2,2-dichloro-1,1-difluoro-1-methoxyethane

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: For isolated mouse aortic rings preparations, male C57 black mice (5-7 weeks of age) were anesthetized with methoxyflurane and sacrificed by exsanguination. The thoracic aorta was dissected and cut into cylindrical segments of three millimeters in length. Using a sterile 48-well tissue culture plate, each ring was incubated in a total volume of one ml with either peptides or vehicle (DMSO) diluted in DMEM supplemented with penicillin (100 units/ml), streptomycin (100 μg/ml) and L-glutamine (1 mM). The plate was incubated with 5% CO2. Following incubation with the peptides (one, six, twelve or twenty hours), rings were washed in oxygenated (95% O2-5% CO2) Krebs-Henseleit bicarbonate buffer solution (KITS) of the following composition (NaCl-118.3 mmol/L, KCl-4.7 mmol/L, CaCl2-2.5 mmol/L, MgSO4-1.2 mmol/L, KH2PO4-1.2 mmol/L; NaHCO3-25 mmol/L, glucose-5.6 mmol/L at 37° C. with ibuprofen (10 μM). The rings were suspended by two tungsten wires (25 mm thickness) inserted into the lumen and mounted in a vessel myograph system (five ml chamber, Kent Scientific). The mouse aortas were submitted to a resting tension of 1.5 grams and isometric tension was recorded using a force transducer coupled to a MacLab data acquisition system. Following a sixty minute equilibration period, with frequent washes, the rings were precontracted with phenylephrine (PE) (10 μM; sub maximal concentration) and concentrations of Ach (1 nM-10 μM), sodium nitroprusside (10 μM) or Nitro-L-Arginine Methyl Ester (L-NAME, 100 mM) were injected at the plateau of the PE contraction. In some experiments concentration-response curves to the contractile effects of PE were monitored. For isolated mouse aortic rings preparations male C57 black mice (5-7 weeks of age) diluted in DMEM were washed in oxygenated (95% O2-5% CO2) Krebs-Henseleit bicarbonate buffer solution (KITS) of the following composition (NaCl-118.3 mmol/L, KCl-4.7 mmol/L, CaCl2-2.5 mmol/L, MgSO4-1.2 mmol/L, KH2PO4-1.2 mmol/L at 37° C. mounted in a vessel Following a sixty minute In some experiments concentration-response curves to the contractile effects of PE

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: For isolated mouse aortic rings preparations, male C57 black mice (5-7 weeks of age) were anesthetized with methoxyflurane and sacrificed by exsanguination. The thoracic aorta was dissected and cut into cylindrical segments of three millimeters in length. Using a sterile 48-well tissue culture plate, each ring was incubated in a total volume of one ml with either peptides or vehicle (DMSO) diluted in DMEM supplemented with penicillin (100 units/ml), streptomycin (100 μg/ml) and L-glutamine (1 mM). The plate was incubated with 5% CO2. Following incubation with the peptides (one, six, twelve or twenty hours), rings were washed in oxygenated (95% O2-5% CO2) Krebs-Henseleit bicarbonate buffer solution (KHS) of the following composition (NaCl—118.3 mmol/L, KCl—4.7 mmol/L, CaCl2—2.5 mmol/L, MgSO4—1.2 mmol/L, KH2PO4—1.2 mmol/L, NaHCO3—25 mmol/L, glucose—5.6 mmol/L at 37° C. with ibuprofen (10 μM). The rings were suspended by two tungsten wires (25 mm thickness) inserted into the lumen and mounted in a vessel myograph system (five ml chamber, Kent Scientific). The mouse aortas were submitted to a resting tension of 1.5 grams and isometric tension was recorded using a force transducer coupled to a MacLab data acquisition system. Following a sixty minute equilibration period, with frequent washes, the rings were precontracted with phenylephrine (PE) (10 μM; sub maximal concentration) and concentrations of Ach (1 nM-10 μM), sodium nitroprusside (10 μM) or Nitro-L-Arginine Methyl Ester (L-NAME, 100 mM) were injected at the plateau of the PE contraction. In some experiments concentration-response curves to the contractile effects of PE were monitored. For isolated mouse aortic rings preparations male C57 black mice (5-7 weeks of age) diluted in DMEM were washed in oxygenated (95% O2-5% CO2) Krebs-Henseleit bicarbonate buffer solution (KHS) of the following composition (NaCl—118.3 mmol/L, KCl—4.7 mmol/L, CaCl2—2.5 mmol/L, MgSO4—1.2 mmol/L, KH2PO4—1.2 mmol/L, NaHCO3—25 mmol/L, glucose—5.6 mmol/L at 37° C. with ibuprofen (10 μM) mounted in a vessel Following a sixty minute In some experiments concentration-response curves to the contractile effects of PE