Search Results (2,321)

The detection of high-consequence viral pathogens is essential for spillover prevention and reduction in transmission but is limited by the low sensitivity of next-generation sequencing technology. Low-titer field samples from a variety of hosts are primarily composed of non-viral genomic material, reducing the probability of obtaining usable sequence data. Targeted enrichment, such as VirCapSeq-VERT, removes background genomic material to improve virus detection but is mainly used for sequencing clinical samples. We customized the VirCapSeq-VERT probe system to aid in the detection of zoonotic viruses of interest and adapted it for use on the Oxford Nanopore sequencing platform. We validated the method on a variety of samples, including a mock virome consisting of seven RNA viruses, samples from an animal laboratory study, and a set of animal field samples. We also developed Nanite, a lightweight bioinformatics pipeline, to perform bioinformatic analyses. Results indicated that the optimized enrichment protocol improved sequencing by enhancing the detection of viruses, increasing read lengths, and, in some cases, improving genomic coverage. Most importantly, the sequencing of zoonotic viruses was improved in field samples with low titers, suggesting that this protocol is a useful tool for increasing the efficacy of Oxford Nanopore sequencing for field-oriented applications. Full article

Stance detection aims to identify whether a text expresses a favorable, opposing, or neutral attitude toward a given target and has become increasingly important for analyzing public discourse on social media. Existing approaches, ranging from supervised neural models to prompt-based large language models (LLMs), face two persistent challenges: the scarcity of annotated stance data across diverse targets and the difficulty of generalizing to unseen targets under pragmatic and rhetorical variation. To address these issues, we propose PAMR (Pragmatic-Aware Multi-Agent Reasoning), a zero-shot stance detection framework that decomposes stance inference into structured reasoning steps. PAMR orchestrates three LLM-driven agents—a linguistic parser that extracts pragmatic markers and canonicalizes claims, an NLI-based estimator that produces calibrated stance probabilities through consensus voting, and a counterfactual and view-switching auditor that probes robustness under controlled rewrites. A stability-aware fusion integrates these signals, conservatively abstaining when evidence is uncertain or inconsistent. Experiments on SemEval-2016 and COVID-19-Stance show that PAMR achieves macro-F1 scores of 71.9% and 73.0%, surpassing strong zero-shot baselines (FOLAR and LogiMDF) by +2.0% and +3.1%. Ablation results confirm that pragmatic cues and counterfactual reasoning substantially enhance robustness and interpretability, underscoring the value of explicit reasoning and pragmatic awareness for reliable zero-shot stance detection on social media. Full article

Radionuclide molecular imaging of epidermal growth factor receptor (EGFR) expression might permit the selection of patients for EGFR-targeting therapies. Designed ankyrin repeat protein (DARPin) E01 with a high affinity to the ectodomain III of the EGFR is a possible EGFR imaging probe. The goal of this study was to evaluate the potential of radiolabeled DARPin E01 for in vivo imaging of EGFR. DARPin E01 containing the (HE)₃-tag was site-specifically labeled with a residualizing ^99mTc (using ^99mTc]Tc(CO)₃). Two methods providing non-residualizing ¹²³I labels, direct electrophilic radioiodination and indirect radioiodination using [¹²³I]I-para-iodobenzoate (PIB), were tested. [^99mTc]Tc-(HE)₃-E01 and [¹²³I]I-(HE)₃-E01-PIB preserved specific binding to EGFR-expressing cells and affinity in the single-digit nanomolar range. Direct labeling with ¹²³I resulted in a substantial loss of binding. In vitro cellular processing studies showed that both [^99mTc]Tc-(HE)₃-E01 and [¹²³I]I-(HE)₃-E01-PIB had rapid binding and relatively slow internalization. Evaluation of [^99mTc]Tc-(HE)₃-E01 biodistribution in normal CD1 mice showed that its hepatic uptake was non-saturable, suggesting that this tracer does not bind to murine EGFR. A side-by-side comparison of biodistribution and tumor targeting of [^99mTc]Tc-(HE)₃-E01 and [¹²³I]I-(HE)₃-E01-PIB was performed in Nu/j mice bearing EGFR-positive A-431 and EGFR-negative Ramos human cancer xenografts. Both radiolabeled DARPins demonstrated EGFR-specific tumor uptake. However, [¹²³I]I-(HE)₃-E01-PIB had appreciably lower uptake in normal organs compared to [^99mTc]Tc-(HE)₃-E01, which provided significantly (p < 0.05) higher tumor-to-organ ratios. Gamma-camera imaging confirmed that [¹²³I]I-(HE)₃-E01-PIB demonstrated a higher imaging contrast in preclinical models than [^99mTc]Tc-(HE)₃-E01. In conclusion, DARPin (HE)₃-E01 labeled using a non-residualizing [¹²³I]I-para-iodobenzoate (PIB) label is the preferred radiotracer for in vivo imaging of EGFR expression in cancer. Full article

