Search Results (1,048)

The epidermal growth factor receptor (EGFR) is a key target for both cancer diagnosis and therapeutic interventions. Assessing EGFR expression before therapy has become routine in clinical practice, yet current methods like biopsy and immunohistochemistry (IHC) have significant limitations, including invasiveness, limited repeatability, and lack of real-time, whole-body data. EGFR-targeted imaging has emerged as a promising alternative. EGFR-targeting peptides, owing to their favorable physicochemical properties and versatility, are increasingly being explored for a variety of applications, including molecular imaging, drug delivery, and targeted therapy. Recent advances have demonstrated the potential of EGFR-targeting peptides conjugated to imaging probes for non-invasive, real-time in vivo tumor detection, precision therapy, and surgical guidance. Here, we provide a comprehensive overview of the latest progress in EGFR-targeting peptides development, with a particular focus on their application in the development of molecular imaging agents, including fluorescence imaging, PET/CT, magnetic resonance imaging, and multimodal imaging. Furthermore, we examine the challenges and future directions concerning the development and clinical application of EGFR-targeting peptide-based imaging probes. Finally, we highlight emerging technologies such as artificial intelligence, mutation-specific peptides, and multimodal imaging platforms, which offer significant potential for advancing the diagnosis and treatment of EGFR-targeted cancers. Full article

Palindromic antimicrobial peptides (PAMs) constitute versatile scaffolds for the design and optimization of anticancer agents with applications in therapy, diagnosis, and/or monitoring. In the present study, fluorolabeled peptides derived from the palindromic sequence RWQWRWQWR containing fluorescent probes, such as 2-Aminobenzoyl, 5(6)-Carboxyfluorescein, and Rhodamine B, were obtained. RP-HPLC analysis revealed that the palindromic peptide conjugated to Rhodamine B (RhB-RWQWRWQWR) exhibited the presence of isomers, likely corresponding to the open-ring and spiro-lactam forms of the fluorescent probe. This equilibrium is dependent on the peptide sequence, as the RP-HPLC analysis of dimeric peptide (RhB-RRWQWR-hF-KKLG)₂K-Ahx did not reveal the presence of isomers. The antibacterial activity of the fluorescent peptides depends on the probe attached to the sequence and the bacterial strain tested. Notably, some fluorescent peptides showed activity against reference strains as well as sensitive, resistant, and multidrug-resistant clinical isolates of E. coli, S. aureus, and E. faecalis. Fluorolabeled peptides 1-Abz (MIC = 62 µM), RhB-1 (MIC = 62 µM), and Abz-1 (MIC = 31 µM) exhibited significant activity against clinical isolates of E. coli, S. aureus, and E. faecalis, respectively. The RhB-1 (IC₅₀ = 61 µM), Abz-1 (IC₅₀ = 87 µM), and RhB-2 (IC₅₀ = 35 µM) peptides exhibited a rapid, significant, and concentration-dependent cytotoxic effect on HeLa cells, accompanied by morphological changes characteristic of apoptosis. RhB-1 (IC₅₀ = 18 µM) peptide also exhibited significant cytotoxic activity against breast cancer cells MCF-7. These conjugates remain valuable for elucidating the possible mechanisms of action of these novel anticancer peptides. Rhodamine-labeled peptides displayed cytotoxicity comparable to that of their unlabeled analogues, suggesting that cellular internalization constitutes a critical early step in their mechanism of action. These findings suggest that cell death induced by both unlabeled and fluorolabeled peptides proceeds predominantly via apoptosis and is likely contingent upon peptide internalization. Functionalization at the N-terminal end of the palindromic sequence can be evaluated to develop systems for transporting non-protein molecules into cancer cells. Full article

