Search Results (295)

Acute kidney injury (AKI) caused by ischemia–reperfusion injury (IRI) involves rapid activation of innate immune responses, in which myeloid-derived immune cells critically shape injury severity. Y-box binding protein 1 (YB-1) regulates pro-inflammatory gene expression intracellularly and can be secreted to function extracellularly, yet how these two compartments jointly influence early IRI pathology remains poorly understood. To dissect the roles of intracellular myeloid versus extracellular YB-1, we subjected myeloid-specific Ybx1 knockout, Ybx1^fl/fl × LysM^cre, mice and wild-type (WT) littermates to unilateral renal IRI following administration of either a neutralizing anti-YB-1 antibody or control IgG. Kidney injury, inflammation, immune cell recruitment, neutrophil extracellular trap (NET) formation, antibody localization, and Fcγ receptor expression were assessed by qRT-PCR, histology, immunostaining, Western blotting, and flow cytometry. Myeloid-specific knockout of Ybx1 markedly reduced renal inflammation, neutrophil infiltration, NET formation, and tubular injury. This protective phenotype was lost when extracellular YB-1 was simultaneously reduced: anti-YB-1 treatment in knockout mice restored pro-inflammatory cytokine expression, increased tubular damage markers such as NGAL and KIM-1, exacerbated neutrophil recruitment and NET formation, and led to luminar accumulation of YB-1/anti-YB-1 immune complexes in tubular cells. Mechanistically, Ybx1-deficient myeloid cells exhibited significantly reduced CD16 expression, pointing to impaired Fcγ receptor-mediated phagocytosis as the cause of defective immune complex clearance. In contrast, wild-type mice efficiently cleared extracellular YB-1 complexes and showed no injury aggravation upon antibody treatment. Our findings identify myeloid YB-1 as a central regulator of early inflammatory injury in renal IRI and reveal that its protective depletion becomes pathogenic when extracellular YB-1 is simultaneously neutralized, likely due to unmasked defects in immune complex clearance. Full article

Background. Newborn ruminants are born agammaglobulinemic, and colostrum quality (IgG concentration) is often insufficient, especially in heifers, leading to high morbidity and mortality from diarrheal infections. Objective. To develop and evaluate colostrum enrichment with specific egg yolk immunoglobulins (IgY) for the prevention of diarrhea in newborn calves. Methods. Hens were hyperimmunized with an inactivated vaccine against rotavirus, coronavirus, and E. coli. Yolk melange was prepared. Total and specific IgY were measured by chromatography and ELISA. Trials were conducted in three farms with high diarrhea incidence: experimental calves (n = 25) received 100 mL of melange (5 yolks) with the first two colostrum feedings; controls (n = 25) received native colostrum. Results. One yolk contained up to 100 mg of polyclonal IgY, with 8% specific antibodies. Diarrhea occurred in 12% of experimental calves (mild, no drugs) vs. 76% in controls (24% required antibiotics/rehydration). Testing in two other farms (n = 42, n = 38) reduced incidence 5.2–6.8-fold compared to the previous period. Conclusions. Enriching colostrum with specific IgY from hyperimmunized hens is highly effective and affordable for preventing diarrheal infections in newborn calves, especially in herds with poor colostrum quality in heifers. Full article

Porcine epidemic diarrhea (PED), caused by the PED virus (PEDV), remains one of the most devastating diseases in the swine industry, with a mortality rate approaching 90–100% in suckling piglets due to severe dehydration and electrolyte imbalances. Passive immunization with egg yolk antibodies (IgY) represents a promising therapeutic strategy. In this study, we developed a novel recombinant adenovirus, rADM-IFN-G-ped, co-expressing selected antigenic regions of the PEDV S protein and chicken interferon-gamma (ChIFN-γ) as a molecular adjuvant. Laying hens were immunized with this construct to produce PEDV-specific IgY, which was subsequently purified from eggs using a polyethylene glycol (PEG-6000) precipitation method. The induced IgY demonstrated potent neutralizing activity against PEDV in vitro, with a neutralization titer (NT₅₀) of 1:96, which was significantly higher than that of IgY derived from hens immunized with a commercial inactivated PEDV G2b vaccine (NT₅₀ = 1:52). In a passive immunization and challenge trial, piglets treated with the rADM-IFN-G-ped-derived IgY exhibited significantly reduced fecal viral RNA shedding following challenge with the virulent PEDV-NX-2022 strain, compared to control groups. Crucially, while all piglets in the challenge control group succumbed to infection within 72 h, a 50% survival rate was achieved in the IgY-treated group. Histopathological examination of intestinal tissues further confirmed the protective efficacy, showing that IgY treatment markedly alleviated villous atrophy, epithelial necrosis, and inflammatory cell infiltration in the small intestine. These findings demonstrate that vaccination of laying hens with the rADM-IFN-G-ped recombinant adenovirus elicits a robust immune response, enabling the production of protective IgY. This proof-of-concept study establishes the viability of the multi-epitope adenoviral IgY platform as a passive immunization strategy against PEDV. Full article

