches. These approaches include molecular docking followed by molecular dynamics simulation for 50 nanoseconds (ns), highlighting the structural stability and compactness of the docked complexes. (3) Results: Molecular docking revealed strong binding affinities for both sofosbuvir and GS441524, particularly with the NA and PB2/CBD protein targets. Among them, GS441524 exhibited superior interaction scores and a greater number of hydrogen bonds with key functional residues of NA and PB2/CBD. The MM-GBSA binding free energy calculations further supported these findings, as GS441524 displayed more favorable binding energies compared to several known standard inhibitors, including F0045S for HA, Zanamivir for NA, Rimantadine and Amantadine for M2, and PB2-39 for PB2/CBD. Additionally, 50 ns molecular dynamics simulations highlighted the structural stability and compactness of the GS441524-PB2/CBD complex, further supporting its potential as a promising antiviral candidate. Furthermore, hydrogen bond monitor analysis over the 50 ns simulation confirmed persistent and specific interactions between the ligand and proteins, suggesting that GS441524 may effectively inhibit the NA, and PB2/CBD might potentially disrupt PB2-mediated RNA synthesis. (4) Conclusions: Our findings are consistent with previous evidence supporting the antiviral activity of certain nucleoside analog inhibitors, including GS441524, against various coronaviruses. These results further support the potential repurposing of GS441524 as a promising therapeutic candidate against H5N1 avian influenza clade 2.3.4.4b. However, further functional studies are required to validate these in silico predictions and support the inhibitory action of GS441524 against the targeted proteins of H5N1, specifically clade 2.3.4.4b. Full article

The Steinernema carpocapsae nematode is known to release several excretory/secretory products (ESPs) in its venom upon contact and during the parasitic infection process of insect hosts. A recurrent family of proteins found in this nematode’s venom is the CAP (cysteine-rich secretory protein/antigen 5/pathogenesis-related 1) protein, but the functional role of these proteins remains unknown. To elucidate the biological function, this study focused on characterising the secreted protein, first identified in the venom of the nematode’s parasitic stage, and the sequence retrieved from transcriptomic analysis. The structural comparisons of the Sc-CAP protein model, as determined by AlphaFold2, revealed related structures from other parasitic nematodes of vertebrates. Some of these closely related proteins are reported to have sterol-binding ability. The Sc-CAP recombinant protein was successfully produced in Escherichia coli in conjunction with a chaperone protein. The results showed that the Sc-CAP protein binds to cholesterol, and docking analyses of sterols on the protein revealed potential molecular interactions. Immunoassays performed in Galleria mellonella larvae revealed that this venom protein has an inhibitory effect against phenoloxidase and the antimicrobial response of insects. This suggests that the venom protein has an immunomodulatory function against insects, emphasising its importance during the parasite–host interaction. Full article

In this review article, statistical mechanisms of oxidative modification reactions in various organic compounds under the influence of reactive oxygen species (ROS) generated by cold atmospheric plasma (CAP) are investigated and analyzed based on reactive molecular dynamics (MD) simulations. As an efficient and hygienic advanced oxidation technology, CAP demonstrates tremendous potential in fields such as biomedicine and environmental protection. Through simulations, this paper provides a detailed analysis of the interaction mechanisms between ROS and components of biological tissues and environmental toxins. In this paper, we review the reactions involving four major ROS (OH radicals, O atoms, ${\mathrm{O}}_3$ molecules, and ${\mathrm{H}}_2{\mathrm{O}}_2$ molecules) and organic compounds, including proteins, DNA, polysaccharides, fatty acids, antibiotics, and mycotoxins. Atomic-level analysis reveals various oxidative modification reactions induced by ROS and their resulting products, including dehydrogenation reactions, bond-formation reactions, oxygen-addition reactions, and bond-cleavage reactions. Additionally, the study elucidates the role of active functional groups in various organic compounds, the presence of special elements, and the specific reactive nature of ${\mathrm{H}}_2{\mathrm{O}}_2$. Furthermore, the influence of different ROS species and concentrations on reaction types is explored, aiming to provide a solid theoretical foundation for the application of CAP technology in biomedicine and environmental remediation. Full article

