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From: TSS ()
Subject: Monthly Creutzfeldt Jakob Disease statistics 2 October 2007
Date: October 2, 2007 at 8:10 am PST

Monthly Creutzfeldt Jakob Disease statistics

DEPARTMENT OF HEALTH News Release issued by The Government News Network on 2 October 2007
The Department of Health is today issuing the latest information about the numbers of known cases of Creutzfeldt Jakob disease. This includes cases of variant Creutzfeldt Jakob disease (vCJD) - the form of the disease thought to be linked to BSE. The position is as follows:
Definite and probable CJD cases in the UK:

As at 28 September 2007
Summary of vCJD cases


Deaths from definite vCJD (confirmed): 114

Deaths from probable vCJD (without neuropathological confirmation): 47

Deaths from probable vCJD (neuropathological confirmation pending): 0

Number of deaths from definite or probable vCJD (as above): 161

Number of definite/probable vCJD cases still alive: 5

Total number of definite or probable vCJD (dead and alive): 166
Following this press notice the Department of Health will stop issuing monthly CJD press notices because the same data is also published by the National CJD Surveillance Unit in Edinburgh. Up to date figures for the number of vCJD cases in the UK can be found on the website of the National CJD Surveillance Unit at:

By Calendar Year

see chart of steady increase of sporadic CJD cases ...tss


* As at 28th September 2007
Referrals: a simple count of all the cases which have been referred to the National CJD Surveillance Unit for further investigation in the year in question. CJD may be no more than suspected; about half the cases referred in the past have turned out not to be CJD. Cases are notified to the Unit from a variety of sources including neurologists, neuropathologists, neurophysiologists, general physicians, psychiatrists, electroencephalogram (EEG) departments etc. As a safety net, death certificates coded under the specific rubrics 046.1 and 331.9 in the 9th ICD Revisions are obtained from the Office for National Statistics in England and Wales, the General Register Office for Scotland and the General Register Office for Northern Ireland.

Deaths: All columns show the number of deaths that have occurred in definite and probable cases of all types of CJD and GSS in the year shown. The figures include both cases referred to the Unit for investigation while the patient was still alive and those where CJD was only discovered post mortem (including a few cases picked up by the Unit from death certificates). There is therefore no read across from these columns to the referrals column. The figures will be subject to retrospective adjustment as diagnoses are confirmed.

Definite cases: this refers to the diagnostic status of cases. In definite cases the diagnosis will have been pathologically confirmed, in most cases by post mortem examination of brain tissue (rarely it may be possible to establish a definite diagnosis by brain biopsy while the patient is still alive).

Probable vCJD cases: are those who fulfil the 'probable' criteria set out in the Annex and are either still alive, or have died and await post mortem pathological confirmation. Those still alive will always be shown within the current year's figures.

Sporadic: Classic CJD cases with typical EEG and brain pathology. Sporadic cases appear to occur spontaneously with no identifiable cause and account for 85% of all cases.

Probable sporadic: Cases with a history of rapidly progressive dementia, typical EEG and at least two of the following clinical features; myoclonus, visual or cerebellar signs, pyramidal/extrapyramidalsigns or akinetic mutism.

Iatrogenic: where infection with classic CJD has occurred accidentally as the result of a medical procedure. All UK cases have resulted from treatment with human derived pituitary growth hormones or from grafts using dura mater (a membrane lining the skull).

Familial: cases occurring in families associated with mutations in the PrP gene (10 - 15% of cases).

GSS: Gerstmann-Straussler-Scheinker syndrome - an exceedingly rare inherited autosomal dominant disease, typified by chronic progressive ataxia and terminal dementia. The clinical duration is from 2 to 10 years, much longer than for CJD.

vCJD: Variant CJD, the hitherto unrecognised variant of CJD discovered by the National CJD Surveillance Unit and reported in The Lancet on 6 April 1996. This is characterised clinically by a progressive neuropsychiatric disorder leading to ataxia, dementia and myoclonus (or chorea) without the typical EEG appearance of CJD. Neuropathology shows marked spongiform change and extensive florid plaques throughout the brain.

Definite vCJD cases still alive: These will be cases where the diagnosis has been pathologically confirmed (by brain biopsy).



