Search Results (10,598)

Nucleosides are extensively employed for the development of pharmaceuticals, chemotherapeutic agents, and bioregulators. Background/Objectives: The introduction of an additional chiral functionality into a carbohydrate or heterocyclic base fragment may increase the selectivity of interactions with nucleos(t)ide-metabolizing enzymes and receptors and, in some cases, lead to more specific physiological activities. Methods: An improvement of the selectivity of nucleoside-based drugs can be achieved either by the chemical modification of a carbohydrate or a base constituent or by a combination of these two approaches. Additionally, stereospecific enzymatic cleavage of nucleos(t)ide prodrugs containing biodegradable substituents can reduce cytotoxicity and enhance bioavailability. Results: A series of enantiomerically pure nucleosides modified at the ribose or heterocyclic base were obtained by chemical and enzymatic methods. Novel antiviral or anticancer active compounds, inhibiting viral or cellular enzymes or activating cellular nucleoside kinases have been found among chemically synthesized derivatives. Some exhibit strengthened “ligand–receptor” interaction, acting on receptors of the purinergic signaling system. During recent extensive structure–activity studies, several drugs and their prototypes have been proposed for the treatment of viral infections: the 2′C-fluoromethyl derivative of sofosbuvir (anti-SARS-CoV, preclinical), VV-261 (SFTSV, phase I clinical), balapiravir (dengue, phase I clinical), mericitabine (approved drug for HCV), and lumicitabine (approved drug for RSV and HMPV). Conclusions: Modern literature data within the scope of the present review suggest that direct modification of nucleosides with various chiral functionalities can be considered as an approach to increase their efficacy, specificity and selectivity. Full article

Myocardial ischemia and reperfusion initiate spatially organized injury–repair programs that subsequently shape ventricular remodeling. Although cardiac fibroblasts are indispensable for scar formation, single-cell and spatial transcriptomic studies reveal temporally dynamic and regionally distinct fibroblast-lineage states. This critical narrative review integrates direct evidence from myocardial ischemia–reperfusion (I/R) with model-labeled evidence from permanent myocardial infarction, clinically heterogeneous human infarction, fibroblast-specific ubiquitin biology, cell-cycle regulation, and cardiac fibroblast atlases. A direct fibroblast I/R study identifies an HSP47–USP10–SMAD4 deubiquitination axis, whereas most other fibroblast ubiquitin–proteasome system (UPS) mechanisms derive from permanent infarction, non-ischemic cardiac stress, or in vitro systems. We propose that CCNB1-associated, G2/M-enriched cycling fibroblast-lineage states may impose heightened proteostatic demands within defined post-ischemic niches. The conceptual novelty is not that CCNB1 turnover or UPS activity is cardiac-specific; both are general features of proliferating cells. Rather, the framework asks whether fibroblast lineage, injury-model provenance, anatomical niche, temporal window, cell state, and substrate-specific UPS nodes jointly define proteostatic dependencies during post-ischemic remodeling. RNA-based ubiquitin-related signatures remain transcriptional proxies and do not directly quantify ubiquitinated substrates, ubiquitin-chain topology, enzyme activity, or proteasome flux. Resolving the proposed relationships will require spatial colocalization, protein-level and ubiquitin-remnant profiling, proteasome and ribosome assays, fibroblast-specific perturbation, and validation in human infarct tissue. Full article

