Whole Blood Gene Expression Profiling in Preclinical and Clinical Cattle Infected with Atypical Bovine Spongiform Encephalopathy
Whole Blood Gene Expression Profiling in Preclinical and Clinical Cattle
Infected with Atypical Bovine Spongiform Encephalopathy
Elena Xerxa, Maura Barbisin, Maria Novella Chieppa, Helena Krmac, Elena
Vallino Costassa, Paolo Vatta, Marion Simmons, Maria Caramelli, Cristina
Casalone, Cristiano Corona, Giuseppe Legname
PLOS
Published: April 13, 2016 • http://dx.doi.org/10.1371/journal.pone.0153425
Abstract
Prion diseases, such as bovine spongiform encephalopathies (BSE), are
transmissible neurodegenerative disorders affecting humans and a wide variety of
mammals. Variant Creutzfeldt-Jakob disease (vCJD), a prion disease in humans,
has been linked to exposure to BSE prions. This classical BSE (cBSE) is now
rapidly disappearing as a result of appropriate measures to control animal
feeding. Besides cBSE, two atypical forms (named H- and L-type BSE) have
recently been described in Europe, Japan, and North America. Here we describe
the first wide-spectrum microarray analysis in whole blood of atypical
BSE-infected cattle. Transcriptome changes in infected animals were analyzed
prior to and after the onset of clinical signs. The microarray analysis revealed
gene expression changes in blood prior to the appearance of the clinical signs
and during the progression of the disease. A set of 32 differentially expressed
genes was found to be in common between clinical and preclinical stages and
showed a very similar expression pattern in the two phases. A 22-gene signature
showed an oscillating pattern of expression, being differentially expressed in
the preclinical stage and then going back to control levels in the symptomatic
phase. One gene, SEL1L3, was downregulated during the progression of the
disease. Most of the studies performed up to date utilized various tissues,
which are not suitable for a rapid analysis of infected animals and patients.
Our findings suggest the intriguing possibility to take advantage of whole blood
RNA transcriptional profiling for the preclinical identification of prion
infection. Further, this study highlighted several pathways, such as immune
response and metabolism that may play an important role in peripheral prion
pathogenesis. Finally, the gene expression changes identified in the present
study may be further investigated as a fingerprint for monitoring the
progression of disease and for developing targeted therapeutic interventions.
snip...
Discussion Whole blood is the most suitable tissue for a prospective rapid
diagnostic test since it minimizes sample handling artifacts and reduces sample
variability due to fractionation. The present study revealed a substantial gene
expression alteration in whole blood from atypical BSE-infected cattle, which
could be investigated in future experiments and, if confirmed, could be
exploited as a signature for the disease. One of the major caveats in using
peripheral blood is that its cellular components may change dramatically during
infections or inflammation. The animals used in the present study did not show
any apparent side pathology, they were monitored daily by the husbandry staff
and their blood was examined for serum aspartate aminotransferase (AST),
creatine kinase (CK) and manganese [23]. Nonetheless, possible interference due
to hidden pathologies or to inter-individual variations in hematocrit and white
blood cell count may affect interpretation of expression data and should be
considered as an important variable in future studies. In light of this, the
present results should be read as a first exploration of whole blood
transcriptomics during a prion infection. To our knowledge, this is the first
microarray study of whole blood from BSE-infected cattle. Indeed, in a study
published by Panelli et al. [36], fractionated white blood cells were analyzed
to detect gene expression changes in L-type infected animals. Very few DEGs are
common between the two studies. However, this discrepancy may be explained by
different infection methods, microarray platforms, statistical analysis
stringency and p value cut-off. Also, the white cells used in the study of
Panelli et al. were isolated from 1 year post-infection animals, while in our
study we used whole blood from preclinical and clinical infected cattle (around
6 months and 22–26 post-infection respectively).
In the present study, 4 statistical comparisons were performed: infected
(preclinical and clinical) versus control (IvsCtrl), preclinical versus control
(PvsCtrl), clinical versus control (CvsCtrl) and clinical versus preclinical
(CvsP) comparisons. Since our goal was to find a common pattern among all the
atypical BSE-infected cattle, we defined them as a single group of infected
(H-type and L-type) animals. Indeed, as published by Konold and colleagues [23],
these animals shared a very similar phenotype in terms of behavioral and
clinical signs. Also, another study by Priemer and colleagues indicated a large
similarity at level of PrPSc anatomical distribution for the atypical strains,
with only slight differences in the overall intensities between H- and L-type
[27]. Nonetheless, even if H- and L-type BSE are reported to share many
similarities, they constitute two distinct BSE variants which are characterized
by a different electrophoretic mobility of PrPSc unglycosylated moiety after
proteinase K (PK) digestion [16, 18]. Statistical comparison between the 4
H-type and the 4 L-type infected animals was carried out in preliminary analyses
(see S4 Table), but only a limited number of DEGs was found. Among them, only 15
had a p value lower than 0.01 and only 16 showed a fold change higher than 3,
indicating that, at least in terms of number of DEGs, these two groups did not
display large differences. For these reasons, we decided to focus our attention
on finding a common gene pattern among all the atypical BSE-infected cattle and
therefore we pooled the two groups. Due to the high inter-animal variability,
which is expected for outbreed animals, further studies in larger animal cohorts
would be required to investigate in detail the strain-specific gene expression
changes occurring during the progression of the disease. Still, the HvsL
analysis can be used as a sort of internal control in this study.
