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The molecular chaperone sigma 1 receptor mediates rescue of retinal cone photoreceptor cells via modulation of NRF2
摘要： Sigma 1 receptor (Sig1R), a putative molecular chaperone, has emerged as a novel therapeutic target for retinal degenerative disease. Earlier studies showed that activation of Sig1R via the high-affinity ligand (+)-pentazocine ((+)-PTZ) induced profound rescue of cone photoreceptor cells in the rd10 mouse model of retinitis pigmentosa; however the mechanism of rescue is unknown. Improved cone function in (+)-PTZ-treated mice was accompanied by reduced oxidative stress and normalization of levels of NRF2, a transcription factor that activates antioxidant response elements (AREs) of hundreds of cytoprotective genes. Here, we tested the hypothesis that modulation of NRF2 is central to Sig1R-mediated cone rescue. Activation of Sig1R in 661W cone cells using (+)-PTZ induced dose-dependent increases in NRF2-ARE binding activity and NRF2 gene/protein expression, whereas silencing Sig1R significantly decreased NRF2 protein levels and increased oxidative stress, although (+)-PTZ did not disrupt NRF2-KEAP1 binding. In vivo studies were conducted to investigate whether, in the absence of NRF2, activation of Sig1R rescues cones. (+)-PTZ was administered systemically for several weeks to rd10/nrf2+/+ and rd10/nrf2-/- mice. Through post-natal day 42, cone function was significant in rd10/nrf2+/+, but minimal in rd10/nrf2-/- mice as indicated by electroretinographic recordings using natural noise stimuli, optical coherence tomography and retinal histological analyses. Immunodetection of cones was limited in (+)-PTZ-treated rd10/nrf2-/-, though considerable in (+)-PTZ-treated rd10/nrf2+/+mice. The data suggest that Sig1R-mediated cone rescue requires NRF2 and provide evidence for a previously-unrecognized relationship between these proteins.
关键词： retinitis pigmentosa,NRF2-KEAP1,retinal neuroprotection,retina,rd10 mouse,NRF2-Neh luciferase assay,oxidative stress
更新于2025-11-21 11:08:12
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Screening of two-photon activated photodynamic therapy sensitizers using a 3D osteosarcoma model
摘要： Photodynamic therapy (PDT) involves a photosensitizing agent activated with light to induce cell death. Two-photon excited PDT (TPE-PDT) offers numerous benefits compared to traditional one-photon induced PDT, including an increased penetration depth and precision. However, the in vitro profiling and comparison of two-photon photosensitizers (PS) are still troublesome. Herein, we report the development of an in vitro screening platform of TPE-PS using a 3D osteosarcoma cell culture. The platform was tested using three different two-photon (2P) active compounds – a 2P sensitizer P2CK, a fluorescent dye Eosin Y, and a porphyrin derivative (TPP). Their 2P absorption cross-sections (σ2PA) were characterised using a fully automated z-scan setup. TPP exhibited a remarkably high σ2PA at 720 nm (8865 GM) and P2CK presented a high absorption at 850 nm (405 GM), while Eosin Y had the lowest 2P absorption at the studied wavelengths (<100 GM). The cellular uptake of PS visualized using confocal laser scanning microscopy showed that both TPP and P2CK were internalized by the cells, while Eosin Y stayed mainly in the surrounding media. The efficiency of the former two TPE-PS was quantified using the PrestoBlue metabolic assay, showing a significant reduction in cell viability after two-photon irradiation. The possibility of damage localization was demonstrated using a co-culture of adipose derived stem cells together with osteosarcoma spheroids showing no signs of damage to the surrounding healthy cells after TPE-PDT.
关键词： two-photon excited photodynamic therapy,PrestoBlue assay,photosensitizers,cellular uptake,localized damage,z-scan,3D osteosarcoma model
更新于2025-11-21 11:08:12
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Gold nanoparticle-based plasmonic probe for selective recognition of adenosine
摘要： Adenosine, as an endogenous molecule in organisms, plays an essential role in biological processes. Here, a plasmonic probe, creatinine-Ag+/gold nanoparticle (AuNPs), is assembled for adenosine detection based on synergistic coordination on AuNPs. The A650 nm/520 nm values of AuNPs system change linearly with adenosine concentration over a range of 1.0–5.0 μM and the detection limits reached 45 nM. The adenosine detection is realized within 4 min. Furthermore, the quantitative detection of adenosine is realized by eyedropper (a function in Microsoft’s PowerPoint) for analyzing RGB value changes of colorimetric assay. Therefore, this sensor can provide accurate and rapid assay of adenosine in patients’ serum sample without complicated instrumentations.