In recent years, a novel class of highly stable multiprotein complexes, with molecular masses ranging from 1 to 2 MDa, has been identified in human milk, placenta, sea urchin eggs, and sea cucumbers. These complexes exhibit extraordinary stability, dissociating only under stringent conditions involving 8 M urea, 3 M MgCl₂, EDTA, and DTT. Previous investigations have demonstrated that complexes derived from different organs of the sea cucumber Paracaudina chilensis differ in size, molecular mass, and protein/peptide composition; however, their enzymatic activities have remained unexplored. In the present work, we performed the first systematic analysis of phosphatase activity associated with highly stable complexes isolated from five organs of P. chilensis: the body wall, gonads, respiratory trees, intestine, and coelomic fluid. Complexes were purified via gel filtration chromatography on Sepharose 4B, followed by ultracentrifugation. Phosphatase activity was determined spectrophotometrically by monitoring the hydrolysis of p-nitrophenyl phosphate. Our results indicate that all five complexes harbor phosphatases with optimal pH values spanning 7.0 to 10.0. Alkaline phosphatases (pH 9.0–10.0) displayed pronounced organ specificity: maximal activity was observed in the intestinal complex, whereas minimal activity was detected in the gonadal complex. Phosphatase activity in complexes from the body wall and respiratory trees exhibited a bell-shaped dependence on Mg²⁺ concentration, with optima at 5 mM and 1 mM, respectively; in contrast, activity in the intestinal and coelomic fluid complexes increased to a plateau at 5–10 mM Mg²⁺. Ca²⁺ ions predominantly inhibited activity, with the notable exception of the intestinal complex, where they exerted no effect on hydrolysis. EDTA treatment resulted in complete inactivation of the enzymes in most complexes; however, intestinal activity was retained at 50% even at high chelator concentrations, suggesting the presence of a metal-independent phosphatase. Collectively, these data indicate that the stable multiprotein complexes of P. chilensis contain an organ-specific repertoire of phosphatases that differ in pH optimum, metal ion dependence, and inhibitor sensitivity. These findings open new avenues for understanding the roles of such complexes in organ-specific physiological functions and regenerative mechanisms in echinoderms. Full article

Drought stress severely limits grapevine (Vitis spp.) productivity, yet the regulatory mechanisms distinguishing drought-tolerant and drought-sensitive cultivars remain insufficiently understood. In this study, we integrated transcriptomic, metabolomic, physiological, and functional analyses to compare drought responses between the tolerant cultivar ‘Miguang’ and the sensitive cultivar ‘Red Globe’, together with previously generated datasets from ‘Shine Muscat’ and ‘Thompson Seedless’. Compared with sensitive cultivars, tolerant cultivars showed stronger antioxidant capacity, lower lipid peroxidation, and more coordinated changes in pathways related to redox balance, osmotic adjustment, and energy metabolism. In ‘Miguang’, drought responses were associated with activation of the pentose phosphate pathway, accumulation of tricarboxylic acid cycle intermediates, and genotype-specific alternative splicing events affecting metabolic and signaling genes. Comparative analysis across four cultivars identified VvGRIK1 as a conserved drought-responsive regulator associated with redox and bioenergetic homeostasis. Heterologous overexpression of VvGRIK1 in tobacco enhanced drought-related physiological performance by increasing antioxidant enzyme activities and reducing membrane damage. Yeast two-hybrid assays and molecular docking further suggested a potential interaction between VvGRIK1 and VvKING1, a SnRK1-related energy sensor. Together, these findings suggest that the VvGRIK1-SnRK1 module may contribute to drought adaptation by coordinating redox protection and energy homeostasis, providing a candidate regulatory target for future functional studies and grapevine molecular breeding. Full article

Oilseed cakes (OC), generated as by-products of mechanical oil extraction, are increasingly recognized as functional feed ingredients in swine production. Besides providing protein and residual lipids, they contain bioactive compounds such as polyphenols, tocopherols, and phytosterols that may exert antioxidant and immunomodulatory effects. This narrative review is based on a narrative literature search conducted in the Web of Science, Scopus, PubMed, and Google Scholar databases, covering publications from 2000 to 2026. This review summarizes current knowledge on the role of OC in pig nutrition, focusing on redox balance, immune modulation, and production performance. Experimental studies indicate that rapeseed, soybean, sunflower, and other oilseed cakes can influence oxidative stress markers and antioxidant enzyme activity, contributing to protection against oxidative damage. Evidence also suggests that these by-products may affect immune responses through improvements in intestinal barrier integrity and modulation of cytokine production. Among the reviewed OC, rapeseed, soybean, sunflower, and flaxseed cakes showed the most consistent evidence of beneficial effects on antioxidant status, immune function, and production performance, particularly when included at moderate dietary levels. In contrast, the available evidence for camelina, cottonseed, Amarula, and high-fiber oilseed by-products remains limited or inconsistent. Their effects appear to depend largely on dietary inclusion level, processing method, and the presence of antinutritional factors. Excessive dietary inclusion may impair nutrient digestibility and growth performance, highlighting the importance of optimizing inclusion levels according to the specific type of OC. Overall, OC represent a promising component of sustainable pig-feeding strategies, supporting animal health and production efficiency while promoting the use of agro-industrial by-products. Full article

