Alterations in glutathione (GSH) homeostasis are associated with a variety of diseases and cellular functions, and therefore, real-time live-cell imaging and quantification of GSH dynamics are important for understanding pathophysiological processes. However, existing fluorescent probes are unsuitable for these purposes due to their irreversible fluorogenic mechanisms or slow reaction rates. In this work, we have successfully overcome these problems by establishing a design strategy inspired by Mayr's work on nucleophilic reaction kinetics. The synthesized probes exhibit concentration-dependent, reversible and rapid absorption/fluorescence changes (t = 620 ms at [GSH] = 1 mM), as well as appropriate K values (1-10 mM: within the range of intracellular GSH concentrations). We also developed FRET-based ratiometric probes, and demonstrated that they are useful for quantifying GSH concentration in various cell types and also for real-time live-cell imaging of GSH dynamics with temporal resolution of seconds.
We studied the trapping of positive and negative charges in the chemical structures of polymers under a high electric field using a space charge measurement system. Positive charges accumulated in low-density polyethylene (LDPE), whereas positive and negative charges accumulated in polyimide (Kapton ® ) and also in ethylene tetrafluoroethylene (ETFE) subjected to electron beam irradiation. To determine the charge-trapping sites in the chemical structures, a quantum chemical calculation was carried out using Density Function Theory (DFT) with Gaussian 09. The relationship between the energy band and the isosurface of orbital electrons at various energy levels was obtained. A threedimensional (3D) electrostatic potential distribution map was obtained for positively and negatively charged polymers to determine the relationship between a trapping site and the charge accumulation center in the 3D potential distribution map. Positive and negative charges in Kapton and ETFE films are trapped in trapping sites in chemical structures and the positive charges in an LDPE film are trapped in physical defects.
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