Search Results (4,494)

Currently, the B-cell response patterns induced by viral antigens in avian disease models and their detailed immunological characteristics still require comprehensive elucidation at the single-cell level. In this study, we employed single-cell sequencing (scRNA-seq) and B cell library technology to conduct an in-depth analysis of B cells in the spleens of mice with inactivated duck hepatitis A virus type 1 (DHAV-1) as model antigen. This study aimed to investigate the immunological characteristics of the virus antigen in the mouse model and characteristics of B-Cell Receptors. The results showed that the DHAV-1 group had distinct changes in splenic B cell subset counts, proportions, and intercellular communication. Additionally, an increased trend in communication strength between Gm26917+B and Gm11837+B cells was observed, with enriched expression of C-X-C motif chemokine ligand (CXCL) and lymphotoxin (LT) detected in the DHAV-1 group. Furthermore, the DHAV-1 group exhibited a prominent combination of the IGHV1 family and IGHV3-1/IGHJ3 in the heavy (H) chain variable region. Compared with the CK group (negative control group), the amino acid sequence length and diversity of the CDR3 region in the DHAV-1 group exhibited a decreasing trend. In summary, our findings characterize the immunological features of splenic B cells in mice after immunization with inactivated DHAV-1, and provide a preliminary characterization of DHAV-1-induced B cell transcriptional states and BCR repertoire features, generating testable hypotheses for subsequent mechanistic investigations of B cell-mediated immune responses to viral antigens. Full article

Pulsed polarization of Au|YSZ gas sensors is examined to clarify the mechanism of NO_x detection under dynamic operation and to disentangle catalytic surface effects from electrochemical relaxation. Using gold electrodes with substantially lower catalytic activity than platinum explicitly enables this mechanistic separation. During pulsed polarization, periodic voltage pulses are followed by self-discharge under open-circuit conditions, and the response is measured based on the self-discharge rate. NO₂ consistently accelerates the self-discharge from the beginning, whereas NO slows the relaxation predominantly at later times. CO and H₂ produce similar delaying effects, and C₃H₆ shows no measurable influence under the tested conditions. Decreasing ambient O₂ slows the discharge and amplifies the NO₂ effect, which indicates that oxygen supply and surface exchange at the triple-phase boundary are rate determining. A Pt-containing catalytic overlayer drives local NO/NO₂ interconversion toward equilibrium so that both gases yield to an accelerated self-discharge. These findings support a mechanistic picture in which NO₂ provides effective oxygen equivalents that accelerate discharge, whereas NO, CO, and H₂ consume oxygen and slow down discharge. Overall, this establishes a materials-based approach for distinguishing between NO and NO₂ and evaluating the underlying mechanism during pulsed polarization. Full article

Influenza viruses are significant causes of acute respiratory infections, often leading to severe health issues and mortality. These viruses undergo continuous mutations and genetic reassortments, resulting in annual epidemics and potential pandemics. The A(H3N2) strains exhibit high genetic and antigenic variability, that influence vaccine efficacy. This study aimed to assess the prevalence of influenza viruses, including A(H3N2) strains, in Kazakhstan during 2020–2025. The study used nasopharyngeal swab and serum samples obtained from patients. The presence of influenza virus antigens in nasopharyngeal swabs was analyzed using real-time polymerase chain reaction. The level of specific antibodies in the blood serum was determined using the hemagglutination inhibition reaction and enzyme-linked immunosorbent assay methods. Influenza A/H1N1, A/H3N2 and B viruses were diagnosed using real-time PCR. Antibodies to A/H1N1pdm09, A/H3N2 and B were detected in serological studies. Our studies revealed a trend toward seasonal patterns in influenza A viruses circulation. Therefore, it was established that the A/H3N2 strains dominated in Kazakhstan during the 2021–2022 and 2023–2024 epidemic seasons. The 2023–2024 strains belong to the specific genetic clade J.2 or 3C.2a1b.2a.2a.3a.1. These studies confirmed the role of influenza viruses in the etiology of respiratory infections and emphasized the need to continue monitoring their spread in Kazakhstan. Full article