Antisense oligonucleotides (ASOs) are short, synthetic DNA fragments able to modulate gene expression. In this work, the cytosolic isoform of glucose-6-phosphate dehydrogenase (Cyt-G6PDH) was selected as a target for ASOs to induce transient gene silencing in barley (Hordeum vulgare). G6PDH is the most important enzyme of the oxidative pentose phosphate pathway (OPPP), supplying NADPH and regulating the entire cycle. Different ASOs were tested at different concentrations on the leaf surface. Their effects were evaluated by measuring enzymatic activity, gene expression, and protein abundance. Treatment with 30 µM ASOs for 6 h represented the optimal condition, inducing a 40–60% reduction in G6PDH total activity in barley leaves, with a specific decrease in the cytosolic isoform, as determined by DTT-sensitive enzymatic assays. ASOs designed on the main regulator of cyt-G6PDH, the shaggy-like kinase (SK11), led to an analogous effect in terms of G6PDH activity, confirming the involvement of HvSK11 in the regulation of cyt-G6PDH in barley. Consistently, qRT-PCR analyses showed that ASO treatment simulates an abiotic stress condition obtained by the down-regulation of Cyt-G6PDH. These results support the use of ASOs as a rapid and efficient method for the functional analysis of key metabolic regulators in plants to overcome complications in recalcitrant organisms or lethal genes. Full article

Background and Objectives: Environmental exposure to endocrine-disrupting chemicals (EDCs) is increasingly recognized as a potential contributor to cancer-related biological variability; however, human biomonitoring data in oncology populations remain limited. The present study aimed to assess urinary concentrations of selected phenolic EDCs and their associations with cardiometabolic, hematological, inflammatory, and survival-related parameters in patients with advanced lung cancer. Materials and Methods: A total of 190 patients diagnosed with stage IIIB/IV lung cancer were included in this study. Urinary concentrations of bisphenol A (BPA), bisphenol S (BPS), triclosan (TCS), and resorcinol (RCO) were determined using validated analytical methods. Associations between exposure biomarkers and clinical laboratory parameters were evaluated using sex-stratified statistical analyses and regression models adjusted for age and body mass index. Results: TCS was the most frequently quantified compound (29.47%), followed by BPS (27.37%), RCO (11.58%), and BPA (7.89%). Higher odds of TCS quantification were observed in patients with lung adenocarcinoma and a higher probability of BPA quantification in patients with squamous-cell carcinoma. Sex-specific exposure patterns were observed, with higher BPA and BPS concentrations measured among female patients. Exposure to phenolic EDCs was associated with alterations in kidney function biomarkers, liver enzyme activity, inflammatory cell profiles, and anthropometric indicators. In particular, BPA and BPS showed associations with renal function markers and systemic inflammatory parameters, while TCS exposure was related to reduced leukocyte subpopulations. Survival analysis demonstrates borderline associations for BPA between exposure groups. Conclusions: These findings provide novel human biomonitoring evidence linking exposure to phenolic endocrine-disrupting chemicals with systemic metabolic and inflammatory variability in patients with advanced lung cancer. The observed associations support the biological plausibility that environmental endocrine disruptors may contribute to interindividual heterogeneity in cancer-related physiological responses. Full article

Morinda citrifolia (Noni) is well known as a plant with therapeutic potential and is also attractive to the food and cosmetic industries. Traditional medicine supports its use, mainly for the treatment of metabolic disorders and chronic inflammation but also for certain types of cancer. Noni has been the subject of considerable interest within the scientific community due to its purported health benefits. However, despite its growing popularity and extensive traditional use, significant gaps remain in the empirical understanding of its properties, mechanisms, and potential applications. To understand its biological activity, particular attention has been given to several chemical compounds present in its leaves and fruits, as they have been identified as bioactive agents. Moreover, several studies support the idea that specific flavonoids and anthraquinones from noni act on enzymes and transporters associated with glucose and lipid metabolism in humans. Its involvement in cardiovascular, neurological, metabolic, and inflammatory regulation across several high-burden diseases expands the potential medical applications of noni. This narrative review presents the current state of knowledge, highlighting preclinical studies that suggest the mechanisms of action underlying the observed effects, including theoretical approaches proposing specific interactions between noni compounds and proteins associated with human diseases as potential therapeutic targets. It also identifies areas where information remains insufficient and proposes future research directions for pharmacological applications, including the need for additional clinical studies and more comprehensive pharmacokinetic and toxicological evaluations. Full article

