# Cwd, Bse, Scrapie, Cjd Update December 2005



## terry (Sep 13, 2002)

CHRONIC WASTING DISEASE OF ELK AND DEER AND CREUTZFELDT-JAKOB DISEASE: COMPARATIVE ANALYSIS OF THE SCRAPIE PRION PROTEIN* 

Zhiliang Xie, Katherine I. ORourke§, Zhiqian Dong, Allen L. Jenny¶, Julie A. Langenberg&#9553;, Ermias D. Belay#, Lawrence B. Schonberger#, Robert B. Petersen, Wenquan Zou, Qingzhong Kong, Pierluigi Gambetti, and Shu G. Chen& 

From the Institute of Pathology and National Prion Disease Pathology Surveillance Center, Case Western Reserve University, Cleveland, OH 44106; §USDA Agricultural Research Services, Animal Disease Research Unit, Pullman, WA 99164; ¶USDA National Veterinary Services Laboratories, Ames, Iowa 50010; &#9553;Wildlife Health Program, Bureau of Wildlife Management, Wisconsin Department of Natural Resources, Madison, WI 53707 and #Centers for Disease Control and Prevention, National Center for Infectious Diseases, Division of Viral and Rickettsial Diseases, Atlanta, GA 30333. 

& To whom correspondence should be addressed: Institute of Pathology, Case Western Reserve University, Cleveland, OH 44106. Tel: (216) 368-8925; Fax: (216) 368-2546; E-mail: [email protected]. 

Running Title: PrPSc in CWD and CJD 

Chronic wasting disease (CWD), a transmissible prion disease that affects elk and deer, poses new challenges to animal and human public health. Although the transmission of CWD to humans has not been proven, it remains a possibility. If this were to occur, it is important to know whether the "acquired" human prion disease would show a phenotype including the scrapie prion protein (PrPSc) features that differ from those associated with human sporadic prion disease. In this study, we have compared the pathological profiles and PrPSc characteristics in brains of CWD-affected elk and deer with that in subjects with sporadic Creutzfeldt-Jakob disease (CJD), as well as CJD-affected subjects who might have been exposed to CWD, using histopathology, immunohistochemistry, immunoblotting, conformation stability assay (CSA) and N-terminal protein sequencing. Spongiform changes and intense PrPSc staining were present in several brain regions of CWD-affected animals. Immunoblotting revealed three proteinase K (PK)-resistant bands in CWD, representing different glycoforms of PrPSc. Following deglycosylation, the unglycosylated PK-resistant PrPSc of CWD migrated at 21kDa with an electrophoretic mobility similar to that of type 1 human PrPSc present in sporadic CJD affecting subjects homozygous for methionine at codon 129 (sCJDMM1). N-terminal sequencing showed that the PK cleavage site of PrPSc in CWD occurred at residues 82 and 78, similar to that of PrPSc in sCJDMM1. Furthermore, CSA also showed no significant difference between elk CWD PrPSc and the PrPSc species associated with sCJDMM1. However, there was a major difference in glycoform ratio of PrPSc between CWD and sCJDMM1 affecting both CWD-exposed and non-exposed subjects. Moreover, PrPSc of CWD exhibited a distinct constellation of glycoforms distinguishable from that of sCJDMM1 in two-dimensional immunoblots. These findings underline the importance of detailed PrPSc characterization in trying to detect novel forms of acquired prion disease. ...

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Coexistence of multiple PrPSc types in individuals with 

Creutzfeldt-Jakob disease 

Magdalini Polymenidou, Katharina Stoeck, Markus Glatzel, Martin Vey, Anne Bellon, and Adriano Aguzzi 

Summary 

Background The molecular typing of sporadic Creutzfeldt-Jakob disease (CJD) is based on the size and glycoform 

ratio of protease-resistant prion protein (PrPSc), and on PRNP haplotype. On digestion with proteinase K, type 1 and 

type 2 PrPSc display unglycosylated core fragments of 21 kDa and 19 kDa, resulting from cleavage around amino 

acids 82 and 97, respectively. 

Methods We generated anti-PrP monoclonal antibodies to epitopes immediately preceding the differential proteinase 

K cleavage sites. These antibodies, which were designated POM2 and POM12, recognise type 1, but not type 2, PrPSc. 

Findings We studied 114 brain samples from 70 patients with sporadic CJD and three patients with variant CJD. 

Every patient classified as CJD type 2, and all variant CJD patients, showed POM2/POM12 reactivity in the 

cerebellum and other PrPSc-rich brain areas, with a typical PrPSc type 1 migration pattern. 

Interpretation The regular coexistence of multiple PrPSc types in patients with CJD casts doubts on the validity of 

electrophoretic PrPSc mobilities as surrogates for prion strains, and questions the rational basis of current CJD 

classifications. 


snip... 


