solution 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.
Subsequently, plates were washed with 1x Tyrode's
buffer (137 mM NaCl, 2.7 mM KCl, 0.4 mM Na[H.sub.2]P[O.sub.4], 0.5 mM Mg[Cl.sub.2] * 6[H.sub.2]O, 1mM Ca[Cl.sub.2], 0.1% BSA, 0.1% glucose, 10 mM HEPES, pH 7.45), 100 [micro]L of the indicated amounts of OVA (grade V, Sigma-Aldrich) in 1x Tyrode's
buffer was added to the cells, and plates were incubated at 37[degrees]C.
The dye solution was dropped into the Tyrode's
solution to be the desired concentration just before the measurement.
Mesenteric rings (1-2 mm length) were obtained and suspended by cotton threads in an organ bath containing 10 mL of Tyrode's
solution, maintained at 37[degrees]C, and gassed with a 95% [O.sub.2] + 5% C[O.sub.2] mixture (pH 7.4).
Platelet-rich plasma (PRP) is then carefully collected and washed through one cycle of resuspension, using three volumes of Tyrode's
buffer, pH 6.5, and one volume of citrate buffer, pH 5.3, and centrifugation at 640 g for 12 minutes at room temperature.
The test solution for AP recording was Tyrode's
solution with 1.8 mM [Ca.sup.2+].
After maturation, Tyrode's
albumin lactate pyruvate (TALP) and synthetic oviductal fluid (SOF) media were used for fertilization and culture environments, respectively.
Rabbits' jejunal preparations were mounted in organ bath containing 10 ml Tyrode's
solution, constantly aerated with carbogen gas.
(a) Washed platelets or (b) the Fenton reaction solution was preincubated with (A) Tyrode's
solution (resting control), (B) 0.1% DMSO, (C) 10 [micro]M Ir-6, or (D) 20 [micro]M Ir-6.
Cells were sensitized with 15 [micro]g/ml myeloma IgE (Athens Research & Technology; Athens, GA) in complete media for 24 h and rinsed three times prior to activation with various concentrations of anti-IgE (Chemicon International Inc.; Temecula, CA) in Tyrode's
solution (Boston BioProducts; Ashland, MA) supplemented with SCF and IL-6 for 1 h.
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).
Fragments were immediately fixed in 100 mL/L formalin and embedded in paraffin or in 25 mL/L glutaraldehyde in Tyrode's
buffer (pH 7.2), postfixed in 10 g/L Os[O.sub.4] in the same buffer, dehydrated in ethanol, and embedded in Spurr resin.