The distribution of white spots on the shell is one of the remarkable characteristics of the appearance of Portunus trituberculatus, but there have been no reports on its characteristics and genetic analysis. In this study, seven parameters of white spots were extracted based on computer vision, including white spot number (WSN), white spot area (WSA), white spot regional area (WSRA), percentage of white spot area (PWSA), percentage of white spot regional area (PWSRA), white spot number of white spot regional area (WSNRA), and white spot number of carapace area (WSNCA). Using 22 full-sib families, we estimated genetic parameters of these white spot traits and their genetic correlations with growth carapace area (CA). Results demonstrated that these parameters exhibit continuous variation, but the performance of these white spot indicators was not consistent. Based on comprehensive principal component analysis (PCA), they could be categorized into three distinct groups within the full-sib families. The heritability estimated under the animal model ranged from 0.00 to 0.35 for white spot traits and was 0.18 for CA. Most white spot traits showed low to moderate heritability. The genetic correlations among WSN, WSA, WSRA, PWSA, and CA were significantly positive, indicating that selection for one of these traits would result in the improvement of others. This study presents the first estimation of characteristics and genetic parameters of white spot traits of P. trituberculatus, providing valuable information for understanding these traits and facilitating the optimization of breeding strategies for this species.
Assessment of cytomegalovirus (CMV) cell-mediated immunity (CMV-CMI) using standardized, commercially available interferon-gamma release assays (IGRAs) may improve the clinical management of CMV infection in allogeneic hematopoietic stem cell transplant recipients (allo-HCT). Studies comparing the performance of CMV IGRA assays have been conducted in this setting, revealing frequent discrepancies between assays. Here, we compared the performance of a newly released VIDAS CMV IGRA (bioMérieux) and the T-SPOT. CMV (Oxford Immunotec) was evaluated in a cohort of 85 allo-HCT recipients who received letermovir prophylaxis. A total of 107 whole blood samples were collected at either day +100 (n = 53) or day +180 (n = 54) after allo-HCT and analyzed in parallel. A moderate qualitative agreement between assays was observed (Cohen's kappa coefficient, 0.47; 95% CI, 0.30-0.63). Discrepant results were obtained in 29 specimens, most of which were collected at day +100. A moderate correlation (rho, 0.54; P < 0.001) was observed between T-SPOT pp65-SPC and IFN-γ levels measured by the VIDAS CMV assay, whereas the correlation was weaker for T-SPOT IE-1 SPC and VIDAS IFN-γ levels (rho, 0.32; P < 0.001). The previously described booster effect of CMV DNAemia on the magnitude of CMV-CMI was captured by the VIDAS CMV assay. Neither the qualitative nor the quantitative results obtained with either IGRA predicted protection from CMV DNAemia following LMV discontinuation. While our data support the validity of the VIDAS CMV assay for the assessment of CMV-CMI in allo-HCT, they reinforce the idea that results from commercially available IGRA assays are not interchangeable, particularly in allo-HCT recipients with impaired CMV-CMI reconstitution. This is the first study comparing a new interferon-gamma release assay (IGRA), the VIDAS cytomegalovirus (CMV), performed on whole blood and based on an enzyme-linked immunofluorescent assay detection technique, with a commercially available ELISpot assay (T-SPOT CMV) for measuring CMV-specific T-cell responses (CMV-CMI). Our cohort included allogeneic hematopoietic stem cell transplant recipients who underwent letermovir prophylaxis. In this clinical setting, tailoring the duration of LMV prophylaxis based on CMV-CMI at the time of drug discontinuation has emerged as a promising application of these assays. Our data revealed substantial qualitative and quantitative differences between the assays, which should be taken into consideration in future observational studies or clinical trials assessing the utility of IGRAs for individualizing LMV prophylaxis duration.
