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HPLC with fluorometric detection was carried out on supernatants of rat brain samples that were homogenized in 0.4 N perchloric acid (PCA) including as additives 0.1% EDTA and 0.05% Na2S2O5.
We examined the effect of specimen size and thickness by comparing rat kidney and brain samples that were either fixed whole (about 1 cm thick), or cut into smaller pieces, around 3 4 mm thick (Fig. 4).
To extend the comparison beyond blood, we compared our results with two studies with brain samples that were also analyzed with the 450-k chip.
Furthermore, the percentage of animal brain samples that were confirmed infected with rabies during the 10 year period remained unchanged, within the range of 23 30%.
Field samples were obtained from regional veterinary laboratories as brain samples that were positive for rabies virus by fluorescent antibody test (FAT) [ 8] with anti-nucleocapsid conjugate (Bio-Rad).
As described below, we finally evaluated the extent of ACh postmortem changes among brain samples that were subjected to ISF, which ensures adequate perfusion until the arrival of the freezing front to avoid unnecessary autolysis, and postmortem freezing (decapitation before freezing).
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The two meta-analyses included several studies that we excluded due to quality control measures, and we included one study on a unique set of brain samples that was not included in theirs.
Such infectious level is approximately equivalent to 1.4 µg of the reference vCJD brain sample that was endpoint-titrated (Table 1).
The infectious titers in both positive plasma samples were estimated to be 2.12 and 3.7 ID/mL of plasma, which is equivalent to 0.3 0.5 µg of the reference sCJD MM1 brain sample that was endpoint titrated (Table 1).
In the brain sample that was obtained by autopsy the Western Blot showed presence of total ERK, but not of pERK, which is in line with the immunohistochemical staining results.
The ileum, colon, spleen and brain stem were examined with RT-PCR, but viral RNA was only detected in the three brain stem samples that were positive in virus titration (Tables 2 and 3).
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