关键词： colorimetric assay,adenosine detection,eyedropper function,gold nanoparticle,plasmonic probe
更新于2025-11-19 16:56:42
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Opposite changing dual-emission luminescence of gold nanoparticles by sulfhydryl to develop a pesticide biosensing strategy
摘要： As the merit of ratiometric assay is impregnable due to potentially interfering processes, a ratiometric method for pesticide detection was developed. By adjusting glutathione : HAuCl4 to an appropriate ratio, dual-emission luminescent ultra-small gold nanoparticles (AuNPs) with a high emission at 800 nm and a low emission at 600 nm were synthesized. Interestingly, the sulfhydryl-containing compounds were found to result in completely opposite changes to strengthen the 600 nm emission and weaken the 800 nm emission. Therefore, dual-emitted AuNPs were engaged to develop a ratiometric pesticide biosensing strategy. In the presence of acetylcholinesterase (AChE), acetylthiocholine can be hydrolyzed into thiocholine, whose newly generated sulfhydryl can interact with AuNPs, resulting in the opposite change of the dual emissions. While adding pesticide as an AChE inhibitor, the catalytic activity of AChE is inhibited and less thiocholine was produced. The biosensing system shows an obvious sensitivity to the pesticide with a limit of detection (LOD) of 0.2 nM for aldicarb and 0.07 nM for chlorpyrifos. Therefore, this simple assay is suitable for AChE activity and pesticide detection, even in vegetable samples.
关键词： sulfhydryl,ratiometric assay,gold nanoparticles,AChE activity,biosensing,pesticide detection
更新于2025-11-19 16:56:35
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Luminescent rhenium(I) carbonyl complex with redox noninnocent ONS donor azo-phenol ligand: Synthesis, X-ray structure, photophysical properties and live cell imaging
摘要： Herein, we have synthesized a new fluorescent rhenium(I) carbonyl complex 1, bearing {Re(CO)3}+ core with ONS donor thioether containing azo-phenol redox noninnocent ligand. The distorted octahedral geometry of the complex is confirmed by single crystal X-ray diffraction method. Cyclic voltammogram in acetonitrile exhibits irreversible oxidation peak (Epa = 1.36 V) along with quasi-reversible reduction peak at E1/2 = -0.92 V (ΔE = 210 mV). The complex exhibits low energy emission band at 525 nm with high emission quantum yield (Φ = 0.115). Cytotoxicity of the complex is studied by MTT method with human breast cancer cell lines (MCF-7) and IC50 value is found to be 23.6 μM. In presence of the complex (10 μM) a bright green fluorescence image of MCF-7 cell lines is observed under fluorescence microscope.
关键词： Electrochemistry,ONS donor ligand,MTT assay,Rhenium(I) carbonyl complex,Live cell imaging
更新于2025-11-19 16:46:39
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Fluorescent turn-on probes for the development of binding and hydrolytic activity assays for mRNA cap-recognising proteins
摘要： The m7G cap is a unique nucleotide structure at the 5' end of all eukaryotic mRNAs. The cap specifically interacts with numerous cellular proteins and participates in biological processes essential for cell growth and function. To provide small molecular probes to study important cap-recognising proteins, we synthesized m7G nucleotides labelled with fluorescent tags via the terminal phosph(on)ate group and studied how their emission properties changed upon protein binding or enzymatic cleavage. Only the pyrene-labelled compounds behaved as sensitive turn-on probes. A pyrene-labelled m7GTP analogue showed up to 8-fold enhanced fluorescence emission upon binding to eukaryotic translation initiation factor 4E (eIF4E) and over 30-fold enhancement upon cleavage by decapping scavenger (DcpS) enzyme. These observations served as the basis for developing binding- and hydrolytic-activity assays. The assay utility was validated with previously characterized libraries of eIF4E ligands and DcpS inhibitors. The DcpS assay was also applied to study hydrolytic activity and inhibition of endogenous enzyme in cytoplasmic extracts from HeLa and HEK cells.
关键词： eIF4E,fluorescence assay,cap,pyrene,DcpS
更新于2025-11-14 15:32:45
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A Simple Colorimetric and Fluorescent Sensor to Detect Organophosphate Pesticides Based on Adenosine Triphosphate-Modified Gold Nanoparticles
摘要： A simple and dual modal (colorimetric and fluorescent) sensor for organophosphate pesticides with high sensitivity and selectivity using adenosine triphosphate (ATP)- and rhodamine B-modified gold nanoparticles (RB-AuNPs), was successfully fabricated. This detection for ethoprophos afforded colorimetric and fluorescence imaging changes visualization. The quantitative determination was linearly proportional to the amounts of ethoprophos in the range of a micromolar scale (4.0–15.0 μM). The limit of detection for ethoprophos was as low as 37.0 nM at 3σ/k. Moreover, the extent application of this simple assay was successfully demonstrated in tap water samples with high reliability and applicability, indicating remarkable application in real samples.