Another aspect to be taken into account when reading the present results is
that additional negative control cattle, aged from 12 to 37 months and derived
from a different herd compared to the Konold’s study groups [23], were
introduced in the analyses. The addition of these controls was useful to balance
the samples from infected animals and allowed a preliminary exploration of the
differentially expressed transcripts. However, age-related and environmental
variability may have affected in some degree the data and need to be considered
for their correct interpretation. Despite some limitations, since several
statistical analyses were performed (including the CvsP analysis, in which all
the animals derived from the same herd) a cross comparison of all them, as we
did with the Venn Diagram, may be very useful in order to define a set of genes
which could be a good starting point for further validation experiments in the
future. In the first statistical analysis we performed (IvsC), we found that
among 101 DEGs, 93 had known functions and were involved in several biological
processes and molecular pathways, such as autoimmune thyroiditis, chemokine and
cytokine activity, regulation of the secretion pathway, the immune system and
antigen presentation[52]. Previous studies on CNS tissues from BSE-infected
animals also showed the involvement of many of these pathways in prion
pathogenesis [4, 35, 52]. This similarity between brain and blood may not be
surprising, since it has been shown in the literature that blood transcriptome
analyses identify genes that are relevant to the pathological processes
occurring in the CNS [53]. Indeed, measuring disease-related gene expression in
peripheral blood may be a useful proxy measure for gene expression in the CNS
[53, 54].
To characterize the gene expression profile in the preclinical and clinical
stages, we performed the PvsCtrl and the CvsCtrl statistical comparisons. We
found that 113 probe sets were differentially regulated in the preclinical stage
of the disease, while 207 probe sets had an altered expression in the clinical
phase. Importantly, the present results indicated that, at least in blood, a
consistent gene expression alteration is present from the early stages of the
disease. This finding is in agreement with microarray analysis carried out by
Tang et al., which revealed the highest degree of differential gene regulation
in brains of cBSE-infected cattle at 21 months post infection, which is prior to
the detection of infectivity [4]. Also, Tortosa and colleagues found a
significant number of DEGs at early stages of the disease in the CNS from
cBSE-infected transgenic mice [52].
Venn diagram analysis revealed that 32 DEGs were in common between the
clinical and preclinical groups and, remarkably, they had a very similar pattern
of expression in both stages of the disease. Since these genes are altered in
both phases, it would be very interesting to confirm their differential
expression in future experiments with additional negative controls, and
eventually in blood from human patients.
Based on GO enrichment analysis, we found that immunity and inflammation
processes were strongly involved during the progression of the disease stages.
Interestingly, we found that antigen processing and presentation via MHC (major
histocompatibility complex) molecules and the autoimmune thyroiditis pathway
were significantly altered in atypical BSE-challenged animals. The majority of
MHC class I molecule coding-genes were down-regulated in infected cattle (three
out of four probes) and, also, MHC class II molecule coding transcripts were
found to be down-regulated during the progression of the clinical signs (four
out of four probes were down-regulated in the CvsP comparison). The involvement
of MHC transcripts in prion pathogenesis is supported by another microarray
study published by Khaniya and colleagues in 2009 [55]. In line with the trend
found by the microarray analysis, the RT-qPCR validation experiments indicated a
downregulation for MHC class I heavy chain (BOLA), even though the results
failed to reach the statistical significance (data not shown).
Regarding the autoimmune thyroiditis pathway, it is well known in the
literature that Hashimoto’s encephalitis, together with the associated
thyroiditis, is a differential diagnosis for CJD, since the two pathologies
share a very similar clinical symptomatology [56]. As hypothesized previously by
Prusiner and colleagues, the clinical and neuropathological similarities between
CJD and Hashimoto’s thyroiditis raise the possibility that protein misprocessing
may underlie both neurodegenerative and autoimmune diseases [57].
