Search Results (572)

Reference-based logo colorization aims to generate controllable color schemes while preserving clear contours and consistent colors within graphic regions. However, existing colorization and style transfer methods often suffer from ambiguous color assignment, inconsistent filling, and cross-boundary color interference when applied to logos with compact structures and nested regions. To address these issues, we propose LogoRef, a reference-based logo colorization framework that integrates sketch structure preservation, reference color extraction, and region-aware color assignment. First, we propose a Hierarchical Region Prior (HRP) to extract region structures at different levels from the input line art. Second, we introduce the Semantic Region Style Attention module, which constructs a reference style library and adaptively selects appropriate color statistics for different logo regions, thereby improving intra-region color consistency and reducing color interference between adjacent regions. In addition, we construct JN-Logo-v2, a high-quality logo image dataset that provides richer and more reliable data support for logo colorization. Finally, we propose two task-specific evaluation metrics, Color Fidelity and Boundary Integrity, to evaluate reference color consistency and boundary-aware colorization quality, respectively. Extensive experiments on JN-Logo-v2 and a public anime sketch colorization dataset demonstrate that our method achieves superior performance in both quantitative and qualitative comparisons. Full article

Bacterial soft rot, caused by Pectobacterium carotovorum subsp. carotovorum, affects numerous vegetable crops. Beneficial microbes like Bacillus amyloliquefaciens PMB05 offer a sustainable alternative to traditional copper- or antibiotic-based controls, which risks the development of pathogen resistance. Bacillus amyloliquefaciens PMB05 enhances disease resistance to soft rot by intensifying PAMP-triggered immunity (PTI). Although our previous research showed that PMB05-intensified PTI correlates with a reduced photosynthetic electron transport rate under high light conditions, it remains unclear whether increasing light intensity can strengthen PMB05-mediated immune signaling to further enhance disease resistance in plant leaves. In this study, Arabidopsis plants were grown under varying light intensities to address this question. Furthermore, expression of the type III elicitor, HrpN protein, was applied to evaluate the impact of PMB05 treatment on immune activation. HrpN acts as a PAMP that can trigger plant defense responses. The results revealed that higher growth light intensity was associated with PMB05-intensified PTI response by increasing reactive oxygen species generation but suppresses callose deposition. In addition, PMB05-facilitated PTI-mediated stomatal closure occurred more rapidly and tightly under high light, thereby restricting pathogen invasion into leaf tissues. Disease assays further confirmed that higher light intensity significantly reduced soft rot severity in A. thaliana, while the combination of high light and PMB05 treatment achieved superior control efficacy. Furthermore, PMB05-associated disease protection under high growth light was not detected in sid2, npr1, and nahG plants, indicating that intact SA-associated functions are required for the detectable protective phenotype under the conditions tested. These findings demonstrate that environmental light management can strategically potentiate immunity-intensifying microbes, establishing a novel paradigm for sustainable disease control through the coordinated manipulation of light conditions and microbial application. Full article

Given the prevalent use of horseradish peroxidase (HRP) as an enzyme label in immunoassays, the practical importance of point-of-care testing for the portable identification of HRP-related biomarkers is evident. Accordingly, this study devised a portable immunoassay platform using a personal glucometer (PGM) to achieve rapid detection of HRP-related biomarkers. 3,3′,5,5′-tetramethylbenzidine (TMB) and dopamine (DA) were used as signal-regulating substrates, and the assay principle was based on the HRP-mediated redox reaction. In the absence of HRP, TMB and DA directly reacted with [Fe(CN)₆]³⁻ on commercial glucose test strips, producing a strong PGM response. When HRP and H₂O₂ were present, HRP facilitated the oxidation of TMB to generate oxidized TMB, which further reacted with DA to produce oxidized products with weak reactivity toward [Fe(CN)₆]³⁻, resulting in a decreased PGM signal. Conducting the quantitative assay involved measuring the HRP-dependent change in the PGM readout. Unlike many PGM immunoassays that rely on additional glucose-producing enzyme labels or amplified nanomaterial tags, the proposed method directly converts the signal of the conventional HRP label into a PGM-readable response. The sensing platform demonstrated a good linear response to HRP concentrations ranging from 156 to 625 ng/mL, with a low detection limit of 26 ng/mL. Interference experiments indicated good tolerance to the tested nonspecific proteins. The preliminary applicability of the platform was further evaluated by detecting cardiac troponin I (cTnI) and heparin-binding protein (HBP) in spiked serum samples, with recoveries ranging from 93.0% to 117.1% for cTnI and from 88.3% to 112.5% for HBP. In summary, the proposed platform provides a portable digital readout for HRP-linked immunoassays and reduces the dependence on dedicated readout instruments. Full article