Background: Reliable platelet (PLT) measurement is crucial for the accurate diagnosis of thrombocytopenia. Several methods exist for automated PLT counting, including the impedance method (PLT-I), as well as optical and fluorescence methods (PLT-F). The impedance method is cost-effective but susceptible to interference from small red blood cells and schistocytes. In contrast, fluorescent assessment offers higher specificity but is more expensive, as it requires additional dyes and detectors. Hybrid platelet counting (PLT-H) combines impedance with measurements from the leukocyte differentiation channel and is available without additional cost. Aim: The aim of this study was to evaluate the accuracy of hybrid PLT counting in anemic samples. Methods: In this retrospective study, PLT counts from 583 unselected anemic samples were analyzed using two different analyzers: the Sysmex XN3500, equipped with fluorescent PLT-F technology, and the Mindray BC6200, which uses both impedance (PLT-I) and hybrid (PLT-H) technologies. Agreement between PLT-I and PLT-F, as well as between PLT-H and PLT-F, was assessed using Bland–Altman plots. Correlation between the methods was evaluated using the Pearson correlation coefficient. Results: The hybrid method demonstrated better accuracy in PLT counting compared to the impedance method. Correlation between PLT-H and PLT-F was excellent, ranging from 0.991 to 0.999. In thrombocytopenic samples (PLT < 50 G/L), the hybrid method also provided more reliable PLT counts than the impedance method, reducing the number of falsely elevated PLT results by nearly fivefold. Conclusions: Hybrid platelet counting yields more accurate results than the impedance method in anemic samples and shows excellent correlation with the fluorescence method. Full article

Theranostic agents enable the simultaneous diagnosis and treatment of diseases, and they are particularly useful in fluorescent imaging and cancer therapies. In this study, carbon quantum dots were synthesized via a microwave-assisted method using citric acid and bovine serum albumin (BSA) as precursors. The resulting CQDs exhibited spherical morphology, an average size of 4 nm, and an amorphous graphitic structure. FT-IR characterization revealed the presence of amide bonds and oxygenated functional groups. At the same time, optical analysis showed excitation at 320 nm and emission between 360 and 400 nm, with fluorescent stability maintained for one month. Furthermore, the CQDs demonstrated good thermal stability and photothermal efficiency, reaching temperatures above 41 °C within 15 min under NIR irradiation, with a mass loss of less than 1%. Their stability was evaluated in media with different pH levels, simulating physiological and tumor environments. While their behavior was affected under acidic conditions, their excellent photothermal conversion capacity and overall stability in triple-distilled water positioned them as promising candidates for theranostic applications in cancer, effectively combining diagnostic imaging and thermal therapy. Full article

Background: Antinuclear antibodies (ANAs) serve as crucial biomarkers for diagnosing systemic autoimmune diseases; however, their interpretation can be complex and may not always correlate with clinical symptoms. Methods: A comprehensive narrative review was conducted to evaluate the peer-reviewed literature published between 1961 and 2025. Databases, including PubMed and Scopus, were searched using combinations of controlled vocabulary and free-text terms relating to antinuclear antibodies and their clinical significance. The objective was to gather and synthesize information regarding the diagnostic utility and interpretation of ANA testing in routine medical practice. Discussion: The indirect immunofluorescence assay (IIF) on HEp-2 cells is established as the gold standard for detecting ANAs, facilitating the classification of various fluorescent patterns. While a positive ANA test can suggest autoimmune disorders, the presence and titre must be interpreted alongside clinical findings, as low titres often lack diagnostic significance. Findings indicate that titres higher than 1:160 may provide greater specificity in differentiating true positives from false positives in healthy individuals. The study also emphasizes the relevance of fluorescence patterns, with specific patterns linked to particular diseases, although many do not have strong clinical correlations. Moreover, certain autoantibodies demonstrate high specificity for diseases like systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD). Ultimately, while ANA testing is invaluable for diagnosing connective tissue diseases, healthcare providers must consider its limitations to avoid misdiagnosis and unnecessary treatment. Conclusions: ANA testing is a valuable tool in the diagnosis of connective tissue diseases, but its interpretation must be approached with caution. Clinical context remains crucial when evaluating ANA results to avoid misdiagnosis and overtreatment. This review is about the diagnostic aspects and clinical consequences of ANA testing, as well as highlighting both the diagnostic benefits and the potential limitations of this procedure in everyday clinical practice. The review fills a gap in the literature by integrating the diagnostic and clinical aspects of ANA testing, with a focus on real-world interpretation challenges. Full article