Finally, a fourth statistical analysis was performed to identify any
specific changes between the clinical and the preclinical stages of disease
(CvsP). Indeed, we found that the last phases of the disease are accompanied by
the overactivation of several genes involved in the immune defense response. In
particular, the shift from the preclinical towards the clinical stage was
characterized by the upregulation of genes involved in B cell proliferation and
the ISG15 (IFN-induced 15-kDa protein) conjugation system. ISG15 is a
ubiquitin-like molecule that is tightly regulated by specific innate immunity
signaling pathways [58]. Interestingly, it has been shown in the literature that
this protein is over-activated in the spinal cord of amyotrophic lateral
sclerosis mice models [59] and it has been indicated as a general marker for
both acute and chronic neuronal injuries [60].
To further analyze the data, we compared the list of DEGs found in PvsCtrl,
CvsP and CvsCtrl and found 22 genes with an oscillatory pattern of expression,
being differentially expressed in the preclinical stage and then going back
roughly to the control level in the clinical stage. Interestingly, some of the
oscillatory DEGs are involved in regulation of transcription, thus suggesting
that the gene expression during atypical BSE infection is tightly regulated.
Venn diagram analysis revealed that one gene, SEL1L3, was down-regulated in all
the comparisons (PvsCtrl, CvsCtrl, CvsP). SEL1L3 codes for a transmembrane
protein whose function is unknown. Interestingly, an important paralog of
SEL1L3, SEL1L, is involved in the retrotranslocation of misfolded proteins from
the lumen of the endoplasmic reticulum to the cytosol, where they are degraded
by the proteasome in an ubiquitin-dependent manner [61]. Therefore, we could
hypothesize that its down-regulation in prion infected animals would lead to a
reduced degradation of PrPSc, thus supporting the progression of the disease. We
validated this gene by RT-qPCR, confirming its downregulation in both the
preclinical and clinical stages of the disease. Further investigation on the
function of SEL1L3L would be of great interest since this gene may play an
important role in prion disease and maybe other neurodegenerative
illnesses.
Besides SEL1L3, five other genes were validated by RT-qPCR; here we will
briefly discuss how these genes may be involved in prion pathogenesis and in
host response to prion infection.
GNLY and CD40L were found to be down-regulated in both preclinical and
clinical stages. GNLY is a powerful antimicrobial protein contained within the
granules of cytotoxic T lymphocyte and natural killer cells. This gene was found
to be downregulated also in a microarray study performed on the medulla
oblongata from sheep with preclinical natural scrapie [62]. Thus, it may be a
good candidate as an early biomarker for atypical BSE but also for other prion
diseases.
CD40–CD40L interactions mediate a broad variety of immune and inflammatory
responses and have been implicated in the pathogenesis of Alzheimer’s disease
(AD) [63, 64]. Although the importance of CD40L in prion disease progression has
not yet been clarified (66–68), its downregulation in blood during both
preclinical and clinical stages of atypical BSE-infection suggests that prion
infection has an impact on the host immune system response and that immune
tolerance may be an active process induced by prions.
Two other downregulated genes were validated by RT-qPCR, namely HBA2 and
XIST. Concerning HBA2, we found a downregulation in preclinical atypical
BSE-infected cattle. Haemoglobins are iron-containing proteins that transport
oxygen in the blood of most vertebrates. Beside blood, HBA and HBB are also
expressed in mesencephalic dopaminergic neurons and glial cells [65] and are
down regulated in AD, PD and other neurodegenerative diseases [66]. Haemoglobin
genes expression alteration during preclinical scrapie was also found in the
spleen and CNS of infected animals [67, 68], as well as in the brains of
nonhuman primates infected with BSE [51]. These findings suggest an involvement
of these genes in the host response to general neurodegenerative processes.
Besides changes in transcript levels, it has been found that both HbA and HbB
protein distribution is altered in mitochondrial fractions from PD degenerating
brain [69]. Moreover, HbA is also expressed in endothelial cells, where it
regulates the nitric oxide signaling [70]. Even though a clear mechanism linking
these molecules to neurodegeneration has not yet been described, taken together
these findings strongly suggest a central role for haemoglobin in
neurodegenerative processes.
A marked downregulation in XIST expression was found in our infected
animals. XIST is a gene located on X chromosomes which codes for a long
non-coding RNA (LncRNA) involved in X-chromosome dosage control [71, 72].
LncRNAs are emerging as useful biomarkers for neurodegenerative diseases such as
AD [73] and other disease processes [74], and they can be easily detected in
blood and urine from patients. In addition, we cannot exclude the possibility
that the alteration in XIST expression may have some role in gender-dependent
response to prion infection [75].
RT-qPCR experiments confirmed the upregulation of PDK4 in clinically
affected animals. PDK4 encodes for a mitochondrial protein involved in glucose
metabolism through the inhibition of pyruvate dehydrogenase complex, which leads
to a reduction in pyruvate conversion to acetyl-CoA [76]. In the literature, a
key role has been suggested for acetyl-CoA fueling for the survival of
cholinergic neurons in the course of neurodegenerative diseases [77]. PDK4
overactivation can lead to a switch from glucose catabolism to fatty acid
utilization [78], thus increasing the production of ketone bodies. Notably, it
has been shown in the literature that these molecules are able to cross the
blood brain barrier. We could speculate that in prion infection (or at least in
atypical BSE infection) the concentration of ketone bodies would rise in blood,
as a consequence of PDK4 upregulation, and act in the brain as neuroprotective
molecules [79, 80]. This would be an attempt by the organism to prevent the
neurodegeneration induced by prions.
snip...