We present a scalable and resource-aware framework for the quantum simulation of large proteins, grounded in systematic molecular fragmentation, analytical Toffoli gate modeling, and empirical validation. The ground-state energy of a target biomolecule is reconstructed from capped amino acid fragments, with fixed corrections to account for artificial boundaries. Analytical cost estimates—derived from reduced Hamiltonians—are benchmarked against empirical Toffoli counts using PennyLane’s resource estimation module. Our model maintains predictive accuracy across biologically relevant systems of up to 1852 electrons, capturing consistent patterns across diverse fragments. This framework enables early-stage feasibility assessments for achieving quantum advantage in biochemical simulation pipelines. Full article

Background/Objectives: Refractory community-acquired pneumonia (r-CAP) has become a thorny issue in clinical practice, especially after the COVID-19 pandemic, even in immunocompetent patients, as conventionally defined. In this study, we aimed to identify the risk factors for immunocompetent patients with r-CAP. Methods: This was a single-center retrospective study. In total, we collected clinical data from 82 patients with r-CAP in whom the first-line antibiotic therapy failed and 82 patients with general CAP (g-CAP) who recovered with first-line antibiotics, matched at a ratio of 1:1, admitted to Beijing Shijitan Hospital, Capital Medical University, from 1 January 2022, to 31 December 2023. The differences between the two groups (clinical characteristics, peripheral blood cell count, lymphocyte subsets, and regular laboratory indicators) were analyzed using paired t, paired Wilcoxon, Chi-square, or Fisher’s exact tests, and univariate and multivariate logistics regression analyses were conducted to identify the independent risk factors. A model for predicting indicators with statistical significance was established and proved with the receiver operating characteristic (ROC) curve. Results: Warm season, a history of chronic obstructive pulmonary disease, longer time from onset to admission (T_O-A), higher percentages of CD4⁺ T, CD8⁺ T, and double-negative T (DNT) lymphocytes, as well as higher levels of C-reactive protein (CRP), low-density lipoprotein cholesterin (LDL-C), serum sodium ion (Na⁺), and free-calcium ion (FCa²⁺) were regarded as independent risk factors, while T lymphocyte percentage (T%) and total cholesterol (TC) were identified as protective factors. The combined multivariate model using all the above factors proved to be sensitive and specific (AUC = 0.8711, p < 0.0001, R² = 0.4235), and thus better than the respective univariate models. Conclusions: Increased CD4⁺ T%Lym, CD8⁺ T%Lym, and DNT%Lym, warm season, a history of COPD, longer T_O-A, and increased levers of CRP, LDL-C, Na⁺, and FCa²⁺ potentially cause CAP to be refractory, while the T lymphocyte count, namely, the overall cellular immunity, was impaired in r-CAP patients, and increased TC levels could be beneficial to pneumonia recovery. Full article

Lake eutrophication, often driving harmful algal blooms (HABs) and ecosystem degradation, involves complex biogeochemical shifts within sediments. Changes in the sedimentary dissolved organic matter (DOM) composition during transitions from macrophyte to algal dominance are thought to critically regulate internal phosphorus (P) loading, yet the underlying mechanisms, especially in vulnerable plateau lakes like Qilu Lake, require further elucidation. This study investigated the coupled cycling of carbon (C) and P in response to historical ecosystem succession and anthropogenic activities using a 0–24 cm sediment core from Qilu Lake. We analyzed the total organic carbon (TOC), total phosphorus (TP), sequential P fractions, and DOM fluorescence characteristics (EEM-PARAFAC), integrated with chronological series data. The results revealed an asynchronous vertical distribution of TOC and TP, reflecting the shift from a submerged macrophyte-dominated, oligotrophic state (pre-1980s; high TOC, low TP, stable Ca-P dominance) to an algae-dominated, eutrophic state. The eutrophication period (~1980s–2010s) showed high TP accumulation (Ca-P and NaOH85 °C-P enrichment), despite a relatively low TOC (due to rapid mineralization), while recent surface sediments (post-2010s) exhibited a high TOC, but a lower TP following input controls. Concurrently, the DOM composition shifted from microbial humic-like dominance (C1) in deeper sediments to protein-like dominance (C3) near the surface. This study demonstrates that the ecosystem shift significantly regulates P speciation and mobility by altering sedimentary DOM abundance and chemical characteristics (e.g., protein-like DOM correlating negatively with Ca-P), reinforcing a positive feedback mechanism that sustains internal P loading and potentially exacerbates HABs. DOM molecular characteristics emerged as a key factor controlling the internal P cycle in Qilu Lake, providing critical insights for managing eutrophication in plateau lakes. Full article

