scholarly journals Rapid-Response and Highly Sensitive Boronate Derivative-Based Fluorescence Probe for Detecting H2O2 in Living Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Muthusamy Selvaraj ◽  
Kanagaraj Rajalakshmi ◽  
Yun-Sik Nam ◽  
Yeonhee Lee ◽  
Byoung Chan Kim ◽  
...  

Intracellular H2O2 monitoring is important and has driven researchers to pursue advancements for the rapid identification of H2O2, since H2O2 is short-lived in cell lines. An arylboronate derivative has been investigated as a chemospecific fluorescence recognition agent for H2O2. Triphenylimidazoleoxadiazolephenyl (TPIOP) boronate was contrived as a novel candidate for the rapid and sensitive recognition of H2O2. The probe was conjugated using the TPIOP functional group acting as an excellent fluorescent enhancer. The TPIOP group stimulated the polarization of C–B bond due to its extended π-conjugation, which included heteroatoms, and induced the production of rapid signal because of the highly polar C–B bond along with the corresponding boronate unit. While H2O2 reacts with TPIOP boronate, its nucleophilic addition to the boron generates a charged tetrahedral boronate complex, and then the C–B bond migrates toward one of the electrophilic peroxide oxygen atoms. The resulting boronate ester is then hydrolyzed by water into a phenol, which significantly enhances fluorescence through aggregation-induced emission. The TPIOP boronate probe responded to H2O2 rapidly, within 2 min, and exhibited high sensitivity with a limit of detection of 8 nM and a 1000-fold selectivity in the presence of other reactive oxygen species. Therefore, the developed TPIOP boronate chemodosimeter was successfully utilized to visualize and quantify intracellular H2O2 from human breast cancer (MCF-7) cells, as well as gaseous and aqueous H2O2 from environmental samples using Whatman paper strips coated with TPIOP boronate.

2022 ◽  
Vol 12 ◽  
Author(s):  
Katharina Radakovics ◽  
Claire Battin ◽  
Judith Leitner ◽  
Sabine Geiselhart ◽  
Wolfgang Paster ◽  
...  

Toll-like receptors (TLRs) are primary pattern recognition receptors (PRRs), which recognize conserved microbial components. They play important roles in innate immunity but also in the initiation of adaptive immune responses. Impurities containing TLR ligands are a frequent problem in research but also for the production of therapeutics since TLR ligands can exert strong immunomodulatory properties even in minute amounts. Consequently, there is a need for sensitive tools to detect TLR ligands with high sensitivity and specificity. Here we describe the development of a platform based on a highly sensitive NF-κB::eGFP reporter Jurkat JE6-1 T cell line for the detection of TLR ligands. Ectopic expression of TLRs and their coreceptors and CRISPR/Cas9-mediated deletion of endogenously expressed TLRs was deployed to generate reporter cell lines selectively expressing functional human TLR2/1, TLR2/6, TLR4 or TLR5 complexes. Using well-defined agonists for the respective TLR complexes we could demonstrate high specificity and sensitivity of the individual reporter lines. The limit of detection for LPS was below 1 pg/mL and ligands for TLR2/1 (Pam3CSK4), TLR2/6 (Fsl-1) and TLR5 (flagellin) were detected at concentrations as low as 1.0 ng/mL, 0.2 ng/mL and 10 pg/mL, respectively. We showed that the JE6-1 TLR reporter cells have the utility to characterize different commercially available TLR ligands as well as more complex samples like bacterially expressed proteins or allergen extracts. Impurities in preparations of microbial compounds as well as the lack of specificity of detection systems can lead to erroneous results and currently there is no consensus regarding the involvement of TLRs in the recognition of several molecules with proposed immunostimulatory functions. This reporter system represents a highly suitable tool for the definition of structural requirements for agonists of distinct TLR complexes.


Nanomaterials ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 496 ◽  
Author(s):  
Xi Zhou ◽  
Yongna Zhang ◽  
Jun Yang ◽  
Jialu Li ◽  
Shi Luo ◽  
...  

Wearable pressure sensors have attracted widespread attention in recent years because of their great potential in human healthcare applications such as physiological signals monitoring. A desirable pressure sensor should possess the advantages of high sensitivity, a simple manufacturing process, and good stability. Here, we present a highly sensitive, simply fabricated wearable resistive pressure sensor based on three-dimensional microstructured carbon nanowalls (CNWs) embedded in a polydimethylsiloxane (PDMS) substrate. The method of using unpolished silicon wafers as templates provides an easy approach to fabricate the irregular microstructure of CNWs/PDMS electrodes, which plays a significant role in increasing the sensitivity and stability of resistive pressure sensors. The sensitivity of the CNWs/PDMS pressure sensor with irregular microstructures is as high as 6.64 kPa−1 in the low-pressure regime, and remains fairly high (0.15 kPa−1) in the high-pressure regime (~10 kPa). Both the relatively short response time of ~30 ms and good reproducibility over 1000 cycles of pressure loading and unloading tests illustrate the high performance of the proposed device. Our pressure sensor exhibits a superior minimal limit of detection of 0.6 Pa, which shows promising potential in detecting human physiological signals such as heart rate. Moreover, it can be turned into an 8 × 8 pixels array to map spatial pressure distribution and realize array sensing imaging.