The above results set the existing CJD classifications 

into debate and introduce interesting questions about 

human CJD types. For example, do human prion types 

exist in a dynamic equilibrium in the brains of affected 

individuals? Do they coexist in most or even all CJD 

cases? Is the biochemically identified PrPSc type simply 

the dominant type, and not the only PrPSc species? 


http://neurology.thelancet.com Published online October 31, 2005 


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HUMAN TSE USA 2005 


Animal Prion Diseases Relevant to Humans (unknown types?)
Thu Oct 27, 2005 12:05
71.248.128.109 


About Human Prion Diseases / 
Animal Prion Diseases Relevant to Humans 

Bovine Spongiform Encephalopathy (BSE) is a prion disease of cattle. Since 1986, when BSE was recognized, over 180,000 cattle in the UK have developed the disease, and approximately one to three million are likely to have been infected with the BSE agent, most of which were slaughtered for human consumption before developing signs of the disease. The origin of the first case of BSE is unknown, but the epidemic was caused by the recycling of processed waste parts of cattle, some of which were infected with the BSE agent and given to other cattle in feed. Control measures have resulted in the consistent decline of the epidemic in the UK since 1992. Infected cattle and feed exported from the UK have resulted in smaller epidemics in other European countries, where control measures were applied later. 

Compelling evidence indicates that BSE can be transmitted to humans through the consumption of prion contaminated meat. BSE-infected individuals eventually develop vCJD with an incubation time believed to be on average 10 years. As of November 2004, three cases of BSE have been reported in North America. One had been imported to Canada from the UK, one was grown in Canada, and one discovered in the USA but of Canadian origin. There has been only one case of vCJD reported in the USA, but the patient most likely acquired the disease in the United Kingdom. If current control measures intended to protect public and animal health are well enforced, the cattle epidemic should be largely under control and any remaining risk to humans through beef consumption should be very small. (For more details see Smith et al. British Medical Bulletin, 66: 185. 2003.) 

Chronic Wasting Disease (CWD) is a prion disease of elk and deer, both free range and in captivity. CWD is endemic in areas of Colorado, Wyoming, and Nebraska, but new foci of this disease have been detected in Nebraska, South Dakota, New Mexico, Wisconsin, Mississippi Kansas, Oklahoma, Minnesota, Montana, and Canada. Since there are an estimated 22 million elk and deer in the USA and a large number of hunters who consume elk and deer meat, there is the possibility that CWD can be transmitted from elk and deer to humans. As of November 2004, the NPDPSC has examined 26 hunters with a suspected prion disease. However, all of them appeared to have either typical sporadic or familial forms of the disease. The NPDPSC coordinates with the Centers for Disease Control and state health departments to monitor cases from CWD-endemic areas. Furthermore, it is doing experimental research on CWD transmissibility using animal models. (For details see Sigurdson et al. British Medical Bulletin. 66: 199. 2003 and Belay et al. Emerging Infectious Diseases. 10(6): 977. 2004.) 


http://www.cjdsurveillance.com/abouthpd-animal.html 


SEE STEADY INCREASE IN SPORADIC CJD IN THE USA FROM 1997 TO 2004. SPORADIC CJD CASES TRIPLED, and that is with a human TSE surveillance system that is terrible flawed. in 1997 cases of the _reported_ cases of cjd were at 54, to 163 _reported_ cases in 2004. see stats here; 

p.s. please note the 47 PENDING CASES to Sept. 2005 

p.s. please note the 2005 Prion D. total 120(8) 8=includes 51 type pending, 1 TYPE UNKNOWN ??? 

p.s. please note sporadic CJD 2002(1) 1=3 TYPE UNKNOWN??? 

p.s. please note 2004 prion disease (6) 6=7 TYPE UNKNOWN??? 


http://www.cjdsurveillance.com/resources-casereport.html 


CWD TO HUMANS = sCJD ??? 


AS implied in the Inset 25 we must not _ASSUME_ that
transmission of BSE to other species will invariably
present pathology typical of a scrapie-like disease. 

snip... 

http://www.bseinquiry.gov.uk/files/yb/1991/01/04004001.pdf 


ATYPICAL TSEs in USA CATTLE AND SHEEP ? 