Nucleic acid aptamers leverage defined tertiary structures for precise molecular recognition, positioning them as transformative biomedical tools. We engineered AP1-F, a G-quadruplex (G4)-structured aptamer that selectively binds membrane-anchored nucleolin (NCL) non-permeabilizing, overcoming a key limitation of conventional probes. Microscale thermophoresis confirmed nanomolar affinity to NCL. By means of rigorous optimization, AP1-F attained a greater than ten-fold fluorescence signal ratio between malignant and normal cells in co-cultures, exceeding the extensively researched AS1411. Dual-channel flow cytometry demonstrated over 98.78% specificity at single-cell resolution within heterogeneous cell populations, owing to AP1-F’s unique membrane localization—unlike AS1411’s intracellular uptake, which elicited erroneous signals from cytoplasmic NCL. Competitive binding experiments and Laser Confocal Imaging confirmed that AP1-F specifically identifies cancer cells by binding to the NCL recognition site on the membrane. In pathological sections, AP1-F exhibited a 40.5-fold fluorescence intensity ratio between tumor and normal tissue, facilitating accurate tissue-level differentiation. Significantly, it delineated molecular subtypes by associating membrane NCL patterns with morphometric analysis: luminal-like MCF-7 displayed consistent staining in cohesive clusters, whereas basal-like MDA-MB-468 revealed sporadic NCL with irregular outlines—characteristics imperceptible to intracellular-targeted antibodies, thus offering subtype-specific diagnostic insights. This combination biochemical–morphological approach accomplished subtype differentiation with a single-step, non-permeabilized process that maintained lower cytotoxicity and tissue integrity. AP1-F enhances diagnostic accuracy by utilizing spatial confinement to eradicate intracellular interference, connecting molecular specificity to intraoperative margin evaluation or biopsy categorization. Full article