Background and Objectives: Conventional assays for M-protein detection in multiple myeloma (MM), including serum immunofixation electrophoresis (sIFE) and serum free light-chain (sFLC) assays, have limitations in selected clinical settings. This pilot study aimed to develop and preliminarily validate a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based workflow using Kunitz trypsin inhibitor (KTI) as an internal standard for patient-specific serum M-protein light-chain tracking, particularly in low-level post-treatment samples in which conventional assays may be negative or difficult to interpret. Materials and Methods: A total of 55 serum samples from 25 patients with MM were analyzed. Serum immunoglobulin light-chain species were enriched using mixed κ/λ affinity beads, followed by reduction, KTI-based calibration, and MALDI-TOF MS analysis. Quantitative performance was evaluated using purified IgG1 κ standards. Time-matched sIFE and sFLC ratio results were used for descriptive comparison. Results: After KTI-based calibration, patient-specific M-protein light-chain molecular masses could be consistently identified. The assay showed good linearity over the range of 0.20–10.00 μg/mL, with a calibration equation of y = 6.0228x + 0.1063 and an R² of 0.9961. The limit of detection and limit of quantification were 0.002 μg/mL and 0.008 μg/mL, respectively. Intra-day and inter-day precision were acceptable, and recovery ranged from 96.0% to 101.2%. In selected low-level or discordant samples, including cases with therapeutic interference, polyclonal background, or non-secretory disease, MALDI-TOF MS provided exploratory complementary monitoring information. Conclusions: This KTI-calibrated MALDI-TOF MS workflow showed preliminary analytical performance within the validated low-concentration range and may serve as a complementary approach for patient-specific serum M-protein light-chain monitoring in selected clinical settings of MM. Larger independent studies are required before its clinical utility can be established. Full article

Heliobacteraceae are unique endospore-forming photosynthetic bacteria that are known for possessing the simplest photosynthetic apparatus of any known organism. More genomic and physiological analysis is needed to further understand the evolution of photosynthesis and the unique metabolic pathways of nitrogen and sulfur metabolism in this family. Here, we present the genome and phylogenetic analysis of “Heliomicrobium sulfidophilum” strain BR4^T, which was isolated previously from an alkaline sulfide-containing hot spring. In addition to the presence of a Type I reaction center, genes for bacteriochlorophyll g synthesis and nitrogenase system, the genomic analysis also explains the need for biotin as a supplied growth factor in Heliomicrobium species. The Heliobacteriaceae genome comparison also revealed a previously unidentified gene cluster of heterodisulfide reductase-like proteins (Hdr genes) and molybdopterin-based enzymes for polysulfide reductase. The whole-genome comparison, including ANI, dDDH, and single-gene phylogenetic analyses, confirms the correct placement of strain BR4 in the Heliomicrobium genus and strengthens the overall phylogenetic distribution of the Heliobacteriaceae. Full article

We extend the classical Euler-type integral representations for the Appell functions $F_1$, $F_2$, and $F_3$, to the appropriate Kampé de Fériet functions by using integration against the Meijer–Nørlund G-function. In particular, these representations provide analytic continuation of the corresponding Kampé de Fériet functions. We further focus on the following two applications. First, we obtain sufficient conditions for complete monotonicity on the positive quadrant for three families of the Kampé de Fériet functions. These conditions can be expressed directly in terms of parameters and imply, among other things, joint log-convexity and related inequalities for partial derivatives of the Kampé de Fériet functions. Second, we show how known reduction and transformation formulas for the Appell and the generalized hypergeometric functions can be lifted to Kampé de Fériet functions by concatenating parameter arrays via the integral representations. This yields several reduction formulas, including extensions of some classical and new product identities. Further combining integration against the Meijer–Nørlund G-function with Slater’s double series transformation we obtain several exotic identities for infinite sums of the generalized hypergeometric functions. Full article