http://www.bseinquiry.gov.uk/files/sc/seac17/tab03.pdf 


Infected and Source Flocks 

As of August 31, 2005, there were 115 scrapie infected and source flocks (figure 3). There were 3 new infected and source flocks reported in August (Figure 4) with a total of 148 flocks reported for FY 2005 (Figure 5). The total infected and source flocks that have been released in FY 2005 are 102 (Figure 6), with 5 flocks released in August. The ratio of infected and source flocks released to newly infected and source flocks for FY 2005 = 0.69 :
1. In addition, as of August 31, 2005, 574 scrapie cases have been confirmed and reported by the National Veterinary Services Laboratories (NVSL), of which 122 were RSSS cases (Figure 7). This includes 55 newly confirmed cases in August 2005 (Figure 8). Fifteen cases of scrapie in goats have been reported since 1990 (Figure 9). The last goat case was reported in May 2005. 

snip... 

full text ; 

http://www.aphis.usda.gov/vs/nahps/scrapie/monthly_report/monthly-report.html 



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Diagnosis and Reporting of Creutzfeldt-Jakob Disease 

Singeltary, Sr et al. JAMA.2001; 285: 733-734. 


http://jama.ama-assn.org/cgi/content/full/285/6/733 




RE-Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob


disease in the United States


Email Terry S. Singeltary:


[email protected]



I lost my mother to hvCJD (Heidenhain Variant CJD). I would like to

comment on the CDC's attempts to monitor the occurrence of emerging

forms of CJD. Asante, Collinge et al [1] have reported that BSE

transmission to the 129-methionine genotype can lead to an alternate

phenotype that is indistinguishable from type 2 PrPSc, the commonest

sporadic CJD. However, CJD and all human TSEs are not reportable

nationally. CJD and all human TSEs must be made reportable in every

state and internationally. I hope that the CDC does not continue to

expect us to still believe that the 85%+ of all CJD cases which are

sporadic are all spontaneous, without route/source. We have many TSEs in

the USA in both animal and man. CWD in deer/elk is spreading rapidly and

CWD does transmit to mink, ferret, cattle, and squirrel monkey by

intracerebral inoculation. With the known incubation periods in other

TSEs, oral transmission studies of CWD may take much longer. Every

victim/family of CJD/TSEs should be asked about route and source of this

agent. To prolong this will only spread the agent and needlessly expose

others. In light of the findings of Asante and Collinge et al, there

should be drastic measures to safeguard the medical and surgical arena

from sporadic CJDs and all human TSEs. I only ponder how many sporadic

CJDs in the USA are type 2 PrPSc?


http://www.neurology.org/cgi/eletters/60/2/176#535



CVM Update
<December 5, 2005

November 2005 Update on Feed Enforcement Activities to Limit the Spread of
BSE

To help prevent the establishment and amplification of BSE through feed in
the United States, FDA implemented a final rule that prohibits the use of
most mammalian protein in feeds for ruminant animals. This rule, Title 21
Part 589.2000 of the Code of Federal Regulations, here called the Ruminant
Feed Ban, became effective on August 4, 1997.

This is an update on FDA enforcement activities regarding the ruminant feed
regulation. FDA's CVM has assembled data from the inspections that have been
conducted AND whose final inspection report has been recorded in the FDA's
inspection database as of November 26, 2005. As of November 26, 2005, FDA
had received over 41,000 inspection reports. The majority of these
inspections (around 68%) were conducted by State officials under contract to
FDA, with the remainder conducted by FDA officials.

Inspections conducted by FDA or State investigators are classified to
reflect the compliance status at the time of the inspection based upon the
objectionable conditions documented. These inspection conclusions are
reported as Official Action Indicated (OAI), Voluntary Action Indicated
(VAI), or No Action Indicated (NAI).

An OAI inspection classification occurs when significant objectionable
conditions or practices were found and regulatory sanctions are warranted in
order to address the establishment's lack of compliance with the regulation.
An example of an OAI inspection classification would be findings of
manufacturing procedures insufficient to ensure that ruminant feed is not
contaminated with prohibited material. Inspections classified with OAI
violations will be promptly re-inspected following the regulatory sanctions
to determine whether adequate corrective actions have been implemented.

A VAI inspection classification occurs when objectionable conditions or
practices were found that do not meet the threshold of regulatory
significance, but do warrant advisory actions to inform the establishment of
findings that should be voluntarily corrected. Inspections classified with
VAI violations are more technical violations of the Ruminant Feed Ban. These
include provisions such as minor recordkeeping lapses and conditions
involving non-ruminant feeds.

An NAI inspection classification occurs when no objectionable conditions or
practices were found during the inspection or the significance of the
documented objectionable conditions found does not justify further actions.

The results to date are reported here both by segment of industry and in
total. NOTE  A single firm can operate as more than one firm type. As a
result, the categories of the different industry segments are not mutually
exclusive.

RENDERERS

These firms are the first to handle and process (i.e., render) animal
proteins and to send these processed materials to feed mills and/or protein
blenders for use as a feed ingredient.