In cotton disease detection, the complex farmland environment and the varying scales of disease spots, especially the presence of small-target disease spots, limit the detection accuracy of lightweight models. To address this issue, an improved YOLOv11n detection algorithm is proposed. First, the backbone network is reconstructed using the GhostConv (G-conv) module, which generates redundant feature maps through linear operations, thereby reducing computational complexity. Second, an Adaptive Calibration and Feature Fusion Architecture Head (ACFFA) with prior calibration and cross-scale fusion capabilities is constructed in the detection stage to handle the problem of varying disease spot scales. Furthermore, the Adaptive Scale-aware Wise Intersection over Union (AS-WIoU) loss function, improved from WIoUv3, is introduced to enhance the stability of bounding box regression and improve detection accuracy for low-resolution, small-target lesions. Experimental results show that on the cotton disease dataset constructed based on the Mendeley Data database, the proposed model achieves mAP50 and mAP50-95 of 90.30% and 73.84%, respectively, with precision and recall of 92.33% and 87.68%, and a parameter count of 3.81 M. The algorithm significantly improves detection accuracy while maintaining efficient inference, making it suitable for real-time monitoring tasks on agricultural embedded terminals.
Corynespora leaf spot, caused by Corynespora cassiicola, is an emerging disease in crops that significantly impacts both yield and quality. Currently, the use of fungicides to control Corynespora leaf spot has led to the development of varying degrees of resistance in the pathogen. Therefore, it is crucial to screen for highly effective fungicides with novel modes of action. This study evaluated the antifungal activity of 1-hydroxyphenazine (1OH-PHZ) against multiple phytopathogenic fungi, with a half-maximal effective concentration (EC50) of 19.23 μg/ml against C. cassiicola hyphae. In vivo assay demonstrated antifungal activity of 67.22 and 45.03% on detached tomato leaves and fruits, respectively, at a dose of 500.0 μg/ml. Microscopic and ultrastructural observations revealed hyphal collapse, surface wrinkling, and indistinct organelle boundaries following treatment. Integrated transcriptomic and metabolomic analyses showed differentially expressed genes and differentially abundant metabolites, primarily affecting amino acid metabolism and biosynthesis pathways. Molecular docking, dynamic simulations, and microscale thermophoresis assays demonstrated that 1OH-PHZ binds to PLP-dependent transferase (PLPDT), exhibiting a binding free energy of -7.2 kcal/mol and a dissociation constant (Kd) value of 1.16 μM. Collectively, these findings suggest that 1OH-PHZ potentially binds to PLPDT, thereby disrupting amino acid metabolism and biosynthesis, which subsequently affects the synthesis and morphological development of the fungal cell wall and cell membrane. Through the combination and screening of highly active fungicidal substances, this study offers mechanistic insights that support the potential development of 1OH-PHZ as a novel agricultural fungicide for managing C. cassiicola infections.
Albashaireh ZS, Altallaq MK. Assessment of patient satisfaction and esthetic outcomes following treatment of post-orthodontic white spot lesions using a bioactive adhesive and resin infiltrant: a randomized split-mouth clinical trial. Journal of Dentistry. 2026;167:106565. Split-mouth randomized clinical trial of 20 orthodontically treated patients (13 female, 7 male) involving 120 teeth with anterior white spot lesions (WSLs). Each participant received ICON resin infiltration or Hi-Bond Universal in contralateral quadrants. Esthetic changes were assessed by the patients and by two independent clinicians using a 100-point visual analogue scale (VAS) at one-month recall. Participants aged 16-30 years and who had completed fixed orthodontic treatment at least one month before enrollment were included. Eligible participants presented with at least one visible WSL on the labial surfaces of the anterior teeth or premolars on contralateral quadrants. All participants were required to have good oral hygiene, clinically healthy gingival tissues, and no evidence of active periodontal disease. Categorical outcome data were presented using descriptive summary measures including frequencies and percentages. The proportion of satisfactory vs. unsatisfactory outcomes (paired binary data) was compared between the Hi-Bond Universal and ICON treatments using the McNemar test. Overall patient satisfaction was equally high (80%) for both treatments, and there were no significant differences between patient or clinician ratings. Inter-assessor agreement was deemed "fair" for both groups (Cohen's kappa 0.318-0.340). Thus, both ICON and Hi-Bond Universal effectively masked WSLs, as deemed by subjective esthetic evaluation. Both bioactive adhesives and resin infiltration achieve satisfactory esthetic outcomes in masking demineralized enamel, at least in the short-term. Further longitudinal studies comparing the outcomes of these materials would be beneficial to determine whether these effects are sustained over time.