Conclusions In conclusion, the present study has led to the identification
of several gene expression changes in whole blood from clinical and preclinical
atypical BSE cattle, which upon further investigation and validation in blood
from human patients, might represent a molecular fingerprint to characterize
this disease. By comparing our results with other studies on various animal
prion diseases, we observed that some of the most significantly altered DEGs we
found in blood were found differentially expressed also in brain tissue from
BSE-infected cattle; this observation indicates that whole blood transcriptome
analyses may serve as a proxy measure for the changes occurring in the CNS of
infected animals. Furthermore, our study underlines the importance of utilizing
whole blood, without any additional manipulation, as a source tissue as it is an
easily accessible body fluid. In addition, the transcription regulation
activated in atypical BSE infections is similar to some extent to the one
observed in the literature for cBSE, even though the clinical characteristics
and biochemical properties are very different. Thus, this gene expression
profile may be investigated in other BSE infections to identify a common
molecular fingerprint.
Overall, our study confirmed the differential expression of 6 genes (XIST,
CD40L, GNLY, PDK4, HBA2 and SEL1L3), which may play several roles in atypical
BSE pathogenesis and, possibly, in other prion infections. Indeed, they are
involved in multiple pathways such as immune response, inflammation, and glucose
catabolism. Even though further studies are required to investigate the specific
involvement of all the identified genes in prion diseases, our data indicate an
important role for immune system regulation in the prion pathogenesis of
atypical BSE and maybe in BSE as well as in other prion diseases in general.
Wednesday, September 23, 2015
NIH Availability for Licensing AGENCY: [FR Doc. 2015–24117 Filed 9–22–15;
8:45 am] Detection and Discrimination of Classical and Atypical L-Type BSE
Strains by RT-QuIC
Monday, March 19, 2012
Infectivity in Skeletal Muscle of Cattle with Atypical Bovine Spongiform
Encephalopathy PLoS One. 2012; 7(2): e31449.
2015-2016
***********OCTOBER 2015*************
*** PRION 2015 ORAL AND POSTER CONGRESSIONAL ABSTRACTS ***
THANK YOU PRION 2015 TAYLOR & FRANCIS, Professor Chernoff, and
Professor Aguzzi et al, for making these PRION 2015 Congressional Poster and
Oral Abstracts available freely to the public. ...Terry S. Singeltary Sr.
O.05: Transmission of prions to primates after extended silent incubation
periods: Implications for BSE and scrapie risk assessment in human populations
Emmanuel Comoy, Jacqueline Mikol, Val erie Durand, Sophie Luccantoni,
Evelyne Correia, Nathalie Lescoutra, Capucine Dehen, and Jean-Philippe Deslys
Atomic Energy Commission; Fontenay-aux-Roses, France
Prion diseases (PD) are the unique neurodegenerative proteinopathies
reputed to be transmissible under field conditions since decades. The
transmission of Bovine Spongiform Encephalopathy (BSE) to humans evidenced that
an animal PD might be zoonotic under appropriate conditions. Contrarily, in the
absence of obvious (epidemiological or experimental) elements supporting a
transmission or genetic predispositions, PD, like the other proteinopathies, are
reputed to occur spontaneously (atpical animal prion strains, sporadic CJD
summing 80% of human prion cases). Non-human primate models provided the first
evidences supporting the transmissibiity of human prion strains and the zoonotic
potential of BSE. Among them, cynomolgus macaques brought major information for
BSE risk assessment for human health (Chen, 2014), according to their
phylogenetic proximity to humans and extended lifetime. We used this model to
assess the zoonotic potential of other animal PD from bovine, ovine and cervid
origins even after very long silent incubation periods. ***We recently observed
the direct transmission of a natural classical scrapie isolate to macaque after
a 10-year silent incubation period, with features similar to some reported for
human cases of sporadic CJD, albeit requiring fourfold longe incubation than
BSE. ***Scrapie, as recently evoked in humanized mice (Cassard, 2014), is the
third potentially zoonotic PD (with BSE and L-type BSE), ***thus questioning the
origin of human sporadic cases. We will present an updated panorama of our
different transmission studies and discuss the implications of such extended
incubation periods on risk assessment of animal PD for human health.
