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Assessing Transmissible Spongiform Encephalopathy Species Barriers with an In Vitro Prion Protein Conversion Assay

Overview of attention for article published in Journal of Visualized Experiments, March 2015
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Title
Assessing Transmissible Spongiform Encephalopathy Species Barriers with an <em>In Vitro</em> Prion Protein Conversion Assay
Published in
Journal of Visualized Experiments, March 2015
DOI 10.3791/52522
Pubmed ID
Authors

Christopher J. Johnson, Christina M. Carlson, Aaron R. Morawski, Alyson Manthei, Neil R. Cashman

Abstract

Studies to understanding interspecies transmission of transmissible spongiform encephalopathies (TSEs, prion diseases) are challenging in that they typically rely upon lengthy and costly in vivo animal challenge studies. A number of in vitro assays have been developed to aid in measuring prion species barriers, thereby reducing animal use and providing quicker results than animal bioassays. Here, we present the protocol for a rapid in vitro prion conversion assay called the conversion efficiency ratio (CER) assay. In this assay cellular prion protein (PrP(C)) from an uninfected host brain is denatured at both pH 7.4 and 3.5 to produce two substrates. When the pH 7.4 substrate is incubated with TSE agent, the amount of PrP(C) that converts to a proteinase K (PK)-resistant state is modulated by the original host's species barrier to the TSE agent. In contrast, PrP(C) in the pH 3.5 substrate is misfolded by any TSE agent. By comparing the amount of PK-resistant prion protein in the two substrates, an assessment of the host's species barrier can be made. We show that the CER assay correctly predicts known prion species barriers of laboratory mice and, as an example, show some preliminary results suggesting that bobcats (Lynx rufus) may be susceptible to white-tailed deer (Odocoileus virginianus) chronic wasting disease agent.

Twitter Demographics

The data shown below were collected from the profile of 1 tweeter who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 12 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 12 100%

Demographic breakdown

Readers by professional status Count As %
Professor 3 25%
Student > Ph. D. Student 2 17%
Researcher 2 17%
Student > Master 2 17%
Student > Bachelor 1 8%
Other 2 17%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 5 42%
Agricultural and Biological Sciences 3 25%
Medicine and Dentistry 2 17%
Neuroscience 1 8%
Unspecified 1 8%
Other 0 0%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 14 April 2015.
All research outputs
#3,544,425
of 5,033,220 outputs
Outputs from Journal of Visualized Experiments
#2,372
of 3,389 outputs
Outputs of similar age
#123,364
of 169,810 outputs
Outputs of similar age from Journal of Visualized Experiments
#161
of 273 outputs
Altmetric has tracked 5,033,220 research outputs across all sources so far. This one is in the 16th percentile – i.e., 16% of other outputs scored the same or lower than it.
So far Altmetric has tracked 3,389 research outputs from this source. They receive a mean Attention Score of 3.0. This one is in the 21st percentile – i.e., 21% of its peers scored the same or lower than it.
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We're also able to compare this research output to 273 others from the same source and published within six weeks on either side of this one. This one is in the 29th percentile – i.e., 29% of its contemporaries scored the same or lower than it.