Passion fruit (Passiflora edulis) leaves displaying yellow spot symptoms were collected in La Réunion. Whilst viral indexing suggested single infections with potyviruses, high-throughput sequencing revealed both single and mixed infections involving cowpea aphid-borne mosaic virus (CABMV; species Potyvirus vignae) and a previously undescribed potyvirus, for which the name passion fruit yellow spot virus (PaYSV; tentative species Potyvirus passifloraflavamaculae) is proposed. Complete genome sequences (9,626 nucleotides) of two PaYSV isolates were obtained from these samples, using RNA sequencing after ribodepletion. The two isolates were highly similar (> 99.3% nucleotide identity) and shared the highest nucleotide (63.6%) and amino acid (71.7%) identities at the polyprotein level with passion fruit woodiness virus (PWV; Potyvirus passiflorae), values that are below the species demarcation thresholds. Our results provide compelling molecular and phylogenetic evidence for the presence of a novel potyvirus species infecting passion fruit in La Réunion.
To evaluate the diagnostic accuracy of glial fibrillary acidic protein (GFAP) measured in dried plasma spots versus conventional plasma- and serum-GFAP testing for assessment of disease severity in aquaporin-4 immunoglobulin G-positive neuromyelitis optica spectrum disorder (AQP4-IgG+ NMOSD). A neuroimmunological prospective cohort of remission samples from 70 participants with the diagnoses AQP4-IgG+ NMOSD (n = 19), myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD; n = 9), relapsing-remitting multiple sclerosis (RRMS; n = 28) and healthy controls (HC; n = 14) from a single center were included. GFAP concentrations were measured in frozen and thawed plasma and paired dried plasma spot (DPS) samples using an ultrasensitive proximity ligation-based assay (nucleic acid-linked immuno-sandwich assay; NULISA). In NMOSD, GFAP was additionally quantified in paired serum samples using a single-molecule array (Simoa) assay as a reference. Cross-matrix correlations, diagnostic performance, and associations with neurological disability were evaluated. GFAP concentrations measured by NULISA correlated strongly between plasma and DPS samples across diagnostic groups and healthy controls. In NMOSD, plasma GFAP measured by NULISA showed strong concordance with serum GFAP quantified by Simoa. DPS-derived and plasma-derived GFAP demonstrated good diagnostic accuracy for AQP4-IgG+ NMOSD and was significantly associated with neurological disability as measured by the Expanded Disability Status Scale (EDSS). Group-wise comparisons across plasma and DPS showed retained elevation of GFAP in AQP4-IgG+ NMOSD compared with the other diagnostic groups and healthy controls. GFAP quantification using the NULISA platform is feasible in plasma and DPS samples and enables reliable biomarker assessment in a potentially remote-compatible setting. DPS-derived GFAP measurements retained meaningful information on disability in NMOSD and may provide an analytical framework for future studies evaluating minimally invasive capillary or self-sampling approaches.
Rapid room-temperature identification of gases and volatile organic compounds remains challenging for compact sensing platforms, particularly when chemically related analytes must be discriminated using accessible sensing materials. In this work, we evaluate whether magneto-optical surface plasmon resonance (MOSPR), combined with multi-spot sensing and conventional SPR readout from the same chip, can provide complementary response features for improved gas/VOC discrimination. The sensing spots are made from accessible chemicals and nanoparticles with plasmonic and magnetic properties. The sensor chip consists of a multilayer structure of metallic materials with both plasmonic and magnetic properties featuring enhanced sensitivity and stability. Measurements are made using a custom-built MOSPR instrument at relevant analyte concentrations. Analyte-specific sensor channels were selected for concentration-dependent calibration while the complete multivariate data were first explored using principal component analysis for supervised analyte classification. The combined 16-feature MOSPR/SPR model achieved an overall accuracy of 88.3% and a balanced accuracy of 87.6% under leave-one-concentration-block-out cross-validation compared with 66.2% and 65.7%, respectively, for the SPR measurement alone. These results show that MOSPR provides response information that encompasses and extends that obtained from conventional SPR measurements, thereby improving analyte discrimination. The proposed approach may provide a basis for future environmental monitoring and industrial process control, including real-time monitoring of harmful gaseous emissions pending further validation under application-specific conditions.
