Carboxylato-Pillar[6]arene-Based Fluorescent Indicator Displacement Assays for Caffeine Sensing

In the present work, we have developed a new indicator displacement system based on pillararene for anionic water-soluble carboxylato pillar [6] arene (WP6) and aromatic fluorescent dye safranine T (ST). A large fluorescence enhancement and colour change of ST were observed after complexation with electron-rich cavity in WP6 because of host-guest twisted intramolecular charge-transfer interactions. The constructed pillararene-indicator displacement system can be applied for caffeine selective detection in water.

Detecting nitric oxide in free-living symbiotic dinoflagellates exposed to nanoparticles v1

Diaminofuorescein-2 diacetate (DAF-2 DA) is a fluorescent indicator of nitric oxide (NO). The DAF reacts with the nitric anhydride (N2O3) which formed by oxidation of NO. Upon crossing the cell membrane, esterases hydrolyse DAF-2 DA to DAF-2, which remains trapped within cells. The DAF-2 reacts to the oxidation of intracellular NO (or more accurately the N2O3) to produce the highly fluorescent triazolofluorescein (DAF-2T) by nitrosation and dehydration (Kojima et al., 1999). Protocol adapted from Bouchard & Yamasaki (2009) and Kojima et al. (1999)

TRPM7 silencing attenuates Mg2+ influx in cardiac myoblasts, H9c2 cells

TRPM7, a member of the melastatin subfamily of transient receptor potential channels, is suggested to be a potential candidate for a physiological Mg2+ channel. However, there is no direct evidence of Mg2+ permeation through endogenous TRPM7. To determine the physiological roles of TRPM7 in intracellular Mg2+ homeostasis, we measured the cytoplasmic free Mg2+ concentration ([Mg2+]i) in TRPM7-silenced H9c2 cells. [Mg2+]i was measured in a cluster of 8–10 cells using the fluorescent indicator, furaptra. TRPM7 silencing did not change [Mg2+]i in Ca2+-free Tyrode’s solution containing 1 mM Mg2+. Increasing the extracellular Mg2+ to 92.5 mM raised [Mg2+]i in control cells (1.56 ± 0.19 mM) at 30 min, while this effect was significantly attenuated in TRPM7-silenced cells (1.12 ± 0.07 mM). The Mg2+ efflux driven by Na+ gradient was unaffected by TRPM7 silencing. These results suggest that TRPM7 regulates the rate of Mg2+ influx in H9c2 cells, although cytoplasmic Mg2+ homeostasis at basal conditions is unaffected by TRPM7 silencing.