Stevia residue (SR), a typical by-product of the stevia industry, is rich in organic matter and has great potential as a feed resource. However, its high fiber content and low utilization efficiency limit its practical application in poultry production. To improve the nutritional value and application potential of SR, this study first optimized the fermentation conditions of SR using response surface methodology (RSM) with chlorogenic acid as the key optimization index and then investigated the effects of different doses of SR and fermented SR (FSR) on laying performance, egg quality, antioxidant capacity, and immunity in laying hens. Six fermentation parameters, including pH, solid-to-liquid ratio, temperature, inoculation quantity, brown sugar addition, and soybean meal addition, were first screened using single-factor experiments and then optimized with RSM. Subsequently, 560 laying hens were randomly divided into seven groups and fed a basal diet supplemented with 0, 0.5%, 1.0%, and 1.5% of either SR or FSR for 28 days. The results showed that the optimal fermentation conditions were a solid-to-liquid ratio of 75.6%, brown sugar addition of 2.7%, temperature of 25 °C, inoculation quantity of 3%, and fermentation time of 9 days. In the animal study, dietary 0.5% SR and FSR reduced average daily feed intake and eggshell strength, whereas the plasma total antioxidant capacity was enhanced in all (SR or FSR) supplemented groups (p < 0.05). Moreover, plasma immunoglobulin (Ig) A level was increased in the 1.0–1.5% SR and 1.5% FSR groups, and plasma IgY was elevated in the 1.0% SR group (p < 0.05). Our results suggested that SR fermentation was effectively optimized through RSM, and dietary FSR supplementation at 1.0% improved the health of laying hens, representing the optimal inclusion level. Full article

Avian influenza infections cause substantial economic losses in the poultry industry and raise public health concerns due to viral adaptation and cross-species transmission. The frequent antigenic drift of influenza viruses further complicates the prevention and treatment of avian respiratory infections. In this study, we generated high-affinity heavy-chain variable domain (VHH) nanobodies from naïve alpaca/camelid VHH libraries using phage display combined with H9N2 influenza A virus (IAV)-infected Madin-Darby Canine Kidney (MDCK) cells. Based on binding affinity and neutralization potential, we identified seven hemagglutinin (HA)-specific and two neuraminidase (NA)-specific VHHs. Molecular docking predicted the interaction sites of HA-specific VHHs (L1-2, L1-4, A5) and NA-specific VHHs (L1-3, L2-2), providing mechanistic insights. Notably, the three HA-specific VHHs (L1-2, L1-4, A5) showed cross-reactivity to representative HA subtypes (H1, H3, and influenza B), indicating recognition of conserved epitopes across divergent influenza strains. For the first time, these camelid nanobodies were fused to the chicken IgY Fc domain, and the expression cassette was integrated into the Saccharomyces cerevisiae genome, achieving a secretion yield of 15–20 mg/L of VHH-Fc antibodies. Experimental validation confirmed that the three HA-specific VHHs-Fc constructs effectively blocked viral infection, while the two NA-specific VHH-Fc constructs (L1-3, L2-2) inhibited NA activity, demonstrating the functional efficacy of the yeast-secreted VHH–IgY Fc platform. This novel IgY Fc fusion approach offers a scalable platform with enhanced stability, extended circulation potential, and applicability in poultry. Full article

Chicken coccidiosis is a parasitic disease caused mainly by Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix, with most cases presenting as mixed infections. Currently, although a subunit vaccine (CoxAbic) targeting Eimeria maxima via maternal immunization is commercially available, no genetically engineered multivalent subunit vaccine exists against mixed infections caused by these four Eimeria species simultaneously. Therefore, we developed a tetravalent recombinant subunit vaccine (designated TEIN) by fusing key antigen genes (TA4, 3-1E, IMP1, NA4) from these four Eimeria species and expressing the construct in Pichia pastoris. A total of 500 chickens were randomly allocated into 25 experimental subgroups (n = 20 each), consisting of five groups (control, challenged, adjuvant, pPIC9K, and TEIN) and five challenge conditions (infection with Eimeria tenella, Eimeria acervulina, Eimeria maxima, Eimeria necatrix, or a mixture of four species). Immunization was performed via leg intramuscular injection at 14 and 21 days of age. At 28 days of age, all chickens except the controls were orally challenged with 1 × 10⁴ sporulated oocysts. Statistical analysis was performed using one-way or two-way ANOVA as appropriate. Results showed that chickens vaccinated with the TEIN subunit vaccine exhibited significantly elevated serum levels of IgY, IL-2, IL-10, and IFN-γ, as well as an increased splenic lymphocyte CD4⁺/CD8⁺ ratio. The anticoccidial indices (ACI) against Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix, and their mixed infection reached 174.82, 174.58, 174.41, 180.61, and 175.95, respectively. Moreover, no significant differences were observed in hematological parameters, serum biochemical markers, or histopathological findings between the vaccinated and control groups. These results demonstrate the vaccine’s potential as a promising candidate for controlling mixed coccidial infections. Full article