Mungbean (Vigna radiata L. Wilczek) is a major cash-generating pulse crop in Ethiopia's lowlands, serving as a key source of plant-based protein and contributing to soil fertility. However, its production is severely constrained by Cercospora Leaf Spot (CLS), a destructive fungal disease. During the 2024 and 2025 cropping seasons, a total of 496 mungbean farms from 33 districts in five regions of Ethiopia were inspected to assess the distribution and occurrence of CLS and its association with agronomic factors. The districts were selected according to their mungbean production potential. Necessary information related to farm field inspected including altitude and absolute location were recorded during field assessments. A logistic regression model was used to analyze the associations between CLS occurrence and agronomic factors. CLS was widespread across all surveyed districts, with prevalence ranging from 72% (Gindo Koisha) to 100% (Sodo), and increased from 85.43% of fields in 2024 to 91.25% in 2025, showing a 5.8% increase in CLS occurrence. High CLS incidence (> 50%) and severity (> 40%) were significantly (p < 0.0001) associated with Sodo districts, followed by Adami Tullu, Awash Melakassa, Kulfo-Halaba, and Lebo Kemkem. These conditions were predominantly observed in the 2025 cropping season at elevations above 1500 m a.s.l. and were linked to high weed infestation, minimal tillage, and infrequent weeding during the grain-filling stage. Therefore, priority should be given to CLS-prone districts through research-based integrated management to ensure sustainable mungbean production. This would enhance nutritional security and recognize mungbean as a future smart food crop.
The fungal pathogen Colletotrichum jiangxiense has a broad host range and high destructive potential. It is a major causal agent of brown spot disease in diverse plants. Antifungal mechanisms used by biocontrol fungi against plant pathogens include disruption of cellular structures and cell wall damage, which can lead to protoplast leakage and hyphal lysis. In this study, we investigated the antifungal mechanism of the endophytic fungus Diaporthe novem DJ13 against C. jiangxiense, the causal agent of rhododendron brown spot. DJ13 is an effective biocontrol strain previously isolated by our research group from healthy leaves of Rhododendron pulchrum. Physiological assays showed that treatment with DJ13 fermentation broth increased membrane permeability, elevated MDA content, reduced TCA cycle enzyme activities, and increased AKP activity. These findings suggest impaired membrane integrity, disrupted energy metabolism, and cell wall damage. Transcriptomic analysis of the treated pathogen identified 1680 significantly differentially expressed genes (DEGs), including 961 up-regulated and 719 down-regulated genes. Among these genes, ABC transporter genes were significantly up-regulated, whereas genes involved in membrane structure metabolism were significantly down-regulated. Chitinase genes were up-regulated, whereas α-glucanase genes were down-regulated. DASH family cryptochrome genes were significantly down-regulated, while genes related to reactive oxygen species (ROS) production, including xanthine dehydrogenase, were significantly up-regulated. In addition, FAD-dependent oxidoreductase genes were up-regulated, while respiratory-metabolism-related genes, including trimethyllysine dioxygenase, were down-regulated. Together, the physiological and transcriptomic data provide a correlative framework supporting the hypothesis that the antifungal mechanism of DJ13 fermentation broth may involve the coordinated action of four processes: cell membrane damage, cell wall disruption, oxidative stress, and inhibition of energy metabolism. These findings also identify candidate genes for future functional validation.