Beyond the PrPres Type1/ Type2 dichotomy in Creutzfeldt-Jakob Disease

Cassard, H1; Uro-Coste, E2; Simon, S3; Bilheude, JM4; Perret-Liaudet, A5;
Ironside, J6; Haik, S7; Basset-Leobon, C2; Lacroux, C1; Peoch’, K8; Stressenberger, N9;
Langeveld, J10; Head, M11; Hauw, JJ12; Schecher, F1; Delisle, MB13; Andreoletti, O1
1Ecole Natinale Vétérinaire de Toulouse, France; 22Service d’Anatomie Pathologique
and INSERM U466 R, France; 3DRM, CEA/Saclay, France; 4 Bio-Rad, R&D, France;
5 Hôpital Neurologique Service de Neurochimie, France; 6School of Molecular
& Clinical Medicine (Pathology),, National Creutzfeldt-Jakob Disease Surveillance Un,
UK; 7Pitié Salpetriere Universitary Hospital, France; 8Hôpital Lariboisière, Service de
Biochimie et Biologie Moléculaire,, France; 9Hôpital Neurologique -Service de
Neurochimie, France; 10CIDC-Lelystad, Netherlands; 11University of Edinburgh, Western General
Hospital, UK; 12Pitié Salpetriere Universitary Hospital,, Laboratoire de Neuropathologie, France;
13Rangueil Universitary Hospital, Service d’Anatomie Pathologique, France

Creutzfeldt-Jakob disease (CJD) cases are currently classified according to
established diagnostic criteria and by the genotype at codon 129 of the PRNP
gene and the Western blotting of the proteinase K digested abnormal prion
protein that distinguishes a type 1 and a type 2 profile. These biochemically
distinct PrPres types have been proposed to represent distinct prion strains.
However, since the cooccurence of type 1 and type 2 PrPres in the same patient
is common, the rationale of this classification and strain concept as applied to
CJD are currently under discussion.

Five different brain areas from of 40 sporadic CJD and 11 iatrogenic CJD
(both dura matter-, and growth hormone-associated) cases, originating from
UK and France, were systematically investigated, using Western blotting typing,
and by two others biochemical assays that depend on the behaviour of PrPSc in
variable PK digestion conditions.

As described previously, co-occurrence of type 1 and type 2 PrPres was
found in 30% of the CJD patients examined. However, our novel PK concentration
dependent assays identified a single uniform PrP type in cases where both type 1 and
type 2 were present. Moreover, in sCJD four distinct biochemical PrPSc
signatures were identified by the PK concentration dependent assays and these
correlated to the current genotype/clinico-pathological sCJD groups. In iCJD the four
similar biochemical signatures were observed, but were not correlated to particular
PRNP 129 polymorphism or Western Blot PrPres patterns. Moreover notable differences
were observed between PrPSc biochemical properties of French and UK GH-CJD cases,
which could reflect, as already suspected, differences in the causative agents.

Identification, in sCJD and iCJD, of four different PrPSc phenotypes
irrespective of patients PRNP polymorphism at codon 129 and Western blot
profile provides new insights into human prion disease aetiology and could reflects an
unsuspected diversity of TSE agents in human disease. Further investigations are
currently underway using animal transmission to correlate agent strain with our new
discriminant biochemical assays.


Diagnostic Utility of CSF Biomarkers and General CSF Findings in a U.S Sporadic CJD Cohort

Geschwind, MD; Haman, A; Torres-Chae, C; Raudabaugh, BJ; Devereux, G;
Miller, BL University of California, San Francisco, USA


The diagnostic utility of CSF biomarkers, including 14-3-3 protein,
neuron-specific enolase (NSE), AB42, total Tau (T-Tau), and phosphorylated
Tau (PTau)/ T-Tau ratio are often are used for sporadic CJD (sCJD) diagnosis
is controversial. We have previously reported the CSF 14-3-3 protein to have
poor sensitivity for CJD diagnosis. In this study, now report the sensitivity and specificity
of several CSF biomarkers and general characteristics in a U.S. cohort of sCJD
and non-prion rapidly progressive dementia (RPD) controls (Non-CJD cohort)


Clinical diagnoses are made through review of medical records, clinical evaluation, and
in many cases pathology. Data is stored in a secure clinical relational database, which was
queried for various CSF findings, including cell count, protein, IgG index, oligoclonal bands
(OCBs), 14-3-3, NSE, T-Tau in patients with probable or definite sporadic CJD and
non-prion rapidly progressive dementias (RPD), most of whom were referred to our center
with a potential diagnosis of CJD. For probable sCJD diagnosis, we used UCSF criteria that
are modified from WHO to substitute MRI for 14-3-3. T-Tau and NSE were considered
positive if > 1300 pg/ml and > 35 ng/ml, respectively. For this analysis, ambiguous 14-3-3
results were considered as negative.