Cold atmospheric plasma (CAP) interacts with tissues, leading to fast wound disinfection. Given the frequent global burden of burn injuries and the risks of infection associated with acute full-thickness burns (FTBs), this investigation examined CAP as a potential therapeutic method for wound healing due to its antimicrobial and pro-healing effects. Here, we examined the impacts of CAP on the healing of wounds resulting from acute FTSBs. We established an animal model that included four groups: (1) healthy control animals without burns, (2) untreated animals with acute FTSBs, (3) animals with acute FTSBs treated with CAP for 5 s per day for 21 days, and (4) animals with acute FTSBs treated with CAP for 10 s per day for 21 days. Wound healing was assessed using immunohistological methods. In animals with FTSBs, CAP therapy was accompanied by (i) accelerated wound closure, (ii) enhanced regeneration of the dermis and epidermis, and (iii) increased protein expression of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF). These changes were more pronounced following CAP treatment for 10 s per day compared to CAP treatment for 5 s per day. Full article

Background/Objectives: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of Coronavirus Disease 2019 (COVID-19), has rapidly evolved, giving rise to multiple Variants of Concern—including Alpha, Beta, Gamma, Delta, and Omicron—which emerged independently across different regions. Licensed COVID-19 vaccines primarily target the highly mutable spike protein, resulting in reduced efficacy due to immune escape by emerging variants. Previously, we developed a live attenuated Francisella tularensis LVS ΔcapB single-vector platform COVID-19 vaccine, rLVS ΔcapB/MN, expressing the conserved membrane (M) and nucleocapsid (N) proteins from the early SARS-CoV-2 WA-01/2020 strain. In this study, we evaluate the efficacy of rLVS ΔcapB/MN and an enhanced version, rLVS ΔcapB::RdRp/MN, which additionally expresses the conserved RNA-dependent RNA polymerase (RdRp) protein from the same strain, in a hamster model. Methods: Both vaccine candidates were administered orally or intranasally to golden Syrian hamsters (equal numbers of males and females) and evaluated against intranasal challenge with SARS-CoV-2 Delta (B.1.617.2-AY.1) and Omicron (BA.5) variants. Results: Vaccinated animals developed robust, TH1-biased IgG responses specific to the nucleocapsid protein. Following SARS-CoV-2 challenge, immunized hamsters exhibited reduced weight loss, lower oropharyngeal and lung viral titers, and improved lung pathology scores compared with unvaccinated controls. Conclusion: These findings support the potential of this universal vaccine to provide broad protection against current and future SARS-CoV-2 variants, with minimal need for updating. Full article

Background and Objectives: Community-acquired pneumonia (CAP) remains a major health burden worldwide, with high morbidity and mortality, particularly among older adults and those with comorbidities. This study aimed to evaluate the etiological factors of CAP and to investigate systemic inflammatory markers (IL-6, IL-8, IFN-γ, and G-CSF) in blood samples collected from CAP patients to identify which markers could be targets for potential etiological, clinical, and therapeutic interventions. Materials and Methods: A prospective study was conducted in 41 patients with confirmed CAP hospitalised during the winter season of 2024–2025. Clinical, demographic, and laboratory data were collected at admission and seven days later. Serum IL-6, IL-8, IFN-γ, and G-CSF concentrations were measured using a multiplex assay. Results: Aetiology was identified in 87.8% of cases, with typical bacterial pathogens being more prevalent among older, smoking patients, while atypical pathogens were more common among younger, non-smoking patients. Hospitalisation and increased inflammatory markers were associated with older age. After seven days of treatment, significant decreases in IL-6, IFN-γ, and G-CSF concentrations were observed. IFN-γ levels were significantly higher in patients with atypical aetiology. Higher concentrations of IL-8 and G-CSF were associated with hospitalisation. IL-6 levels were positively correlated with age, C-reactive protein (CRP), and pneumonia severity index (PSI) scores. Conclusions: Systemic inflammatory markers, especially IL-6, IL-8, IFN-γ, and G-CSF, may be valuable tools in managing generalised pneumonia. They can help to differentiate etiologically, assess disease severity, and make treatment decisions. Full article