Zymoseptoria tritici (Desm. Quaedvlieg & Crous), known as the wheat leaf spot disease agent, is a highly virulent fungus that induces blotch and necrosis on leaves. Although it is known to cause severe infections under humid conditions, recent observations suggest that it can also infect wheat leaves under drought and high-temperature conditions, possibly influenced by global warming. Recent findings showed that Z. tritici could easily tolerate various abiotic stresses, including drought, water stress, salinity, and temperature. It has been evident that the fungus can tolerate pesticide stress, as indicated by the increased frequency and number of pesticide applications throughout the growing season. Under stress conditions, the fungi, unlike crop plants, could easily tolerate stress by rapidly modifying gene expression and reducing spore production and mycelial growth without downregulating major biochemical components that play significant roles in pathogenicity and virulence. Z. tritici can accumulate melanin under stress conditions; therefore, an increase in pathogenicity under drought or salinity stress is not unexpected. Recent studies have shown that the pathogenicity of the fungus is increasing, and more virulent, toxin-producing pathogens might emerge in the future. Since drought and high-temperature stresses significantly affect crop plants, the adaptation of pathogenic microorganisms to these conditions could be inevitable if abiotic stress persists. Under these circumstances, the crop loss would be more pronounced. A critical aspect of this process is the assessment of DNA integrity in both wheat and the pathogen under drought stress conditions. The organism that better maintains DNA integrity is considered to exhibit greater drought tolerance. Therefore, our main target should be DNA health when developing or breeding new wheat varieties, considering double- or even multiple-stress conditions. We should finally state that we are very optimistic about generating highly stress-tolerant wheat varieties via metabolomic and proteomic approaches without compromising quality. However, the impact and combination of stress factors are becoming increasingly complex.
Soybean (Glycine max) is a major economic and food crop whose yield is severely affected by frogeye leaf spot (FLS), caused by Cercospora sojina. Current knowledge of resistance genes remains insufficient for effective molecular breeding. In this study, a recombinant inbred line (RIL) population derived from a cross between the resistant parent, Henong 60 (H60), and the susceptible parent, Dongnong L13 (DN L13), was evaluated under field conditions in Acheng (AC) and Xiangyang (XY). Plants were artificially inoculated with physiological race 7 of C. sojina, and disease severity at the R3 growth stage was recorded. Genotyping using the SoySNP660K chip yielded 54,836 high-quality single-nucleotide polymorphism (SNP) markers. A genome-wide association study (GWAS) was performed using the 3VmrMLM model by integrating dual-environment phenotypic data, and four quantitative trait nucleotides (QTNs) significantly associated with resistance to FLS were identified on chromosomes 8 (1), 17 (1), and 20 (2). By the analysis of genomic annotation, functional enrichment, metabolic pathway analyses, haplotype-phenotype association and quantitative real-time PCR (qRT-PCR), Glyma.20G155700 and Glyma.17G070500 are intended to be candidate genes related to soybean resistance to race 7 of FLS. The findings of this study provide insights into the genetic mechanisms underlying resistance to FLS in soybean. The identified molecular markers and candidate genes may provide useful resources for marker-assisted breeding and the development of disease-resistant germplasm.
Marine shrimp are among the most economically significant aquatic animals facing serious challenges from pathogen infections, particularly the white spot syndrome virus (WSSV), which leads to massive mortality in shrimp aquaculture. An understanding of shrimp defense mechanisms is crucial for developing effective protection strategies. Long noncoding RNA (lncRNA), one of the potential non-coding RNAs, plays regulatory roles in controlling several physiological processes, particularly immune responses. However, the role of lncRNA in WSSV infection in shrimp remains poorly understood. In this study, the putative lncRNAs and differentially expressed lncRNAs (DELs) from the gills of WSSV-infected Pacific white shrimp, Litopenaeus vannamei were identified. The total of 184 DELs were found among 2135 differential expressed genes (DEGs), and the expression of six selected DELs in different shrimp tissues and in the gills of shrimp infected with WSSV were confirmed. Among these DELs, lnc17924 was chosen to investigate its role in WSSV infection due to its significant response to WSSV administration. The lnc17924 knockdown prolonged shrimp survival following WSSV infection suggesting its role in regulation of WSSV infection. Investigation of the interaction network of lnc17924-miRNA-mRNA revealed that lnc17924 interacted with only miRNA_5514, which subsequently interacted with putative 925 differentially expressed mRNAs. Seven lnc17924-miRNA regulated mRNAs, including LvCLEC, LvKAT6A, LvNRE75, LvKLF10, LvHSP90, LvSPARC, and LvPDCD7, exhibited distinct responses following lnc17924 knockdown and WSSV infected shrimp. This study provides lncRNA profiling and comprehensive involvement of lncRNA, especially lnc17924, in the control of WSSV infection, thereby offering molecular insight for shrimp antiviral strategies.
