The p38 MAPK and heat shock protein 27 (hsp27) form a signaling complex with serine/threonine kinase Akt and MAPK-activated protein kinase-2 (MK2), which plays an important role in controlling stress-induced apoptosis and reorganizing actin cytoskeleton. However, regulation of the complex is poorly understood. In this study, the interaction between p38 and hsp27 was visualized in single living L929 cells using fluorescence resonance energy transfer technology, while their association with Akt was examined by immunoprecipitation analysis. Under normal growth conditions, p38 kinase constitutively interacts with hsp27. When cells were exposed to H 2 O 2 or stimulated by arachidonic acid, this interaction was disrupted. However, inhibition of the activation of p38 and Akt by selective inhibitors or overexpression of the kinase-dead mutant of p38 diminished such effects. Furthermore, mutation of phosphorylation sites of hsp27 renders the interaction resistant to H 2 O 2 and arachidonic acid. It was interesting to find that the interaction disappeared in the cells from MK2-knock-out mice or the cells treated with lemptomycin B that blocks export of MK2 from nucleus to cytosol. However, MK2 is not required for the association of hsp27 with Akt. This study suggests that MK2 mediates the incorporation of p38 into the pre-existing complex of hsp27 with Akt. Phosphorylation of hsp27 finally breaks the signaling complex.
Recent years have witnessed thriving progress of flexible and portable electronics, with very high demand for cost-effective and tailor-made multifunctional devices. Here, we report on an ingenious origami hierarchical sensor array (OHSA) written with a conductive ink. Thanks to origami as a controllable hierarchical framework for loading ink material, we have demonstrated that OHSA possesses unique time-space-resolved, high-discriminative pattern recognition (TSR-HDPR) features, qualifying it as a smart sensing device for simultaneous sensing and distinguishing of complex physical and chemical stimuli, including temperature, relative humidity, light and volatile organic compounds (VOCs). Of special importance, OSHA has shown very high sensitivity in differentiating between structural isomers and chiral enantiomers of VOCs – opening a door for wide variety of unique opportunities in several length scales.
We present a novel sampling imaging technique capable of performing simultaneous two-dimensional measurements of the temporal and spectral characteristics of light-emission processes by use of a specially designed streak camera. A proof-of-principle experiment was performed with a homemade multifocal multiphoton fluorescence microscope. The system was calibrated with a Fabry-Perot etalon and a standard fluorophore solution (rhodamine 6G in ethanol) and was shown to have temporal and spectral resolution of 6.5 ps and 3 nm, respectively, as well as high accuracy and reproducibility in lifetime and spectrum measurement. Temporally and spectrally resolved images of 4 x 4 foci on the sample can be obtained with a snapshot.
There is a close correlation between body health and the level of biofluid-derived metal ions, which makes it an attractive model analyte for noninvasive health monitoring. The present work has developed a novel nose/tongue-mimic chemosensor array based on bioinspired polydopamine/polyethylenimine copolymers (PDA/PEI ) for label-free fluorescent determination of metal ions in biofluids. Three types of PDA/PEI (PDA/PEI, PDA/PEI, and PDA/PEI) were prepared by using different concentrations of PEI to construct the proposed sensor array, which would lead to unique fluorescence response patterns upon challenged with metal ions for their pattern discrimination. The results show that as few as 3 PDA/PEI sensors can successfully realize the largescale sensitive detection of metal ions in biofluids. Moreover, we have demonstrated that PDA/PEI sensors are qualified for lifetime-based pattern discrimination application. Furthermore, the sensors can distinguish between different concentrations of metal ions, as well as a mixture of different metal ions in biofluids, even the mixtures with different valence states. The method promises the simple, rapid, sensitive, and powerful discrimination of metal ions in accessible biofluids, showing the potential applications in the diagnosis of metal ion-involved diseases.
It is shown experimentally for small spectral bandwidth that spiral phase plates (SPPs) can generate optical vortices in femtosecond beams without the need for additional dispersion compensation elements. An autocorrelation measurement performed on the obtained optical vortex (OV) beam showed that the SPP did not significantly change the pulse duration. Hence the single element SPP, already an established beam shaping technique in continuous wave and high power regimes, is presented as an efficient and practical means to obtain OVs that are free from spatial chirp in femtosecond beams.
A protein/lanthanide complex (BSA/Tb 3+ )-based sensor array in two different pH buffers has been designed for high-throughput recognition and time-resolved fluorescence (TRF) detection of metal ions in biofluids. BSA, which acted as an antenna ligand, can sensitize the fluorescence of Tb 3+ (i.e., antenna effect), while the presence of metal ions would lead to the corresponding conformational change of BSA for altering the antenna effect accompanied by a substantial TRF performance of Tb 3+ . This principle has also been fully proved by both experimental characterizations and coarse-grained molecular dynamics (CG-MD) studies. By using Tris-HCl buffer with different pHs (at 7.4 and 8.5), 17 metal ions have been welldistinguished by using our proposed BSA/Tb 3+ sensor array. Moreover, the sensor array has the potential to discriminate different concentrations of the same metal ions and a mixture of metal ions. Remarkably, the detection of metal ions in biofluids can be realized by utilizing the presented sensor array, verifying its practical applications. The platform avoids the synthesis of multiplex sensing receptors, providing a new method for the construction of convenient and feasible lanthanide complex-based TRF sensing arrays.
Recent years have witnessed the rapid development of pattern-based sensors due to their potential to detect and differentiate a wealth of analytes with only few probes. However, no one has found or used the combination of DNA and terbium(III) (Tb) as a pattern recognition system for large-scale mix-and-measure assays. Here we report for the first time that DNA-sensitized Tb (DNA/Tb), as a label-free and versatile "chemical nose/tongue", can be employed for wide-scale time-gated luminescent (TGL) monitoring of metal ions covering nearly the entire periodic table in a cost-effective fashion. A series of guanine/thymine (G/T)-rich DNA ligands was screened to sensitize the luminescence of Tb (referring to the antenna effect) as smart pattern responders to metal ions in solution, and metal ion-DNA interactions can differentially alter the antenna effect of DNA toward Tb as pattern signals. Our results show that as few as 3 DNA/Tb label-free sensors could successfully discriminate 49 analytes, including alkali-metal ions, alkaline-earth-metal ions, transition/post-transition metal ions, and lanthanide ions. A blind test with 49 metals further confirmed the discriminating power of DNA/Tb sensors. Moreover, the lifetime-based pattern recognition application using DNA/Tb sensors was also demonstrated. This DNA/Tb pattern recognition strategy could be extended to construct a series of "chemical noses/tongues" for monitoring various biochemical species by using different responsive DNA ligands, thus promising a versatile and powerful tool for a sensing application and investigation of DNA-involving molecular interactions.
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