The present study investigates the role of royal jelly as a natural endocrine modulator of serum estradiol levels in juvenile sterlets (Acipenser ruthenus), a species of major interest for sustainable aquaculture. The experiment was conducted over a period of 85 days under controlled recirculating system conditions, using four dietary treatments (n = 30 fish per group): a control group and three groups supplemented with 1%, 3%, and 5% royal jelly. Serum estradiol concentrations were determined by high-performance liquid chromatography (HPLC), while biometric assessment included the determination of total length (L), standard length (Sl), maximum body height (H), body circumference (C), and body mass (BM). Royal jelly supplementation significantly increased serum estradiol levels in a dose-dependent manner (p < 0.05), with the highest values recorded in the 5% group compared to the control. The proportion of individuals with non-detectable estradiol levels decreased progressively with increasing supplementation level. Biometric analysis revealed moderate effects on growth parameters, with no statistically significant differences among groups for most traits (p > 0.05), except for maximum body height, which showed a significant overall effect (ANOVA, p = 0.0089). Principal Component Analysis (PCA) highlighted the relative independence between endocrine variability and growth dynamics. Overall, the findings support the potential of royal jelly as a natural endocrine modulator of serum estradiol, representing a promising and environmentally friendly alternative to synthetic hormonal substances used in aquaculture. This approach may contribute to the development of innovative nutritional strategies for endocrine control and the optimization of biological performance in sturgeons, in accordance with the principles of sustainable aquaculture. Full article

Background: Rabies is a highly fatal zoonotic disease that causes approximately 59,000 human deaths worldwide each year. Current inactivated rabies vaccines require multiple doses and are associated with high costs. The full-length rabies virus glycoprotein (RVG), a membrane protein, exhibits substantial instability in its trimeric structure during recombinant expression. This instability makes it difficult to obtain high-purity, correctly folded antigens. Objectives: This study focuses on the preparation of a full-length recombinant RVG subunit vaccine candidate expressed in a glycoengineered Pichia pastoris system with mammalian-like glycosylation. Methods: The full-length RVG gene (including the transmembrane domain and cytoplasmic tail) from the Challenge Virus Standard-11 (CVS-11) strain was codon-optimized and inserted into the pPICZαA vector to construct the recombinant expression plasmid pPICZαA-RVG. The plasmid was transformed into glycoengineered Pichia pastoris X33-7 (low-mannose type) by electroporation for inducible expression. The target protein was purified by nickel affinity chromatography, anion-exchange chromatography, and Superdex-200 size-exclusion chromatography. The structural characteristics of the purified protein were analyzed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The purified antigen was formulated with the adjuvants AS03 or MF59. BALB/c mice (n = 5 per group) were immunized intramuscularly following a four-dose schedule (days 0, 7, 14, and 28). Antigen-specific IgG antibody titers were measured by ELISA, and neutralizing antibody titers were determined using the rapid fluorescent focus inhibition test (RFFIT). Results: Glycoengineered Pichia pastoris yeast strains expressing wild-type RVG (RVG-WT) or a mutant variant (RVG-M6: R84S, R199S, H270P, R279S, K300S, and R463S) were successfully constructed. The purified RVG antigen formed nanoparticles with an average particle size of approximately 75 nm. Immunized mice generated robust RVG-specific IgG responses, with titers reaching approximately 6.31 × 10⁵ for RVG-WT after the fourth immunization, compared to 3.16 × 10³ for RVG-M6 and 5.62 × 10³ for the RVG-WT-PEG control. Two weeks after the fourth immunization, RVG-WT formulated with AS03 or MF59 induced significant neutralizing antibody responses compared with the control group (p < 0.0001 and p < 0.01, respectively). The neutralizing antibody titers reached 1:79.43 in the AS03 group and 1:33.11 in the MF59 group, whereas the WT-PEG + AS03 control group showed a low titer of 1:3.72. In contrast, RVG-M6 formulated with MF59 failed to induce detectable neutralizing antibodies (1:3.02). Furthermore, RVG-WT + AS03 induced significantly higher neutralizing antibody responses than the WT-PEG + AS03 control group (p < 0.0001), and a significant difference was also observed between the RVG-WT + MF59 and RVG-M6 + MF59 groups (p < 0.01). Conclusions: The glycoengineered Pichia pastoris expression system successfully produced uniform full-length rabies virus glycoprotein nanoparticles with high purity. When formulated with the AS03 adjuvant, RVG-WT induced high-titer neutralizing antibodies in mice, suggesting a promising strategy for the development of recombinant subunit vaccines against rabies. However, this study is limited by the absence of challenge studies and validation in target animal species, which will be further investigated in future work. Full article