Long-acting spot-on formulations are widely used for parasite control in cats, yet the pharmacokinetics of combined fluralaner and moxidectin remain incompletely defined beyond the recommended dose. This study evaluated the pharmacokinetics, absolute bioavailability, and dose proportionality of a novel fixed-combination spot-on formulation in healthy cats after topical administration of fluralaner/moxidectin at 40/2, 80/4, and 120/6 mg/kg BW, and compared intravenous administration of each compound alone or in combination. Forty-eight healthy cats were allocated to six groups of eight cats each. Non-compartmental analysis and a power model were used to characterize systemic exposure and dose proportionality. After intravenous co-administration, fluralaner exposure increased whereas moxidectin exposure decreased compared with single-agent administration, indicating a potential pharmacokinetic interaction. Topical administration produced slow absorption and sustained systemic persistence, with Tmax values occurring approximately 10-18 days after dosing and quantifiable concentrations maintained throughout the 160 days sampling period. Systemic exposure increased non-proportionally across the tested dose range, and absolute bioavailability was limited and dose-dependent, ranging from 9.7% to 26.9% for fluralaner and from 10.22% to 21.28% for moxidectin. No adverse reactions were observed. These findings demonstrate nonlinear topical absorption, and prolonged systemic persistence, supporting further dose optimization and formulation development of this long-acting feline antiparasitic delivery system.
This study reports the optimization of a cost-effective one-step PCR assay for the detection of White Spot Syndrome Virus (WSSV) in wild shrimp populations, providing an accessible alternative to expensive commercial nested PCR kits in Mozambique. Primer sets targeting VP19, VP24, VP26 and VP28 were evaluated, with the VP28_2 primer set demonstrating the highest sensitivity and no cross-reactivity with Infectious Hypodermal and Haematopoietic Necrosis Virus (IHHNV) or Decapod Iridescent Virus 1 (DIV1). Cost analysis showed that the optimized assay is approximately three times less expensive than commercial nested PCR (USD 5.57 vs. USD 15.75 per reaction). The assay was applied to 300 wild shrimp specimens (Penaeus indicus and Penaeus monodon) collected during a cross-sectional survey conducted in the rainy season (October-November 2022) at 15 landing sites across six provinces. WSSV was detected in 60.7% of samples, with significant regional variation: 78.0% in the South, 58.0% in the Centre and 48.1% in the North. Although the findings indicate widespread WSSV occurrence in wild shrimp populations, the cross-sectional design, single-season sampling and sample size limit inference on temporal trends. Nevertheless, the optimized assay provides a practical and affordable tool for routine WSSV surveillance under existing resource constraints.
Congenital cytomegalovirus infection (cCMV) is the most common congenital infection and an important cause of sensorineural hearing loss and neurodevelopmental impairment, yet many affected infants remain undetected under selective screening approaches. We conducted a prospective national pilot study to evaluate the feasibility and diagnostic yield of universal dried blood spot (DBS)-based screening for cCMV within the Slovenian newborn screening program. DBS samples collected within 72 h of life were tested by polymerase chain reaction (PCR), and screen-positive newborns underwent confirmatory urine PCR within 21 days together with standardized clinical evaluation. Among 5556 screened newborns, 13 (0.23%) screened positive and cCMV was confirmed in 10, corresponding to a lower-bound birth prevalence of 1.80 per 1000 live births (95% confidence interval, 0.98-3.31), because confirmatory testing was limited to DBS-positive newborns. None of the confirmed cases were clinically suspected at birth, and all passed newborn hearing screening. Six infants met protocol-defined criteria for symptomatic cCMV and received valganciclovir. Historical registry-based clinical case ascertainment in Slovenia corresponded to 0.09 detected cases per 1000 live births. These findings demonstrate the feasibility of universal DBS-based cCMV screening within an established newborn screening infrastructure and suggest substantial under-ascertainment under selective clinical detection pathways.
