1. + High CJD infectivity remains after the prion protein is destroyed By Sylvester Gates

2. + High CJD infectivity remains after prion protein is destroyed (2011) - Kohtaro Miyazawa, Kaitlin Emmerling and Laura Manuelidis Goals: Determine if infectivity is linked to PrP load. Infectivity decreases as direct proportion to decrease of PrP with proteinase K. Experiments: Mouse passaged FU-CJD PrP Protease digestion (Keratinase = NAP and proteinase K = PK) at different concentrations of Tx100 (detergent) http://onlinelibrary.wiley.com/doi/10.1002/jcb.23286/full

3. + High CJD infectivity remains after prion protein is destroyed (2011) - Kohtaro Miyazawa, Kaitlin Emmerling andLaura Manuelidis Tested NAP effectiveness under specific conditions Digestion of FU-CJD brain with NAP Ln5&6 NAP digestion Ln7 under PK digestion – reveals max PrP-res

4. + High CJD infectivity remains after prion protein is destroyed (2011) - Kohtaro Miyazawa, Kaitlin Emmerling andLaura Manuelidis 99.5% digested 99.8% digested PrP takes longer to digest in cells 95.5% digested

5. + High CJD infectivity remains after prion protein is destroyed (2011) - Kohtaro Miyazawa, Kaitlin Emmerling andLaura Manuelidis Tissue Culture Infectious Dose

6. + High CJD infectivity remains after prion protein is destroyed (2011) - Kohtaro Miyazawa, Kaitlin Emmerling andLaura Manuelidis Tissue Culture Infectious Dose 2hrs PK at 6 and 8 hrs only yield <2 log reduction NAP reduced infectious dosage by >3.5 logs

7. + High CJD infectivity remains after prion protein is destroyed (2011) - Kohtaro Miyazawa, Kaitlin Emmerling andLaura Manuelidis Showed no loss of infectivity after virtually complete PK digestion of PrP Suggests PrP-res regenerates to normal levels after p13 99.5% digested in 10x 99.8% digested 97.9% digested

8. + High CJD infectivity remains after prion protein is destroyed (2011) - Kohtaro Miyazawa, Kaitlin Emmerling andLaura Manuelidis “Virtually complete digestion of all PrP with preservation of infectivity lead… to the conclusion that no form of prion protein is infectious” PrP-res levels climb back up to normal even after PrP digestion by PK digestion left ≤0.3% PrP after 2hr, yet there was no reduction in titer. NAP digestion left 0.8% residual PrP after 2hr, yet decreased titer by >2.5logs “GdnSCN… shown to reduce infectivity by >4logs in brain [Manuelidis, 1997], practical and complete sterilization of precious instruments should be further effected by a subsequent digestion with NAP, and probably other keratinases.”

9. + Exposure of RML scrapie agent to a sodium percarbonate- based product and sodium dodecyl sulfate renders PrP Sc protease sensitive but does not eliminate infectivity (2012) - Jodi D Smith, Eric M Nicholson, Gregory H Foster and Justin J Greenlee Goals: Evaluate effectiveness of a commercial product containing sodium percarbonate to inactivate prions. Experiments: Mouse brain with mouse-adapted scrapie agent (RML) Exposed to sodium percarbonate-based product (SPC-P). Western blots to test immunoreactivity for abnormal prion protein Residual infectivity tested by mouse bioassay

10. + Exposure of RML scrapie agent to a sodium percarbonate- based product and sodium dodecyl sulfate renders PrP Sc protease sensitive but does not eliminate infectivity (2012) - Jodi D Smith, Eric M Nicholson, Gregory H Foster and Justin J Greenlee http://www.biomedcentral.com/1746-6148/9/8 Ladder RML +ctrl 30 min 90 min 180 min 2.5% SDS

11. + Exposure of RML scrapie agent to a sodium percarbonate- based product and sodium dodecyl sulfate renders PrP Sc protease sensitive but does not eliminate infectivity (2012) - Jodi D Smith, Eric M Nicholson, Gregory H Foster and Justin J Greenlee ladder ctrl 30m 90m 180m ladder ctrl 30m 90m 180m Brain treatment with 0.35 M sodium hydrogen phosphate buffered solution PrP Sc undetectable after PK digestion

12. + Exposure of RML scrapie agent to a sodium percarbonate- based product and sodium dodecyl sulfate renders PrP Sc protease sensitive but does not eliminate infectivity (2012) - Jodi D Smith, Eric M Nicholson, Gregory H Foster and Justin J Greenlee RML ctrl Avg. survival time

13. + Exposure of RML scrapie agent to a sodium percarbonate- based product and sodium dodecyl sulfate renders PrP Sc protease sensitive but does not eliminate infectivity (2012) - Jodi D Smith, Eric M Nicholson, Gregory H Foster and Justin J Greenlee Exposure of RML (scrapie agent) to an SPC-containing product alone or in combination with SDS does not eliminate prion infectivity Small effect of SPC-P alone, but an 2–3 log10 reduction observed with the addition of SDS exposure to SDS alone resulted in an approximate 2 log10 reduction.

14. + BSE infectivity in the absence of detectable PrPSc accumulation in the tongue and nasal mucosa of terminally diseased cattle (2010) - Anne Balkema-Buschmann, Martin Eiden, Christine Hoffmann, Martin Kaatz, Ute Ziegler, Markus Keller and Martin H. Groschup “Challenged transgenic mice overexpressing the bovine prion protein with homogenates prepared from a wide variety of tissue samples collected from BSE-infected cattle” Various detection methods: purification, immunohistochemistry, and the protein misfolding cyclic amplification technique http://vir.sgmjournals.org/content/92/2/467.full

15. + BSE infectivity in the absence of detectable PrPSc accumulation in the tongue and nasal mucosa of terminally diseased cattle (2010) - Anne Balkema-Buschmann, Martin Eiden, Christine Hoffmann, Martin Kaatz, Ute Ziegler, Markus Keller and Martin H. Groschup http://vir.sgmjournals.org/content/92/2/467.full

16. + BSE infectivity in the absence of detectable PrPSc accumulation in the tongue and nasal mucosa of terminally diseased cattle (2010) - Anne Balkema-Buschmann, Martin Eiden, Christine Hoffmann, Martin Kaatz, Ute Ziegler, Markus Keller and Martin H. Groschup http://vir.sgmjournals.org/content/92/2/467.full protein misfolding cyclic amplification (PMCA)

17. + BSE infectivity in the absence of detectable PrPSc accumulation in the tongue and nasal mucosa of terminally diseased cattle (2010) - Anne Balkema-Buschmann, Martin Eiden, Christine Hoffmann, Martin Kaatz, Ute Ziegler, Markus Keller and Martin H. Groschup Detect BSE infectivity in tongue and nasal mucosa of terminally diseased BSE cases as well as experimentally challenged cattle by transgenic-mouse bioassay. This shows that BSE infectivity can be present in the peripheral tissues. http://vir.sgmjournals.org/content/92/2/467.full