Oxidative stress is one of the major contributors to DNA damage and genomic instability, emphasizing the importance of identifying natural compounds with antioxidant genoprotective potential. Cedrus atlantica essential oil (EO) has been widely reported to possess antioxidant properties and potential genoprotective effects due to the presence of a cohort of antioxidant compounds, including polyphenols and terpenes. Nevertheless, its effects on DNA integrity remain poorly understood. The present study aimed to evaluate the genotoxic and antigenotoxic effects of C. atlantica EO in human peripheral blood mononuclear cells (PBMCs) using the alkaline Comet assay. PBMCs were exposed to increasing concentrations of the EO (0.2–3% w/v) under basal conditions and in the presence of hydrogen peroxide (H₂O₂, 25 µM) as an oxidative DNA-damaging agent. Genetic damage was quantified by visual score, and arbitrary units were converted into a percentage of DNA in the comet tail. The EO was characterized by gas chromatography–mass spectrometry. The results showed that C. atlantica EO did not induce detectable genotoxic effects under the experimental conditions and within the tested concentration range (0.2–3% w/v). H₂O₂ exposure markedly increased DNA strand breaks, whereas co-treatment with the EO significantly attenuated H₂O₂-induced oxidative DNA damage, particularly at intermediate concentrations. The chemical characterization analysis revealed a sesquiterpene-rich profile dominated by cedrene- and himachalene-type compounds. Overall, these findings indicate that C. atlantica EO exerts antigenotoxic effects against oxidative DNA damage, supporting its genoprotective potential in moderate concentrations. Full article

Life as we know it depends on peptide and nucleic acid polymers built from a limited set of backbone residues, yet planetary environments beyond Earth motivate consideration of alternative chemical frameworks for genetic- and protein-like polymers. In this context, we synthesize four azatide dipeptide analogs (Ala^a-Gly^a (1), Gly^a-Ala^a (2), Gly^a-Gly^a (3), and Ala^a-Ala^a (4)) as candidate backbone motifs for non-standard biologically relevant polymers. We then systematically assess their stability and reactivity in 98% w/w sulfuric acid, a solvent relevant to Venusian cloud chemistry. We assess the stability of the azatides via ¹H and ¹³C NMR spectroscopy supported with ELSD-LCMS. We monitor the stability of the compounds over periods from hours to two weeks at room temperature and at elevated temperatures (50–80 °C). All four azatides readily dissolve in 98% w/w D₂SO₄ and are generally stable at room temperature. Gly^a-Ala^a (2) shows no detectable degradation over a two-week incubation in 98% w/w sulfuric acid. The other three azatide analogs display only minor decomposition. ELSD-LCMS measurements qualitatively confirm the NMR results, revealing only minor-to-moderate loss of parent compounds after two weeks at room temperature. At higher temperatures, representative of the lower Venusian cloud deck, the stability of the azatides decreases dramatically. All four compounds undergo significant decomposition at 50 °C and completely degrade within one to two weeks at 80 °C. Our findings indicate that azatides, despite being generally stable in concentrated sulfuric acid at room temperature, lack the thermal stability that might be required to serve as viable backbone motifs for biological polymers in environments spanning the full temperature range of Venusian clouds. Full article

Highly pathogenic avian influenza virus (HPAIV) H5N1 has been detected in dairy cattle in the United States, with high viral loads observed in milk from infected animals. This raises public health concerns regarding potential transmission through exposure to raw milk. The sale of raw milk via vending machines represents a well-established distribution model in many European countries, including Switzerland. Although a notice must be posted on these milk vending machines stating that it is raw milk, together with appropriate processing instructions (heating to over 70 °C required, storage below 5 °C, consumption within 3 days), these notices are sometimes missing, and consumers often do not follow these guidelines. Over a four-month period, spanning from June 2025 to September 2025, 124 raw milk samples were collected from vending machines across Switzerland. Samples were screened for influenza A using reverse-transcription quantitative PCR (RT-qPCR). No samples tested positive for influenza A virus. The data from this study demonstrate the feasibility of implementing a sampling and detection system for HPAIV H5N1 in direct-to consumer raw milk samples and highlight the currently very low risk of HPAIV in raw milk samples sold via vending machines in Switzerland. Full article