14-3-3 protein (n=149) sensitivity was only 48% (47% for definite; 50% for
probable sCJD). NSE (n=49) sensitivity was 63% (66% for definite; 59% for probable
sCJD). T-Tau (n=28) was the most sensitive at 68% (69% for definite; 67% for
probable sCJD). The specificity of these biomarkers among our CJD and RPD controls
(n=72) was 66% for 14-3-3 (n=47), 81% for NSE (n=21), and 100% for T-Tau (n=7). The
14-3-3 had the lowest sensitivity and specificity. Mildly elevated CSF protein
(<100 mg/dl) is common in sCJD; High WBC; >20, is uncommon in sCJD.


In a cohort from a major U.S. CJD referral center, the 14-3-3
is neither sensitive nor specific for sCJD. NSE and T-Tau also show poor sensitivity for
sCJD. T-Tau may be more specific than 14-3-3 and NSE, but our “n” is small.
WHO sCJD criteria should be revised; by eliminating 14-3-3 and including brain
MRI into the criteria. We are currently analyzing the effects of disease duration and
codon 129 polymorphism on these CSF biomarker results.

see full text 143 pages ;

Date: September 24, 2007 at 6:52 pm PST

Molecular Features of the Protease-resistant Prion Protein (PrPres) in
H-type BSE

Biacabe, A-G1; Jacobs, JG2; Gavier-Widén, D3; Vulin, J1; Langeveld, JPM2;
Baron, TGM1 1AFSSA, France; 2CIDC-Lelystad, Netherlands; 3SVA, Sweden

Western blot analyses of PrPres accumulating in the brain of BSE-infected cattle have
demonstrated 3 different molecular phenotypes regarding to the apparent molecular
masses and glycoform ratios of PrPres bands. We initially described isolates (H-type
BSE) essentially characterized by higher PrPres molecular mass and decreased
levels of the diglycosylated PrPres band, in contrast to the classical type of BSE.
This type is also distinct from another BSE phenotype named L-type BSE, or
also BASE (for Bovine Amyloid Spongiform Encephalopathy), mainly characterized by a
low representation of the diglycosylated PrPres band as well as a lower PrPres
molecular mass. Retrospective molecular studies in France of all available BSE cases
older than 8 years old and of part of the other cases identified since the beginning of the
exhaustive surveillance of the disease in 20001 allowed to identify 7 H-type
BSE cases, among 594 BSE cases that could be classified as classical, L- or H-type BSE.
By Western blot analysis of H-type PrPres, we described a remarkable
specific feature with antibodies raised against the C-terminal region of PrP that
demonstrated the existence of a more C-terminal cleaved form of PrPres (named PrPres#2 ),
in addition to the usual PrPres form (PrPres #1). In the unglycosylated form,
PrPres #2 migrates at about 14 kDa, compared to 20 kDa for PrPres #1. The proportion
of the PrPres#2 in cattle seems to by higher compared to the PrPres#1. Furthermore
another PK–resistant fragment at about 7 kDa was detected by some more N-terminal
antibodies and presumed to be the result of cleavages of both N- and C-terminal parts
of PrP. These singular features were maintained after transmission of the disease to
C57Bl/6 mice. The identification of these two additional PrPres fragments (PrPres #2 and
7kDa band) reminds features reported respectively in sporadic Creutzfeldt-Jakob disease
and in Gerstmann-Sträussler-Scheinker (GSS) syndrome in humans.

BASE Transmitted to Primates and MV2 sCJD Subtype Share PrP27-30 and PrPSc
C-terminal Truncated Fragments

Zanusso, G1; Commoy, E2; Fasoli, E3; Fiorini, M3; Lescoutra, N4; Ruchoux,
MM4; Casalone, C5; Caramelli, M5; Ferrari, S3; Lasmezas, C6; Deslys, J-P4;
Monaco, S3 1University of Verona, of Neurological and Visual Sciences,
Italy; 2CEA, IMETI/SEPIA, France; 3University of Verona, Neurological and
Visual Sciences, Italy; 4IMETI/SEPIA, France; 5IZSPLVA, Italy; 6The Scripps
Research Insitute, USA

The etiology of sporadic Creutzfeldt-Jakob disease (sCJD), the most frequent human
prion disease, remains still unknown. The marked disease phenotype heterogeneity
observed in sCJD is thought to be influenced by the type of proteinase K-resistant
prion protein, or PrPSc (type 1 or type 2 according to the electrophoretic mobility of
the unglycosylated backbone), and by the host polymorphic Methionine/Valine
(M/V) codon 129 of the PRNP. By using a two-dimensional gel electrophoresis
(2D-PAGE) and imunoblotting we previously showed that in sCJD, in addition to the
PrPSc type, distinct PrPSc C-terminal truncated fragments (CTFs) correlated with
different sCJD subtypes. Based on the combination of CTFs and PrPSc type, we distinguished
three PrPSc patterns: (i) the first was observed in sCJD with PrPSc type 1 of all