===============
***thus questioning the origin of human sporadic cases...
===============
***Our study demonstrates susceptibility of adult cattle to oral
transmission of classical BSE. ***
***our findings suggest that possible transmission risk of H-type BSE to
sheep and human. ***
P.86: Estimating the risk of transmission of BSE and scrapie to ruminants
and humans by protein misfolding cyclic amplification
Morikazu Imamura, Naoko Tabeta, Yoshifumi Iwamaru, and Yuichi Murayama
National Institute of Animal Health; Tsukuba, Japan
To assess the risk of the transmission of ruminant prions to ruminants and
humans at the molecular level, we investigated the ability of abnormal prion
protein (PrPSc) of typical and atypical BSEs (L-type and H-type) and typical
scrapie to convert normal prion protein (PrPC) from bovine, ovine, and human to
proteinase K-resistant PrPSc-like form (PrPres) using serial protein misfolding
cyclic amplification (PMCA).
Six rounds of serial PMCA was performed using 10% brain homogenates from
transgenic mice expressing bovine, ovine or human PrPC in combination with PrPSc
seed from typical and atypical BSE- or typical scrapie-infected brain
homogenates from native host species. In the conventional PMCA, the conversion
of PrPC to PrPres was observed only when the species of PrPC source and PrPSc
seed matched. However, in the PMCA with supplements (digitonin, synthetic polyA
and heparin), both bovine and ovine PrPC were converted by PrPSc from all tested
prion strains. On the other hand, human PrPC was converted by PrPSc from typical
and H-type BSE in this PMCA condition.
Although these results were not compatible with the previous reports
describing the lack of transmissibility of H-type BSE to ovine and human
transgenic mice, ***our findings suggest that possible transmission risk of
H-type BSE to sheep and human. Bioassay will be required to determine whether
the PMCA products are infectious to these animals.
================
==========================================
***our findings suggest that possible transmission risk of H-type BSE to
sheep and human. Bioassay will be required to determine whether the PMCA
products are infectious to these animals.
==========================================
Research Project: TRANSMISSION, DIFFERENTIATION, AND PATHOBIOLOGY OF
TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHIES
Title: Transmission of scrapie prions to primate after an extended silent
incubation period
Authors
item Comoy, Emmanuel - item Mikol, Jacqueline - item Luccantoni-Freire,
Sophie - item Correia, Evelyne - item Lescoutra-Etchegaray, Nathalie - item
Durand, Valérie - item Dehen, Capucine - item Andreoletti, Olivier - item
Casalone, Cristina - item Richt, Juergen item Greenlee, Justin item Baron,
Thierry - item Benestad, Sylvie - item Hills, Bob - item Brown, Paul - item
Deslys, Jean-Philippe -
Submitted to: Scientific Reports Publication Type: Peer Reviewed Journal
Publication Acceptance Date: May 28, 2015 Publication Date: June 30, 2015
Citation: Comoy, E.E., Mikol, J., Luccantoni-Freire, S., Correia, E.,
Lescoutra-Etchegaray, N., Durand, V., Dehen, C., Andreoletti, O., Casalone, C.,
Richt, J.A., Greenlee, J.J., Baron, T., Benestad, S., Brown, P., Deslys, J.
2015. Transmission of scrapie prions to primate after an extended silent
incubation period. Scientific Reports. 5:11573.
Interpretive Summary:
The transmissible spongiform encephalopathies (also called prion diseases)
are fatal neurodegenerative diseases that affect animals and humans. The agent
of prion diseases is a misfolded form of the prion protein that is resistant to
breakdown by the host cells. Since all mammals express prion protein on the
surface of various cells such as neurons, all mammals are, in theory, capable of
replicating prion diseases. One example of a prion disease, bovine spongiform
encephalopathy (BSE; also called mad cow disease), has been shown to infect
cattle, sheep, exotic undulates, cats, non-human primates, and humans when the
new host is exposed to feeds or foods contaminated with the disease agent. The
purpose of this study was to test whether non-human primates (cynomologous
macaque) are susceptible to the agent of sheep scrapie. After an incubation
period of approximately 10 years a macaque developed progressive clinical signs
suggestive of neurologic disease. Upon postmortem examination and microscopic
examination of tissues, there was a widespread distribution of lesions
consistent with a transmissible spongiform encephalopathy. This information will
have a scientific impact since it is the first study that demonstrates the
transmission of scrapie to a non-human primate with a close genetic relationship
to humans. This information is especially useful to regulatory officials and
those involved with risk assessment of the potential transmission of animal
prion diseases to humans.
Technical Abstract:
Classical bovine spongiform encephalopathy (c-BSE) is an animal prion
disease that also causes variant Creutzfeldt-Jakob disease in humans. Over the
past decades, c-BSE's zoonotic potential has been the driving force in
establishing extensive protective measures for animal and human health. In
complement to the recent demonstration that humanized mice are susceptible to
scrapie, we report here the first observation of direct transmission of a
natural classical scrapie isolate to a macaque after a 10-year incubation
period. Neuropathologic examination revealed all of the features of a prion
disease: spongiform change, neuronal loss, and accumulation of PrPres throughout
the CNS.
