Search Results (839)

Background: Streptococcus equi subspecies equi (S. equi) is the cause of strangles, one of the most prevalent diseases of horses worldwide. The disease is characterised by fever and the formation of abscesses in the lymph nodes of the head and neck, which can restrict the airway. A multicomponent subunit vaccine, Strangvac, has been shown to effectively reduce clinical signs of strangles and to reduce its incidence. Objective: The aim of this study was to determine the immune response against the immunoglobulin-cleaving endopeptidase IdeE, a key protective component within the vaccine and the ability of antibodies to neutralize the proteolytic activity of IdeE. Methods: An in vitro assay was developed to measure the functional inhibition of recombinant IdeE by horse sera pre- and post-vaccination. The IdeE-neutralising titres were compared to the corresponding IdeE-specific antibody titres measured by iELISA (indirect Enzyme-Linked Immunosorbent Assay). Results: A significant IdeE-specific antibody response in blood serum collected from ponies was induced after Strangvac vaccinations. Concomitantly, significant increases in the neutralising activity of IdeE occurred, persisting for at least 12 months post-second vaccination. IdeE-neutralising activity was further increased significantly after a third vaccination, even when the third dose was administered 12 months after the second dose, demonstrating that immunological memory to the vaccine persisted for 12 months. There was a significant correlation between the IdeE-neutralising activity of blood sera and the level of IdeE-specific antibodies. Conclusions: These data provide insights into one potential mechanism by which this vaccine protects Equids against or during S. equi infection. Full article

Psoriasis is a chronic inflammatory skin disease characterized by keratinocyte hyperactivation and dysregulated cytokine signaling, with nuclear factor kappa B (NF-κB), a master transcription factor that regulates immune and inflammatory gene expression, playing a central role. Adalimumab, a monoclonal antibody that inhibits tumor necrosis factor alpha (TNF-α), is widely used in psoriasis therapy, yet its molecular effects on NF-κB-associated genes and microRNAs (miRNAs) in keratinocytes remain insufficiently defined. In this study, immortalized human keratinocytes (HaCaT cells) were exposed to lipopolysaccharide (LPS) to induce inflammatory stress and treated with adalimumab for 2, 8, and 24 h. Transcriptome-wide profiling was performed using messenger RNA (mRNA) and miRNA microarrays, followed by validation with reverse transcription quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). Bioinformatic analyses included prediction of miRNA–mRNA interactions, construction of protein–protein interaction (PPI) networks, and gene ontology (GO) enrichment. Adalimumab reversed LPS-induced upregulation of NF-κB-associated genes, including inhibitor of nuclear factor kappa-B kinase subunit beta (IKBKB), interleukin-1 receptor-associated kinase 1 (IRAK1), TNF receptor-associated factor 2 (TRAF2), mitogen-activated protein kinase kinase kinase 7 (MAP3K7), and TNF alpha-induced protein 3 (TNFAIP3), with concordant changes observed at the protein level. Several regulatory miRNAs, notably miR-1297, miR-30a, miR-95-5p, miR-125b, and miR-4329, showed reciprocal expression changes consistent with anti-inflammatory activity. STRING analysis identified IKBKB as a central hub in the PPI network, while GO enrichment highlighted immune regulation, apoptosis, and NF-κB signaling. These findings demonstrate that adalimumab modulates NF-κB activity in keratinocytes through coordinated regulation of gene, protein, and miRNA expression, providing mechanistic insight into TNF-α blockade in psoriasis. Full article