Background/Objectives: The autoimmune disorder Systemic Lupus Erythematosus (SLE) is characterized by increased titers of autoantibodies with different specificities against autoantigens, including the complement proteins C1q, C3 and Factor H. SLE is characterized by chronic inflammation and tissue damage due to the secretion of pro-inflammatory molecules and a tissue deposition of immune complexes formed by autoantibodies and their target antigens. The inflammatory process in SLE is maintained by the phospholipase A2 (PLA2) enzymes generating pro-inflammatory lipid mediators. Hereditary and environmental factors trigger SLE, with increased genetic heritability in first-degree relatives of SLE patients. Methods: A cohort of 48 healthy FDRs of SLE patients was analyzed with the ELISA method for the presence of antibodies to complement proteins C1q, C3 and Factor H, with a focus on detecting autoepitopes both on immobilized and soluble C1q and its globular fragments ghA, ghB and ghC. The total serum PLA2 activity of FDRs was measured using the chromogenic substrate 4-nitro-3-octanoyloxy-benzoic acid (NOBA). Results: Only C1q, and specifically its globular domains in both an immobilized and soluble state, was targeted by antibodies in the healthy FDRs similarly to the pattern established in SLE patients. In contrast, C3 and Factor H which are known autoantigens in SLE were not found as targets for the antibodies in the analyzed FDRs. Some of the FDRs showed increased serum PLA2 activity, which correlated weakly with anti-C1q antibodies. Conclusions: C1q and its globular domains are estimated as autoantigenic molecules for binding in the analyzed FDRs of SLE patients. Full article

Iron (Fe) is an essential micronutrient for plants. However, both iron deficiency and excess can severely inhibit plant growth and productivity. Arbuscular mycorrhizal (AM) fungi have been shown to improve plant mineral nutrition and stress tolerance, yet the integrated physiological and molecular mechanisms underlying AM-mediated iron homeostasis regulation, particularly in woody tree species, remain poorly understood. In this study, we investigated the effects of inoculating Eucalyptus grandis seedlings with the AM fungus Rhizophagus irregularis on their growth, photosynthetic performance, antioxidant defense, and transcriptional responses under varying Fe supply levels (5, 25, and 200 µM). Our results demonstrated that AM symbiosis significantly alleviated the growth inhibition induced by both low-Fe (5 µM) and high-Fe (200 µM) stress, enhanced photosynthetic capacity, as evidenced by increased net photosynthetic rate (Pn), stomatal conductance (Gs), and PSII photochemical efficiency; under low-Fe, Pn, Gs, and Fv/Fm increased by 33.7%, 42.4%, and 25.2%, respectively. AM symbiosis also significantly enhanced the activities of antioxidant enzymes (POD, SOD, and CAT) under high-iron stress, accompanied by reduced accumulation of reactive oxygen species and lipid peroxidation; specifically, POD, SOD, and CAT activities increased by 79.3%, 88.7%, and 87.0%, respectively. Transcriptomic analysis identified 44 MYB transcription factors that were differentially induced by AM symbiosis under iron stress, among which six genes (EgMYB-2, EgMYB-3, EgMYB315-1, EgMYB315-2, EgMYB61, and EgMYB306) were significantly upregulated by AM under both low- and high-iron conditions, as verified by qRT-PCR. Correlation analysis revealed strong positive associations between the expression of these EgMYB genes and antioxidant enzyme activities as well as photosynthetic parameters, suggesting their potential involvement in coordinating iron stress responses. Overall, our findings indicate that AM fungus enhances iron stress tolerance in E. grandis through a multilevel strategy that includes photosynthetic protection, antioxidant defense activation, and transcriptional reprogramming involving MYB transcription factors as potential regulators. This study provides integrated physiological-molecular analysis and novel insights into AM-mediated Fe homeostasis regulation in the woody tree species E. grandis. Full article

