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Protein staining was carried out on embedded cross sections following a method detailed by Schäfer [8].
PAHs in fronds were extracted and quantified according to the method detailed by Sun et al. [ 13].
After 24 h of toxin exposure, cells were harvested and washed with PBS at 2000× g, 4 °C for 5 min (Eppendorf 5424, Hamburg, Germany) based on the method detailed by Dong et al. [ 14].
Hippocampus and amygdala were manually traced in their entirety according to the method detailed by Pruessner et al. [ 71], which used the coronal view as the default orientation but also employed saggital and horizontal views to determine specific boundaries.
For neuron isolation the forebrain of ipsilateral side was dissected and processed according to the method detailed by Rathore et al (2008), Dohare et al (2008) [ 6, 20] and Rastogi et al (2006) [ 22].
Citrate bicarbonate dithionite (CBD) extractions were performed in duplicate on both the <2 mm (5 g) and fines (2 g) of each sample per the modified method detailed by Loppert and Inskeep,1996 at 80 °C and pH 7. (33) The <2 mm size fraction was also ground to pass through a 0.063 mm sieve (2 g) and extracted with CBD.
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In summary, despite applying the rigorous methods detailed by Scott et al. [9] to our therapeutic trial, we were unable to show efficacy of chronic lithium treatment in the SOD1G93A mouse ALS model.
Hair cortisol extraction procedures were similar to the methods detailed by Stalder et al. [ 32] with some modifications as below.
Interviewer administered Baseline Dyspnea Index (BDI) and Transition Dyspnea Index (TDI) were assessed using the methods detailed by Mahler et al. [ 18].
Relative mRNA expression was determined using the ΔΔCT method, as detailed by manufacturer guidelines (Bio-Rad).
Relative mRNA expression was determined with the ΔΔ C t method, as detailed by manufacturer guidelines (Applied Biosystems).
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