Abdominal aortic aneurysms (AAAs) are progressive, life-threatening vascular disorders characterized by focal dilation of the abdominal aorta due to chronic weakening of the arterial wall. The condition often remains asymptomatic until rupture, which carries mortality rates exceeding 70–85%. Among the various etiological theories of AAA development, degradation of the extracellular matrix (ECM) has emerged as the most widely accepted paradigm, with the breakdown of elastin representing a central and irreversible hallmark event. Elastin, a highly cross-linked and durable structural protein, provides elasticity and recoil to the aortic wall. In human AAA specimens, reduced elastin content, impaired cross-linking, and extensive fiber fragmentation are consistently observed, while experimental studies across multiple animal models confirm that elastin degradation directly correlates with aneurysm initiation, expansion, and rupture risk. Elastin loss is driven by a complex interplay of proteolytic enzymes coupled with inflammatory cell infiltration and oxidative stress. Furthermore, elastin-derived peptides perpetuate immune cell recruitment and matrix degradation, creating a vicious cycle of wall injury. Genetic and epigenetic factors, including variants in ECM regulators and dysregulation of non-coding RNAs, further modulate elastin homeostasis in AAA pathobiology. Clinically, biomarkers of elastin turnover and elastin-targeted molecular imaging techniques are emerging as tools for risk stratification. Therapeutically, novel strategies aimed at stabilizing elastin fibers, enhancing cross-linking, or delivering drugs directly to sites of elastin damage have shown promise in preclinical models and early translational studies. In parallel, regenerative approaches employing stem cells, exosomes, and bioengineered elastin scaffolds are under development to restore structural integrity. Collectively, these advances underscore the pivotal roles of elastin not only as a structural determinant of aneurysm development but also as a diagnostic and therapeutic target. This review summarizes and integrates recent discoveries on elastin biology in AAA, with a particular emphasis on molecular mechanisms of elastin degradation and the translational potential of elastin-centered interventions for the prevention and treatment of AAA. Full article

Background: Peste des petits ruminants (PPR) is an acute or subacute contagious trans-boundary viral disease causing high morbidity and mortality in domestic and wild small ruminants. The national UAE-PPR control and eradication plan follows the PPR Global Control and Eradication Strategy (PPR GCES) and relies on the annual mass vaccination of small ruminants to eradicate the disease from the country by 2030. Despite the immunization effort against PPR, the vaccination coverage reached 65% at maximum, which necessitates conducting a post-vaccination evaluation (PVE) study at the national level. Methods: Using multistage random sampling to assess the PPR vaccine and vaccination effectiveness, protocol (2) of the PPR GCES, using two serosurveys; serosurvey (1) (pre-vaccination) at day 0 before vaccination, to assess the primary PPR serological investigation, and serosurvey (2) at (30–90) days post-PPR vaccination, to evaluate the immune response, were carried out from September to December 2024 across the seven Emirates of the UAE. The nucleoprotein-based competitive enzyme-linked immunosorbent assay (c-ELISA) was used to detect PPR antibodies in a total of 1592 and 1589 sera samples collected, respectively, before and after vaccination from different (n = 163) sheep and goats holdings (epi-unit) distributed in the different Emirates of the UAE. Results: In serosurvey (1). prior to vaccination, out of the total 1592 samples tested (839 goats and 753 sheep), 833 animals (52.32%) were found to be seropositive for PPR antibodies. In contrast, in serosurvey (2), after vaccination, 1490 (93.77%) animals were found to be seropositive out of the total 1589 small ruminants (825 goats and 764 sheep) tested by c-ELISA. A statistically significant increase (41.45%) in the overall seroprevalence from (52.32%) pre-vaccination to (93.77%) post-vaccination was observed. Post-vaccination, 93.87% (n = 153) of the vaccinated epi-units achieved more than 70% seroprevalence compared to 43.56% (n = 71) before vaccination. Prediction analysis showed that all the seven UAE Emirates require 1.2 years maximum to reach 100% immune-protection levels. Conclusions: An efficient PPR vaccine was used to immunize small ruminants in the UAE. Higher (89.47–100%) post-vaccination herd immunity than the threshold recommended by the PPR GCES (>80% immunity) was attained, which can efficiently break the spread of PPRV within the UAE. To enhance the eradication of PPR I the UAE, conducting mass vaccination campaigns targeting over the (95%) immunization coverage of eligible animals for the next three years is recommended to attain the requested sustained (>80%) immunity at the animals holding level. Full article

