As can be seen in Figure 4A, a rapid uptake by the kidneys occurs during the first 2 min followed by a slow decline up to 11.7 min. After 2 min, the radioactivity was eliminated from the kidneys and accumulated in the bladder.
We found that LTP evoked at parallel fibre synapses by 1 Hz stimulation or by NO donors was not sustained beyond 30 min when P-Rex was eliminated or Rac inhibited, suggesting that cerebellar LTP exhibits a late phase analogous to hippocampal LTP.
After a deoxygenation procedure by purging buffer with nitrogen for 30 min, this peak was eliminated.
Sample B, containing the culture medium with viable cells was treated as follows: cells treated with trypsin were centrifuged at 4,000 rpm for 15 min, the supernatant was eliminated and the pellet was treated with ZnSO4 as above described.
The filtrate was then centrifuged at 3,000 rpm for 10 min. The supernatant was eliminated, and the sediment was mixed with 5 mL of phosphate buffered saline.
Before immunostaining, the slides were dryed for 30 min at room temperature and then fixed in acetone for 10 min. Endogenous peroxidase activity was eliminated by incubation in 0.3% hydrogen peroxide for 3 min. For alfa-smooth muscle actin (α-SMA) determination, we used monoclonal mouse anti α-SMA (cat. no. A 2547; Sigma-Aldrich, St . Louis MO).
Following this, cells were re-suspended in 1.5 mM borate buffer (pH 8.5), sonicated for 4 min and, finally, cellular debris was eliminated by centrifugation (20,000 r.p.m. for 30 min).
Briefly, the crude extract was obtained by turbo-extraction (Skymsen) of 100 g of the bark with 70% acetone in water for 20 min. The organic solvent was eliminated by reduced pressure and lyophilized to yield a crude extract (F1).
It was found that when the catalysts were oxidized with an oxygen stream more than 6 h were needed to eliminate the coke at 400 °C while at 500 °C the coke was eliminated in 30 min.
After gentle mixing for 30 min at 25°C, the solid bead was eliminated by centrifugation.