Green undersown crops (GUCs) are increasingly promoted as a sustainable management strategy to improve soil quality and ecosystem services in Mediterranean orchards, particularly in the calcareous, low-organic-matter soils typical of Eastern Spain. In this context, soil quality is understood as the soil’s overall functional capacity, integrating physical structure, chemical balance, and biological activity, whereas soil fertility refers specifically to the soil’s ability to supply nutrients to plants; soil quality therefore encompasses a broader set of ecosystem functions beyond nutrient provision. However, their effects on the calcareous, low-organic-matter soils typical of Eastern Spain remain insufficiently quantified. This study evaluates the effects of seeded undersown (grasses, legumes, and flower mixtures), spontaneous vegetation, and herbicide-managed bare soil on topsoil (0–15 cm) physicochemical and biological indicators in 54 plots across three irrigated persimmon orchard sites (Granja, Cargol, and Alginet) over 18 months of treatment in the València region (Eastern Spain). Seasonal sampling was conducted at the START (early winter) and END (late spring) of the experiment period. Soil measurements at both sampling times included soil organic matter (SOM), nitrogen (N), C/N ratio, pH, electrical conductivity (EC), soil respiration rate (RR), collembolan abundance, mite abundance, and the QBS-ar index of soil arthropods. Legumes increased SOM by +1.12%, grasses by +0.22%, whereas flower mixtures (−0.44%) and spontaneous vegetation (−1.36%) showed SOM reductions associated with rapid biomass turnover. RR increased under all GUCs (+0.06 to +0.16 g CO₂ m⁻² h⁻¹), and QBS-ar improved markedly under grasses (+26.6) and spontaneous vegetation (+36.7). EC decreased across all treatments (−16 to −84 µS cm⁻¹). These results were analysed using principal component analysis (PCA). Four PCA components explained 74% of the total variance, revealing functional gradients driven by SOM, N, EC, RR and mesofauna. After 18 months, microbial biomass carbon (MBC) increased by +45–60% under legumes, water-soluble organic carbon (WSOC) by +30–50% under legumes and flower mixtures, and the enzyme activities (EA) by +20–40% under all GUCs. Herbicide-managed soils showed reduced biological activity and detectable residues of glyphosate and oxyfluorfen. PCA and linear discriminant analysis (LDA) were used to identify functional gradients and treatment separation. GUCs significantly increased SOM, MBC, EA, and mesofauna abundance compared with herbicide treatments, which showed reduced biological activity and detectable residues of glyphosate and oxyfluorfen. Legumes and flower mixtures produced the strongest improvements in biological functioning due to higher MBC, WSOC, EA and RR. PCA and LDA confirmed clear separation between GUCs and herbicide-managed soils based on multivariate differences in SOM, N, EC, RR, MBC, WSOC and mesofauna indicators. Overall, GUCs modulated soil chemistry and biodiversity and enhanced soil functioning and biological quality, supporting their adoption as a sustainable management strategy in Mediterranean orchards. Legume-based covers are recommended for rapid biological activation, whereas grass-based covers favour longer-term SOM stabilisation. These findings highlight their role as key tools for improving soil resilience in Mediterranean persimmon orchards. Full article

Baijiu brewing generates large quantities of wastewater sludge, which presents major disposal challenges but contains abundant organic matter and nutrients, indicating great potential as a soil amendment. This study investigated the soil amendment effects of granulated and coated Baijiu brewing sludge through a 112-day soil column leaching experiment. Sludge particles were coated with sodium alginate (SA), polyvinyl alcohol (PVA), and ester gum (EG), and compared with untreated air-dried sludge (CK₁), uncoated granulated sludge (CK₂), compound fertilizer (F), and a blank control (B). The results showed that coated sludge treatments significantly increased soil leachate pH, total organic carbon, total nitrogen, total phosphorus, and total potassium, and exhibited obvious controlled-release effects on nitrogen, phosphorus, and potassium nutrients compared with uncoated sludge. Soil enzyme analysis indicated that SA treatment increased catalase activity, acid phosphatase activity was generally enhanced by sludge addition, and urease activity was reduced in coated sludge treatments. FTIR and BET analysis showed that both coated and uncoated sludge increased soil-specific surface area and changed soil pore structure. These findings confirm that granulated and coated Baijiu brewing sludge can be used as an effective slow-release soil amendment, and sodium alginate coating shows the most comprehensive improvement in multiple soil health indicators, with high application value for resource utilization of Baijiu sludge and soil quality improvement. Full article

