Neutrophil recruitment and abundance in the lungs of people with cystic fibrosis (CF) perpetuates a cycle of chronic pulmonary inflammation. Phospholipids comprise the majority of pulmonary surfactant and, in a disease state such as those in the CF lung, altered phospholipid distribution and abundance implicates metabolic and cellular dysregulation. We explored the impact of neutrophils on the lung lipidomic landscape utilizing the β-ENaC transgenic mouse (as a phenocopy model of the CF lung environment) and infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging (MSI). Histology-driven MSI, supported by localization-dependent tissue isolation of epithelial-lined airways and parenchyma, revealed significant lipid remodeling associated with transgenic lungs and neutrophil load. For the first time, IR-MALDESI MSI identified 85 differential lipids and metabolites, including the critical neutrophil recruitment factor Leukotriene B4 (LTB4). Phosphatidylglycerols (PG) and diacylglycerols (DG) were differentially upregulated in β-ENaC tissues, while PAs were downregulated relative to WT tissue. These alterations coincided with significant mucus plugging, supporting the broad remodeling of lung lipid composition that occurs in CF. Notably, induced neutropenia in the transgenic lung shifted or completely restored the β-ENaC lipidomic profiles to the wildtype baseline across both the parenchyma and epithelial-lined airway features. These findings identify the consequences of sodium/chloride transport imbalance and neutrophil loading on spatial lipidomic remodeling in lung tissue.
Closed-tube, one-pot CRISPR diagnostics based on light-triggered crRNA uncaging offer a practical strategy for contamination-free nucleic acid detection, yet buffer incompatibility, reagent costs, and instrument dependence remain limiting factors. Here, we report a light-responsive, dual-modal, one-pot platform that integrates ERA (enzymatic recombinase amplification) with CRISPR-Cas12a for rapid Mycoplasma pneumoniae (MP) detection. A photocleavable linker-modified blocking strand suppresses Cas12a activity during amplification and releases functional crRNA upon brief UV irradiation, enabling on-demand activation within a sealed tube. A unified buffer was developed to support both ERA and Cas12a activity without compromising either component. Under the specific procurement conditions of this study, the per-reaction reagent cost was USD 3.21 for bulk-purchased ERA reagents, compared with USD 7.47 for RPA reagents. The assay achieves limits of detection of 5 copies/μL by real-time fluorescence and 10 copies/μL by LFA (lateral flow assay). Within this study cohort of 71 clinical nasopharyngeal swab specimens, the fluorescence readout yielded 100
Sulfur dioxide (SO2) derivatives, mainly bisulfite (HSO3−) and sulfite (SO32−), are widely used in food processing as preservatives, antioxidants, and antibrowning agents, but excessive residues may pose health risks to sensitive individuals. Ratiometric fluorescent probes offer a self-calibrated strategy for SO2 derivative detection by recording two emission signals, thereby reducing errors from probe concentration, excitation fluctuation, and complex food backgrounds. This review summarizes recent advances in ratiometric fluorescent probes for SO2 derivatives in food analysis. Reaction mechanisms, including addition to electron-deficient C=C bonds, aldehyde addition, deprotection, and cleavage reactions, are discussed with design strategies based on intramolecular charge transfer (ICT), Förster resonance energy transfer (FRET), excited-state intramolecular proton transfer (ESIPT), and aggregation-induced emission (AIE). Applications in alcoholic beverages, sugar products, other foods, traditional Chinese medicines, and food-related plant tissues are highlighted. Current challenges and future directions for water-compatible probes, portable and real-time readout, matrix-resilient analysis, and standardized validation are also discussed. This review aims to provide guidance for developing robust ratiometric fluorescent probes suitable for complex food systems.
Kahweol (KAH), a bioactive diterpene abundant in unfiltered coffee, has drawn growing pharmacological interest for its anticancer, anti-inflammatory, anti-angiogenic, and metabolic regulatory properties. However, sensitive and rapid methods for its determination in biological matrices remain scarce. This study developed a molecularly imprinted polymer (MIP)-based electrochemical sensor for selective KAH detection using a glassy carbon electrode modified with a sodium alginate-multi-walled carbon nanotube nanocomposite. The imprinted film was formed by electropolymerizing methacrylic acid (MA) with aniline (ANI) as a co-monomer around the KAH template, producing a poly(MA@ANI) recognition layer. Fabrication was optimized for nanocomposite composition, monomer-to-template ratio, polymerization cycles, template removal, and rebinding time. Under optimized conditions, differential pulse voltammetry (DPV) showed a linear range of 2.5 × 10–12 to 2.5 × 10–11 M, with a limit of detection (LOD) of 1.17 × 10–13 M and a quantification limit (LOQ) of 3.51 × 10–13 M. Selectivity tests, including imprinting factor calculations and competitive binding against caffeic acid, caffeine, and interferents, confirmed high specificity toward KAH. Recovery experiments in real rabbit serum gave an LOD of 2.88 × 10–13 M, and favorable AGREE, AGREE-MIP, and ComplexMoGAPI scores supported the method’s greenness. Computational modeling (MD, RDF, and DFT) supported the MIP design, revealing stable KAH–monomer complexation through strong hydrogen bonding. This is the first electrochemical determination of KAH in a biological matrix and offers a rapid, cost-effective, green alternative for bioanalytical use.
Dysregulated amino acid (AA) profiles are closely associated with diseases. Comprehensive and quantitative profiling of AA is therefore essential. This study developed a simple, direct, and high-throughput serum AA profiling method and evaluated its auxiliary diagnostic potential for type 2 diabetes mellitus (T2DM). A cold-induced phase separation strategy coupled with ultra-performance liquid chromatography–tandem mass spectrometry was developed, enabling comprehensive serum amino acid profiling of 27 AAs and related metabolites across branched-chain, aromatic, and tryptophan metabolic pathways. The optimized sample preparation used acetonitrile (ACN) at a serum-to-solvent ratio of 1:3, 50 min of cold-induced phase separation, and reconstitution in ACN/H2O (v/v, 1:4). The method showed good linearity (R2 ≥ 0.9929), limits of detection of 0.005–5.00 ng/mL, limits of quantification of 0.01–10.00 ng/mL, recoveries of 82.6