Epstein-Barr virus (EBV) is highly prevalent worldwide and has been linked to different cancers and autoimmune disorders, including Multiple sclerosis (MS). Primary infection typically occurs during early childhood or adolescence and often goes undetected. Serological studies allow the detection of EBV-specific antibodies after infection, but venous blood collection is resource-intense and thus limits sample size. To overcome these challenges, we developed a high-throughput, semi-automated serology protocol targeting antibodies against EBV in capillary blood samples (Dried Blood Spots, DBS). Paired serum and DBS samples were obtained from 417 participants aged 18-25 years. We used Roche Elecsys® EBV nuclear antigen-1 EBNA IgG and Elecsys® EBV viral capsid antigen VCA IgG assays, established for serum, to detect antibodies against EBV anti-EBNA‑1 IgG and anti-VCA IgG in DBS. Cut-off Indices (COI) for the DBS assays were determined using a classification tree. Of the 416 valid paired serum samples, 78.6% (327/416) were anti-VCA-positive and 76.4% (318/416) anti-EBNA-positive. Discrepant results were observed in 3.6% (15/416) of participants. Based on combined assay results, 20.7% (86/416) of the samples were classified as EBV-negative. The serum COI values revealed that most of the negative cases clustered well below the cutoff, while positive cases spanned a broad range of higher values. Overall, 98.8% (326/330) of participants classified as positive based on serum testing (n = 330) were also classified as positive in DBS (n = 326), corresponding to the sensitivity of DBS relative to serum results. Likewise, 96.5% (83/86) of participants classified as serum-negative participants (n = 86) were also negative in DBS (n = 83), corresponding to the specificity of DBS relative to serum results. Among the 416 participants, only 4/330 (1.2%) were false negatives and 3/86 (3.5%) were false positives in DBS. The newly established assays were validated in a self-sampling cohort of 295 participants. We established DBS-specific cutoff values to detect antibodies against EBV in DBS, achieving high sensitivity (98.8%) and specificity (96.5%) relative to the corresponding venous blood assay. Furthermore, we derived a correction formula to convert semi-quantitative DBS values to serum-equivalents, enabling comparison with other studies and standardized datasets. The DBS-based approach allows sero-status assessment in a large population and thus e.g. simplifies the identification of suitable participants for clinical trials.
Alternaria brassicicola is a necrotrophic fungal pathogen causing black spot disease on cruciferous crops worldwide. In this study, we comprehensively characterized the pathogenic isolate Ab0920a from diseased broccoli in Shanghai using morphological, phylogenetic, host range, fungicide sensitivity, genomic, and functional analyses. Pathogenicity tests on 27 cruciferous varieties revealed a broad host range with varying resistance levels. Fungicide sensitivity assays showed that fluxapyroxad (EC50 = 0.0695 µg/mL), prochloraz (0.0711 µg/mL), and difenoconazole (0.0863 µg/mL) were highly effective, whereas fluazinam was least effective. Genome-wide annotation identified 941 secreted proteins (8.95% of the proteome) and 237 candidate effectors, including 31 small cysteine-rich secreted proteins and 68 homologs of known virulence factors. Conserved effector-associated motifs (e.g., RXLR, [Y/F/W]xC) were detected, and carbohydrate-active enzyme CAZyme annotation revealed diverse families potentially involved in plant cell wall degradation. Functional validation using the pSUC2 yeast system confirmed that N-terminal signal peptides of tested effectors are competent for secretion. A PVX-based transient expression assay in Nicotiana benthamiana identified two effectors that suppress Bax-induced programmed cell death, suggesting their potential roles in modulating host immunity. Overall, this study provides comprehensive insights into the pathogen Ab0920a, offering resources for disease management and functional studies on necrotrophic fungal pathogenesis.
暂无摘要（点击查看详情）
暂无摘要（点击查看详情）