Background/Objectives: The role of α-synuclein (α-syn) in the pathogenesis of Parkinson’s disease (PD) or neurodegenerative diseases such as Lewy body dementia (LBD) and multiple system atrophy (MSA) is commonly accepted. Through different physiological dysfunctions, abnormal forms of α-syn are generated. These abnormal aggregates accumulate and alter pre- and postsynaptic transmission, in particular that of dopamine. Thus, the development of a diagnostic biomarker of synucleinopathies remains crucial and challenging. The development of an α-syn positron emission tomography (PET) radiopharmaceutical may be suitable to early diagnose and stratify patients, follow up disease progression, and evaluate future therapies. Methods: To develop a selective α-syn PET tracer, we synthesized an original series based on alkynyl(aza)indoles. Fifteen final ligands were synthesized bearing indoles or azaindoles from one side of the alkyne and a substituted phenyl ring for the opposite side of the alkyne. The final ligands were tested to determine Ki and/or Kd toward α-syn, tau, and Aβ. Results: The SAR showed that the indole series exhibited moderate to low affinity for α-syn and, moreover, lower Ki toward Aβ and tau (i.e., compound 39, Ki_(αsyn) 21.7 nM, Ki_(Aβ) 64.4 nM, Ki_(Tau) 27.6 nM), highlighting the low potency of these series to afford an α-syn tracer. The introduction of a nitrogen on the different positions of the phenyl to obtain the corresponding azaindoles resulted for most of the compounds in better affinity for α-syn and selectivity towards Aβ compared to the indole analogs (i.e., compound 43, Ki_(αsyn) 4.7 nM, Ki_(Aβ) 24.4 nM, and Ki_(Tau) 4.61 nM). A fluorinated azaindole derivative was prepared with a view to obtaining a ¹⁸F tracer and exhibited the highest affinity for α-syn but without selectivity against tau and Aβ. The radiosynthesis of [¹⁸F]45 was performed in a two-step procedure starting from the tosylated and protected precursor. [¹⁸F]45 was obtained in 85 ± 5 min with a radiochemical yield of 32 ± 3%. Molar activity, determined from a calibration with stable 45, was around 130 GBq/µmole. The dynamic PET imaging showed that [¹⁸F]45 was able to cross the blood–brain barrier, but non-specific uptake was observed, confirming the in vitro results. Conclusions: Although promising nanomolar affinity for the target, the new tracer showed mainly non-specific in vivo uptake in the rat brain, indicating that further pharmacomodulations on the azaindole series are required. Full article

Gliomas, the most common primary brain tumors, present significant diagnostic and treatment challenges due to their infiltrative nature and heterogeneity. Our previous research revealed that glioma tumors in both animals and humans accumulate beta-amyloid protein (Aβ), detectable through immunohistochemical methods or staining with amyloid-specific dyes. We hypothesize that beta-amyloid-specific dyes could serve as glioma markers, potentially enabling the delineation of glioma tumors or targeted therapeutics delivery. In this study, the specificity and blood-brain barrier permeability of two fluorescent beta-amyloid-specific dyes, Brilliant Blue G (BBG) and BODIPY-based Amyloid Probe-1 (BAP-1), were evaluated in C57Bl/6 mouse glioma implantation models using GL261 and KR158 glioma cells. The findings demonstrate that both BBG and BAP-1 selectively stain gliomas, providing a clear contrast from normal brain tissue. The study results open avenues for further development of glioma visualization methods and targeted therapeutic delivery strategies for clinical applications. Full article

Accurate phase identification is essential for characterising complex mineral systems but remains a challenge in SEM-based automated mineralogy (AM) for compositionally variable rock-forming or accessory minerals. While platforms such as the Tescan Integrated Mineral Analyzer (TIMA) offer high-resolution phase mapping through BSE-EDS data, classification accuracy depends on the quality of the user-defined phase library. Generic libraries often fail to capture site-specific mineral compositions, resulting in misclassification and unclassified pixels, particularly in systems with solid solution behaviour, compositional zoning, and textural complexity. We present a refined approach to developing and validating custom TIMA phase libraries. We outline strategies for iterative rule refinement using mineral chemistry, textures, and BSE-EDS responses. Phase assignments were validated using complementary microanalytical techniques, primarily electron probe microanalysis (EPMA) and laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). Three Queensland case studies demonstrate this approach: amphiboles in an IOCG deposit; cobalt-bearing phases in a sediment-hosted Cu-Au-Co deposit; and Li-micas in an LCT pegmatite system. Targeted refinement of phases improves identification, reduces unclassified phases, and enables rare phase recognition. Expert-guided phase library development strengthens mineral systems research and downstream applications in geoscience, ore deposits, and critical minerals while integrating datasets across scales from cores to mineral mapping. Full article

