A model of STAT1-p53 coordination: Regulating p21 and PUMA to program cell fate under stress.

Neuronal nitric oxide synthase in hypothalamic leptin signaling.

Why radical oxidation prevails: Dual advantages in sludge valorization and ARG suppression over nonradical pathways.

A second near-infrared light-activated nanoplatform with spatiotemporal signal transduction for simultaneous enrichment and portable detection of circulating tumor cells.

RACK1: A multifunctional scaffold protein involved in signal transduction, ribosomal regulation, and Cancer pathogenesis.

Here, we report a simple and rapid extracellular vesicle (EV)-based membrane-type immunoglobulin-directed hybridoma screening (MIHS) platform that enables efficient isolation of monoclonal antibodies (mAbs) recognizing three-dimensional conformations of transmembrane proteins. Transmembrane proteins are essential membrane components involved in signal transduction, transport, and energy conversion. As they function in native conformations, isolating mAbs that recognize conformational epitopes is critical for both research and therapeutic use. However, traditional methods, such as the enzyme-linked immunosorbent assay (ELISA), can disrupt the antigen structure, making it challenging to screen for such mAbs. To overcome this limitation, we developed a screening strategy combining MIHS with EVs as antigen-presenting tools. EVs, which are naturally bounded by lipid bilayers, display transmembrane proteins in native conformations and can be easily isolated. As a model, we engineered cells co-expressing alkaline phosphatase (ALP) and green fluorescent protein, generating fluorescent EVs displaying ALP. Hybridomas were screened by MIHS using these EVs, followed by ELISA without direct antigen immobilization. All isolated mAbs recognized conformational ALP epitopes, demonstrating the structural selectivity of this method. These results demonstrate that the EV-based MIHS approach provides a convenient and structurally faithful strategy for isolating conformation-specific mAbs, offering broad utility in basic biology and antibody development.

A simple and instrument-free colorimetric method for quantitative analysis is reported, in which analyte concentration is determined by measuring the length of a colored adsorption band formed in a packed silica gel column. The proposed method employs a miniature silica gel column that acts as a signal transducer after it adsorbs the colored compound from a solution during its flow in the column. The length of this band increases proportionally with analyte concentration, which enables quantitative detection via simple distance measurement. A theoretical model was developed to describe the relationship between solute concentration, adsorption behavior, and band propagation along the column. The principle was validated via the detection of both iron ions and enzyme-mediated glutamic acid. For Fe2+ analysis, the o-phenanthroline complexation method shows a level of sensitivity comparable to that of conventional spectrophotometry, which enables an almost quantitative recovery of trace iron in tap water. The limit of detection (LOD) and the limit of quantification (LOQ) were estimated to be 0.20μM and 0.60μM for the proposed method and 0.23μM and 0.70μM for spectrophotometry, respectively. The approach was further extended to glutamate detection using a cascade reaction involving glutamate oxidase and horseradish peroxidase with N-benzoyl leucomethylene blue as the chromogenic substrate. The LOD and the LOQ of the proposed method were 0.08 and 0.25μM, and both values are superior to the 0.24μM and 0.73μM obtained using a microplate reader. By integrating preconcentration with a distance-based readout, this method provides a simple yet highly sensitive analytical platform and establishes distance as a quantitative signal for colorimetric detection.

In silico exploration of the RAPTOR-STAT3 interface reveals potential crosstalk between mTORC1 and JAK-STAT pathways in Sjogren's syndrome and its linked cancer.