***This observation strengthens the questioning of the harmlessness of
scrapie to humans, at a time when protective measures for human and animal
health are being dismantled and reduced as c-BSE is considered controlled and
being eradicated. Our results underscore the importance of precautionary and
protective measures and the necessity for long-term experimental transmission
studies to assess the zoonotic potential of other animal prion strains.
***This information will have a scientific impact since it is the first
study that demonstrates the transmission of scrapie to a non-human primate with
a close genetic relationship to humans. This information is especially useful to
regulatory officials and those involved with risk assessment of the potential
transmission of animal prion diseases to humans.
***This observation strengthens the questioning of the harmlessness of
scrapie to humans, at a time when protective measures for human and animal
health are being dismantled and reduced as c-BSE is considered controlled and
being eradicated. Our results underscore the importance of precautionary and
protective measures and the necessity for long-term experimental transmission
studies to assess the zoonotic potential of other animal prion strains.
PRION 2015 CONFERENCE FT. COLLINS CWD RISK FACTORS TO HUMANS
*** LATE-BREAKING ABSTRACTS PRION 2015 CONFERENCE ***
O18
Zoonotic Potential of CWD Prions
Liuting Qing1, Ignazio Cali1,2, Jue Yuan1, Shenghai Huang3, Diane Kofskey1,
Pierluigi Gambetti1, Wenquan Zou1, Qingzhong Kong1 1Case Western Reserve
University, Cleveland, Ohio, USA, 2Second University of Naples, Naples, Italy,
3Encore Health Resources, Houston, Texas, USA
*** These results indicate that the CWD prion has the potential to infect
human CNS and peripheral lymphoid tissues and that there might be asymptomatic
human carriers of CWD infection.
==================
***These results indicate that the CWD prion has the potential to infect
human CNS and peripheral lymphoid tissues and that there might be asymptomatic
human carriers of CWD infection.***
==================
P.105: RT-QuIC models trans-species prion transmission
Kristen Davenport, Davin Henderson, Candace Mathiason, and Edward Hoover
Prion Research Center; Colorado State University; Fort Collins, CO USA
Conversely, FSE maintained sufficient BSE characteristics to more
efficiently convert bovine rPrP than feline rPrP. Additionally, human rPrP was
competent for conversion by CWD and fCWD.
***This insinuates that, at the level of protein:protein interactions, the
barrier preventing transmission of CWD to humans is less robust than previously
estimated.
================
***This insinuates that, at the level of protein:protein interactions, the
barrier preventing transmission of CWD to humans is less robust than previously
estimated.***
================
*** PRICE OF CWD TSE PRION POKER GOES UP 2014 ***
Transmissible Spongiform Encephalopathy TSE PRION update January 2, 2014
*** chronic wasting disease, there was no absolute barrier to conversion of
the human prion protein.
*** Furthermore, the form of human PrPres produced in this in vitro assay
when seeded with CWD, resembles that found in the most common human prion
disease, namely sCJD of the MM1 subtype.
*** These results would seem to suggest that CWD does indeed have zoonotic
potential, at least as judged by the compatibility of CWD prions and their human
PrPC target. Furthermore, extrapolation from this simple in vitro assay suggests
that if zoonotic CWD occurred, it would most likely effect those of the PRNP
codon 129-MM genotype and that the PrPres type would be similar to that found in
the most common subtype of sCJD (MM1).***
*** The potential impact of prion diseases on human health was greatly
magnified by the recognition that interspecies transfer of BSE to humans by beef
ingestion resulted in vCJD. While changes in animal feed constituents and
slaughter practices appear to have curtailed vCJD, there is concern that CWD of
free-ranging deer and elk in the U.S. might also cross the species barrier.
Thus, consuming venison could be a source of human prion disease. Whether BSE
and CWD represent interspecies scrapie transfer or are newly arisen prion
diseases is unknown. Therefore, the possibility of transmission of prion disease
through other food animals cannot be ruled out. There is evidence that vCJD can
be transmitted through blood transfusion. There is likely a pool of unknown size
of asymptomatic individuals infected with vCJD, and there may be asymptomatic
individuals infected with the CWD equivalent. These circumstances represent a
potential threat to blood, blood products, and plasma supplies.