(ii) the second was found in M/M-2 (cortical form); (iii) the third in amyloidogenic M/V-
2 and V/V-2 subtypes (Zanusso et al., JBC 2004) . Recently, we showed that sCJD
subtype M/V-2 shared molecular and pathological features with an atypical form of
BSE, named BASE, thus suggesting a potential link between the two conditions. This
connection was further confirmed after 2D-PAGE analysis, which showed an
identical PrPSc signature, including the biochemical pattern of CTFs. To pursue this
issue, we obtained brain homogenates from Cynomolgus macaques intracerebrally
inoculated with brain homogenates from BASE. Samples were separated by using
a twodimensional electrophoresis (2D-PAGE) followed by immunoblotting. We
here show that the PrPSc pattern obtained in infected primates is identical to
BASE and sCJD MV-2 subtype. These data strongly support the link, or at least a
common ancestry, between a sCJD subtype and BASE. This work was supported by




USA BASE CASE, (ATYPICAL BSE), AND OR TSE (whatever they are calling it
today), please note that both the ALABAMA COW, AND THE TEXAS COW, both were
''H-TYPE'', personal communication Detwiler et al Wednesday, August 22, 2007
11:52 PM. ...TSS


FC5.5.2 Transmission of Italian BSE and BASE Isolates in Cattle Results into
a Typical BSE Phenotype and a Muscle Wasting Disease

Zanusso, G1; Lombardi, G2; Casalone, C3; D’Angelo, A4; Gelmetti, D2; Torcoli, G2;
Barbieri, I2; Corona, C3; Fasoli, E1; Farinazzo, A1; Fiorini, M1; Gelati, M1; Iulini, B3;
Tagliavini, F5; Ferrari, S1; Monaco, S1; Caramelli, M3; Capucci, L2
1University of Verona, Neurological and Visual Sciences, Italy; 2IZSLER, Italy; 3IZSPLVA,
Italy; 4University of Turin, Animal Pathology, Italy; 5Isituto Carlo Besta, Italy

The clinical phenotype of bovine spongiform encephalopathy has been extensively
reported in early accounts of the disorder. Following the introduction of statutory active
surveillance, almost all BSE cases have been diagnosed on a pathological/molecular
basis, in a pre-symptomatic clinical stage. In recent years, the active surveillance
system has uncovered atypical BSE cases, which are characterized by distinct
conformers of the PrPSc, named high-type (BSE-H) and low-type (BSE-L), whose
clinicopathological phenotypes remain unknown. We recently reported two
Italian atypical cases with a PrPSc type similar to BSE-L, pathologically
characterized by PrP amyloid plaques. Experimental transmission to TgBov mice has
recently disclosed that BASE is caused by a distinct prion strain which is extremely
virulent. A major limitation of transmission studies to mice is the lack of reliable
information on clinical phenotype of BASE in its natural host. In the present study, we
experimentally infected Fresian/Holstein and Alpine/Brown cattle with Italian BSE and
BASE isolates by i.c. route. BASE infected cattle showed survival times significantly
shorter than BSE, a finding more readily evident in Fresian/Holstein, and in keeping with
previous observations in TgBov mice. Clinically, BSE-infected cattle developed a
disease phenotype highly comparable with that described in field BSE cases
and in experimentally challenged cattle. On the contrary, BASE-inoculated cattle
developed an amyotrophic disorder accompanied by mental dullness.
The molecular and neuropathological profiles, including PrP deposition
pattern, closely matched those observed in the original cases. This study further confirms
that BASE is caused by a distinct prion isolate and discloses a novel disease
phenotype in cattle, closely resembling the phenotype previous reported in scrapie-inoculated
cattle and in some subtypes of inherited and sporadic Creutzfeldt-Jakob disease.
Oral Abstracts

Transmission Results in Squirrel Monkeys Inoculated with Human sCJD, vCJD,
and GSS Blood Specimens: the Baxter Study

Brown, P1; Gibson, S2; Williams, L3; Ironside, J4; Will, R4; Kreil, T5;
Abee, C3 1Fondation Alliance BioSecure, France; 2University of South Alabama, USA;
3University of Texas MD Anderson Cancer Center, USA; 4Western General
Hospital, UK; 5Baxter BioSience, Austria

Background: Rodent and sheep models of Transmissible Spongiform Encephalopathy
(TSE) have documented blood infectivity in both the pre-clinical and clinical phases of
disease. Results in a (presumably more appropriate) non-human primate model
have not been reported.

Objective: To determine if blood components (red cells, white cells, platelets, and
plasma) from various forms of human TSE are infectious.

