containing 3.5 mg/mL BSA was used to prepare the final suspension of washed human platelets.
After the electrophysiological study (EPS), the hearts were perfused on a Langendorff apparatus at 37[degrees]C by pumping with Tyrode's solution
(calcium, 1.8 mmol/L).
Rabbits' jejunal preparations were mounted in organ bath containing 10 ml Tyrode's solution
, constantly aerated with carbogen gas.
Cells were allowed to settle in Tyrode's solution
. The solution was subsequently removed and replaced with 10 mL Tris (20 mmol, pH 7.5), then centrifuged at 94g for 3 min.
The test solution for AP recording was Tyrode's solution
with 1.8 mM [Ca.sup.2+].
EOPh was dissolved in Tyrode's solution
for the in vitro protocols using cremophor (0.1% v/v) as the eluent.
The dye solution was dropped into the Tyrode's solution
to be the desired concentration just before the measurement.
was used for the study as a nutrient solution, the composition of which was NaCl: 137 mM (08.00 g), KCl: 2.7 mM (0.20 g), CaCl2:1.8 mM (0.20 g), MgCl2: 1.05 mM (0.10 g), NaHCO3: 12.0 mM (1.00 g), NaH2PO4: 0.42 mM (0.05 g), Glucose: 5.6 mM (1.00 g) dissolved in 1 L of distilled water.
The ileum was cut into 2 inches pieces6 and transferred to an organ bath of 50ml capacity containing tyrode's solution
(in mM: NaCl, 136.8mM; KCl, 2.7mM; MgCl2, 0.5mM; CaCl2, 1.3mM; NaH2PO4, 0.14mM; NaHCO3, 12.0mM, Dextrose, 5.5mM) and aerated continuously with 95% oxygen and 5% carbondioxide.5 One end of the ileal strip was attached to the bottom of the oxygen tube in tissue bath and the other end was connected to a research grade force Displacement transducer.2 After equilibration the isotonic ileal smooth muscle activity was recorded through the Displacement Transducer on Power lab.7
The bath perfusate for sarc[K.sub.ATP] channel and AP recording was the [Ca.sup.2+] containing tyrode's solution
consisting of (in mM) NaCl 137, KCl 5.4, Mg[Cl.sub.2] 1.2, Ca[Cl.sub.2] 1, Na[H.sub.2]P[O.sub.4] 1.2, HEPES 20, and glucose 10, with the pH controlled to 7.4 with NaOH.
To check whether an active acid-extruding mechanism exists in the HUASMCs, we firstly performed the experiments in HEPES-buffered Tyrode's solution
, that is, nominally free of CO[sub]2 /HCO[sub]3 [sup]− .
Briefly, the hearts were perfused with a Tyrode's solution
containing 0.7 mg/mL type II collagenase (Worthington Biochem).