Tuesday, April 12, 2016
The first detection of Chronic Wasting Disease (CWD) in Europe Norway
Thursday, January 29, 2015
Atypical H-TYPE BSE Case Confirmed in Norway
Monday, April 11, 2016
***DECLARATION OF EXTRAORDINARY EMERGENCY DUE TO A FOREIGN ANIMAL DISEASE
TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY TSE PRION CHRONIC WASTING DISEASE CWD IN
THE UNITED STATES AND NORTH AMERICA ? ***
MAD SHEEP OF MAD RIVER VALLEY AND THE USDA ‘FALSE FLAG’ WAR ON THAT FAMILY,
AND WHY TODAY A DECLARATION OF EXTRAORDINARY EMERGENCY DUE TO A FOREIGN ANIMAL
DISEASE TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY TSE PRION CHRONIC WASTING
DISEASE CWD IN THE UNITED STATES AND NORTH AMERICA MUST BE CALLED FOR !!!
Friday, February 20, 2015
***APHIS Freedom of Information Act (FOIA) Appeal Mouse Bio-Assays
2007-00030-A Sheep Imported From Belgium and the Presence of TSE Prion Disease
Kevin Shea to Singeltary 2015
http://transmissiblespongiformencephalopathy.blogspot.com/2015/02/aphis-freedom-of-information-act-foia.html
http://www.amazon.com/Mad-Sheep-Story-behind-Family/dp/1933392762
Friday, March 18, 2016 CFSAN
Constituent Update: FDA Announces Final Rule on Bovine Spongiform
Encephalopathy BSE MAD COW TSE PRION Center for Food Safety and Applied
Nutrition - Constituent Update
Tuesday, March 15, 2016
*** Docket No. FDA-2016-N-0321 Risk Assessment of Foodborne Illness
Associated with Pathogens from Produce Grown in Fields Amended with Untreated
Biological Soil Amendments of Animal Origin; Request for Comments, Scientific
Data, and Information Singeltary Submission ***
Monday, March 28, 2016
National Scrapie Eradication Program February 2016 Monthly Report
*** Docket No. APHIS-2007-0127 Scrapie in Sheep and Goats Terry Singeltary
Sr. Submission ***
Monday, November 16, 2015
*** Docket No. APHIS-2007-0127 Scrapie in Sheep and Goats Terry Singeltary
Sr. Submission ***
Saturday, July 18, 2015
SPONTANEOUS TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY TSE PRION AKA MAD COW
TYPE DISEASE, DOES IT EXIST NATURALLY IN THE FIELD?
SPONTANEOUS TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY TSE PRION AKA MAD COW
TYPE DISEASE, DOES IT EXIST NATURALLY IN THE FIELD?
*** We describe the transmission of spongiform encephalopathy in a
non-human primate inoculated 10 years earlier with a strain of sheep c-scrapie.
Because of this extended incubation period in a facility in which other prion
diseases are under study, we are obliged to consider two alternative
possibilities that might explain its occurrence.
We first considered the possibility of a sporadic origin (like CJD in
humans). Such an event is extremely improbable because the inoculated animal was
14 years old when the clinical signs appeared, i.e. about 40% through the
expected natural lifetime of this species, compared to a peak age incidence of
60–65 years in human sporadic CJD, or about 80% through their expected
lifetimes.
***Moreover, sporadic disease has never been observed in breeding colonies
or primate research laboratories, most notably among hundreds of animals over
several decades of study at the National Institutes of Health25, and in nearly
twenty older animals continuously housed in our own facility.***
Epidemiology | Neurological manifestations | Prion diseases Received: 16
February 2015 Accepted: 28 May 2015 Published online: 30 June 2015
Transmission of scrapie prions to primate after an extended silent
incubation period
SPONTANEOUS ATYPICAL BSE (LOL!)
Thursday, March 24, 2016
FRANCE CONFIRMS BOVINE SPONGIFORM ENCEPHALOPATHY BSE MAD COW (ESB) chez une
vache dans les Ardennes
Sunday, October 5, 2014
France stops BSE testing for Mad Cow Disease
***atypical spontaneous BSE in France LOL***
FRANCE STOPS TESTING FOR MAD COW DISEASE BSE, and here’s why, to many
spontaneous events of mad cow disease $$$
If you Compare France to other Countries with atypical BSE, in my opinion,
you cannot explain this with ‘spontaneous’.
Table 1: Number of Atypical BSE cases reported by EU Member States in the
period 2001–2014 by country and by type (L- and H-BSE) (extracted from EU BSE
databases on 1 July 2014). By 2015, these data might be more comprehensive
following a request from the European Commission to Member States for re-testing
and retrospective classification of all positive bovine isolates in the EU in
the years 2003–2009
BSE type
Country 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013(a)
2014(a) Total
H-BSE Austria 1 1
France(b) 1 2 3 1 2 2 2 2 15
Germany 1 1 2
Ireland 1 1 2 1 5
The Netherlands 1 1
Poland 1 1 2
Portugal 1 1
Spain 1 1 2
Sweden 1 1
United Kingdom 1 1 1 1 1 5
Total 2 3 3 1 1 2 2 2 4 4 5 1 4 1 35
L-BSE Austria 1 1 2
Denmark 1 1
France(b) 1 1 1 1 2 1 3 2 1 1 14
Germany 1 1 2
Italy 1 1 1 1 1 5
The Netherlands 1 1 1 3
Poland 1 2 2 1 2 1 2 1 12
Spain 2 2
United Kingdom 1 1 1 1 4
Total 0 5 3 4 3 3 6 3 3 4 3 6 1 1 45
Total Atypical cases (H + L)
2 8 6 5 4 5 8 5 7 8 8 7 5 2 80
(a): Data for 2013-2014 are incomplete and may not include all
cases/countries reported.