Verticillium wilt is characterized by chlorosis in leaves and is a devastating disease in eggplant. Early diagnosis, prior to the manifestation of symptoms, enables targeted management of the disease. In this study, we aim to detect early leaf wilt in eggplant leaves caused by Verticillium dahliae by integrating multispectral imaging with machine learning and deep learning techniques. Multispectral and chlorophyll fluorescence images were collected from leaves of the inbred eggplant line 11-435, including data on image texture, spectral reflectance, and chlorophyll fluorescence. Subsequently, we established a multispectral data model, fusion information model, and multispectral image–information fusion model. The multispectral image–information fusion model, integrated with a two-dimensional convolutional neural network (2D-CNN), demonstrated optimal performance in classifying early-stage Verticillium wilt infection, achieving a test accuracy of 99.37%. Additionally, transfer learning enabled us to diagnose early leaf wilt in another eggplant variety, the inbred line 14-345, with an accuracy of 84.54 ± 1.82%. Compared to traditional methods that rely on visible symptom observation and typically require about 10 days to confirm infection, this study achieved early detection of Verticillium wilt as soon as the third day post-inoculation. These findings underscore the potential of the fusion model as a valuable tool for the early detection of pre-symptomatic states in infected plants, thereby offering theoretical support for in-field detection of eggplant health. Full article

Amyloid-β (Aβ) aggregates are considered as the important factors of Alzheimer’s disease (AD). Multifunctional materials have shown significant effects in the diagnosis and treatment of AD by modulating the aggregation of Aβ and production of reactive oxygen species (ROS). Compared to traditional surgical treatment and radiotherapy, phototherapy has the advantages, including short response time, significant efficacy, and minimal side effects in disease diagnosis and treatment. Recent studies have shown that local thermal energy or singlet oxygen generated by irradiating certain organic molecules or nanomaterials with specific laser wavelengths can effectively degrade Aβ aggregates and depress the generation of ROS, promoting progress in AD diagnosis and therapy. Herein, we outline the development of photothermal therapy (PTT) and photodynamic therapy (PDT) strategies for the diagnosis and therapy of AD by modulating Aβ aggregation. The materials mainly include organic photothermal agents or photosensitizers, polymer materials, metal nanoparticles, quantum dots, carbon-based nanomaterials, etc. In addition, compared to traditional fluorescent dyes, aggregation-induced emission (AIE) molecules have the advantages of good stability, low background signals, and strong resistance to photobleaching for bioimaging. Some AIE-based materials exhibit excellent photothermal and photodynamic effects, showing broad application prospects in the diagnosis and therapy of AD. We further summarize the advances in the detection of Aβ aggregates and phototherapy of AD using AIE-based materials. Full article