Mollusks, such as bivalves, face increasing threats, such as disease, in aquaculture. Exosomes, widely derived from living cells carrying diverse bioactive molecules, affect the immune response. To overcome these challenges, bivalves utilize exosomal miRNAs as critical regulators of immune responses. This study investigates the role of exosomal miRNAs in modulating immune and metabolic responses in Pinctada fucata martensii following lipopolysaccharide (LPS) stimulation. Exosomes (75–150 nm) were isolated from hemolymph and characterized. High-throughput sequencing identified 30 differentially expressed miRNAs (DEMs) and 1349 differentially expressed genes (DEGs) in LPS-treated oysters, with significant enrichment in TNF, TLR/NF-κB, and metabolic pathways. This study revealed exosomal miRNA-mediated regulation of immune genes (IκBα, TRAF6, IRAK1, and BIRC2/3) and metabolic enzymes (PCK and CYP2J), demonstrating their role in apoptosis, inflammation, and metabolic reprogramming. Network analysis highlighted miRNA–mRNA interactions, including miR-7/IκBα (TNF pathway) and miR-34_5/IRAK1 (TLR pathway). Additionally, exosomal miRNAs (miR-92_2 and novel_mir5) were found to regulate oxidative stress (SOD1) and gluconeogenesis (PCK), linking immune defense with metabolic adaptation. These findings provide novel insights into exosomal miRNA-mediated immune regulation in bivalves, revealing conserved mechanisms with potential implications for molluscan health and disease management. Full article

Treatment options for major depression are limited: only about one-third of patients achieve remission with first line treatments with no established predictive markers. Parameters associated with treatment refractory depression, including metabolic markers (increased BMI, increased triglyceride levels), inflammation markers (C-reactive protein, CRP), autonomic disturbances (reduced blood pressure, reduced heart rate variability), and brain morphology changes (increased volume of the choroid plexus and brain ventricle volumes), may serve such purpose. These features can be linked mechanistically to an increase in aldosterone plasma concentration due to a reduced mineralocorticoid receptor (MR) sensitivity. The primary CNS target of aldosterone is the nucleus of the solitary tract (NTS), which is also the entry point of the vagus nerve. This nucleus integrates signals from endocrine, inflammatory, chemoreceptive, and physiological parameters, including blood pressure. In search of a mechanism to overcome this pathology, we identified a molecule which is derived from the licorice plant glycyrrhiza glabra, namely glycyrrhizin and its biologically active metabolite enoxolone. These molecules potentially reverse the above-described pathology. They inhibit the enzyme 11beta hydroxysteroid-dehydrogenase type 2 (11betaHSD2) and the toll-like receptor 4 (TLR4). 11betaHSD2 regulates the activity of the mineralocorticoid receptor (MR) by degrading cortisol/corticosterone, which allows aldosterone to bind to the MR. TLR4 is the ligand for lipopolysaccharide (LPS, endotoxin) and trigger of innate immunity. Consequently, patients with increased inflammation markers, increased aldosterone, or low blood pressure may preferentially benefit from the treatment with glycyrrhizin/enoxolone. Importantly, these patients can be identified BEFORE treatment is initiated. Clinically, patients sharing these biological indicators are primarily young females or patients with a history of childhood trauma. A combination of enoxolone with standard antidepressants may therefore avoid a trial-and-error approach and allow to achieve recovery faster. Full article

Protein homeostasis is a dynamic process essential for cellular function and survival, tightly controlled by the ubiquitin–proteasome system. Within this system, ubiquitin-specific protease 7 (USP7) plays a key role as a deubiquitinating enzyme, thus modulating the stability, localization, and activity of a wide variety of substrates. USP7 is involved in critical cellular processes such as DNA repair, apoptosis, immune response, and epigenetic regulation. The dysregulation of USP7 expressions or activity has been linked to several pathological conditions, including cancer, neurodegenerative and inflammatory diseases, and viral infections. This enzyme exerts its biological functions through the stabilization of both oncogenic and tumor suppressor proteins, highlighting its sensitive role in tumorigenesis. Despite the identification of selective USP7 inhibitors with promising preclinical activity, the development of clinically effective compounds remains a major challenge. This review summarizes the current understanding of USP7 structure, function, and biological relevance, with a particular emphasis on its potential as a therapeutic target in oncology. Full article

