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From: TSS ()
Subject: Inactivation of amyloid-enhancing factor (AEF): study on experimental murine AA amyloidosis
Date: June 24, 2007 at 1:11 pm PST

Masatoshi Omoto · Tadaaki Yokota · Dan Cui
Yoshinobu Hoshii · Hiroo Kawano · Toshikazu Gondo
Tokuhiro Ishihara · Takashi Kanda

Inactivation of amyloid-enhancing factor (AEF): study on experimental
murine AA amyloidosis

Abstract It is known that amyloid-enhancing factor (AEF)
shortens the preamyloid phase in experimentally induced
AA amyloidosis in mice. Because it is reported that AEF
serves as both a nidus and a template for amyloid formation,
AA amyloidosis may have transmissibility by a prionlike
mechanism. It has been shown that amyloid fi brils also
have AEF activity, and amyloid fi brils with AEF activity
were named fi bril-amyloid enhancing factor (F-AEF). In
this study, we investigated methods to inactivate the AEF
activity. AEF was extracted from the thyroid gland obtained
at autopsy of a patient with AA amyloidosis. Before
injection into mice, AEF was treated with several methods
for inactivation. Of all the tested treatments, 1 N NaOH,
0.1 N NaOH, and autoclaving consistently demonstrated
complete inactivation of AEF. Heat treatment led to incomplete
inactivation, but 0.01 N NaOH, 0.001 N NaOH,
pepsin, trypsin, pronase, and proteinase K treatment had
no effect on AEF activity. By analysis with transmission
electron microscopy, the AEF preparation contains amyloid
fi brils, and a change of ultrastructure was shown after
1 N NaOH, 0.1 N NaOH, and autoclaving treatment. Furthermore,
immunoblotting of AEF with antihuman AA
antibody revealed that the protein band was scarcely found
after autoclaving, 1 N NaOH, and 0.1 N NaOH treatment.
Our results suggest that, similar to Creutzfeldt–Jakob
M. Omoto (*) · T. Kanda
Department of Neurology and Clinical Neuroscience, Yamaguchi
University School of Medicine, 1-1-1 Minamikogushi, Ube City,
Yamaguchi 755-8505, Japan
Tel. +81-836-22-2719; Fax +81-836-22-2364
T. Yokota
Department of Pathology, Kokura Memorial Hospital, Yamaguchi,
D. Cui · Y. Hoshii · H. Kawano · T. Ishihara
Department of Radiopathological and Science, Yamaguchi
University School of Medicine, Fukuoka, Japan
T. Gondo
Department of Surgical Pathology, Yamaguchi University Hospital,
Yamaguchi, Japan
disease (CJD), amyloidosis may require chemical or autoclaving
Key words Amyloid-enhancing factor · Amyloidosis ·
Creutzfeldt–Jakob disease · Prion · Transmission electron



