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Brain samples were then cut using a vibratome (section thickness 30 40 µm).
The brain samples were then immediately frozen on dry ice and stored at −80 °C.
Brain samples were then embedded in paraffin and sectioned into 4- μm slides.
Brain samples were then mounted using Vectashield and were visualized by a Leica TCS SP5 confocal microscope with a 20×, 40×, or 63× objective lens.
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AChE enzymatic activity in the brain samples was then calculated as micromoles of substrate hydrolyzed per minute per gram wet weight (ww) (Padilla et al. 1998).
Extraction of blood and brain samples was then performed by a method based on protein precipitation using acetonitrile containing a structural analogue of -U50,488 as an internal standard.
Blood samples were then transferred in culture bottles of brain heart infusion broth (Hi Media, Mumbai, India).
Tissue samples were then dried in an oven for 24 h at 100 °C and brain water content was calculated as [ wet weight-dry weight)/wet weight)] × 100.
The samples were then combined.
The two samples were then combined.
The samples were then counterstained with hematoxylin.
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