Methods: Blood components were inoculated intra-cerebrally (0.1 ml) and
intravenously (0.5 ml) into squirrel monkeys from 2 patients with sporadic
Creutzfeldt- Jakob disease (sCJD) and 3 patients with variant Creutzfeldt-Jakob disease
(vCJD). Additional monkeys were inoculated with buffy coat or plasma samples from
chimpanzees infected with either sCJD or Gerstmann-Sträussler-Scheinker
disease (GSS). Animals were monitored for a period of 5 years, and all dying
or sacrificed animals had post-mortem neuropathological examinations and
Western blots to determine the presence or absence of the misfolded ‘prion’
protein (PrPTSE).

Results: No transmissions occurred in any of the animals inoculated with
blood components from patients with sporadic or variant CJD. All donor chimpanzees
(sCJD and GSS) became symptomatic within 6 weeks of their pre-clinical phase
plasmapheresis, several months earlier than the expected onset of illness.
One monkey inoculated with purified leukocytes from a pre-clinical GSS
chimpanzee developed disease after 36 months.

Conclusion: No infectivity was found in small volumes of blood components
from 4 patients with sporadic CJD and 3 patients with variant CJD.

***However, a single transmission from a chimpanzee-passaged strain of GSS
shows that infectivity may be present in leukocytes, and the ‘shock’ of
general anaesthesia and plasmspheresis appears to have triggered the onset
of illness in pre-clinical donor chimpanzees.

Interim Transmission Results in Cynomolgus Macaques Inoculated with BSE and

vCJD Blood Specimens

Lasmezas, C1; Lescoutra, N2; Comoy, E2; Holznagel, E3; Loewer, J3; Motzkus,
D4; Hunsmann, G4; Ingrosso, L5; Bierke, P6; Pocchiari, M5; Ironside, J7; Will,
R7; Deslys, JP2 1Scripps Florida, Infectology, USA; 2CEA, France; 3PEI,
Germany; 4DPZ, Germany; 5Istituto Superiore di Sanita, Italy; 6SMI, Sweden;
7CJD Surveillance Unit, UK

BSE and vCJD transmitted to cynomolgus macaques reproduce many features of
human vCJD, including clinical symptoms, neuropathological hallmarks of vCJD,
PrPres electrophoretical pattern and, most importantly, the wide distribution of
infectivity in peripheral organs. The latter characteristic distinguishes vCJD from sCJD
in both humans and cynomolgus macaques, and prompted us to use this non-human
primate model for further investigations of vCJD and its risk for human health. The
occurrence of four vCJD infections in humans transfused with blood from patients who
later developed vCJD has raised concern about blood transfusion safety in
countries with vCJD.
In this collaborative European study, we investigated the infectivity of
blood components and whole blood administered by intracerebral (ic) and
intravenous (iv) routes. Buffy-coat and whole blood was inoculated by ic and
iv route, respectively, from two vCJD patients and from two clinical vCJD-inoculated
macaques. Transfusions were also performed from whole blood and blood leucodepleted
according to hospital practice standards from two clinical BSE inoculated
macaques. Blood infectivity during the preclinical phase is being examined in orally
infected macaques. Whole blood was collected and transfused from one such animal
two years after oral challenge, whereas buffy-coat and plasma from two animals
at 2 and 4.5 years post-challenge, respectively, have been inoculated by the
ic route.
This is an ongoing study in which recipient animals continue to be observed
at various times post-inoculation. So far, we have had one positive transmission in one
animal transfused 65 months earlier with 40 ml of whole blood from a vCJD macaque
(the characteristics of the disease in this animal will be shown in a separate poster by E.
Comoy). This positive transmission reproduces transfusion transmission of vCJD in
humans, with an incubation of 5.5 years compatible with incubation periods
observed in humans.

Assessing the Risk of vCJD Transmission by Dentistry; Distribution of
Infectivity in Oral Tissues of VM Mice after Simulated Oral Feeding of

Sutton, JM1; Kirby, E1; Dickinson, J1; Dennis, M1; Cornwall, M1; Vassey,
MJ1; Smith, A2; Marsh, PD3; Walker, JT1; Raven, NDH1
1Health Protection Agency, Centre for Emergency Preparedness and Response,,
TSE Research group, UK; 2University of Glasgow, Dental School, UK; 3Health
Protection Agency, Centre for Emergency Preparedness and Response,, UK