Some pancreatic ductal-type (PDADK) and lung adenocarcinomas (LADK) lacking other molecular drivers are reported to harbor NRG1 fusions as potential novel therapeutic targets. We investigated the feasibility of a fluorescent in situ hybridization (FISH)-based diagnosis of NRG1 fusions in a case series of PDADK and LADK lacking other identified oncogenic drivers. First, among a case series of PDADK, KRAS analyses (PCR followed in PCR-negative cases by RNA sequencing—RNAseq) found 27/162 (16.7%) KRAS wild-type cases, among which 1/162 (0.6%) NRG1 fusion was diagnosed using FISH. Secondly, among a case series of LDAK, 191/446 (42.8%) cases had no molecular alterations in EGFR, KRAS, BRAF, HER2, MET, ALK, ROS1 and RET according to NGS and FISH analyses and, among them, 4/446 (0.9%) cases had NRG1 fusions using FISH. Finally, four additional cases out of the two previously mentioned cases series (1 PDADK and 3 LADK) with NRG1 fusions diagnosed by first-line RNAseq were also concluded as NRG1 FISH-positive. The NRG1 FISH tests for the nine NRG1 FISH-positive cases resulted in 50% to 80% of positive tumor nuclei, all with single 3′-NRG1 FISH signals. In our series, of the 22 cases analyzed with both NRG1 FISH (positivity criterion of at least 15% of tumor nuclei with a split between the 5′- and the 3′- parts of the probes and/or isolated single 3′-NRG1 signal) and RNAseq, 17 cases were FISH– RNAseq– and 5 cases were FISH+ RNAseq+ (no FISH+ RNAseq– or FISH– RNAseq+ cases in our study) resulting in 100% sensibility and specificity for the NRG1 FISH test. In the case of no access to RNAseq, NRG1 FISH consists of a valuable tool searching for NRG1 fusions in patients with advanced cancers. Full article

Förster resonance energy transfer (FRET)-based biosensors are versatile tools for obtaining insights into various biological processes. Their working principles are based on nonradiative energy transfer from donor to acceptor fluorophores. This energy transfer is responsible for a change in fluorescence intensity, which provides a basis for the detection of biomolecules. Advantageous features of FRET biosensors include their high sensitivity and specificity. Recently, there have been notable developments to extend the usage of FRET biosensors for diverse applications. In this review, we briefly summarize the state-of-the-art developments of FRET biosensors for cellular imaging, drug discovery, pathogen detection, and cancer diagnosis. Continued research on biosensor design, donor acceptor pair optimization, and integration of innovative materials can further extend the applications of FRET biosensors across health care settings. Full article

Blood analogs are widely employed in in vitro experiments such as particle image velocity (PIV) to secure hemodynamics, assisting pathophysiological diagnoses of neurovascular and cardiovascular diseases, as well as pre-surgical planning and intraoperative orientation. To obtain accurate physical parameters, which are critical for diagnosis and treatment, blood analogs should exhibit realistic non-Newtonian shear-thinning features. In this study, two types of blood analogs working under room temperature (293.15 K) were created to mimic the steady-state shear-thinning features of blood over a temperature range of 295 to 312 K and a shear range of 1~250 s⁻¹ at a hematocrit of ~40%. Type I was a general-purpose analog composed of deionized (DI) water and xanthan gum (XG) powder, while Type II was specially designed for PIV tests, incorporating DI water, XG, and fluorescent microspheres. By minimizing the root mean square deviation between generated blood analogs and an established viscosity model, formulas for both blood analogs were successfully derived for the designated temperatures. The results showed that both blood analogs could replicate the shear-thinning viscosities of real blood, with the averaged relative discrepancy < 5%. Additionally, a strong linear correlation was observed between body temperature and XG concentration in both blood analogs (coefficient of determination > 0.96): for Type I, 295–312 K correlates with 140–520 ppm, and for Type II, 295–315 K correlates with 200–560 ppm. This work bridges the gap between idealized steady-state non-Newtonian viscosity models of blood and the complexities of real-world physiological conditions, offering a versatile platform for advancing particle image velocimetry tests and hemodynamics modeling, optimizing therapeutic interventions, and enhancing biomedical technologies in temperature-sensitive environments. Full article