Omsk hemorrhagic fever is an acute zoonotic disease caused by Omsk hemorrhagic fever virus, a member of the genus Flavivirus (family Flaviviridae), with a reported case-fatality rate of approximately 3%. Historically confined to southwestern Siberia, ecological changes raise concerns about possible spread to non-endemic regions. Although no Omsk hemorrhagic fever cases have been reported in the Republic of Korea, the risk of accidental importation highlights the importance of establishing a reliable diagnostic protocol. We established and validated an institutionally developed diagnostic protocol employing real-time reverse transcription polymerase chain reaction targeting the NS2A and C genes of Omsk hemorrhagic fever virus. Primers and probes were designed from all available genomes to ensure broad strain coverage. Human ribonuclease P was used as an internal control to verify nucleic acid extraction and amplification. Using synthetic deoxyribonucleic acid fragments and in vitro-transcribed ribonucleic acid, assay performance was optimized, and analytical sensitivity was determined using probit analysis. The limits of detection were 74.50 copies/µL (threshold cycle 32.99) for NS2A and 70.41 copies/µL (threshold cycle 35.38) for C. Specificity testing using representative flaviviruses (West Nile virus, Yellow fever virus, Zika virus, St. Louis encephalitis virus, and Tick-borne encephalitis virus) and an alphavirus (Venezuelan equine encephalitis virus) demonstrated no cross-reactivity. The assay demonstrated high sensitivity, specificity, and reproducibility, supporting its potential application in national and international Omsk hemorrhagic fever virus surveillance systems. Full article

Identifying age-appropriate books for children is a complex task that requires balancing linguistic, cognitive, and thematic factors. This study introduces an artificial intelligence–supported framework to estimate the Suitable Age Value (SAV) of Turkish children’s books targeting the 2–18-year age range. We employ repeated, stratified $5\times 5$ cross-validation and report out-of-fold (OOF) metrics with 95% confidence intervals for a dataset of 300 Turkish children’s books. As classical baselines, linear/ElasticNet, SVR, Random Forest (RF), and XGBoost are trained on the engineered features; we also evaluate a rule-based Ateşman readability baseline. For text, we use a frozen dbmdz/bert-base-turkish-uncased encoder inside two hybrid variants, Concat and Attention-gated, with fold-internal PCA and metadata selection; augmentation is applied only to the training folds. Finally, we probe a few-shot LLM pipeline (GPT-4o-mini) and a convex blend of RF and LLM predictions. A few-shot LLM markedly outperforms the zero-shot model, and zero-shot performance is unreliable. Among hybrids, Concat performs better than Attention-gated, yet both trail our best classical baseline. A convex RF + LLM blend, learned via bootstrap out-of-bag sampling, achieves a lower RMSE/MAE than either component and a slightly higher QWK. The Ateşman baseline performance is substantially weaker. Overall, the findings were as follows: feature-based RF remains a strong baseline, few-shot LLMs add semantic cues, blending consistently helps, and simple hybrid concatenation beats a lightweight attention gate under our small-N regime. Full article

Capsaicinoids (CPCs) are regarded as a typical marker of waste oil, which has emerged as a serious food safety issue in developing countries, necessitating the development of rapid, sensitive, and specific detection methods. In this study, a novel hapten was synthesized to generate a high-affinity monoclonal antibody (mAb) targeting CPCs. Subsequently, aggregation-induced emission fluorescent microspheres (AIEFMs), known for their superior fluorescence intensity, were utilized as an enhanced probe to develop a lateral flow immunoassay (LFIA) based on mAb 8B4 for CPC detection. For comparison, a traditional gold nanoparticle (AuNP)-LFIA was also constructed using the corresponding mAb. The AIEFM-LFIA demonstrated a limit of detection (LOD) of 0.33 µg/kg for CPCs in edible oil samples, which is 4.21 times lower than the LOD of 1.39 µg/kg achieved by the AuNP-LFIA. And the assay effectively identified three additional CPCs, with LODs ranging from 0.26 to 0.99 µg/kg, while exhibiting minimal cross-reactivity with CPC analogs, indicating high specificity. The recovery rates of the AIEFM-LFIA in oil samples ranged from 75.0% to 106.0%, with coefficients of variation ≤ 8.3%, exhibiting excellent accuracy and precision. Furthermore, the results of the AIEFM-LFIA demonstrated a strong degree of correlation with liquid chromatography–tandem mass spectrometry, with a correlation coefficient (R²) of 0.978. Consequently, the developed AIEFM-LFIA shows significant promise as a rapid, sensitive, specific, and reliable method for detecting CPCs in oil samples. Full article