To investigate the tumor-intrinsic lymphocyte cytosolic protein 2 (LCP2) in esophageal squamous cell carcinoma (ESCA) and its molecular mechanisms in mediating resistance to programmed death-1 (PD-1) therapy. The expression of LCP2 in ESCA was analyzed using bioinformatics databases and further verified in clinical specimens. Functional studies employed patient-derived organoid models, xenograft models, and molecular assays to assess the impact of LCP2 knockout or overexpression on macrophage polarization, CD8 + T cell exhaustion, and PD-1 therapy response. Mechanistic investigations included nuclear factor-κB (NF-κB) inhibition, signal transducer and activator of transcription 5A (STAT5A) knockdown, chromatin immunoprecipitation, and dual-luciferase reporter assays. LCP2 was markedly upregulated in ESCA and correlated with advanced stage, lymph node metastasis, and poor survival. Tumor-intrinsic LCP2 expression positively correlated with M2 macrophage polarization and sorted CD8+ T cell exhaustion. Mechanistically, this association depended on NF-κB pathway activation in EpCAM+ tumor fractions, while STAT5A transcriptionally regulates tumor-intrinsic LCP2 expression as an upstream transcription factor. Knockdown of tumor-intrinsic LCP2 or STAT5A in EpCAM+ tumor fractions suppressed the secretion of immunosuppressive cytokines and restored effector T cell function of sorted CD8+ T cells. In vivo, LCP2 depletion significantly inhibited tumor growth and synergized with PD-1 blockade. This synergistic effect was characterized by reduced tumor volume and increased CD8+ T cell infiltration. Overexpression of LCP2 reversed these effects, confirming its central role in immune escape. The STAT5A-LCP2-NF-κB axis remodels the immunosuppressive tumor microenvironment to mediate ESCA immune escape and PD-1 resistance. Targeting this regulatory axis provides a novel strategy to overcome immunotherapy resistance in esophageal cancer. Antioxid. Redox Signal. 45, 242-264.

4-Octyl itaconate blocks STAT3-induced ACE2 expression to reduce uptake of SARS-CoV-2.

Bifunctional DNA multivalent structure integrating stable capture of urothelial carcinoma cells with CRISPR/Cas12a signal amplification for bladder cancer detection.

ZmCIPK14 acts as a positive regulator of the response of Arabidopsis to low-temperature stress.

Bioelectronic interface for real-time brain biomarker analysis.

Brassinosteroid regulation of quality in horticultural crops: Physiological and molecular mechanisms from preharvest development to postharvest preservation.

Endogenous enzyme-driven unarming of spherical nucleic acid in a divide-and-conquer pattern for amplified imaging of microRNA in tumor cells.

Mice with tamoxifen-inducible endothelium-restricted human endothelin-1 (ET-1) overexpression (ieET-1) exhibited blood pressure elevation after 3 weeks and 3 months, and vascular injury after 3 months. We hypothesised that ET-1 overexpression-induced vascular injury is mediated by gene expression dysregulation. Male ieET-1 mice and mice expressing a tamoxifen-inducible Cre recombinase in endothelial cells were induced with tamoxifen and studied 3-week and 3-month post-treatment. MicroRNA and gene expression were profiled in mesenteric arteries by RNA-sequencing and validated by RT-qPCR. Aorta was used for immunofluorescence microscopy. KH domain containing RNA binding signal transduction associated 3 ( Khdrbs3 ) was the top upregulated gene identified by RNA-sequencing ( P = 8x10 -12 ) validated by RT-qPCR ( P < 0.001) after 3-week and 3-month ET-1 overexpression. KHDRBS3 protein was increased in aortic endothelial and vascular smooth muscle cells (VSMCs) after 3-month ET-1 overexpression. KHDRBS3 is a splicing regulator of various mRNAs, including human vascular endothelial growth factor A ( VEGFA ) 165 isoform with exon 7 retention. Three-month ET-1 overexpression upregulated isoforms Vegfa164 containing exon 7 and Vegfa188 retaining exons 6 and 7, and downregulated Vegfa120 isoform lacking both exons ( P < 0.05). Analysis of RNA-immunoprecipitation-sequencing of myocardium of patients with dilated cardiomyopathy identified VEGFA and four key VSMC-specific pre-mRNAs as KHDRBS3 targets. Reanalysis of RNA-sequencing data of angiotensin II-infused mice revealed a high correlation between ET-1 ( Edn1 ) and Khdrbs3 mRNAs in mesenteric arteries ( r2 = 0.82, P = 3.7 x 10 -9 ). Endothelial human ET-1 overexpression induced RNA splicing via enhanced Khdrbs3 expression, which may play a role in vascular injury, and could represent a therapeutic target in hypertension.

Polyhistidine-functionalized phosphorene-ZnO nanorods as an EGFET urea biosensor via enzymatically induced local pH modulation.

The complex mechanistic network of periprosthetic osteolysis: Integration from inflammatory response and tissue stress to signaling pathways.

Comparative biomechanical and molecular mechanisms shaping erect and prostrate winter growth habits in wheat.

MOF-based composite sensors for nitrogen-containing compounds: From selective capture to signal transduction