P-glycoprotein (P-gp) is a key element of cancer treatment resistance, actively extruding cytotoxic drugs from cells and diminishing their efficacy. While its role at the plasma membrane is well established, its intracellular localization, particularly on lysosomes, is increasingly recognized as a critical contributor to drug resistance. This study investigates four innovative LightSpot fluorescent compounds to detect and quantify both membrane and lysosomal P-gp in Triple-Negative Breast Cancer (TNBC) SUM1315 and DU4475 cell lines. Results highlighted lysosomal P-gp staining by the LightSpot-FL-1, LightSpot-BrX-1, and LightSpot-BdO-1 fluorescent compounds (Mander’s coefficients > 0.8 overlapping with LAMP2 immunostaining). After both cell lines were exposed to Olaparib, a significant increase in P-gp expression level and lysosomal distribution of P-gp was detected. Indeed, after 100 µM Olaparib exposure, LightSpot-FL-1 allowed us to quantify an increase in P-gp-positive lysosome number of 1293 and 334% for SUM1315 and DU4475 cells, respectively, compared to the control. Findings suggest that P-gp may relocate to lysosomes upon drug exposure, highlighting a dual resistance mechanism involving both membrane and lysosomal P-gp. This study demonstrated the potential of LightSpot fluorescent compounds to evaluate P-gp-mediated cell resistance to treatment and emphasized the need to assess global cell P-gp expression to improve cancer diagnosis. Full article

Background/Objectives: The clinically validated multiplex Oncuria bladder cancer (BC) assay quickly and noninvasively identifies disease risk and tracks treatment success by simultaneously profiling 10 protein biomarkers in voided urine samples. Oncuria uses paramagnetic bead-based fluorescence multiplex technology (xMAP^®; Luminex, Austin, TX, USA) to simultaneously measure 10 protein analytes in urine [angiogenin, apolipoprotein E, carbonic anhydrase IX (CA9), interleukin-8, matrix metalloproteinase-9 and -10, alpha-1 anti-trypsin, plasminogen activator inhibitor-1, syndecan-1, and vascular endothelial growth factor]. Methods: In a pilot study (N = 36 subjects; 18 with BC), Oncuria performed essentially identically across three different common analyzers (the laser/flow-based FlexMap 3D and 200 systems, and the LED/image-based MagPix system; Luminex). The current study compared Oncuria performance across instrumentation platforms using a larger study population (N = 181 subjects; 51 with BC). Results: All three analyzers assessed all 10 analytes in identical samples with excellent concordance. The percent coefficient of variation (%CV) in protein concentrations across systems was ≤2.3% for 9/10 analytes, with only CA9 having %CVs > 2.3%. In pairwise correlation plot comparisons between instruments for all 10 biomarkers, R² values were 0.999 for 15/30 comparisons and R² ≥ 0.995 for 27/30 comparisons; CA9 showed the greatest variability (R² = 0.948–0.970). Standard curve slopes were statistically indistinguishable for all 10 biomarkers across analyzers. Conclusions: The Oncuria BC assay generates comprehensive urinary protein signatures useful for assisting BC diagnosis, predicting treatment response, and tracking disease progression and recurrence. The equivalent performance of the multiplex BC assay using three popular analyzers rationalizes test adoption by CLIA (Clinical Laboratory Improvement Amendments) clinical and research laboratories. Full article