This study examined child outcomes for five minimally verbal (or non-speaking) autistic preschoolers who participated in cascading coaching programs in which naturalistic developmental behavioral intervention (NDBI) techniques were taught to graduate student clinicians and Hispanic caregivers (three who primarily spoke English, and two who spoke Spanish). While prior studies reported on adult participant outcomes, this study analyzed child multimodal communication outcomes, using multiple baselines/probes single case experimental designs across contexts. Neurodiversity-affirming and culturally responsive principles were embedded within the intervention procedures. Following the introduction of a coached NDBI, all five children (three who received the intervention in English and two who received the intervention in Spanish) demonstrated increased use of (a) the total targeted communicative responses and (b) the targeted unprompted communicative responses, across both student clinician-led and caregiver-led play sessions. The Tau-U effect size measures revealed large-to-very large effects across all of the variables. Overall, higher rates of communication responses were observed during student clinician-led sessions than in caregiver-led sessions. Additionally, behavioral coding of the multimodal response forms (e.g., gestures, aided augmentative and alternative communication, signs, vocal words) using the Communication Matrix revealed that the children used a variety of response topographies during the intervention sessions beyond their preferred communication mode (e.g., signs for three participants). Four of the five children used symbolic communication forms consistently across both caregiver and student clinician-led sessions. Importantly, adults’ reinforcement of pre-symbolic or less advanced communication forms during the intervention did not inhibit the use of more advanced forms. Full article

Tourism performance often depends on the joint provision of built (“hard”) and environmental (“green”) infrastructure, yet their combined effects are not well established. Using official data for Saudi Arabia’s 13 regions (2023–2024), this study constructs composite hard and green indices, estimates ordinary least squares models with heteroskedasticity-consistent inference, and probes spatial heterogeneity using geographically weighted regression (exploratory) alongside k-means/hierarchical clustering. Hard infrastructure is the strongest and most consistent correlate of overnight visitors and spending, whereas green infrastructure exhibits non-positive marginal effects over the observed range of hard capacity; a negative, statistically significant Hard × Green interaction indicates diminishing returns to greening as built capacity increases. Clustering differentiates metropolitan hubs from nature-oriented regions, underscoring place-specific policy needs. Practically, results support sequencing prioritizing foundational access and basic accommodation in under-served regions, quality upgrades and public-realm enhancement in mature centers, and targeted green interventions where marginal gains are greatest. Key limitations (cross-sectional design; coarse green metrics) motivate richer environmental indicators and longitudinal data to clarify dynamics and thresholds over time. Full article

Background/Objectives: Glypican-3 (GPC3) is a cell surface oncofetal protein that is highly expressed in hepatocellular carcinoma (HCC) but absent in normal liver tissue, making it an attractive target for molecularly targeted diagnosis and therapy. To support GPC3-targeted treatment strategies, there is a need for a non-invasive imaging tool capable of detecting GPC3-positive tumors. Methods: We conjugated a commercially available murine anti-GPC3 antibody (1G12), or a proprietary chimeric anti-GPC3 antibody (ET58) to the standard magnetic resonance imaging (MRI) contrast agent, gadolinium, via a DOTA chelator. The resulting probes, 1G12-DOTA-Gd or ET58-DOTA-Gd, respectively, were assessed for in vitro relaxivity and binding specificity to GPC3-positive HCC cells, as well as for in vivo imaging performance in mouse xenograft models bearing GPC3-positive or GPC3-negative HCC tumors. Conclusions: ET58-DOTA-Gd shows high specificity, imaging efficacy, and a favorable immunogenicity profile, thereby making it a promising candidate for clinical translation as a GPC3-targeted MRI probe. It holds potential as a non-invasive companion diagnostic for identifying GPC3-positive HCC patients who may benefit from GPC3-targeted therapies. Full article