Animal venoms are rich in bioactive molecules with promising biotechnological potential. They comprise both protein and non-protein toxins. Among the protein toxins are enzymes, such as phospholipases A₂, proteases and L-amino acid oxidases (LAAOs). LAAOs exhibit antimicrobial, antiparasitic, antiviral, and anticancer effects, making them potential candidates for biotechnological applications. These activities are linked to their ability to catalyze oxidative reactions that convert L-amino acids into α-keto acids, releasing ammonia and hydrogen peroxide, which contribute to the immune response, pathogen elimination, and oxidative stress. However, in snakes of the Micrurus genus, LAAOs generally represent a small portion of the venom (up to ~7%), which limits their isolation and study. To overcome this, the present study aimed to produce Ml-LAAO, the enzyme from Micrurus lemniscatus, through heterologous expression in mammalian cells. The gene sequence was inferred from its primary structure and synthesized into the pSecTag2B vector for expression in HEK293T cells. After purification using a His Trap-HP column, the presence of recombinant Ml-LAAO (Ml-LAAOrec) was confirmed by Western blot and mass spectrometry, validating its identity. These results support successful recombinant expression of Ml-LAAO and highlight its potential for scalable production and future biotechnological applications. Full article

Biodegradable plastics are not a primary solution to plastic pollution, and empirical evidence on whether they are environmentally friendly remains lacking. In this study, we systematically compared the toxic effects of traditional microplastics (polypropylene, PP; polystyrene, PS) with biodegradable microplastics (polylactic acid, PLA; polyhydroxyalkanoates, PHA) on the haplic phaeozem ecosystem. Through mathematical modeling analysis, it was found that earthworms initially rely on antioxidant enzymes to resist stress, mid-term activation of detoxifying enzymes to repair damage, and maintaining physiological balance through metabolic regulation and immune enhancement in later stages. We elucidated their mechanism differences: PLA and PP caused severe damage to the antioxidant system and cell membrane, with PLA mainly relying on POD to clear peroxides and PP relying on GST. In addition, PLA and PS can induce early neurotoxicity (AChE), while PHA induces late neurotoxicity. Furthermore, this study provides direct evidence proving that biodegradable microplastics are not environmentally friendly by breaking through the one-way research framework of “microplastic biotoxicity” and innovatively constructing a path analysis model that links biological physiological responses with soil ecological functions. We also provide a scientific basis to evaluate the ecological risks of microplastic pollution in soil and the whether biodegradable plastics are truly environmentally friendly. Full article

Background: The hepatitis A virus (HAV) is a major cause of acute viral hepatitis and a significant global health concern. This study provides a pre-vaccination baseline for Oman, enabling longitudinal comparison with post-hepatitis A vaccination cohorts. This study aimed to determine the pre-vaccination seroprevalence of HAV antibodies (anti-HAV) in Oman and explore the associated demographic factors. Methods: A cross-sectional study was conducted from April 2014 to August 2015 among patients attending the medical outpatient clinic of the Medical City Hospital for Military and Security Services. Demographic data were collected via a structured questionnaire, and serum samples were tested for anti-HAV immunoglobulin IgG and IgM using enzyme-linked immunosorbent assays. Multivariate analysis was performed to identify the predictors of anti-HAV seroprevalence. Results: Among 1975 participants, 88.1% were positive for anti-HAV IgG. The mean age was 37.4 ± 16.1 years; however, those negative for anti-HAV IgG were considerably younger (mean age: 24.8 ± 15.7 years). Anti-HAV IgG seroprevalence was 37% in individuals aged ≤18 years and 91% in those >18 years (p < 0.001). The factors associated with seropositivity included older age (p < 0.001), consuming food prepared outside the home (p < 0.001), occupation (p < 0.001), and education level (p = 0.003). In the multivariable analysis, only age showed a strong independent association with serostatus: per 10-year increase, the aOR for anti-HAV IgG seropositivity was 2.87 (95% CI 2.25–3.63; p < 0.001). Conclusions: Our study estimates show high anti-HAV IgG seroprevalence and serve as a pre-vaccination baseline for evaluating the hepatitis A vaccination program in Oman over time. Given the lower natural exposure among younger cohorts, continued routine vaccination, scheduled serosurveys, and strengthened surveillance are required to identify emerging immunity gaps and prevent future HAV outbreaks. Full article