Microsatellite instability (MSI) testing is frequently used to screen patients for the early detection of Lynch syndrome, the most common hereditary colorectal cancer syndrome. MSI testing compares microsatellite repeat lengths in tumor DNA with those in matched normal tissue from the same patient. Therefore, precise sample identification is critical for obtaining reliable test results. The Penta-C and Penta-D pentanucleotide markers are widely used for sample identification in MSI testing. We investigated instability, defined as allelic mismatches or shifts, discordant fragment sizes, or the appearance of alleles in tumor DNA that were absent in the corresponding normal DNA, in the Penta-C and Penta-D loci across 2609 paired colorectal tumor and matched normal tissue or blood DNA samples. The allele sizes of both markers did not match in 0.3% of microsatellite-stable (MSS) and 12.3% of microsatellite instability-high (MSI-H) patients (p < 0.001, difference in proportions, 12.0% (95% CI, 8.9–15.1%)). Non-matching allele sizes in 12.3% of the MSI-H tumors suggest that other repeat markers may also be unstable and not suitable for sample identification in these tumors. Full article

This study addresses the inefficiencies, subjectivity, and poor adaptability to lighting variations inherent in traditional candling methods used in large-scale egg incubation. We developed a high-throughput transmissive imaging system capable of capturing 30 eggs simultaneously. Based on this system, we propose PDA-YOLO, an enhanced YOLOv8-based object detection model featuring a position-decoupled attention strategy. Specifically, a lightweight C2f-SE module is integrated into the backbone to amplify subtle feature responses in low-contrast regions, while a CBAM is deployed prior to the detection head to mitigate background clutter through precise spatial attention. Experimental results on a self-constructed Hailan White egg dataset show that at the critical 60 h incubation stage, PDA-YOLO achieves a Recall of 91.5% and an mAP@0.5 of 97.4%, outperforming the YOLOv8 baseline while maintaining a real-time inference speed of 62.1 FPS. Grad-CAM visualizations confirm the model’s ability to focus on vascular textures and suppress noise. Furthermore, the model demonstrates robust performance under varying illumination (180–540 lumens), effectively mitigating missed detections in low light and recognition degradation from overexposure. This work provides a scalable, real-time solution for non-destructive, early-stage detection of poultry health and fertilization status in commercial hatcheries. Full article

Background: Crimean-Congo hemorrhagic fever (CCHF) is a severe tick-borne viral disease with a high fatality rate, and no licensed vaccines are currently available. The nucleoprotein (NP) of the Crimean-Congo hemorrhagic fever virus (CCHFV) plays a critical role in viral replication and immune recognition, making it a promising target for vaccine development. This study aimed to design and evaluate a multiepitope recombinant DNA vaccine targeting the NP of CCHFV. Methods: Cytotoxic T lymphocyte (CTL) epitopes from the NP were predicted via immunoinformatics approaches and systematically assessed for antigenicity, allergenicity, toxicity, hydrophobicity, and global population coverage. The selected epitopes were incorporated into four DNA vaccine constructs driven by a cytomegalovirus promoter, adjuvanted with human β-defensin 3 (hBD3), and fused to the reporter protein mRuby3. The constructs were evaluated in vitro using a fluorescent reporter system designed to provide a readout of TCR signaling upon the co-culture of T lymphocytes with differentiated monocytic cells expressing antigens. In vivo immunogenicity and protective efficacy were assessed in BALB/c (exploratory pilot) and IFNAR^−/− mice, a highly susceptible model for viral infection. Cytokine responses were measured to assess immunogenicity. Results: In vitro assays showed predominantly antigen-independent T-cell activation, suggesting that nonspecific stimulation inherent to the reporter co-culture system likely obscured the detection of antigen-specific TCR signaling. In vivo analyses in BALB/c mice revealed that the constructs elicited only modest systemic cytokine profiles while CCHFV-specific IgG and IFN-γ secretion remained undetectable, indicating that antigen-specific T-cell and antibody responses were limited. In the IFNAR^−/− challenge model, several peptide groups achieved significant 2–3 log reductions in tissue viral RNA and infectious titers (p < 0.05 vs. sham). However, the observed viral modulations were insufficient to reach the protective threshold and did not translate to a survival benefit (0%). Conclusion: Despite a rational in silico foundation, the multiepitope DNA vaccine constructs demonstrated limitations in inducing potent, antigen-specific immunity across both mouse models. The lack of antigen-specific responses indicates limitations in epitope selection, construct design, and delivery strategies, requiring optimization of next-generation epitope-based vaccines. These findings highlight the complexity of translating computational epitope predictions into functional vaccines, and provide benchmark data as a framework to guide future optimizations. Full article