Secondary amyloidosis occurs in individuals with longstanding
infl ammatory diseases. Since the incidence of
chronic infl ammatory diseases such as tuberculosis and leprosy
has decreased in recent years, rheumatoid arthritis
(RA) is now the most common disease involving secondary
AA amyloidosis, especially in elderly patients with a long
history of RA. Autopsy studies indicate that the incidence
of secondary amyloidosis in RA patients may be between
20% and 25%.21 Generally, treating the underlying disease
is the conventional approach in AA amyloidosis, because
no specifi c treatment exists. It is not yet certain whether
preventing amyloid proteins from aggregating will be therapeutically
An essential factor for the development of AA amyloidosis
is a continual high plasma concentration of SAA.
However, it is still unclear why only a subset of such individuals
develops AA amyloidosis. Therefore, in addition to
high concentrations of an amyloidogenic protein, other
factors are thought to be necessary in the pathogenesis of
AA amyloidosis. AEF is thought to be one of the essential
factors for the development of amyloidosis, although the
mechanism responsible for the formation of amyloid fi brils
is still unclear. Many studies have identifi ed that AEF activity
is a large macromolecular complex, and all tissue extracts
studied to date appear to contain glycoprotein with
an approximate size of 10–15 kD.4,22–24
Most strains of mice are susceptible to developing AA
amyloid deposition following chronic administration of
infl ammatory stimuli such as casein or azocasein.25 The
prolonged preamyloid phase in experimentally induced AA
amyloidosis can be dramatically shortened by intravenous
or intraperitoneal administration of AEF with the infl am-
matory stimuli.2–4 All mice that were exposed to AEF and
injected with silver nitrate developed amyloidosis by day
7.16 Amyloid fi brils extracted from different types of amyloidosis
from a wide variety of species display biologically
similar AEF activity to that in experimental animals. In fact,
Niewold et al.26 showed that intravenous and intraperitoneal
injection of hamster AA amyloid fi brils, bovine AA
amyloid fi brils, and human light chain-derived (A?) amyloid
fi brils markedly accelerated hamster amyloidosis.
The AEF activity found in the amyloid fi bril preparation
was named F-AEF.6 Furthermore, intravenous injection
of amyloid-like fi brils made from synthetic peptides of
transthyretin27 or denatured silk28 accelerates murine AA
amyloidosis. By double immunogold labeling and microautoradiographic
methods, Johan et al. reported that intravenously
administered, radiolabeled, heterologous,
amyloid-like, synthetic fi brils reached the lung and spleen,
accelerated amyloidosis, and were associated with topographical
deposition of murine protein AA fi brils in the recipient
Drastic structural changes of amyloid protein from the
normal and soluble forms to the unique ß-sheet fi brils may
be the most important event in amyloidosis. Recently, it was
reported that AEF serves as both a nidus and a template
for amyloid formation,8 and AA amyloidosis may show
transmissibility as a prion-like mechanism.9 Johan et al. also
suggested that amyloid-like synthetic fi brils had a nidus activity,
29 and amyloid-enhancing activity may occur through
the mechanism of amyloid-like fi brils serving as seed for
fi bril formation. Moreover, by using the method of negative
staining with TEM, electron micrographs of amyloid fi brils
were observed under some conditions. O’Nuallain et al.
revealed that electron micrographs of islet amyloid polypeptide
showed aggregates initially and then grew into fi bril
formations after incubation.30 In contrast, Santhoshkumar
et al. revealed that, using TEM, amyloid ß-peptide showed
signifi cantly decreased fi bril formation and some amorphous
aggregates after incubation with aA-crystallin in
their in vitro study.31 They suggested that aA-crystallin had
the ability to inhibit amyloid fi bril formation. Similarly, in
our study, electron micrographs of F-AEF revealed some
amorphous aggregates after autoclaving, 1 N NaOH, and
0.1 N NaOH treatment and short and transformed fi brils
after heat treatment. Furthermore, immunoblotting of AEF
with antihuman AA antibody revealed that a protein band
was scarcely found after autoclaving, 1 N NaOH, and 0.1 N
NaOH treatments and that weak protein bands were found
after heat treatment. We suggested that these results indicated
the activity of F-AEF disappeared after autoclaving,
1 N NaOH, and 0.1 N NaOH treatments and that the activity
of F-AEF was decreased after heat treatment.
Prion diseases are associated with the accumulation of a
conformational isomer (PrPSc) of host-derived prion protein
(PrPC), and PrPSc forms amyloid fi brils. The exogenous abnormal
form of the prion protein is generally regarded as a
seed that promotes the association of cellular proteins.
Prions are very resistant to inactivation, and accidental
transmission has occurred through the use of inadequate
decontamination procedures.

In our experiments with mice, the activity of F-AEF was
markedly decreased after autoclaving treatment under conditions
of 132°C for 1 h and 1 N NaOH and 0.1 N NaOH
treatment for 1 h. For CJD materials, the Committee on
Health Care Issues of the American Neurological Association
recommended treatment with 1 N NaOH as a standard
sterilization procedure.18 Heat treatment led to substantial
but incomplete inactivation in this study. The Committee
on Health Care Issues of the American Neurological Association
reported that boiling was an ineffective procedure
for CJD tissues and contaminated materials.18 Tateishi et al.
showed that heat treatment with SDS was effective.17
The acceleration of amyloid deposition may be a primary
event in disease, CJD, bovine spongiform encephalopathy
(BSE), familial amyloid polyneuropathy, and AA and
human senile systemic amyloidosis.32 Walker et al. reported
Aß amyloid extracted from an Alzheimer disease brain may
have potential of prion protein.33

The property described for F-AEF is similar to that
of prion reported in CJD. Chemical or autoclaving decontamination
for CJD is necessary for most items associated
with surgery or autopsy.34 We suggest that amyloidosis may
need chemical or autoclaving decontamination similar to

Acknowledgments We thank Mr. Jitsuo Kash!tani for excellent technical
assistance. This work was supported by a grant from the Intractable
Disease Division, the Ministry of Health and Welfare, a Research
Committee for Epochal Diagnosis and Treatment of amyloidosis in
Japan, and a Research Committee for amyloidosis.