In plants, pathogen-associated molecular patterns (PAMPs) activate pattern-triggered immunity and can contribute to resistance against phytopathogenic infections. Identifying novel microbial elicitors is important for developing resistance-inducing strategies that reduce reliance on pesticides. This study evaluated BV801-derived protein preparations obtained from Xanthomonas euvesicatoria strain BV801 grown in host-associated media containing lyophilized Solanum lycopersicum or Capsicum annuum tissue. Six treatments were evaluated in C. annuum against bacterial spot caused by X. euvesicatoria (BSX) under in vitro conditions: SP-AWSL and SP-AWCA (obtained from BV801 grown in the host-associated AWSL and AWCA media), SP-NYGA and SP-M9 (obtained from BV801 grown in rich and minimal control media), and two commercial reference inducers, Actigard^® 50 GS (acibenzolar-S-methyl) and Messenger Gold^® (commercial harpin formulation). Plants were treated 48 h before pathogen inoculation. SDS-PAGE revealed protein bands in the 12–26 kDa range that were enriched in SP-AWSL and SP-AWCA relative to the control-media preparations. Several BV801-derived preparations produced localized necrosis in N. tabacum leaves consistent with a hypersensitive-response-like reaction. SP-AWSL and SP-AWCA reduced BSX severity in chili pepper by up to 67% compared with the diseased control (Kruskal–Wallis, p ≤ 0.05) and significantly reduced bacterial proliferation (Tukey, p ≤ 0.05). At the final sampling point, plants pretreated with BV801-derived protein preparations and subsequently challenged with X. euvesicatoria showed higher CaBPR1 expression than corresponding pretreated but unchallenged plants, a pattern consistent with a priming-like defense response. These findings support the potential of BV801-derived protein preparations obtained under putative Hrp-inducing conditions as candidates for sustainable resistance-inducing strategies against bacterial spot in chili pepper. Full article

Food is always at risk of becoming waste, with household refrigerator practices either slowing or hastening this process. While household food waste has received growing scholarly attention, the interplay of human behavior, refrigerator use, and packaging (HRP) remains underexplored, as research typically examines these elements in isolation rather than their everyday entanglement. To address this gap, this study conceptualizes HRP as a co-constitutive design system and integrates a scoping literature review with qualitative fieldwork in 24 Swedish and international households. Photo-elicitation was used to document real refrigerator organization and storage conditions, while semi-structured interviews (n = 12) captured participants’ routines, decision-making, and interpretations of stored food and packaging. The empirical material was analyzed using thematic analysis to identify recurring patterns in domestic food storage practices. The analysis reveals two key findings: (1) the refrigerator must be recognized as a critical contextual and ethical factor in both packaging design and domestic food waste studies; and (2) packaging’s communicative functions often collapse after purchase, limiting its ability to guide storage, visibility, and timely consumption. By framing HRP as dynamically interlinked, the study introduces a new analytical lens on household food waste and calls for sustainable design strategies that better support waste-reducing practices. Full article

A horseradish peroxidase (HRP)-based electrochemical biosensor was developed for melatonin (MLT) detection using a graphene-modified screen-printed electrode (G/SPE). HRP was immobilized at different temperatures (4 °C and 20 °C) and enzyme amounts (10 and 20 µL of 5 mg/mL HRP solution) to optimize the performance of the biosensors. FTIR analysis confirmed successful enzyme immobilization, while differential pulse voltammetry (DPV) showed a clear oxidation peak of MLT at 1.1 V with enhanced current responses for HRP-modified electrodes. The optimal biosensor, prepared at 4 °C with 10 µL enzyme of 5 mg/mL HRP solution, exhibited the best analytical performance, with a linear range of 0.4–3.6 µM, and a detection limit of 1.02 µM. Kinetic studies confirmed strong enzyme-substrate affinity, while the biosensors demonstrated excellent repeatability and a good stability. The method was successfully validated on pharmaceutical products, proving to be a reliable and an accurate tool for MLT quantification in real samples. Full article

