Brain specimens from all animals with suspected rabies in South Africa were submitted to the Rabies Reference Laboratory at Onderstepoort Veterinary Institute, Pretoria.
Brain specimens from TgOvPrP4 mice in which a TSE developed were subsequently passaged by intracerebral inoculation of a 1% (wt/vol) homogenate into a second group of transgenic mice.
Therefore, we first examined gross brain specimens from control and Gcdh−/− mice placed on a standard or protein diet (60% protein) in order to identify hemorrhage locations.
A total of 80 serum or brain specimens from horses with unexplained fever (n = 48) and/or neurologic symptoms (n = 32) were tested for WNV over the 16-month period.
Complete gene sequences of the nucleoprotein gene were obtained from 52 RABV-positive specimens: brain specimens from 42 dogs, brain specimens from 3 persons who died, 1 specimen from a cow that died, 5 human saliva specimens, and 1 human CSF specimen.
Additionally, neuropathological postmortem examinations of brain specimens from primates of the NIH study did not provide evidence for a pre- or subclinical disease transmission, according to a personal communication by members of the former NIH research team (P. Brown, D. Asher) reported by Irwin et al. [ 42].
Brain specimens from one 6-week-old goose, which died during the encephalitis outbreak in a Hungarian goose flock, and brain samples from a goshawk, which also died from encephalitis, were used for WNV nucleic acid determination.
The combination of these 3 techniques represents a novel method for exploring brain connectivity in postmortem brain specimens from virtually any species, although it is currently most practical in small animal models due to imaging hardware and scan time requirements.
Briefly, brain specimens from infected and uninfected mice were homogenized and lysed in radioimmunoprecipitation assay buffer containing a protease inhibitor cocktail (Sigma, Saint Louis, MO, USA) on ice for at least 1 h, after which the proteins were harvested by centrifugation at 10,000 g at 4°C for 10 min, and stored at -80°C for further studies.
These methods allow the investigation of mRNA-based transcriptosomal profiles of brain specimens from AD humans and transgenic animals.