2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Aitor Urrutia ◽  
Kartheka Bojan ◽  
Leonel Marques ◽  
Kevin Mullaney ◽  
Javier Goicoechea ◽  
...  

Novel protein sensors based on tapered optical fibres modified with Au coatings deposited using two different procedures are proposed. Au-based coatings are deposited onto a nonadiabatic tapered optical fibre using (i) a novel facile method composed of layer-by-layer deposition consisting of polycation (poly(allylamine hydrochloride), PAH) and negatively charged SiO2nanoparticles (NPs) followed by the deposition of the charged Au NPs and (ii) the sputtering technique. The Au NPs and Au thin film surfaces are then modified with biotin in order to bind streptavidin (SV) molecules and detect them. The sensing principle is based on the sensitivity of the transmission spectrum of the device to changes in the refractive index of the coatings induced by the SV binding to the biotin. Both sensors showed high sensitivity to SV, with the lowest measured concentration levels below 2.5 nM. The calculated binding constant for the biotin-SV pair was2.2×10-11 M−1when a tapered fibre modified with the LbL method was used, with a limit of detection (LoD) of 271 pM. The sensor formed using sputtering had a binding constant of1.01×10-10 M−1with a LoD of 806 pM. These new structures and their simple fabrication technique could be used to develop other biosensors.


2019 ◽  
Vol 19 (6) ◽  
pp. 3637-3642 ◽  
Author(s):  
Yas Al-Hadeethi ◽  
Ahmad Umar ◽  
Kulvinder Singh ◽  
Ahmed A Ibrahim ◽  
Saleh. H Al-Heniti ◽  
...  

Herein, we report the synthesis, characterization and picric acid chemical sensing application of samarium (Sm) doped ZnO nanorods. The Sm-doped ZnO nanorods were synthesized by facile hydrothermal process and characterized using various analytical methods which confirmed the large-scale synthesis and wurtzite hexagonal crystal structure for the synthesized nanorods. The doping of Sm ions in the lattices of the synthesized nanorods was evaluated by the energy dispersive X-ray spectroscopy (EDS). The synthesized Sm-doped ZnO nanorods were used as potential scaffold to fabricate high sensitive and reproducible picric acid chemical sensor based on I–V technique. The fabricated picric acid chemical sensor based on Sm-doped ZnO nanorods exhibited a high sensitivity of 213.9 mA mM−1 cm−2 with the limit of detection of ∼0.228 mM and correlation coefficient of R═0.9889. The obtained results revealed that the facile grown Sm-doped ZnO nanorods can efficiently be used to fabricate high sensitive and reproducible chemical sensors.


Author(s):  
Zhong-Zheng Ding ◽  
Guang-Song Zheng ◽  
Qing Lou ◽  
Jiang-Fan Han ◽  
Meng-Yuan Wu ◽  
...  

Abstract Excellent luminescent materials are essential for high-performance fluorescent nanosensors. Here, a new-type self-calibrated humidity sensor has been established through monitoring the fluorescent color change of carbon dots (CDs) confined in sodium hydroxide (CDs@NaOH). The CDs are prepared by a facile and rapid microwave assisted heating method using citric acid, urea, and NaOH as precursors. The confinement effect from NaOH has reduced the nonradiative transition and suppressed the aggregation-induced quenching of the CDs in solid. Compared with other sensors based on CD fluorescent visualization, the sensor has good linearity and wide humidity detection range from 6.9% to 95.4%. With the increased relative humidity, the fluorescence color of the sensor change from green to blue. The proposed sensing mechanism is due to the breaking and reforming of hydrogen bonds and proton transfer occurring at the CD-NaOH matrix interfaces. This finding suggests a potential role for the spatial confinement effect and may provide an avenue for developing highly sensitive humidity readouts.


2020 ◽  
Vol 21 (24) ◽  
pp. 9522
Author(s):  
Azusa Terasaki ◽  
Hiromi Kurokawa ◽  
Hiromu Ito ◽  
Yoshiki Komatsu ◽  
Daisuke Matano ◽  
...  

Hyperthermia (HT) treatment is a noninvasive cancer therapy, often used with radiation therapy and chemotherapy. Compared with 37 °C, 42 °C is mild heat stress for cells and produces reactive oxygen species (ROS) from mitochondria. To involve subsequent intracellular accumulation of DOX, we have previously reported that the expression of ATP-binding cassette sub-family G member 2 (ABCG2), an exporter of doxorubicin (DOX), was suppressed by a larger amount of intracellular mitochondrial ROS. We then hypothesized that the additive effect of HT and chemotherapy would be induced by the downregulation of ABCG2 expression via intracellular ROS increase. We used human breast cancer cell lines, MCF-7 and MDA-MB-453, incubated at 37 °C or 42 °C for 1 h to clarify this hypothesis. Intracellular ROS production after HT was detected via electron spin resonance (ESR), and DOX cytotoxicity was calculated. Additionally, ABCG2 expression in whole cells was analyzed using Western blotting. We confirmed that the ESR signal peak with HT became higher than that without HT, indicating that the intracellular ROS level was increased by HT. ABCG2 expression was downregulated by HT, and cells were injured after DOX treatment. DOX cytotoxicity enhancement with HT was considered a result of ABCG2 expression downregulation via the increase of ROS production. HT increased intracellular ROS production and downregulated ABCG2 protein expression, leading to cell damage enhancement via DOX.


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