After collection, samples were processed for determination of heavy metals i.e. cobalt (Co), chromium (Cr), nickel (Ni), lead (Pb) and zinc (Zn).
Following sacrifice, the cecal materials, livers and spleens were collected and processed for determination of Salmonella titre as described below.
Samples of the ventrolateral medulla were processed for determination of ATP concentration as described previously [17], using an ATP bioluminescence assay (Roche Diagnostics GmbH, Mannheim, Germany).
The remaining half of each kidney was processed for determination of Hyp and Pro levels.
A portion of the liver was immediately fixed in formalin for histological analyses, and another portion was processed for determination of oxidative-stress-related products (see below).
Venous blood samples were taken at 5, 15, 30 and 60 min postinjection and processed for determination of metabolic stability in vivo.
The resultant supernatant media were processed for determination of hepatitis B surface antigen (HBsAg) expression, HBV DNA polymerase activity and HBV DNA level as below.
Owing to histological problems, brain sections from a single mouse were not processed for determination of BBB injury (Fig. 1E, KO to KO group).
Triplicate vials for each sample and blank were retrieved from the incubator after 6 h and 72 h of incubation and processed for determination of VFAs, ammonia and IVDMD as previously described [ 48].
This tissue was then processed for determination of choline acetyltransferase (ChAT) activity in order to provide a quantitative assessment of the degree to which cholinergic denervation had been achieved.