Number of active firms whose initial inspection has been reported to FDA 
274

Number of active firms handling materials prohibited from use in ruminant
feed  185 (68% of those active firms inspected)

Of the 185 active firms handling prohibited materials, their most recent
inspection revealed that:

1 firm (0.5%) was classified as OAI

11 firms (5.9%) were classified as VAI

LICENSED FEED MILLS

FDA licenses these feed mills to produce medicated feed products. The
license is required to manufacture and distribute feed using certain potent
drug products, usually those requiring some pre-slaughter withdrawal time.
This licensing has nothing to do with handling prohibited materials under
the feed ban regulation. A medicated feed license from FDA is not required
to handle materials prohibited under the Ruminant Feed Ban.

Number of active firms whose initial inspection has been reported to FDA 
1,079

Number of active firms handling materials prohibited from use in ruminant
feed  426 (39% of those active firms inspected)

Of the 426 active firms handling prohibited materials, their most recent
inspection revealed that:

0 firm (0%) was classified as OAI

8 firms (1.9%) were classified as VAI

FEED MILLS NOT LICENSED BY FDA

These feed mills are not licensed by the FDA to produce medicated feeds.

Number of active firms whose initial inspection has been reported to FDA 
5,165

Number of active firms handling materials prohibited from use in ruminant
feed  2,036 (39% of those active firms inspected)

Of the 2,036 active firms handling prohibited materials, their most recent
inspection revealed that:

2 firms (0.1%) were classified as OAI

24 firms (1.2%) were classified as VAI

PROTEIN BLENDERS

These firms blend rendered animal protein for the purpose of producing
quality feed ingredients that will be used by feed mills.

Number of active firms whose initial inspection has been reported to FDA --
340

Number of active firms handling materials prohibited from use in ruminant
feed  147 (43% of those active firms inspected)

Of the 147 active firms handling prohibited materials, their most recent
inspection revealed that:

0 firms (0%) were classified as OAI

7 firms (4.8%) were classified as VAI

RENDERERS, FEED MILLS, AND PROTEIN BLENDERS

This category includes only those firms that actually use prohibited
material to manufacture, process, or blend animal feed or feed ingredients.

Number of active renderers, feed mills, and protein blenders whose initial
inspection has been reported to FDA  6,576

Number of active renderers, feed mills, and protein blenders processing with
prohibited materials  539 (8.2% of those active firms inspected)

Of the 539 of active renderers, feed mills, and protein blenders processing
with prohibited materials, their most recent inspection revealed that:

3 firms (0.6%) were classified as OAI

23 firms (4.3%) were classified as VAI

OTHER FIRMS INSPECTED

Examples of such firms include ruminant feeders, on-farm mixers, pet food
manufacturers, animal feed salvagers, distributors, retailers, and animal
feed transporters.

Number of active firms whose initial inspection has been reported to FDA 
13,477

Number of active firms handling materials prohibited from use in ruminant
feed  3,748 (28% of those active firms inspected)

Of the 3,748 active firms handling prohibited materials, their most recent
inspection revealed that:

8 firms (0.2%) were classified as OAI

95 firms (2.5%) were classified as VAI

TOTAL FIRMS

Note that a single firm can be reported under more than one firm category;
therefore, the summation of the individual OAI/VAI firm categories will be
more than the actual total number of OAI/VAI firms, as presented below.

Number of active firms whose initial inspection has been reported to FDA 
16,476

Number of active firms handling materials prohibited from use in ruminant
feed  4,553 (27% of those active firms inspected)

Of the 4,553 active firms handling prohibited materials, their most recent
inspection revealed that:

9 firms (0.2%) were classified as OAI

107 firms (2.4%) were classified as VAI


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continued...


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## terry (Sep 13, 2002)

Issued by:
FDA, Center for Veterinary Medicine,
Communications Staff, HFV-12
7519 Standish Place, Rockville, MD 20855
Telephone: (240) 276-9300 FAX: (240) 276-9115
Internet Web Site: http://www.fda.gov/cvm

=======================================


Semiannual Report to Congress FY - 2005 - First Half 

snip... 


Stopping BSE at the BorderUSDA Needs To 

Strengthen Controls Over Canadian Beef Imports 

Following the detection of a Canadian cow with bovine 

spongiform encephalopathy (BSE or mad cow disease) 

in May 2003, we examined the Animal and Plant Health 

Inspection Services (APHIS) oversight of the importation 

of beef products from Canada. Following requests from 

four U.S. Senators, we began several reviews in June 

2004 to explore whether USDA did not follow appropriate 

safety measures, beginning sometime in the fall of 2003, 

in allowing expanded Canadian beef imports into the 

United States. 

