Unrestrained, free-moving animals were placed in individual chambers and allowed 1 min to settle down followed by 5 min of monitoring.
However, for statistical purposes, we assigned latency to 15 min and GTCS duration to 0 sec in animals that did not have a GTCS during the 15 min of monitoring.
As indicated by the black line in Figure 3,[ 22] no photobleaching was observed in the first 5 min and even after 20 min of monitoring the bleaching was still below 7%%, which is insignificant compared to the loss of fluorescence by quenching caused by transport of chloride.
The amount of release of curcuminoids was determined at 15 min intervals, followed by 30 min intervals of monitoring the variation of absorbance in the UV Vis absorption spectroscopy, with the peak at 427 nm (λmax of curcuminoids).
A respiratory gas facemask was worn throughout the trial and respiratory gases were analysed in the last 2 min of every 10 min to monitor exercise intensity; HR was recorded every 10 min.
After 15 min of monitoring, the abdominal wall was closed with a Vicryl 4/0 and the rat was monitored during the acute post-surgery without any evidence of a bleeding syndrome.
At pre-chosen time intervals post-infection mice were imaged for 1 min to monitor dissemination of pneumococci into the lungs.
The spectra were obtained every 1 min to monitor the time course of arrestin-induced fluorescence quenching (if any), until there was no further reduction in the bimane fluorescence.
At intervals of 10 min, we monitored the number of progeny produced per individual of the cohort.
The device was connected to the constant current generator for 5 minutes (I-ON XL, Sooft, Montegiorgio, FM, Italy) set at 1 mA (the total dose of 5 min5 min was monitored by the generator).
