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Blood samples for serum preparation were allowed to clot at room temperature for 15 min, centrifuged at 3,000 rpm for 10 min then immediately frozen at −80 °C.
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While investigations were under way, all work had to stop and no food preparation was allowed to restart until yesterday morning.
The resting tension was fixed at 1 g and the preparation was allowed to stabilization for 30 min.
Live attenuated influenza vaccine is not indicated in the immune-compromised host and only the inactivated preparation is allowed [ 54].
The preparation was allowed to stabilize for about 30 min and was then intermittently challenged with hypoxic solutions (Krebs bubbled with 5%O2+5%CO2+90%0% N2).
The preparation was allowed to equilibrate for about 1 h to reach a stable beating rate before APs were recorded with sharp microelectrodes.
The preparation was allowed to equilibrate for about 1 h to reach a stable beating rate before responses to CCh or Iso were determined.
Each preparation was allowed to equilibrate for 60 minutes with the changing of the K-H solution every 15 minutes before the start of the experiments.
After preparation of the exteriorized cremaster muscle, the tissue was superfused with low oxygen Krebs' solution and the preparation was allowed to rest for the next 15-20 minutos tobtainin steady state.
For selected experiments, the sympathomimetic inotrope dobutamine was added (final concentration 300 nM) and the preparation was allowed to stabilise for 10 min before new measurements of developed pressure, heart rate and coronary flow measured.
Then, matrigel containing preparations were allowed to solidify at 37°C and cells loading were formed a compact crowd of cells.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com