Intracellular sodium changes in cancer cells using a microcavity array-based bioreactor system and sodium triple-quantum mr signal

The sodium triple-quantum (TQ) magnetic resonance (MR) signal created by interactions of sodium ions with macromolecules has been demonstrated to be a valuable biomarker for cell viability. The aim of this study was to monitor a cellular response using the sodium TQ signal during inhibition of Na/K-ATPase in living cancer cells (HepG2). The cells were dynamically investigated after exposure to 1 mM ouabain or K$^{+}$-free medium for 60 min using an MR-compatible bioreactor system. An improved TQ time proportional phase incrementation (TQTPPI) pulse sequence with almost four times TQ signal-to-noise ratio (SNR) gain allowed for conducting experiments with 12–14 × 10$^{6}$ cells using a 9.4 T MR scanner. During cell intervention experiments, the sodium TQ signal increased to 138.9 ± 4.1% and 183.4 ± 8.9% for 1 mM ouabain (n = 3) and K$^{+}$-free medium (n = 3), respectively. During reperfusion with normal medium, the sodium TQ signal further increased to 169.2 ± 5.3% for the ouabain experiment, while it recovered to 128.5 ± 6.8% for the K$^{+}$-free experiment. These sodium TQ signal increases agree with an influx of sodium ions during Na/K-ATPase inhibition and hence a reduced cell viability. The improved TQ signal detection combined with this MR-compatible bioreactor system provides a capability to investigate the cellular response of a variety of cells using the sodium TQ MR signal

The cellular heat shock response monitored by chemical exchange saturation transfer MRI

CEST-MRI of the rNOE signal has been demonstrated in vitro to be closely linked to the protein conformational state. As the detectability of denaturation and aggregation processes on a physiologically relevant scale in living organisms has yet to be verified, the aim of this study was to perform heat-shock experiments with living cells to monitor the cellular heat-shock response of the rNOE CEST signal. Cancer cells (HepG2) were dynamically investigated after a mild, non-lethal heat-shock of 42 °C for 20 min using an MR-compatible bioreactor system at 9.4 T. Reliable and fast high-resolution CEST imaging was realized by a relaxation-compensated 2-point contrast metric. After the heat-shock, a substantial decrease of the rNOE CEST signal by 8.0 ± 0.4% followed by a steady signal recovery within a time of 99.1 ± 1.3 min was observed in two independent trials. This continuous signal recovery is in coherence with chaperone-induced refolding of heat-shock induced protein aggregates. We demonstrated that protein denaturation processes influence the CEST-MRI signal on a physiologically relevant scale. Thus, the protein folding state is, along with concentration changes, a relevant physiological parameter for the interpretation of CEST signal changes in diseases that are associated with pathological changes in protein expression, like cancer and neurodegenerative diseases

The cellular heat shock response monitored by chemical exchange saturation transfer MRI

Chemical exchange saturation transfer of the relayed nuclear Overhauser effect(rNOE) enables detection of mobile protein protons via their exchange with the water signal. Recent studies have shown a close relationship between the rNOE-signal and the protein conformation. This study used the -3.5ppm rNOE-signal to monitor the heat shock response of HepG2 cells in a microcavity array-based bioreactor system. A significant drop of the rNOE-signal to 92.4±1.3% and a slow, steady increase of the rNOE-signal for 136.1±13.6min after heat shock were observed. Therefore, the rNOE-signal in CEST spectroscopic imaging is a sensitive readout for the cellular heat shock response

Degradation Kinetics during Oxygen Electrocatalysis on Perovskite-Based Surfaces in Alkaline Media

The oxygen evolution reaction (OER) during alkaline water electrolysis is the bottleneck of water splitting. Perovskite materials have been particularly proposed as good and economically reasonable electrocatalysts for the OER, showing promise and advantages with respect to classic metallic electrodes. However, the degradation of perovskites during catalysis limits their service lifetime. Recently, the material BaCo0.98Ti0.02O3−δ:Co3O4 was shown to be electrocatalytically and chemically stable during water electrolysis even under industrially relevant conditions. The lifetime of this perovskite-based system is prolonged by a factor of 10 in comparison to that of Pr0.2Ba0.8CoO3−δ and is comparable to that of industrially applied electrodes. Here we report on the degradation kinetics of several OER catalysts at room temperature, comparatively studied by monitoring the oxygen evolution at microelectrodes. A decrease in the reaction rate within a maximum of 60 s is observed, which is related to chemical and/or structural changes at the oxide surface