Tryptophan (Trp) and tryptamine (Tryp), critical biomarkers in mood regulation, immune function, and metabolic homeostasis, are increasingly recognized for their roles in both oral and systemic pathologies, including neurodegenerative disorders, cancers, and inflammatory conditions. Their rapid, sensitive detection in biofluids such as saliva—a non-invasive, real-time diagnostic medium—offers transformative potential for early disease identification and personalized health monitoring. This review synthesizes advancements in electrochemical sensor technologies tailored for Trp and Tryp quantification, emphasizing their clinical relevance in diagnosing conditions like oral squamous cell carcinoma (OSCC), Alzheimer’s disease (AD), and breast cancer, where dysregulated Trp metabolism reflects immune dysfunction or tumor progression. Electrochemical platforms have overcome the limitations of conventional techniques (e.g., enzyme-linked immunosorbent assays (ELISA) and mass spectrometry) by integrating innovative nanomaterials and smart engineering strategies. Carbon-based architectures, such as graphene (Gr) and carbon nanotubes (CNTs) functionalized with metal nanoparticles (Ni and Co) or nitrogen dopants, amplify electron transfer kinetics and catalytic activity, achieving sub-nanomolar detection limits. Synergies between doping and advanced functionalization—via aptamers (Apt), molecularly imprinted polymers (MIPs), or metal-oxide hybrids—impart exceptional selectivity, enabling the precise discrimination of Trp and Tryp in complex matrices like saliva. Mechanistically, redox reactions at the indole ring are optimized through tailored electrode interfaces, which enhance reaction kinetics and stability over repeated cycles. Translational strides include 3D-printed microfluidics and wearable sensors for continuous intraoral health surveillance, demonstrating clinical utility in detecting elevated Trp levels in OSCC and breast cancer. These platforms align with point-of-care (POC) needs through rapid response times, minimal fouling, and compatibility with scalable fabrication. However, challenges persist in standardizing saliva collection, mitigating matrix interference, and validating biomarkers across diverse populations. Emerging solutions, such as AI-driven analytics and antifouling coatings, coupled with interdisciplinary efforts to refine device integration and manufacturing, are critical to bridging these gaps. By harmonizing material innovation with clinical insights, electrochemical sensors promise to revolutionize precision medicine, offering cost-effective, real-time diagnostics for both localized oral pathologies and systemic diseases. As the field advances, addressing stability and scalability barriers will unlock the full potential of these technologies, transforming them into indispensable tools for early intervention and tailored therapeutic monitoring in global healthcare. Full article

Background/Objectives: Calsequestrin (CSQ) is a calcium-binding protein that is highly soluble and can serve as a solubility-enhancing fusion tag in recombinant protein expression. Its unique property of calcium-induced precipitation followed by EDTA-mediated resolubilization enables efficient purification. However, the broader application of CSQ-tagged proteins in research have been hampered by the lack of reliable anti-CSQ detection reagents. This study aimed to develop single-domain antibodies (sdAbs) against CSQ for use in diverse immunoassays and cell-based analyses. Methods: Single-domain antibodies were selected from phage-displayed chicken VH libraries generated from CSQ-immunized chickens. After biopanning, CSQ-specific VH sdAb clones were isolated and expressed as VH–human kappa light chain constant region (VH-Cκ) fusion proteins in E. coli. The PE06 clone was chosen for further characterization and conjugated to horseradish peroxidase (HRP) and Alexa Fluor 647 for assay applications. Results: PE06 VH-Cκ fusion protein demonstrated specific binding to CSQ-tagged proteins and enabled reliable detection in enzyme-linked immunosorbent assay (ELISA), immunoblotting, and flow cytometry. These results validated its utility as a chemically defined detection reagent for CSQ fusion proteins expressed in E. coli. Conclusions: This study establishes a CSQ-specific chicken VH sdAb as a versatile detection tool for CSQ-tagged proteins. The approach expands the utility of CSQ as a protein fusion tag and enables the development of recombinant antibodies fused with CSQ, such as scFv-CSQ constructs, for broad application in research and assay systems. Full article

Organophosphorus flame retardants (OPFRs) are emerging alternatives to halogenated compounds, yet their environmental toxicity remains underexplored. This study evaluated the developmental toxicity of two aryl-OPFRs, triphenyl phosphate (TPP) and tricresyl phosphate (TCP), in zebrafish (Danio rerio) from 2 h to 5 days post fertilization (hpf–dpf). Survival, hatching rate, and malformations were assessed across concentrations of 250–1000 µg/L, alongside with gene expression analysis at 5 dpf (250 and 500 µg/L) targeting detoxification (ces2), immune responses (il1β, casp9), and epigenetic markers (dnmt1, dnmt3). In vitro enzymatic assays evaluated interactions of both aryl-OPFRs with carboxylesterase (CE) and acetylcholinesterase (AChE) enzymes. While no significant morphological effects were observed, TPP showed higher toxicity than TCP. Notably, TCP (500 µg/L) downregulated genes linked to metabolism and immunity. CE activity and ces2 modulation may suggest CE as a potential biomarker for aryl-OPFR exposure. These findings, although at concentrations above the environmental ones, may be valuable for mechanistic purposes and underscore the need for further investigation in developmental toxicity given their lipophilic nature and distinct molecular responses. Full article