关键词： multimodal assay,ethoprophos detection,gold nanoparticles,colorimetric and fluorescent sensor,organophosphate pesticides
更新于2025-09-23 15:23:52
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A smartphone-based system for fluorescence polarization assays
摘要： This paper demonstrates the use of a smartphone-based sensor for fluorescence polarization (FP) analysis of biomolecules. The FP detection can rapidly sense ligand-analyte bindings by measuring molecule mobility, and thus, FP-based assays have been widely used for rapid diagnostics in clinics. Here, we implemented the FP detection apparatus using a 3D-printed compact holder and the built-in camera of a smartphone. The system offers accurate measurements of the degree of polarization by simultaneously detecting the fluorescence intensities parallel and perpendicular to the polarization of the excitation. The fluorescence signal of the sample is excited by a laser or light-emitting diode and separated by a polarization beam cube depending on the polarization. Parallel and perpendicular polarized emissions are projected onto two different regions of the sensor chip in the smartphone camera. A custom software app was developed to count the average intensity in the areas of interest and compute the degree of polarization. We validated the system by measuring the polarization of dye molecules dissolved in solutions with different viscosities. As an example of biomolecule sensing, a competitive FP immunoassay of Prostaglandin E2 was demonstrated using the developed system and exhibited the limit of detection of 1.57 ng/mL. The smartphone-based FP assay platform can also be implemented for the detection of toxins, disease biomarkers, and pathogens in resource-limited settings.
关键词： immunoassay,mobile sensor,fluorescence polarization assay,smartphone
更新于2025-09-23 15:23:52
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NIR-Fluorescent Multidye Silica Nanoparticles with Large Stokes Shifts for Versatile Biosensing Applications
摘要： We have synthesized and characterized of a series of single and multidye copolymerized nanoparticles with large to very large Stokes shifts (100 to 255 nm) for versatile applications as standalone or multiplexed probes in biological matrices. Nanoparticles were prepared via the St?ber method and covalently copolymerized with various combinations of three dyes, including one novel aminocyanine dye. Covalently encapsulated dyes exhibited no significant leakage from the nanoparticle matrix after more than 200 days of storage in ethanol. Across multiple batches of nanoparticles with varying dye content, the average yields and average radii were found to be highly reproducible. Furthermore, the batch to batch variability in the relative amounts of dye incorporated was small (relative standard deviations <2.3%). Quantum yields of dye copolymerized nanoparticles were increased 50% to 1000% relative to those of their respective dye-silane conjugates, and fluorescence intensities were enhanced by approximately three orders of magnitude. Prepared nanoparticles were surface modified with polyethylene glycol and biotin and bound to streptavidin microspheres as a proof of concept. Under single wavelength excitation, microsphere-bound nanoparticles displayed readily distinguishable fluorescence signals at three different emission wavelengths, indicating their potential applications to multicolor sensing. Furthermore, nanoparticles modified with polyethylene glycol and biotin demonstrated hematoprotective qualities and reduced nonspecific binding of serum proteins, indicating their potential suitability to in vivo imaging applications.
关键词： Fluorescent silica nanoparticles,Biocompatible nanoparticles,Large stokes shift,Near-infrared fluorescence,Multicolor assay,Resonance energy transfer
更新于2025-09-23 15:23:52
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[IEEE 2018 IEEE Life Sciences Conference (LSC) - Montreal, QC, Canada (2018.10.28-2018.10.30)] 2018 IEEE Life Sciences Conference (LSC) - Assay Development and Storage for Fluorescence-Based Lateral Flow Immunoassay
摘要： Point-of-care medical diagnostics can provide efficient, cost-effective medical care, and have the potential to fundamentally change our current approach to global health. There have been substantial efforts in developing lateral flow assays for serologic testing, but most of the existing approaches have limited portability, are expensive, and offer limited analytical sensitivity. In this paper, we demonstrated an assay for the detection of antibodies in plasma to Epstein-Barr Nuclear Antigen-1 (EBNA-1) protein and optimization of the assay including washing and blocking conditions. We also investigated the effect of the storage on the assay strips. Using our optimized conditions, we were able to detect anti-Human Papilloma Virus (HPV)-16 E7 antibodies after three weeks of storage. Our goal is to adapt this system to detect HPV biomarkers for cervical cancers in low and middle-income countries.
关键词： storage,immunoassay,sensitivity,Lateral flow assay,fluorescence,point-of-care
更新于2025-09-23 15:22:29