To investigate how the content of silica fume (SF) influences the performance of foam concrete (FC) after high-temperature exposure and the underlying mechanisms, this study prepared standard FC cube specimens with SF contents of 0%, 0.15%, 0.2%, 0.25%, and 0.3%. The working properties of the material at room temperature were systematically tested, and the mass loss, residual compressive strength, failure mode, microstructure and acoustic emission (AE) data at different temperatures (100 °C, 200 °C, 300 °C and 400 °C) were analyzed. The test results indicate that increasing the SF content reduces the fluidity of the fresh paste yet significantly enhances the compressive strength and lowers the water absorption of FC at room temperature. After high-temperature exposure, the effect of SF exhibits a dual character: at 200 °C and below, SF effectively mitigates the performance degradation of FC. However, when the temperature reaches 300–400 °C, specimens with an excessively high SF content (e.g., 0.3%) experience rapidly built-up internal steam pressure that cannot escape in time, which triggers the formation and propagation of a microcrack network and leads to a sharp drop in strength. Based on AE detection and scanning electron microscopy (SEM) image analysis, the failure process of silica fume foam concrete (SFFC) proceeds through three stages: free water evaporation at low temperatures, dehydration shrinkage of the C-S-H gel at medium temperatures, and finally, structural failure marked by the collapse of the C-S-H gel network at high temperatures. This study indicates that an SF content of 0.25% allows FC to achieve an optimal balance between mechanical properties and high-temperature stability. The findings provide a theoretical basis for optimizing FC mix proportions and enhancing fire prevention design. Full article

In the present study, Shuidong mustard (Brassica juncea) produced in Maoming City, Guangdong Province, was fermented at 25 °C for 15 days, using wild microorganisms and 20 g/kg table salt in water. The results showed that this fermentation endowed Shuidong mustard with acid production via utilizing reduced sugar as the fermentation substrate, causing the fermented Shuidong mustard to have a decreased pH value and increased total titratable acidity. Partly as the result of NaCl usage or fermentation, the fermented Shuidong mustard had enhanced NaCl or ash contents, decreased contents in nitrite/nitrate, vitamin C, total phenols, and total carotenoids, and altered textural features reflected as reduced hardness, chewiness, springiness, and fracturability. Moreover, 90 volatile compounds, including 2-butyl, 3-butenyl, isobutyl, and ethyl isothiocyanates, were detected in the fermented Shuidong mustard after the 15-day fermentation, while 21 members comprised 95% (w/w) of total volatiles. Additionally, the analysis results revealed that the microorganism community of fermented Shuidong mustard was structured at respective phylum, genera, or species levels by Firmicutes and Proteobacteria, or Lactiplantibacillus, Enterobacteriaceae, Lactococcus, and Pediococcus, or Lactiplantibacillus, Enterobacteriaceae, Pediococcus, and Lactococcus. It is thus concluded that this explored fermentation induced both acid production and, more importantly, compositional and textural changes in Shuidong mustard, which had production potential at an industrial scale as part of a healthy diet because these bioactive compounds include isothiocyanates, polyphenols, and carotenoids. Overall, this study focused on the Shuidong mustard fermentation using 20 g/kg table salt to fill a research gap in low-salt fermentation, showing its significance by providing a scientific basis for product development. Full article