The Dingtao M2 Tomb, the largest, highest-specification, and best-preserved “Huangchangticou” tomb currently discovered in China, is of great significance for cultural relic conservation. During its dismantling and protection, extensive white filamentous fungal contamination was observed on the surface of sand-buried wood components. To clarify the dominant fungal species and its impact on wood cultural relics, the dominant fungus was isolated and purified from contaminated wood samples, and identified as Serpula himantioides (designated as DTW) via molecular and morphological methods. Systematic studies were conducted on DTW’s wood degradation capacity, cellulase activities, regulatory effects of Fe³⁺ and Ca²⁺ on cellulase activities, whole-genome characteristics, and sensitivity to fungistatic agents. The results indicated that DTW exhibited strong wood degradation ability: after 60 days of inoculation, the maximum force, elongation at break, and tensile strength of wood chips decreased significantly, by 88.0%, 54.6%, and 88.6%, respectively, compared with the control group. The cultural relic microenvironment colonized by strain DTW is rich in Fe³⁺ and Ca²⁺. Both ions can significantly suppress the activity and specific activity of cellulase from strain DTW at elevated concentrations. Whole-genome sequencing revealed that DTW had a genome size of 68,519,434 bp with a Guanine–Cytosine (GC) content of 44.01% and 18,373 genes; Carbohydrate-Active Enzyme (CAZy) and Cell Wall-Degrading Enzyme (CWDE) database annotations confirmed its strong plant cell wall degradation ability. Additionally, DTW was sensitive to screened green fungistatic agents, which effectively inhibited its growth. This study clarifies the species and wood degradation mechanism of the dominant fungus on Dingtao M2 Tomb’s sand-buried wood, providing theoretical and technical support for the protection of the tomb’s wood cultural relics. Full article

Cardiac disease is a leading cause of morbidity and early death across several lysosomal storage disorders (LSDs); however, the cardiomyopathies of Fabry, Pompe, and Danon disease are still largely treated as separate, substrate-specific disorders. We argue that they are better understood as variations on a single theme: the breakdown of the autophagy–lysosome system within cardiomyocytes. In the healthy heart, this system clears damaged proteins and organelles and is regulated by mTORC1 and the master regulator TFEB. Once lysosomal degradation or autophagosome–lysosome fusion fails, undegraded substrates and defective mitochondria accumulate, driving hypertrophy, interstitial fibrosis, and conduction disease. Danon disease, resulting from the loss of LAMP2, is the clearest example of a primary defect in autophagic flux, whereas the glycogen storage of Pompe disease and the globotriaosylceramide accumulation of Fabry disease impair flux through different upstream mechanisms that converge on the same downstream injury. The same framework extends to other storage disorders with cardiac involvement, such as mucopolysaccharidosis (MPS). We trace this shared pathobiology from molecule to bedside, examine biomarkers that reflect lysosomal and autophagic dysfunction rather than storage alone, and re-examine treatment in that light: why enzyme replacement therapy corrects substrate accumulation but leaves much of the autophagic and mitochondrial damage unresolved, and why gene therapy—particularly AAV9-LAMP2B for Danon disease—together with autophagy- and TFEB-directed strategies may help close that gap. Viewing these disorders through a single mechanistic lens reshapes how we monitor them and where future therapies should be directed. Full article

Tomato yellow leaf curl virus (TYLCV) is a major threat to tomato production, creating a need for sensitive, low-cost detection methods that can be applied to early symptomatic or low-viral-load samples. Recombinant antigen configuration may influence serological assay development, although the specific contribution of multiple-cloning-site (MCS)-derived intermediate sequences remains uncertain. In this study, a recombinant Trx-His-coat protein (CP) fusion antigen was produced using an MCS-free direct-fusion construct that retained the Trx-His tag while removing the MCS-derived intermediate sequence, followed by gradient refolding. No direct comparison with linker-containing, tag-cleaved, or tag-free antigen constructs was performed. The purified antigen was used to immunize rabbits and generate a high-titre polyclonal antibody (pAb). The resulting indirect enzyme-linked immunosorbent assay (ELISA) achieved a theoretical limit of detection of 1.8 ng/mL and an estimated pre-dilution equivalent the limit of detection (LOD) of 72 ng/mL after sample dilution. The assay showed favourable tolerance to crude tomato leaf matrices, with spike-recovery rates of 95.45–100.40%. In a preliminary evaluation using a balanced panel of 32 field-collected samples, ELISA absorbance correlated with droplet digital PCR quantification (R² = 0.9819) and plant disease index values (R² = 0.9774). Liquid chromatography–tandem mass spectrometry (LC-MS/MS) peptide mapping and AlphaFold2-based modelling were used only to provide preliminary computational context for antigen interpretation. The assay showed cross-recognition toward Tobacco curly shoot virus (TbCSV), indicating that it should not be considered strictly TYLCV species-specific. Therefore, this assay may support preliminary serological screening under the tested conditions, whereas molecular confirmation remains necessary when species-level identification is required. Full article