Optical imaging technologies expand gastrointestinal endoscopy beyond white-light endoscopy (WLE), improving visualization of mucosal, vascular, and subsurface features. They are applied to the detection of neoplastic and premalignant lesions, inflammatory diseases, and small bowel and pancreatic disorders, though their validation and readiness for routine practice vary. This review critically evaluates both guideline-endorsed and investigational optical imaging techniques across major gastrointestinal indications, highlighting diagnostic performance, level of validation, current guideline recommendations, and practical challenges to adoption. In Barrett’s esophagus, narrow-band imaging (NBI) is guideline-endorsed, while acetic acid chromoendoscopy is validated in expert centers. For gastric intestinal metaplasia and early gastric cancer, magnifying NBI achieves diagnostic accuracies exceeding 90% and is guideline-recommended, with acetic acid chromoendoscopy aiding in margin delineation. In inflammatory bowel disease, dye-spray chromoendoscopy is the reference standard for dysplasia surveillance, with virtual methods such as NBI, FICE, and i-SCAN serving as practical alternatives when dye application is not feasible. In the colorectum, NBI supports validated optical diagnosis strategies (resect-and-discard, diagnose-and-leave), while dye-based chromoendoscopy improves detection of flat and serrated lesions. Capsule endoscopy remains the standard for small bowel evaluation of bleeding, Crohn’s disease, and tumors, with virtual enhancement, intelligent chromo capsule endoscopy, and AI-assisted interpretation emerging as promising adjuncts. Pancreaticobiliary applications of optical imaging are also advancing, though current evidence is still preliminary. Investigational modalities including confocal laser endomicroscopy, optical coherence tomography, autofluorescence, Raman spectroscopy, and fluorescence molecular imaging show potential but remain largely restricted to research or expert settings. Guideline-backed modalities such as NBI and dye-based chromoendoscopy are established for clinical practice and supported by robust evidence, whereas advanced techniques remain investigational. Future directions will rely on broader validation, integration of artificial intelligence, and adoption of molecularly targeted probes and next-generation capsule technologies, which together may enhance accuracy, efficiency, and standardization in gastrointestinal endoscopy. Full article

In the context of the study of G-quadruplex (G4), the main field of focus is usually referred to binding molecules able to interact with these non-canonical conformations and stabilizing them leading to diverse biological effects. Although the cellular events triggered by these ligands are useful for potential anticancer applications, the development of innovative molecular tools to gain new information about G4 has become more urgent. The concept of G4-interacting molecular tools refers to chemical entities that can bind and interact with G-rich sequences of the genome—like traditional ligands—but simultaneously provide external outputs that can be interpreted and studied to obtain insights on their dynamics, position in the cellular context and more. Starting from traditional chemical approaches, researchers have worked to produce sophisticated and complex synthetic strategies in order to introduce more accurate instruments for their aims. This review provides a comprehensive and up-to-date overview of this research area by detailing the major classes of molecular tools, describing the latest updates about three main classes: small-molecules fluorescence probes, G4-binding metal complexes, and products of conjugation strategies. Overall, advancements in molecular tools targeting G4s have made the study of G4 formation, dynamics, and functions much easier; thus, increasing the knowledge of G4 biology, and creating new opportunities for biomedical and therapeutic applications, ultimately highlighting the importance of the development of molecular tools in G4 research. Full article