After the initial halt of imports, in August 2003 the 

Secretary announced a list of low-risk products that would 

be allowed from Canada. APHIS also allowed an 

expansion in the type of Canadian facilities that could 

produce items for export to the United States. The 

gradual expansion occurred because agency employees 

included products similar to those on the published lowrisk 

list, but APHIS did not communicate this broadly. 

As a result, from August 2003 to April 2004, APHIS issued 

permits for products with questionable eligibility. Contrary 

to publicly stated policy, the agency allowed the import of 

products from Canadian facilities that produced both 

eligible and ineligible products, increasing the possibility 

that higher-risk product could be inadvertently imported. 

APHIS also issued permits to allow the import of more 

than 63,000 pounds of beef cheek meat with questionable 

eligibility because the agency did not establish a clear 

definition for boneless beef. Further, we found that 

FSIS did not always communicate effectively about the 

eligibility status of beef cheek meat, specifically to import 

inspectors. In addition, APHIS issued 1,155 permits for 

the importation of ruminant (e.g., cow, goat) products 

from Canada without ensuring that the agency had an 

appropriate system of internal controls to manage the 

process for a suddenly overwhelming volume of requests. 

From May through September 2004, we identified more 

than 42,000 pounds of product with questionable 

eligibility. 

APHIS generally agreed to institute procedures for 

communicating changes in policy and monitoring the 

consistency between agency practice and publicly stated 

policy, as well as to strengthen controls and finalize 

procedures to issue and monitor permits. FSIS generally 

agreed to implement controls to communicate the specific 

eligibility of product when its eligibility status changes and 

to implement an edit check in its import information 

system to identify ineligible product. (Audit Report No. 

33601-1-Hy, APHIS Oversight of the Importation of Beef 

Products from Canada) 


snip... 


Restaurant Owner Sentenced for Smuggling Beef 

from Japan, Importation of Which Is Prohibited Due 

to Disease Concerns 

In January 2005, a Los Angeles restaurant owner was 

placed on probation for 60 months, to include 800 hours 

of community service, after he pled guilty to smuggling 

beef from Japan. Under 9 Code of Federal Regulations 

(C.F.R.) § 94, beef from Japan is a prohibited product 

for United States importation due to disease. On two 

occasions in 2001 and 2002, inspectors in Anchorage, 

Alaska, intercepted shipments sent from Japan that 

were manifested as book, but upon inspection by 

USDA and the United States Customs Service, were 

found to contain approximately 25 kilograms of beef 

inside a Styrofoam ice chest. Both shipments were 

addressed to the restaurant owner. Shipping records 

showed that the restaurant owner had received 13 

shipments manifested as book from the same sender 

in Japan in 2001 and 2002. All but one of the shipments 

were in the same weight range as the two intercepted 

shipments. The shipper and the restaurant owner were 

subsequently indicted for various charges including 

conspiracy and smuggling. An arrest warrant was 

issued for the shipper, who is still in Japan. 



http://www.usda.gov/oig/webdocs/SarcFirstHalf05.pdf



[Docket No. 03-025IFA] FSIS Prohibition of the Use of Specified Risk Materials for Human Food and Requirement for the Disposition of Non-Ambulatory Disabled Cattle

03-025IFA
03-025IFA-2
Terry S. Singeltary


Page 1 of 17

From: Terry S. Singeltary Sr. [[email protected]]

Sent: Thursday, September 08, 2005 6:17 PM

To: [email protected]

Subject: [Docket No. 03-025IFA] FSIS Prohibition of the Use of Specified Risk Materials for Human Food and Requirements

for the Disposition of Non-Ambulatory Disabled Cattle

Greetings FSIS,

I would kindly like to submit the following to [Docket No. 03-025IFA] FSIS Prohibition of the Use of Specified Risk Materials for Human Food and

Requirements for the Disposition of Non-Ambulatory Disabled Cattle

THE BSE/TSE SUB CLINICAL Non-Ambulatory Disabled Cattle

Broken bones and such may be the first signs of a sub clinical BSE/TSE Non-Ambulatory Disabled Cattle ;

SUB CLINICAL PRION INFECTION

MRC-43-00

Issued: Monday, 28 August 2000

NEW EVIDENCE OF SUB-CLINICAL PRION INFECTION: IMPORTANT RESEARCH

FINDINGS RELEVANT TO CJD AND BSE

A team of researchers led by Professor John Collinge at the Medical

Research Council Prion Unit1 report today in the Proceedings of the

National Academy of Sciences, on new evidence for the existence of a

?sub-clinical? form of BSE in mice which was unknown until now.