(b): France has performed extensive retrospective testing to classify BSE
cases, which is probably the explanation for the higher number of Atypical BSE
cases reported in this country.
The number of Atypical BSE cases detected in countries that have already
identified them seems to be similar from year to year. In France, a
retrospective study of all TSE-positive cattle identified through the compulsory
EU surveillance between 2001 and 2007 indicated that the prevalence of H-BSE and
L-BSE was 0.35 and 0.41 cases per million adult cattle tested, respectively,
which increased to 1.9 and 1.7 cases per million, respectively, in tested
animals over eight years old (Biacabe et al., 2008). No comprehensive study on
the prevalence of Atypical BSE cases has yet been carried out in other EU Member
States. All cases of Atypical BSE reported in the EU BSE databases have been
identified by active surveillance testing (59 % in fallen stock, 38 % in healthy
slaughtered cattle and 4 % in emergency slaughtered cattle). Cases were reported
in animals over eight years of age, with the exception of two cases (one H-BSE
and one L-BSE) detected in Spain in 2011/2012. One additional case of H-BSE was
detected in Switzerland in 2012 in a cow born in Germany in 2005 (Guldimann et
al., 2012).
Wednesday, July 15, 2015
Additional BSE TSE prion testing detects pathologic lesion in unusual brain
location and PrPsc by PMCA only, how many cases have we missed?
*** URGENT UPDATE ON FEED IN THE USA WITH HIGH RISK CWD TSE PRION DEER
***
Sunday, March 20, 2016
Docket No. FDA-2003-D-0432 (formerly 03D-0186) Use of Material from Deer
and Elk in Animal Feed ***UPDATED MARCH 2016*** Singeltary Submission
Friday, February 05, 2016
*** Report of the Committee on Wildlife Diseases FY2015 CWD TSE PRION
Detections in Farmed Cervids and Wild
Saturday, February 6, 2016
*** Secretary's Advisory Committee on Animal Health; Meeting [Docket No.
APHIS-2016-0007] Singeltary Submission
Thursday, January 14, 2016
EMERGING ANIMAL DISEASES Actions Needed to Better Position USDA to Address
Future Risks Report to the Chairman, Committee on Energy and Commerce, House of
Representatives December 2015 GAO-16-132
GAO
Saturday, January 9, 2016
Transmission of sheep-bovine spongiform encephalopathy to pigs
Research article
Long live the OIE, or time to close the doors on a failed entity?
Long live the OIE
By Dr. Bernard Vallat, OIE Director General December 29, 2015 | 10:08 am
EST
Wednesday, April 25, 2012
4th MAD COW DISEASE U.S.A. CALIFORNIA ATYPICAL L-TYPE BSE 2012
Saturday, January 29, 2011
Atypical L-Type Bovine Spongiform Encephalopathy (L-BSE) Transmission to
Cynomolgus Macaques, a Non-Human Primate
Monday, September 13, 2010
atypical BSE strains and sporadic CJD strains, is there a connection and
why shouldn't there be $
Sunday, April 12, 2009
TRANSMISSION OF ATYPICAL BOVINE SPONGIFORM ENCEPHALOPATHY (BSE) IN
HUMANIZED MOUSE MODELS
Wednesday, March 26, 2008
MAD COW DISEASE terminology UK c-BSE (typical), atypical BSE H or L, and or
Italian L-BASE
Thursday, January 3, 2008
ANIMAL HEALTH REPORT 2006 (BSE h-BASE EVENT IN ALABAMA, Scrapie, and CWD)
Thursday, April 14, 2016
Arizona 22 year old diagnosed with Creutzfeldt Jakob Disease CJD
Saturday, January 16, 2016
Revised Preventive Measures to Reduce the Possible Risk of Transmission of
Creutzfeldt-Jakob Disease and Variant Creutzfeldt-Jakob Disease by Blood and
Blood Products Guidance for Industry
Thursday, September 10, 2015
25th Meeting of the Transmissible Spongiform Encephalopathies Advisory
Committee Food and Drug Administration Silver Spring, Maryland June 1, 2015
Terry S. Singeltary Sr.
posted by Terry S. Singeltary Sr. at
12:23 PM
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