Background/Objectives: In children, gastroesophageal reflux disease (GERD) may lead to epithelial barrier dysfunction and the release of thymic stromal lymphopoietin (TSLP), interleukin-25 (IL-25), interleukin-33 (IL-33) and periostin, known as alarmins. These cytokines are associated with type 2 inflammation and may contribute to respiratory and allergic conditions. The main purpose of this study is to evaluate serum concentrations of TSLP, IL-25, IL-33, and periostin in children with and without GERD and to assess their relationships with bronchial hyperresponsiveness (BHR) and sensitization to inhaled allergens. Methods: The study included 93 children aged 7–17 years. GERD was diagnosed based on 24-h esophageal pH impedance monitoring. Serum levels of TSLP, IL-25, IL-33, and periostin were measured using enzyme-linked immunosorbent assay (ELISA). It should be noted that the assay used does not distinguish between TSLP isoforms, which represents a limitation of the study. BHR was assessed via a methacholine challenge test, and allergen sensitization was determined using skin prick tests and allergen-specific immunoglobulin E (asIgE). Results: Serum TSLP levels were significantly higher in children with GERD compared to those without, whereas IL-25, IL-33 and periostin did not differ notably between groups. Periostin was associated with the degree of sensitization to inhalant allergens, but no significant links were found between cytokine levels and bronchial hyperresponsiveness. Conclusions: Significantly higher TSLP levels were noted in children with GERD than in those without. Hence, TSLP may have a potential role as a biomarker of epithelial immune activation in pediatric GERD. In addition, periostin was associated with sensitization to inhalant allergens, although it did not differentiate between children with and without GERD. Full article

Background and Objectives: The pathogenesis of thyroid eye disease (TED) is driven by interactions between orbital fibroblasts and immune cells. Lymphocyte-activation gene 3 (LAG-3) is an immune checkpoint molecule with a similar structure to the T lymphocyte CD4 receptor but with higher affinity for MHC class II, and LAG-3–MHC class II interaction inhibits T lymphocyte activity. Lymphocytes shed LAG-3, generating soluble LAG-3 (sLAG-3), whose function is unclear. We investigated sLAG-3 involvement in Graves’ disease (GD) and GD-associated TED pathogenesis. Materials and Methods: Patients with GD-associated TED (n = 47) and GD without TED (n = 35) were enrolled alongside 37 healthy controls (HCs). Peripheral blood serum sLAG-3 levels were measured using enzyme-linked immunosorbent assays and compared across the three groups. The effect of intravenous glucocorticosteroid (IVGC) treatment (12 weeks) on sLAG-3 concentrations in patients with GD-associated TED was monitored, and associations of sLAG-3 levels with clinical characteristics were analyzed. Disease activity before and after IVGC treatment was assessed using Clinical Activity Score. Results: Relative to those in HCs, serum sLAG-3 levels were significantly higher in GD patients both with (p < 0.001) and without (p = 0.0129) TED. No significant difference in sLAG-3 levels was observed between the two patient groups (p = 1.000), and no significant change in sLAG-3 levels was detected in patients with TED after IVGC therapy (p = 0.0536). Conclusions: The higher sLAG-3 levels in patients compared to HCs suggest that sLAG-3 dysregulation may contribute to GD and GD with orbitopathy development and the pathomechanisms underlying these conditions. Metalloproteinase-mediated cleavage of LAG-3 from the lymphocyte surface enables T lymphocyte proliferation and activation, while released sLAG-3 may enhance the immune response. Further studies of sLAG-3’s mechanisms of action are needed to establish clear cut-off values and to define the diagnostic role of sLAG-3 in GD diagnosis. Full article