Background: Ongoing concerns about the prevalence of variant Creutzfeldt Jakob
Disease (vCJD) in the UK population has heightened concerns about the risks
of iatrogenic transmission of the disease. Although there have been no cases
to date of transmission by surgery there have been 4 cases involving blood
transfusion. This study aims to assess the potential of transmission of the disease by
dental procedures. Whilst the risks are undoubtably low the very large numbers of
procedures carried out annually have the potential to amplify the risks considerably
and there is very little data in this area to form the basis for accurate risk
assessments. Aim(s)/Objective(s): To assess the relative levels of infectivity in oral
tissues from a murine model following exposure to BSE-301V through the small

Methods. The study uses a BSE-301V, VM mouse model as a clinically relevant
model for assessing iatrogenic vCJD transmission between humans. Infectious mouse
brain homogenate was prepared and inoculated into a loop of the duodenum, to
prevent direct contamination of the oral tissues. Mice were sacrificed at 3-weekly
intervals and at appearance of clinical symptoms. A range of oral tissues,
including dental pulp, gingival margin, salivary gland, saliva, lingual tonsil and
trigeminal ganglia, together with brain and spleen tissues were removed, processed as
homogenates and reinoculated intracranially (ic.) into indicator mice.

Results: The primary challenge proved to be a very efficient route of
infection with a 100% attack rate and a mean incubation to clinical disease
of 157 ± 17 days (compared to 120 days for the same titre inoculum ic.).
Infectivity was observed in all oral and control tissues with varying time-courses
and titres estimated from incubation period.

Discussion: The results throw new light on the potential routes of
dissemination and spread of infectivity from the small intestine to the oral
cavity and its implications for possible iatrogenic transmission of vCJD via dental,
endoscopic or other forms of surgery.

Conclusion: The data generated from the study provides support for ongoing
risk assessments to look at the potential for vCJD transmission via dental
procedures alongside other elements of studies looking at effectiveness of
decontamination and re-use of dental instruments.

From: "Terry S. Singeltary Sr."
Subject: CWD UPDATE 88 AUGUST 31, 2007

Date: Wed, 29 Aug 2007 21:13:08 -0500
From: "Terry S. Singeltary Sr."
Subject: CWD NEW MEXICO RECORDS IT'S 19 CASE (near Texas border again)

Monitoring the Potential Transmission of Chronic Wasting Disease to Humans
Using a Hunter Registry Database in Wyoming (405 lines)
From: Terry S. Singeltary Sr. <[log in to unmask]>
Date: Thu, 30 Aug 2007 21:23:42 -0500


Quantifying the Species Barrier in Chronic Wasting Disease by a Novel in vitro Conversion Assay

Li, L1; Coulthart, MB2; Balachandran, A3; Chakrabartty, A4; Cashman, NR1
1University of British Columbia, Brain Research Centre, Canada; 2Public Health Agency
of Canada, National Microbiology Laboratory, Canada; 3Animal Diseases Research
Institute, Canada Food Inspection Agency, National Reference Laboratory for Scrapie
and CWD, Canada; 4Ontario Cancer Institute and Department of Medical
Biophysics, University of Toronto, Canada

Background: Chronic wasting disease (CWD) is a transmissible spongiform
encephalopathy that can affect North American cervids (deer, elk, and moose).
Although the risk of CWD crossing the species barrier and causing human disease is
still unknown, however, definite bovine spongiform encephalopathy (BSE) transmission
to humans as variant CJD (vCJD), it would seem prudent to limit the exposure of
humans to CWD.

Aim: In view of the fact that BSE can be readily transmitted to non-bovid species, it is
important to establish the species susceptibility range of CWD.

Methods: In vitro conversion system was performed by incubation of prions with
normal brain homogenates as described before, and protease K (PK) resistant
PrP was determined by immunoblotting with 6H4 monoclonal prion antibody.

Results: Our results demonstrate that PrPC from cervids (including moose) can be
efficiently converted to a protease-resistant form by incubation with elk CWD prions,
presumably due to sequence and structural similarities between these species.
Interestingly, hamster shows a high conversion ratio by PrPCWD. Moreover, partial
denaturation of substrate PrPC can apparently overcome the structural barriers
between more distant species.

Conclusions: Our work correctly predicted the transmission of CWD to a wild
moose. We find a species barrier for prion protein conversion between cervids and
other species, however, this barrier might be overcome if the PrPC substrate has
been partially denatured in a cellular environment. Such an environment might
also promote CWD transmission to non-cervid species, *** including humans.
Acid/GdnHCl-treated brain PrPC was a superior substrate for the in vitro conversion
than PrPC treated at physiological pH. This has implications for the process by which
the prion protein is converted in disease.