Tetracycline (TC), a widely used broad-spectrum antibiotic, was experimentally and theoretically investigated for its potential effect on thyroid hormone biosynthesis through interaction with thyroid peroxidase (TPO). Horseradish peroxidase (HRP) was used as a model of TPO, while FeTPPCl was employed as a model of the active site of the enzyme. The inhibitory effect of TC on the catalytic oxidation of iodide to active iodine was studied spectrophotometrically through the formation of triiodide (I₃⁻), and the IC₅₀ value was determined. Kinetic studies using Michaelis–Menten and Lineweaver–Burk analysis demonstrated that TC acts as a reversible non-competitive inhibitor of HRP, leading to decreased Km and Vmax values and the formation of the ESI complex. Molecular docking calculations further supported the experimental findings, indicating that TC binds at a site different from the active center of the enzyme. The results suggest that tetracycline may interfere with thyroid hormone biosynthesis by inhibiting TPO-mediated iodide oxidation, revealing a possible mechanism associated with thyroid-related side effects during prolonged antibiotic administration. Full article

Amplex Red is a widely used fluorogenic probe for hydrogen peroxide (H₂O₂) detection. In the presence of horseradish peroxidase (HRP), it reacts with H₂O₂ to form fluorescent resorufin. However, during high-voltage electric pulse treatment, fluorescence readouts may be influenced not only by H₂O₂ generation but also by the medium composition, pulse conditions, pH-dependent effects, and metal ions released as a result of electrode corrosion. In this study, we examined how medium composition and selected metal ions relevant to electroporation conditions, such as Fe²⁺, Fe³⁺, Al³⁺, and Cr⁶⁺, affect the Amplex Red/HRP/H₂O₂ fluorescence signal. Fluorescence intensity differed markedly among media, with the highest signal in phosphate-buffered saline (PBS, ~34,000 a.u.), followed by HB1 buffer (~19,000 a.u.), cell culture medium (~9000 a.u.), and distilled water (~4100 a.u.). To model ion-mediated interference, metal ions were added directly to the assay mixture. All tested ions reduced fluorescence in a concentration-dependent manner. At 0.4 µM H₂O₂, Fe²⁺ reduced fluorescence by up to 92%, Fe³⁺ by 60%, Al³⁺ by 35%, and Cr⁶⁺ by 40%. At 10 µM H₂O₂, the quenching effect was reduced but remained evident. In addition, the fluorescence readout showed strong dependence on medium composition and its pH, further demonstrating its vulnerability to the chemical environment. These findings show that the Amplex Red assay is highly sensitive to the assay environment and metal-ion interference. Therefore, it should not be used as a standalone, interference-free measure of H₂O₂ in electroporation-related experiments without appropriate validation controls. Full article

We developed a site-directed immobilization strategy exploiting the competitive inhibitor Remazol Brilliant Blue R (RB) to orient horseradish peroxidase (HRP) on functionalized glass surfaces. RB was covalently bound to poly(ethylene-alt-maleic anhydride) (PEMA)-coated substrates, enabling HRP binding via its active site. Subsequent contact with a second PEMA-functionalized surface facilitated covalent immobilization of HRP with controlled orientation. Interfacial layer assembly and the comparative random control were characterized by AFM and XPS analyses, while successful RB acetylation was confirmed by NMR and ATR-IR spectroscopy. Directed contact approaches using stepwise RB and PEMA+RB functionalization yielded HRP loadings with specific activities approximately twofold higher than non-directed immobilization. Our inhibitor-mediated method preserves the native enzyme structure without genetic modification, offering a straightforward platform for enhancing enzyme performance. This strategy is promising for applications in biosensors, biocatalysis, and enzyme-based devices requiring efficient enzyme immobilization. Full article

Mycoplasma bovis (M. bovis) is a major pathogen in cattle, primarily responsible for bovine respiratory disease complex (BRDC). Characterized by high infectivity and morbidity, M. bovis spreads rapidly within herds and is challenging to control and eradicate, underscoring the need for early and accurate diagnosis. To address this, we developed a competitive enzyme-linked immunosorbent assay (cELISA) for detecting M. bovis-specific antibodies in bovine sera. The assay uses recombinant MbovP579 protein (1 μg/mL) as the coating antigen and an HRP-conjugated 1A2 monoclonal antibody against MbovP579 (mAb, 0.35 μg/mL) as the competitor. With a cut-off value of 40.69% (percentage inhibition, PI), the cELISA demonstrated apparent sensitivity (90%; 95% CI: 78.64–95.65%) and specificity (96.15%; 95% CI: 90.53–98.49%) using the known background positive and negative sera. In addition, it yielded 100% positivity (9/9) against sera collected from 21 days post-immunization. The assay showed no cross-reactivity with other Mycoplasma species or two common bovine bacterial pathogens, confirming its high specificity. The intra- and inter-assay coefficients of variation (CVs) were below 10%. In clinical evaluations, this cELISA generated a 55.57% positivity rate for field samples (n = 1069) and exhibited 90.46% overall agreement (967/1069; 95% CI: 88.54–92.15%) with the commercial indirect ELISA kit, demonstrating strong diagnostic consistency. Discrepant specimens were further analyzed by a metabolic inhibition test (MIT), which suggested improved specificity, although further validation is required of this cELISA. Full article