Recent reports have advocated the global resurgence of human metapneumovirus (hMPV) infections in the post-COVID-19 era. Considering the absence of relevant data from Greece, the present study aimed to explore the molecular epidemiology and possible resurgence of significant genotypic variants. Whole genome and F gene-specific sequencing were implemented in order to obtain complete information on the genotype and lineage distribution of circulating hMPV strains in Greece. The results showed a statistically significant increase in monthly positivity rates in 2025, especially from February to April, compared to the respective period in 2024. Overall, 21 strains were classified as genotype A2.2.2 (42.9%) and 27 as genotype B2 (55.1%), whereas only one strain belonged to genotype B1. The G gene of most completely sequenced A2.2.2 strains harbored a 111nt duplication sequence and a genotype-specific pattern of N-glycosylation sites. Maximum likelihood and time-scaled, Bayesian phylogenetic analyses demonstrated the dominance of specific sub-lineages at the regional level and international transmission events. This complex epidemic pattern in conjunction with the differential evolutionary pressure exerted on the hMPV genes, advocates continuous surveillance of hMPV epidemiology with multiple genes, or complete genome sequencing methodologies. Full article

Avian colibacillosis, caused by Escherichia coli, remains a significant threat to poultry health and production, particularly in the context of rising antimicrobial resistance. Efficient and scalable vaccination strategies are needed to reduce economic losses and reliance on antibiotics. This study investigated the safety and immunogenicity of a novel single-dose in ovo vaccine candidate based on three inactivated E. coli strains formulated with cationic maltodextrin nanoparticles. The vaccine was evaluated in broilers under commercial hatchery conditions. In ovo administration was well tolerated and did not adversely affect hatchability, survival, growth performance, or feed efficiency. Vaccinated birds mounted a measurable serum immunoglobulin Y (IgY) response against E. coli from 14 days post-hatch, which persisted until slaughter age. Furthermore, when co-administered with routinely used live-attenuated viral vaccines, no interference with the immunogenicity of these vaccines was observed. These results demonstrate that the inactivated nanovaccine is safe, immunogenic, and compatible with an industrial-scale in ovo vaccination. The findings support its potential as a practical prophylactic approach to prevent avian colibacillosis in broiler production. Full article

Approximately 10–24% of people suffer from a cat allergy. Fel d 1, the major allergen, triggers reactions in approximately 94% of sensitized individuals. Current therapeutic strategies for allergic diseases primarily involve medication or immunotherapy to alleviate symptoms, which are often burdened by low efficacy, high cost, and extended duration, posing significant challenges for patients. In contrast, IgY antibodies offer a promising alternative by reducing the level of allergens produced by cats. In this study, T-cell epitopes of Fel d 1 were predicted using ProPred/CTLpred, connected via a suitable linker (GGGGS), and expressed in E. coli. Immunization of mice and hens with recombinant cFel d 1 yielded high-titer specific antibodies (IgG: 1:301,500; IgY: 1:4,194,304). Cats administered anti-cFel d 1 IgY-enriched yolk powder (1–3% of diet) for four weeks exhibited a 30–71% reduction in salivary Fel d 1. These findings indicate that the allergen epitope-targeted IgY strategy effectively reduces allergen levels in cats, providing a promising basis for preventing and treating allergic conditions. Full article