Subject: Cross-sequence transmission of sporadic Creutzfeldt-Jakob disease
creates a new prion strain

Date: August 25, 2007 at 12:42 pm PST

Subject: Cross-sequence transmission of sporadic Creutzfeldt-Jakob disease
creates a new prion strain
Date: August 25, 2007 at 12:42 pm PST

J Biol Chem. 2007 Aug 20; : 17709374

Cross-sequence transmission of sporadic Creutzfeldt-Jakob disease creates a
new prion strain.

[My paper] Atsushi Kobayashi , Masahiro Asano , Shirou Mohri , Tetsuyuki

The genotype (methionine or valine) at polymorphic codon 129 of the human
prion protein (PrP) gene and the type (type 1 or type 2) of abnormal isoform
of PrP (PrP(Sc)) are major determinants of the clinicopathological
phenotypes of sporadic Creutzfeldt-Jakob disease (sCJD). Here we found that
transmission of sCJD prions from a patient with valine homozygosity (129V/V)
and type 2 PrP(Sc) (sCJD-VV2 prions) to mice expressing human PrP with
methionine homozygosity (129M/M) generated unusual PrP(Sc) intermediate in
size between type 1 and type 2. The intermediate type PrP(Sc) was seen in
all examined dura mater graft-associated CJD cases with 129M/M and
plaque-type PrP deposits (p-dCJD). p-dCJD prions and sCJD-VV2 prions
exhibited similar transmissibility and neuropathology, and the identical
type of PrP(Sc) when inoculated into PrP-humanized mice with 129M/M or
129V/V. These findings suggest that p-dCJD could be caused by cross-sequence
transmission of sCJD-VV2 prions.


In this study, the strain-dependent traits of sCJDMM1
prions were inherited through cross-sequence
transmission without any modification. The
humanized mice with 129V/V produced type 1 PrPres
after inoculation with sCJD-MM1 prions. Because
sCJD-VV1 cases are extremely rare (at most 1-2%
of the total number of sCJD cases) and characterized
by early onset (mean age at onset: 39.3 years) (5),


our results raise the possibility that CJD cases
classified as VV1 may include cases caused by
iatrogenic transmission of sCJD-MM1 prions or
food-borne infection by type 1 prions from animals,
e.g., chronic wasting disease prions in cervid. In fact,
two CJD-VV1 patients who hunted deer or
consumed venison have been reported (40, 41). The
results of the present study emphasize the need for
traceback studies and careful re-examination of the
biochemical properties of sCJD-VV1 prions.


In conclusion, cross-sequence transmission of
sCJD-VV2 prions generates a new prion strain with
altered conformational properties and disease
phenotypes as p-dCJD prions. Furthermore, the
newly generated prions have unique transmissibility
including the traceback phenomenon. In the future, if
atypical prion strains emerge through cross-sequence
transmission, especially from animals, traceback
studies will enable us to identify the origin of the



Re: Colorado Surveillance Program for Chronic Wasting Disease
Transmission to Humans (TWO SUSPECT CASES)

Subject: Aspects of the Cerebellar Neuropathology in Nor98
Date: September 26, 2007 at 4:06 pm PST


Aspects of the Cerebellar Neuropathology in Nor98

Gavier-Widén, D1; Benestad, SL2; Ottander, L1; Westergren, E1
1National Veterinary Insitute, Sweden; 2National Veterinary Institute,

Nor98 is a prion disease of old sheep and goats. This atypical form of
scrapie was first described in Norway in 1998. Several features of Nor98
were shown to be different from classical scrapie including the distribution
of disease associated prion protein (PrPd) accumulation in the brain. The
cerebellum is generally the most affected brain area in Nor98. The study here
presented aimed at adding information on the neuropathology in the cerebellum
of Nor98 naturally affected sheep of various genotypes in Sweden and Norway.
A panel of histochemical and immunohistochemical (IHC) stainings such as IHC
for PrPd, synaptophysin, glial fibrillary acidic protein, amyloid, and cell markers
for phagocytic cells were conducted. The type of histological lesions and tissue
reactions were evaluated. The types of PrPd deposition were characterized. The
cerebellar cortex was regularly affected, even though there was a variation
in the severity of the lesions from case to case.
Neuropil vacuolation was more marked in the molecular layer, but affected
also the granular cell layer. There was a loss of granule cells. Punctate
deposition of PrPd was characteristic. It was morphologically and in distribution
identical with that of synaptophysin, suggesting that PrPd accumulates in
the synaptic structures. PrPd was also observed in the granule cell layer and in the
white matter.
*** The pathology features of Nor98 in the cerebellum of the affected sheep
showed similarities with those of sporadic Creutzfeldt-Jakob disease in

From: "Terry S. Singeltary Sr."
Sent: Tuesday, August 21, 2007 9:50 AM
Subject: TWO MORE Nor98 atypical Scrapie cases detected in USA bringing
total to 3 cases to date

Infected and Source Flocks

As of June 30, 2007, there were .....