The scientists took a closer look at what is known as the ?species

barrier? - the main protective factor which limits the ability of

prions2 to jump from one species to infect another. They found the mice

had a ?sub-clinical? form of disease where they carried high levels of

infectivity but did not develop the clinical disease during their normal

lifespan. The idea that individuals can carry a disease and show no

clinical symptoms is not new. It is commonly seen in conventional

infectious diseases.

Researchers tried to infect laboratory mice with hamster prions3 called

Sc237 and found that the mice showed no apparent signs of disease.

However, on closer inspection they found that the mice had high levels

of mouse prions in their brains. This was surprising because it has

always been assumed that hamster prions could not cause the disease in

mice, even when injected directly into the brain.

In addition the researchers showed that this new sub-clinical infection

could be easily passed on when injected into healthy mice and hamsters.

The height of the species barrier varies widely between different

combinations of animals and also varies with the type or strain of

prions. While some barriers are quite small (for instance BSE easily

infects mice), other combinations of strain and species show a seemingly

impenetrable barrier. Traditionally, the particular barrier studied here

was assumed to be robust.

Professor John Collinge said: "These results have a number of important

implications. They suggest that we should re-think how we measure

species barriers in the laboratory, and that we should not assume that

just because one species appears resistant to a strain of prions they

have been exposed to, that they do not silently carry the infection.

9/13/2005

2

Page 2 of 17

This research raises the possibility, which has been mentioned before,

that apparently healthy cattle could harbour, but never show signs of, BSE.

"This is a timely and unexpected result, increasing what we know about

prion disease. These new findings have important implications for those

researching prion disease, those responsible for preventing infected

material getting into the food chain and for those considering how best

to safeguard health and reduce the risk that theoretically, prion

disease could be contracted through medical and surgical procedures."

ISSUED FRIDAY 25 AUGUST UNDER EMBARGO. PLEASE NOTE THAT THE EMBARGO IS

SET BY THE JOURNAL.

FOR FURTHER INFORMATION CONTACT THE MRC PRESS OFFICE ON 020 7637 6011

(OFFICE HOURS) OR 07818 428297 OR 0385 774357 (OUT-OF-OFFICE-HOURS) OR

PROFESSOR JOHN COLLINGE ON 020 7594 3760. PLEASE NOTE THAT OWING TO

TRAVEL COMMITMENTS PROFESSOR COLLINGE WILL ONLY BE AVAILABLE UNTIL 16.30

ON FRIDAY 25 AUGUST AND CONTACTABLE AGAIN ON MONDAY 28 AUGUST VIA THE

MRC PRESS OFFICE. DR ANDREW HILL (A CO-AUTHOR ON THE PAPER) FROM THE

DEPARTMENT OF PATHOLOGY AT THE UNIVERSITY OF MELBOURNE WILL BE AVAILABLE

ON 00 61 3 8344 3995 (DURING OFFICE HOURS) OR 00 61 3 9443 0009

(OUT-OF-OFFICE HOURS). PLEASE NOTE THAT AUSTRALIA IS TEN HOURS AHEAD OF

UK TIME.

NOTES FOR EDITORS

Professor Collinge is a consultant neurologist and Director of the newly

formed MRC Prion Unit based at The Imperial College School of Medicine

at St Mary?s Hospital. He is also a member of the UK Government?s

Spongiform Encephalopathy Advisory Committee (SEAC). The MRC prion unit

is was set up in 1999, and its work includes molecular genetic studies

of human prion disease and transgenic modelling of human prion diseases.

Prions are unique infectious agents that cause fatal brain diseases such

as Creutzfeldt-Jakob disease (CJD) in humans and scrapie and BSE (mad

cow disease) in animals. In some circumstances prions from one species

of animals can infect another and it is clear that BSE has done this to

cause the disease variant CJD in the UK and France. It remains unclear

how large an epidemic of variant CJD will occur over the years ahead.

The strain of prion used here to infect the mice is the Sc237 strain

(also known as 263K) which infects hamsters, and until now was assumed

not to infect mice.

This research was funded by the Medical Research Council and Wellcome

Trust.

The Medical Research Council (MRC) is a national organisation funded by

the UK tax-payer. Its business is medical research aimed at improving

human health; everyone stands to benefit from the outputs. The research

it supports and the scientists it trains meet the needs of the health

services, the pharmaceutical and other health-related industries and the

academic world. MRC has funded work which has led to some of the most

significant discoveries and achievements in medicine in the UK. About

half of the MRC?s expenditure of £345 million is invested in over 50 of

its Institutes and Units, where it employs its own research staff. The

remaining half goes in the form of grant support and training awards to

individuals and teams in universities and medical schools.

The Wellcome Trust is the world's largest medical research charity with

a spend of some £600 million in the current financial year 1999/2000.