Enzyme immobilisation within Metal–organic Frameworks (MOFs) provides a promising strategy for improving enzyme dispersion and local environment control, although the resulting performance depends strongly on the host materials, enzyme type and immobilisation conditions. Conventional in situ biomimetic mineralisation typically produces enzyme–MOF composites (enzyme@MOFs) with irregular morphologies, broad particle size distributions and aggregation, which can compromise catalytic performance and reproducibility. This study presents a flow-based microfluidic biomimetic mineralisation strategy for preparing horseradish peroxidase-encapsulated ZnBDC-NH₂ MOF composites. A flow-focusing microfluidic chip containing multiple rectangular baffle structures was designed to enhance transverse mixing, extend the effective residence time, and mitigate clogging during particle formation. Under the selected conditions, homogeneous HRP@ZnBDC-NH₂ particles with an average hydrodynamic diameter of 868.5 nm and a polydispersity index of 0.266 were obtained. The homogeneous HRP@ZnBDC-NH₂ showed an encapsulation efficiency of 56.17% and a loading content of 1.49%. Michaelis–Menten analysis gave a Km value of 52.49 μM for HRP@ZnBDC-NH₂, suggesting improved apparent substrate affinity compared with the corresponding bulk-synthesised sample. The results support the use of baffle-structured microfluidics as a controllable platform for enzyme@MOF synthesis, while further studies on enzyme leaching, reusability, long-term stability and extended chip operation are required to evaluate its operational robustness. Full article

Background: In 2022, we adapted the World Health Organization Digital Adaptation Kit (DAK) for antenatal care (ANC) into the BornFyne digital health platform. Because ANC services in many facilities relied on fragmented paper-based guidance, the DAK represented the first exposure for many providers to a structured, comprehensive ANC guideline framework integrated into a digital system. Understanding providers’ motivation, capabilities, and readiness to adopt guideline-driven digital care was essential to inform implementation strategies. We therefore conducted a workshop to explore healthcare providers’ expectations, perceived challenges, and readiness to integrate a comprehensive digital ANC system into routine practice. Methods: A prospective mixed-methods pre–post implementation pilot workshop was conducted between June and August 2023 across four districts. The 5-h workshop targeted ANC providers and included exposure to the WHO ANC DAK and demonstration of its integration into the BornFyne platform. Survey tools assessed perceived knowledge, attitudes, perceived behavioural control, intention, and social norms using Likert scales. Constructs were mapped to the COM-B framework. Paired student’s t-tests assessed pre–post changes (p < 0.05), and Cohen’s d estimated effect sizes. Qualitative responses were thematically analysed. Results: Nineteen participants, including doctors, nurses, midwives, and data managers from public, private, and faith-based facilities, completed the surveys. Most participants were women (90%). Following the workshop, perceived knowledge of behaviour change and digital guideline use significantly increased (t(19) = 3.22, p = 0.004, d = 0.78). Although no statistically significant changes were observed in attitudes, perceived behavioural control, intentions, or social norms, descriptive trends suggested improved confidence and openness toward digital ANC systems. Qualitative findings identified increased knowledge as the most important outcome, with 80% of participants emphasizing its role in strengthening capability. Participants also reported greater confidence in integrating digital tools into routine care but highlighted contextual and motivational barriers as ongoing challenges. Acceptability was high: 90% expressed satisfaction, and all participants rated the workshop as useful, relevant, engaging, and worth their time. Conclusions: The workshop improved providers’ perceived knowledge and demonstrated high acceptability of DAK-aligned digital ANC systems. While changes in attitudes and behavioural control were not statistically significant, the findings support the feasibility of behaviour change-based digital ANC interventions. Addressing contextual and motivational barriers will be essential for sustained adoption in resource-limited settings. Full article