Ultra-high dose-rate (FLASH) irradiation can transiently deplete oxygen and modulate radical-mediated chemistry in irradiated cells. Cellular antioxidants also contribute to mitigating oxidative damage in a manner dependent on linear energy transfer (LET), as suggested by recent experimental studies. In this work, we employed our multi-track Monte Carlo simulation framework (IONLYS-IRT) to investigate how LET influences transient radiation-induced oxygen depletion (ROD) in a cell-like aqueous environment under FLASH irradiation conditions. FLASH exposures were modeled as single, instantaneous pulses of protons with energies from 300 MeV to 150 keV, corresponding to LET values of ~0.3 to 71 keV/μm. Our simulations revealed a marked decline in oxygen depletion with increasing LET, in agreement with experimental observations. For an intracellular O₂ concentration of 30 μM, the oxygen consumption yield, G(–O₂), decreased from ~4.0 molecules/100 eV at low LET (~0.3 keV/μm) to ~1.6 molecules/100 eV at high LET (~71 keV/μm), representing a ~60% reduction. To assess whether ROD depends solely on LET or is also governed by ion track structure, we systematically compared multiple ion species (protons, ⁴He²⁺, ¹⁰B⁵⁺, ¹²C⁶⁺, ¹⁶O⁸⁺, ²⁰Ne¹⁰⁺, ²⁸Si¹⁴⁺, ³²S¹⁶⁺, and ⁴⁰Ar¹⁸⁺) at comparable LET values. At ~70 keV/μm, heavier ions produced significantly higher G(−O₂) values than protons—though still below those at low LET—suggesting that track structure plays a key role beyond LET alone. These findings highlight the dual importance of LET and ion-specific track structure in modulating ROD under FLASH conditions. Notably, enhanced ROD in surrounding normal tissues (low-LET plateau regions) may contribute to radioprotective effects, whereas reduced ROD in tumor tissues (high-LET Bragg peak regions) would be expected to preserve tumoricidal efficacy. Together, these results provide a mechanistic framework for optimizing proton and heavy-ion approaches in FLASH radiotherapy. Full article

Background: African animal trypanosomosis, caused by Trypanosoma vivax, remains a significant challenge to cattle health and productivity in regions where it is endemic. The development of vaccines against this parasite is particularly challenging due to its highly effective immune evasion mechanisms. Methods: An immunoproteomic approach was employed to identify T. vivax antigens through the immunocapture of parasite proteins using purified IgG from naturally infected sera. The objective of this strategy was to identify novel vaccine candidates, evaluated in a BALB/c murine model, aimed at promoting the induction of trypanotolerance. Results: An invariant surface glycoprotein (Uniprot code: F9WVM3, Tritryps code: TvY486_0045500), here designated TvISGAf, was selected based on its reported diagnostic relevance and its classification within the vivaxin antigen family. The protective potential of TvISGAf was evaluated in a murine model of T. vivax infection. Immunization with TvISGAf induced a robust antigen-specific humoral response, accompanied by a substantial cellular immune response. Following challenge, mice immunized with TvISGAf formulated with the ISPA adjuvant demonstrated enhanced control of body weight and hematocrit, and improved survival during the acute phase of infection in comparison to control group. Cytokine profiling revealed elevated levels of IFN-γ and TNF-α, accompanied by increased IL-10 production. Conclusions: Collectively, these findings demonstrate that TvISGAf formulated with ISPA confers partial protection during acute phase of infection, consistent with the induction of trypanotolerance. These results support its potential as a promising component of a multivalent vaccine strategy against T. vivax, and highlight the need for further evaluation prior to assessment in the bovine host. Full article

Bovine respiratory syncytial virus (BRSV) is a major viral pathogen associated with bovine respiratory disease (BRD), a leading cause of illness and economic loss in cattle worldwide. In June 2022, an acute respiratory outbreak occurred in a dairy herd in Photharam District, Ratchaburi Province, Thailand, affecting 25 of 103 cows (24.3%) and resulting in three deaths (2.9%). This study aimed to confirm BRSV as the etiological agent of the outbreak and to genetically and phylogenetically characterize Thai BRSV strains using partial G gene sequencing. Clinical signs included fever, nasal discharge, coughing, and subcutaneous emphysema. Nested reverse transcription-polymerase chain reaction (nested RT-PCR), a sensitive method for detecting viral RNA and targeting the F and G genes, confirmed BRSV in all samples. At the same time, bovine parainfluenza virus type 3 (BPIV-3), bovine viral diarrhea virus (BVDV), and bovine herpesvirus 1 (BoHV-1) were not detected. Phylogenetic analysis of partial G gene sequences showed that all Thai isolates clustered closely within subgroup III, with 100% nucleotide identity among themselves and 85.9–97.7% similarity to subgroup III strains from other countries. Amino acid alignment indicated conservation of key antigenic motifs, including the cysteine noose, with only minor substitutions compared to some foreign strains. This study provides the first genetic and phylogenetic characterization of BRSV in Thailand, highlighting the genetic stability of subgroup III and providing baseline molecular data to support regional surveillance, diagnostics, and vaccine strategies. Full article