The Wellcome Trust supports more than 5,000 researchers, at 400

locations, in 42 different countries to promote and foster research with

the aim of improving human and animal health. As well as funding major

initiatives in the public understanding of science, the Wellcome Trust

is the country's leading supporter of research into the history of

medicine.

http://www.mrc.ac.uk/index/public_i...blic-press_releases_2000/public-mrc-43-00.htm

SNIP...FULL TEXT;

9/13/2005

Page 3 of 17

https://web01.aphis.usda.gov/regpublic.nsf/0/eff9eff1f7c5cf2b87256ecf000df08d?OpenDocument



Merry Christmas. ...TSS


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## terry (Sep 13, 2002)

Research Project: Transmission, Differentiation, and Pathobiology of Transmissible Spongiform Encephalopathies

Location: Virus and Prion Diseases of Livestock

Title: Experimental Second Passage of Chronic Wasting Disease (Cwd-Mule Deer) Agent to Cattle

Authors
item Hamir, Amirali
item Kunkle, Robert - bob
item Miller, Janice - ARS RETIRED
item Greenlee, Justin
item Richt, Juergen

Submitted to: Journal Of Comparative Pathology
Publication Acceptance Date: July 25, 2005
Publication Date: N/A

Interpretive Summary: To compare the findings of experimental first and second passage of chronic wasting disease (CWD) in cattle, 6 calves were inoculated into the brain with CWD-mule deer agent previously (first) passaged in cattle. Two other uninoculated calves served as controls. Beginning 10-12 months post inoculation (PI), all inoculates lost appetite and weight. Five animals subsequently developed clinical signs of central nervous system (CNS) abnormality. By 16.5 months PI, all cattle had been euthanized because of poor prognosis. None of the animals showed microscopic lesions of spongiform encephalopathy (SE) but the CWD agent was detected in their CNS tissues by 2 laboratory techniques (IHC and WB). These findings demonstrate that inoculated cattle amplify CWD agent but also develop clinical CNS signs without manifestation of microscopic lesions of SE. This situation has also been shown to occur following inoculation of cattle with another TSE agent, namely, sheep scrapie. The current study confirms previous work that indicates that the diagnostic tests currently used for confirmation of bovine spongiform encephalopathy (BSE) in the U.S. would detect CWD in cattle, should it occur naturally. Furthermore, it raises the possibility of distinguishing CWD from BSE in cattle due to the absence of microscopic lesions and a unique multifocal distribution of PrPres, as demonstrated by IHC, which in this study, appears to be more sensitive than the WB.

Technical Abstract: To compare clinicopathological findings of first and second passage of chronic wasting disease (CWD) in cattle, a group of calves (n=6) were intracerebrally inoculated with CWD-mule deer agent previously (first) passaged in cattle. Two other uninoculated calves served as controls. Beginning 10-12 months post inoculation (PI), all inoculates lost appetite and lost weight. Five animals subsequently developed clinical signs of central nervous system (CNS) abnormality. By 16.5 months PI, all cattle had been euthanized because of poor prognosis. None of the animals showed microscopic lesions of spongiform encephalopathy (SE) but PrPres was detected in their CNS tissues by immunohistochemistry (IHC) and Western blot (WB) techniques. These findings demonstrate that intracerebrally inoculated cattle not only amplify CWD PrPres but also develop clinical CNS signs without manifestation of morphologic lesions of SE. This situation has also been shown to occur following inoculation of cattle with another TSE agent, scrapie. The current study confirms previous work that indicates the diagnostic techniques currently used for confirmation of bovine spongiform encephalopathy (BSE) in the U.S. would detect CWD in cattle, should it occur naturally. Furthermore, it raises the possibility of distinguishing CWD from BSE in cattle due to the absence of neuropathologic lesions and a unique multifocal distribution of PrPres, as demonstrated by IHC, which in this study, appears to be more sensitive than the WB. 


http://www.ars.usda.gov/research/publications/publications.htm?seq_no_115=178318

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## terry (Sep 13, 2002)

Journal of Virology, January 2006, p. 596-604, Vol. 80, No. 2
0022-538X/06/$08.00+0 doi:10.1128/JVI.80.2.596-604.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved. 