Objectives: To evaluate the physicochemical properties of a novel strontium silicate-based root canal sealer (C-Root SP) in comparison with a calcium silicate-based sealer (TotalFill BC) and an epoxy resin-based sealer (AH Plus). Methods: Setting time, net mass change (apparent solubility behavior), pH changes, and surface characteristics were assessed based on ISO 6876 and ANSI/ADA Specification No. 57, with minor methodological modifications. Net mass change and pH were evaluated over 28 days. Surface morphology and elemental composition were analyzed after dry and aqueous aging in deionized water using scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy. Data were analyzed using one-way and repeated-measures ANOVA with Tukey’s post hoc test (α = 0.05). Results: AH Plus exhibited the longest initial and final setting times (10.93 ± 0.65 h and 37.33 ± 0.13 h), whereas TotalFill BC showed the shortest (7.98 ± 0.32 h and 30.18 ± 0.20 h); C-Root SP demonstrated intermediate values (9.35 ± 0.38 h and 32.75 ± 0.57 h) (p < 0.001). C-Root SP exhibited positive net mass change values (indicative of net mass loss), ranging from 5.32 ± 4.72% at 24 h to 6.83 ± 5.55% at 28 days, significantly higher than AH Plus and TotalFill BC (p < 0.001), which showed negative values indicative of apparent mass gain. All sealers demonstrated alkaline conditions, with C-Root SP maintaining the highest apparent pH values throughout the evaluation period (p < 0.001). Surface and compositional changes were observed in the bioceramic sealers following aqueous aging, with increased detectable strontium content in C-Root SP. Conclusions: C-Root SP exhibited physicochemical behavior consistent with a strontium-modified calcium silicate-based sealer, characterized by hydration-driven hydroxyl ion release resulting in apparent alkalinity and ion exchange-associated behavior, and dynamic surface changes consistent with those reported for bioceramic materials. Clinical Significance: Strontium incorporation may influence hydration-mediated physicochemical behavior; however, further in vitro and in vivo studies are required to determine its clinical relevance. Full article

Background: Stable isotope-based analytical methods have brought about a significant transformation in the study of energy nutrient metabolism, enabling precise in vivo measurement of metabolic fluxes at systemic, tissue, and organ-specific levels in both healthy and diseased states. The regulation of these metabolic fluxes is governed by dynamic interactions between proteins, lipids, carbohydrates, and their precursors—such as glucose, fatty acids, and amino acids—as well as final metabolic products. Discussion: Advanced analytical technologies, including nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS), which can offer enhanced precision, have been developed for investigating nutrient metabolism and fluxes in humans, providing precise information on metabolic pathways. These techniques have primarily utilized stable isotopes, such as ²H, ¹³C, ¹⁵N, and ¹⁸O, which have largely replaced radioactive isotopes and are now central to metabolic research. These isotopes have been used to label glucose, fatty acids, or amino acids—the main biomolecular precursors—enabling detailed investigation at systemic, tissue, and organ-specific levels of carbohydrate, lipid, and protein metabolism, and revealing pathway alterations associated with diseases conditions, such as diabetes, non-alcoholic fatty liver disease, cardiovascular disorders, and cancer. The use of deuterium oxide (D₂O) has allowed for long-term metabolic studies, providing a cost-effective and less invasive means to monitor metabolic changes over days to months. Total daily energy expenditure can be measured in free living conditions by the doubly stable isotopes ²H- and ¹⁸O-labeled water method. Stable isotope tracing, combined with advanced imaging and modeling, has also been instrumental in assessing body composition, energy expenditure, and nutrient bioavailability. Collectively, these methods have expanded our understanding of human physiology and disease, supporting the development of novel diagnostic tools, the identification of new biomarkers, and the tailoring of nutritional and therapeutic interventions. Conclusions: This review aimed to provide an overview of the applications of stable isotopes for the study of energy nutrient metabolic pathways. The ongoing integration of stable isotope approaches with artificial intelligence, omics technologies, and miniaturized detection techniques could promise to further refine our understanding of human metabolism and drive advances in personalized medicine. Full article