One field case and one validation case were consistent with Nor-98 scrapie.

IN the February 2007 Scrapie report it only mentions ;

''One case was consistent with Nor98 scrapie.''

(please note flocks of origin were in WY, CO, AND CA. PERSONAL COMMUNCATIONS


An evaluation of scrapie surveillance in the United States


From: "Terry S. Singeltary Sr."
Reply-To: Sustainable Agriculture Network Discussion Group
Date: Fri, 2 Feb 2007 17:32:58 -0600

1997 TO 2006. SPORADIC CJD CASES TRIPLED, with phenotype
of 'UNKNOWN' strain growing. ...

There is a growing number of human CJD cases, and they were presented last
week in San Francisco by Luigi Gambatti(?) from his CJD surveillance

He estimates that it may be up to 14 or 15 persons which display selectively
SPRPSC and practically no detected RPRPSC proteins.

Diagnosis and Reporting of Creutzfeldt-Jakob Disease

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


MARCH 26, 2003

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

disease in the United States

Email Terry S. Singeltary:

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?

Copyright © 2003 Published by Elsevier Ltd.

Tracking spongiform encephalopathies in North America

Xavier Bosch

Available online 29 July 2003.

Volume 3, Issue 8, August 2003, Page 463

“My name is Terry S Singeltary Sr, and I live in Bacliff, Texas. I lost my
mom to hvCJD (Heidenhain variant CJD)
and have been searching for answers ever since. What I have found is that we
have not been told the truth. CWD
in deer and elk is a small portion of a much bigger problem.”

see history of cjd questionnaire

Sent: Monday May 28, 2007


Subject: HUMAN and ANIMAL TSE Classifications i.e. mad cow disease and the UKBSEnvCJD only theory
Date: January 29, 2006 at 9:03 am PST

Comments sent via JAMA Feedback Page
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HUMAN and ANIMAL TSE Classifications i.e. mad cow disease and the UKBSEnvCJD
only theory

TSEs have been rampant in the USA for decades in many species, and they all
have been rendered and fed back to animals for human/animal consumption.
propose that the current diagnostic criteria for human TSEs only enhances
and helps the spreading of human TSE from the continued belief of the
UKBSEnvCJD only theory in 2007. With all the science to date refuting it, to
continue to validate this myth, will only spread this TSE agent through a
multitude of potential routes and sources i.e. consumption, surgical, blood,
medical, cosmetics etc. I propose as with Aguzzi, Asante, Collinge, Caughey,
Deslys, Dormont, Gibbs, Ironside, Manuelidis, Marsh, et al and many more,
that the world of TSE Transmissible Spongiform Encephalopathy is far from an
exact science, but there is enough proven
science to date that this myth should be put to rest once and for all, and
that we move forward with a new classification for human and animal TSE that
would properly identify the infected species, the source species, and then
the route.

This would further have to be broken down to strain of species and then the
route of transmission would further have to be broken down. Accumulation and
Transmission are key to the threshold from sub-clinical to clinical disease,
and key to all this, is to stop the amplification and transmission of this
agent, the spreading of, no matter what strain. In my opinion, to continue
with this myth that the U.K. strain of BSE (one strain TSE in cows), and the
nv/v CJD (one strain TSE humans) and that all the rest of human TSE are just
one single strain i.e. sporadic CJD (when to date there are 6 different
phenotypes of sCJD, and growing per Gambetti et al), and that no other
animal TSE transmits to humans, to continue with this masquerade will only
continue to spread, expose, and kill, who knows how many more in the years
and decades to come. ONE was enough for me, My Mom, hvCJD i.e. Heidenhain
Variant CJD, DOD 12/14/97 confirmed, which is nothing more than another mans
name added to CJD, like CJD itself, Jakob and Creutzfeldt, or
Gerstmann-Straussler-Scheinker syndrome, just another CJD or human TSE,
named after another human.

WE are only kidding ourselves with the current diagnostic criteria for human
and animal TSE, especially differentiating between the nvCJD vs the sporadic
strains and then the GSS strains and also the FFI fatal familial insomnia
strains or the ones that mimics one or the other of those TSE? Tissue
infectivity and strain typing of the many variants of the human and animal
TSEs are paramount in all variants of all TSE. There must be a proper
classification that will differentiate between all these human TSE in order
to do this. With the CDI and other more sensitive testing coming about, I
only hope that my proposal will some day be taken seriously. ...

Terry S. Singeltary Sr.
P.O. Box 42
Bacliff, Texas USA 77518


CJD VOICE (voice for _all_ victims of human TSE)


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