Inhibition of Protease-Resistant Prion Protein Formation in a Transformed Deer Cell Line Infected with Chronic Wasting Disease 
Gregory J. Raymond,1 Emily A. Olsen,1 Kil Sun Lee,1 Lynne D. Raymond,1 P. Kruger Bryant III,2 Gerald S. Baron,1 Winslow S. Caughey,1 David A. Kocisko,1 Linda E. McHolland,2 Cynthia Favara,1 Jan P. M. Langeveld,3 Fred G. van Zijderveld,3 Richard T. Mayer,2 Michael W. Miller,4 Elizabeth S. Williams,5, and Byron Caughey1* 
Laboratory of Persistent Viral Diseases, NIAID, NIH, Rocky Mountain Laboratories, Hamilton, Montana 59840,1 USDA/ARS/ABADRL, Laramie, Wyoming 82071,2 Department for Bacteriology and TSEs, CIDC-Lelystad, 8203 AA 2004, Lelystad, The Netherlands,3 Colorado Division of Wildlife, Wildlife Research Center, Fort Collins, Colorado 80526-2097,4 Department of Veterinary Sciences, University of Wyoming, Laramie, Wyoming 820705

Received 3 August 2005/ Accepted 17 October 2005

Chronic wasting disease (CWD) is an emerging transmissible spongiform encephalopathy (prion disease) of North American cervids, i.e., mule deer, white-tailed deer, and elk (wapiti). To facilitate in vitro studies of CWD, we have developed a transformed deer cell line that is persistently infected with CWD. Primary cultures derived from uninfected mule deer brain tissue were transformed by transfection with a plasmid containing the simian virus 40 genome. A transformed cell line (MDB) was exposed to microsomes prepared from the brainstem of a CWD-affected mule deer. CWD-associated, protease-resistant prion protein (PrPCWD) was used as an indicator of CWD infection. Although no PrPCWD was detected in any of these cultures after two passes, dilution cloning of cells yielded one PrPCWD-positive clone out of 51. This clone, designated MDBCWD, has maintained stable PrPCWD production through 32 serial passes thus far. A second round of dilution cloning yielded 20 PrPCWD-positive subclones out of 30, one of which was designated MDBCWD2. The MDBCWD2 cell line was positive for fibronectin and negative for microtubule-associated protein 2 (a neuronal marker) and glial fibrillary acidic protein (an activated astrocyte marker), consistent with derivation from brain fibroblasts (e.g., meningeal fibroblasts). Two inhibitors of rodent scrapie protease-resistant PrP accumulation, pentosan polysulfate and a porphyrin compound, indium (III) meso-tetra(4-sulfonatophenyl)porphine chloride, potently blocked PrPCWD accumulation in MDBCWD cells. This demonstrates the utility of these cells in a rapid in vitro screening assay for PrPCWD inhibitors and suggests that these compounds have potential to be active against CWD in vivo. 



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* Corresponding author. Mailing address: Rocky Mountain Labs, 903 S. 4th St., Hamilton, MT 59840. Phone: (406) 363-9264. Fax: (406) 363-9286. E-mail: [email protected]. 

We dedicate this paper to the memory of Elizabeth S. Williams, a pioneer of CWD research. 

Deceased. 



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Journal of Virology, January 2006, p. 596-604, Vol. 80, No. 2
0022-538X/06/$08.00+0 doi:10.1128/JVI.80.2.596-604.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved. 


http://jvi.asm.org/cgi/content/abst...IRSTINDEX=0&volume=80&issue=2&journalcode=jvi




Journal of Virology, January 2006, p. 1044-1046, Vol. 80, No. 2
0022-538X/06/$08.00+0 doi:10.1128/JVI.80.2.1044-1046.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved. 

Mefloquine, an Antimalaria Drug with Antiprion Activity In Vitro, Lacks Activity In Vivo 
David A. Kocisko1* and Byron Caughey1 
National Institute of Allergy & Infectious Diseases, National Institutes of Health, Hamilton, Montana 598401

Received 25 August 2005/ Accepted 27 October 2005

In view of the effectiveness of antimalaria drugs inhibiting abnormal protease-resistant prion protein (PrP-res) formation in scrapie agent-infected cells, we tested other antimalarial compounds for similar activity. Mefloquine (MF), a quinoline antimalaria drug, was the most active compound tested against RML and 22L mouse scrapie agent-infected cells, with 50% inhibitory concentrations of 0.5 and 1.2 µM, respectively. However, MF administered to mice did not delay the onset of intraperitoneally inoculated scrapie agent, the result previously observed with quinacrine. While most anti-scrapie agent compounds inhibit PrP-res formation in vitro, many PrP-res inhibitors have no activity in vivo. This underscores the importance of testing promising candidates in vivo. 



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* Corresponding author. Mailing address: Rocky Mountain Laboratories, 903 S. 4th Street, Hamilton, MT 59840. Phone: (406) 375-9692. Fax: (406) 363-9286. E-mail: [email protected]. 



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Journal of Virology, January 2006, p. 1044-1046, Vol. 80, No. 2
0022-538X/06/$08.00+0 doi:10.1128/JVI.80.2.1044-1046.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved. 



http://jvi.asm.org/cgi/content/abst...IRSTINDEX=0&volume=80&issue=2&journalcode=jvi


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