First, the definition of isotonicity
is given below .
To ensure thorough homogenization, the homogenate was subjected to two round of freeze-thaw and centrifuged at 1,000 g for 10 min at 4[degrees]C, the suspension was collected and pellet was homogenized again, then supernatants from each centrifugation-step were pooled and equal volume of 2xPBS was added to restore isotonicity
. Sequently, membrane fractions were collected by ultracentrifugation at 120,000 g for 2 h at 4[degrees]C using an Optima[TM] Max Ultracentrifuge with the MLA-80 rotor (Beckman-Coulter, USA).
While considering input and output factors the isotonicity
relations are assumed for DEA (i.e., an increase in any input should not result in a decrease in any output).
To estimate the construct validity, we used correlation analysis to check the isotonicity
(Golany and Roll, 1985; Golany, 1988) among the inputs and outputs.
The resulting pellet of white ghosts was incubated in 10 mmol/L 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ; Molecular Probes, Eugene, Ore) for 5 min, and sufficient NaCl was added to restore isotonicity
. The ghosts were then resealed by incubation at 37 [degrees] C for 1 hour and washed 3 more times in phosphate-buffered saline (PBS) to remove extracellular SPQ.
To maintain the intracellular isotonicity
, an IV fluid with 1000 cc D5 1/3 NS with 10 meq KCI will be ordered to infuse at 50 cc/hr.