Bovine respiratory disease complex (BRDC) is one of the primary causes of morbidity, mortality, and economic loss in cattle worldwide. Accurate and rapid identification of causative pathogenic agents is essential for effective disease management and control. In this study, a novel multiplex fluorescence-based quantitative polymerase chain reaction (qPCR) assay was developed for the simultaneous detection of eight major pathogens associated with BRDC. The targeted pathogens included the following: bovine viral diarrhea virus (BVDV), bovine parainfluenza virus type 3 (BPIV3), bovine respiratory syncytial virus (BRSV), bovine coronavirus (BcoV), Mycoplasma bovis (M.bovis), Pasteurella multocida (PM), Mannheimia haemolytica (MH), and infectious bovine rhinotracheitis virus (IBRV). The assay was rigorously optimized to ensure high specificity with no cross-reactivity among targets. The limit of detection (LOD) was determined to be as low as 5 copies per reaction for all target pathogens. The coefficient of variation (CVs) for both intra-assay and inter-assay measurements were consistently below 2%, demonstrating excellent reproducibility. To validate the clinical utility of the assay, a total of 1012 field samples were tested, including 504 nasal swabs from Farm A and 508 from Farm B in Jiangsu Province. BVDV, BcoV, PM, and MH were detected from Farm A, with a BVDV-positive rate of 21.63% (109/504), BcoV-positive rate of 26.79% (135/504), PM-positive rate of 28.77% (145/504), and MH-positive rate of 15.08% (76/504). Also, BcoV, PM, MH, and IBRV were detected from Farm B, with a BcoV-positive rate of 2.36% (12/508), PM-positive rate of 1.38% (7/508), MH-positive rate of 14.76% (75/508), and IBRV-positive rate of 5.51% (28/508). Notably, a significant proportion of samples showed evidence of mixed infections, underscoring the complexity of BRDC etiology and the importance of a multiplex diagnostic approach. In conclusion, the developed multiplex qPCR assay provides a reliable, rapid, and cost-effective tool for simultaneous detection of multiple BRDC-associated pathogens, which will hold great promise for enhancing disease surveillance, early diagnosis, and targeted intervention strategies, ultimately contributing to improved BRDC management and cattle health outcomes. Full article

In this study, we describe the preparation of carbon dots (CDs) from natural charcoal by laser ablation in a liquid. A continuum wave (CW) laser diode operating at a wavelength of 450 nm, hitting a solid carbon target placed into a biocompatible liquid, constituted of a phosphate-buffered saline (PBS) solution and distilled water, was used for the generation of the CDs suspension. Exploring the practical applications of carbon dots, it was observed that the luminescence of the produced CDs can be used as bioimaging in living organisms, environmental monitoring, chemical analysis, targeted drug delivery, disease diagnosis, therapy, and others. The CDs’ luminescence can be induced by UV irradiation and, as demonstrated in this study, by energetic MeV proton beams. The fluorescence was revealed mainly at 480 nm when UV illuminated the CDs, and also in the region at 514–642 nm when the CDs were irradiated by energetic proton ions. Atomic force microscopy (AFM) of the CD films revealed their spherical shape with a size of about 10 nm. The significance of the manuscript lies in the use of CDs produced by laser ablation exhibiting luminescence under irradiation of an energetic proton beam. Full article

The petiole nitrate–nitrogen concentration (PNNC) has been an industry standard indicator for in-season potato (Solanum tuberosum L.) nitrogen (N) status diagnosis. Leaf sensors can be used to predict the PNNC and other N status indicators non-destructively. The SPAD meter is a common leaf chlorophyll (Chl) meter, while the Dualex is a newer leaf fluorescence sensor. Limited research has been conducted to compare the two leaf sensors for potato N status assessment. Therefore, the objectives of this study were to (1) compare SPAD and Dualex for predicting potato N status indicators, and (2) evaluate the potential prediction improvement using multi-source data fusion. The plot-scale experiments were conducted in Becker, Minnesota, USA, in 2018, 2019, 2021, and 2023, involving different cultivars, N treatments, and irrigation rates. The results indicated that Dualex’s N balance index (NBI; Chl/Flav) always outperformed Dualex Chl but did not consistently perform better than the SPAD meter. All N status indicators were predicted with significantly higher accuracy with multi-source data fusion using machine learning models. A practical strategy was developed using a linear support vector regression model with SPAD, cultivar information, accumulated growing degree days, accumulated total moisture, and an as-applied N rate to predict the vine or whole-plant N nutrition index (NNI), achieving an R² of 0.80–0.82, accuracy of 0.75–0.77, and Kappa statistic of 0.57–0.58 (near-substantial). Further research is needed to develop an easy-to-use application and corresponding in-season N recommendation strategy to facilitate practical on-farm applications. Full article