Background and Objectives: The assessment of HER2 status in invasive breast carcinomas (IBCs) is critical for determining treatment strategies. The aim of this study was to evaluate the FICTION technique as a potential method for assessing HER2 status and to compare it with the standard sequential immunohistochemistry (IHC)–in situ hybridization (ISH) assays. Materials and Methods: This study included 49 patients diagnosed with invasive breast carcinomas. HER2 status was assessed using both IHC+FISH and FICTION techniques, and the results were compared. Results: Comparative analysis demonstrated an 83.67% categorical agreement between IHC and IF using the ASCO/CAP system. The percentage of cells showing any degree of HER2 protein expression was higher with IF (73.77%) than with IHC (60.71%) (p = 0.00026). The in situ hybridization assays showed an excellent agreement, with a 90% or higher concordance. The concordance of the ASCO/CAP group classification of cases using both ISH assays (FICTION and standard FISH) was high (85, 7%). Agreement was 100% for the final classification of cases (Her2 positive/negative). Conclusions: We compared standard tests for Her2 protein expression and the gene copy number with a modified FICTION protocol. The study showed moderate agreement between IHC and IF for Her2 protein and excellent agreement between FISH and FICTION ISH for the gene copy number. Final Her2 status was unaffected by low IF IHC concordance. Optimizing the FICTION protocol could improve results. Combining protein and gene assays may enhance IBC patient stratification. Full article

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a common cause of chronic liver disease and is closely associated with metabolic abnormalities and cardiovascular risks. Butyrate, a short-chain fatty acid produced by gut microbiota, has the potential to enhance liver health by modulating inflammation and supporting gut barrier integrity. This study aimed to investigate and compare the effects of sodium butyrate and calcium butyrate in patients with MASLD. In this single-center, randomized clinical trial, 181 patients with MASLD were enrolled and assigned to receive either sodium butyrate (n = 121) or calcium butyrate (n = 60) supplementation at a daily dose of 1000 mg. The primary endpoint was the change in liver steatosis, measured using the Controlled Attenuation Parameter (CAP) via FibroScan^®. Secondary endpoints included liver stiffness, biochemical parameters, hepatic steatosis and fatty liver indices, fecal calprotectin levels, stool short-chain fatty acid levels, and microbiome composition. A subgroup analysis compared responders (a ≥ 5% reduction in CAP) to non-responders. There were no significant changes in CAP values for either group (ΔCAP: sodium butyrate, 0.84; calcium butyrate, −0.23; p = 0.70). Sodium butyrate significantly reduced serum trimethylamine N-oxide and fatty liver index, while calcium butyrate led to a decrease in fecal calprotectin levels. Responders demonstrated a lower body mass index, higher levels of high-sensitivity C-reactive protein and HbA1c, and distinct microbiome profiles, characterized by lower abundance of Subdoligranulum and higher abundance of Catenibacterium. Although butyrate supplementation did not significantly improve liver steatosis as measured by CAP, the differing effects on metabolic and inflammatory markers suggest that there may be potential benefits for specific subgroups of patients with MASLD. Full article

In cattle, the transfer of passive immunity from mother to calf across the placenta is limited. Therefore, providing quality colostrum or supplementing with additives that enhance this attribute is crucial to optimise the transfer of passive immunity (TPI). The objective of the present study was to investigate the effect of capsaicin on the absorption of immunoglobulins, metabolites, and enzymes to optimise TPI in neonatal calves, and to assess its impact on selected physiological parameters. Two experimental groups were established: a control group (CON; n = 8), which received colostrum in three feedings (at 1, 12, and 20 h after calving), and a capsaicin group (CAP; n = 8) which received 40 mg of capsaicin per kilogram of body weight added to the colostrum. Birth weight (CON = 36.7 ± 1.10 kg; CAP = 36.2 ± 2.64 kg) and weight at 48 h of age (CON = 38.5 ± 1.11 kg; CAP = 38.0 ± 2.82 kg) were similar between the two experimental groups. At 48 h post-colostrum feeding, the CAP group exhibited significantly higher serum concentrations of IgG, total protein, and albumin—up to 23.4 times greater than those observed in the CON group (p < 0.001). Moreover, levels of glucose, cholesterol, and triglycerides were significantly elevated in the CAP group compared to the CON group (p < 0.05). Similarly, higher concentrations of alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase were observed in the CAP group. These findings suggest that the inclusion of 40 mg of capsaicin per kilogram of body weight in colostrum is safe and contributes positively to improving TPI in crossbred Holstein calves raised at altitudes higher than 2500 m above sea level. Full article