Background: Ambulatory blood pressure monitoring (ABPM) offers a comprehensive assessment of mean blood pressure (BP), circadian patterns, and out-of-office hypertension phenotypes. We evaluated the association of ABPM parameters with left ventricular hypertrophy (LVH) and hypertensive retinopathy (HRP) in children with hypertension. Methods: This retrospective analysis included 269 children who underwent ABPM at a tertiary care facility. Multivariable logistic regression was used to evaluate associations between BP Z scores and target organ damage (TOD), adjusting for age, sex, and body mass index standard deviation score. Machine learning techniques (logistic regression, random forest, and multilayer perceptron) were also evaluated using 10-fold cross-validation across exploratory combinations of the 90th or 95th percentiles and BP-load thresholds of 25% or 50%. Results: LVH was detected in 45/247 patients (18%) and HRP was observed in 22/240 patients (9%). The 24 h systolic BP Z score was independently associated with LVH (adjusted OR 1.44; 95% CI 1.16–1.80; p = 0.001). Cohort-derived systolic Z-score thresholds for LVH were 1.31 for 24 h systolic BP and 1.47 for nighttime systolic BP, corresponding to approximately the 90.6th and 93rd percentiles, respectively. No variable evaluated was independently associated with retinopathy. Logistic regression using the 90th percentile/25% load dataset showed the highest performance among the evaluated classifiers, but discrimination was limited (ROC area 0.603; MCC 0.041). Conclusions: Higher ambulatory systolic BP was independently associated with LVH but not with clinically detected hypertensive retinopathy. ABPM values at or above the 90th percentile and a systolic load of ≥25% may help identify children at increased risk of TOD, although these thresholds remain exploratory and require prospective external validation. Full article

Olive tree (Olea europaea L.) cultivation is increasingly challenged by climate change and the necessity for more sustainable crop management strategies. In this context, plant biostimulants have emerged as inputs of high potential for improving crop performance and resilience. Nonetheless, knowledge of their underlying biochemical mechanisms remains largely fragmented. Here, an untargeted GC/EI/MS-based metabolomics approach was employed to investigate the early metabolic responses of young olive trees to foliar application of two commercial biostimulants: a seaweed (SE)- and a harpin (HRP)-based formulation. Leaf samples were collected seven days post-treatment and were subjected to photosynthetic-content and metabolomics analyses. The former revealed no statistically significant differences in chlorophyll and carotenoid contents among treatments, indicating that photosynthetic pigment homeostasis was maintained. In contrast, metabolomics analysis demonstrated extensive and coordinated metabolic reprogramming induced by both products. SE treatments resulted in a primarily promoted growth-oriented metabolic profile, characterized by enhanced carbon utilization, modulation of antioxidant metabolism, activation of plastidial isoprenoid biosynthesis, and remodeling of the shikimate–phenylpropanoid pathway. Conversely, HRP treatments elicited a distinct stress-priming signature involving the accumulation of α,α-trehalose, phytol, and α-ketoglutaric acid, together with membrane lipid remodeling associated with improved osmoprotection and stress acclimation. These findings provide novel insights into the mode(s) of action of olive biostimulants and highlight the importance of metabolomics for identifying biochemical markers associated with enhanced plant resilience and sustainable olive production. Full article

Biolayer Interferometry (BLI) has traditionally been used for characterization of protein–protein interactions (PPI) with proteins, such as antibodies and their antigens, through kinetic and quantitation assays. Limitations, for example in sensitivity and the availability of established assay formats, have restricted its adoption across other analytical applications. This article highlights three case studies which demonstrate the expansion of BLI into novel applications, spanning the areas of protein detection and viral vector characterization. The first case study details the use of a multi-step signal amplification assay to enable the detection of low abundant molecules, such as cytokines, at lower concentrations than can be detected using the standard one-step binding approach. Cytokines are sandwiched between biotinylated and HRP-conjugated antibodies, then dipped into 3-amino-9-ethylcarbazole (AEC) reagent, resulting in an enhancement of the cytokine detection sensitivity. The second case study uses BLI for the quantitation of mixed populations of a bispecific antibody (bsAb). Bridging and dual binding assay formats are evaluated for their assessment of bsAb antigen binding kinetics and ability to determine the ratio of correctly assembled bsAb within a sample. In the third case study, BLI detection principles are used to estimate the percentage full capsids in a mixed population of AAV particles. Collectively, these case studies demonstrate the versatility of Octet^® BLI and highlight its potential to support an increasing range of analytical workflows beyond its traditional applications. Full article