Alzheimer-type neuropathology 28 year old patient with idCJD
Sun Feb 19, 2006 11:14

Alzheimer-type neuropathology in a 28 year old patient with iatrogenic
Creutzfeldt-Jakob disease after dural grafting
M Preusser1, T Ströbel1, E Gelpi1,2, M Eiler3, G Broessner4, E Schmutzhard4
and H Budka1,2
1 Institute of Neurology, Medical University Vienna, Austria
2 Austrian Reference Centre for Human Prion Diseases (OERPE), General
Hospital Vienna, Austria
3 Department of Neurology, LKH Rankweil, Austria
4 Department of Neurology, Medical University Innsbruck, Innsbruck, Austria

Correspondence to:
Dr H Budka
Institute of Neurology, Medical University of Vienna, Waehringer Guertel
18-20, 4J, 1097 Vienna, Austria;

We report the case of a 28 year old man who had received a cadaverous dura
mater graft after a traumatic open skull fracture with tearing of the dura
at the age of 5 years. A clinical suspicion of Creutzfeldt-Jakob disease
(CJD) was confirmed by a brain biopsy 5 months prior to death and by
autopsy, thus warranting the diagnosis of iatrogenic CJD (iCJD) according to
WHO criteria. Immunohistochemistry showed widespread cortical depositions of
disease associated prion protein (PrPsc) in a synaptic pattern, and western
blot analysis identified PrPsc of type 2A according to Parchi et al.
Surprisingly, we found Alzheimer-type senile plaques and cerebral amyloid
angiopathy in widespread areas of the brain. Plaque-type and vascular
amyloid was immunohistochemically identified as deposits of beta-A4 peptide.
CERAD criteria for diagnosis of definite Alzheimer’s disease (AD) were met
in the absence of neurofibrillar tangles or alpha-synuclein immunoreactive
inclusions. There was no family history of AD, CJD, or any other
neurological disease, and genetic analysis showed no disease specific
mutations of the prion protein, presenilin 1 and 2, or amyloid precursor
protein genes. This case represents (a) the iCJD case with the longest
incubation time after dural grafting reported so far, (b) the youngest
documented patient with concomitant CJD and Alzheimer-type neuropathology to
date, (c) the first description of Alzheimer-type changes in iCJD, and (d)
the second case of iCJD in Austria. Despite the young patient age, the
Alzheimer-type changes may be an incidental finding, possibly related to the
childhood trauma.


CJD1/9 0185

Ref: 1M51A


Dr McGovern From: Dr A Wight

Date: 5 January 1993

Copies: Dr Metters

Dr Skinner

Dr Pickles

Dr Morris

Mr Murray


1. CMO will wish to be aware that a meeting was held at DH yesterday,
4 January, to discuss the above findings. It was chaired by Professor
Murray (Chairman of the MRC Co-ordinating Committee on Research in
the Spongiform Encephalopathies in Man), and attended by relevant
experts in the fields of Neurology, Neuropathology, molecular biology,
amyloid biochemistry, and the spongiform encephalopathies, and by
representatives of the MRC and AFRC.

2. Briefly, the meeting agreed that:

i) Dr Ridley et als findings of experimental induction of p amyloid
in primates were valid, interesting and a significant advance in the
understanding of neurodegeneradve disorders;

ii) there were no immediate implications for the public health, and no
further safeguards were thought to be necessary at present; and

iii) additional research was desirable, both epidemiological and at the
molecular level. Possible avenues are being followed up by DH
and the MRC, but the details will require further discussion.


BSE101/1 0136


5 NOV 1992

CMO From: Dr J S Metters DCMO 4 November 1992


1. Thank you for showing me Diana Dunstan's letter. I am glad that MRC have
recognised the public sensitivity of these findings and intend to report
them in their
proper context. This hopefully will avoid misunderstanding and possible
distortion by
the media to portray the results as having more greater significance than
the findings
so far justify.

2. Using a highly unusual route of transmission (intra-cerebral injection)
researchers have demonstrated the transmission of a pathological process
from two
cases one of severe Alzheimer's disease the other of Gerstmann-Straussler
disease to
marmosets. However they have not demonstrated the transmission of either
condition as the "animals were behaving normally when killed'. As the report
emphasises the unanswered question is whether the disease condition would
revealed itself if the marmosets had lived longer. They are planning funher
to sec if the conditions, as opposed to the partial pathological process, is

What are the implications for public health?

3. . The route of transmission is very specific and in the natural state of
highly unusual. However it could be argued that the results reveal a
potential risk,
in that brain tissue from these two patients has been shown to transmit a
process. Should therefore brain tissue from such cases be regarded as
infective? Pathologists, morticians, neuro surgeons and those assisting at
surgical procedures and others coming into contact with "raw" human brain
could in theory be at risk. However, on a priori grounds given the highly
route of transmission in these experiments that risk must be negligible if
the usual
precautions for handling brain tissue are observed.


BSE101/1 0137

4. The other dimension to consider is the public reaction. To some extent
the GSS
case demonstrates little more than the transmission of BSE to a pig by
injection. If other prion diseases can be transmitted in this way it is
little surprise that
some pathological findings observed m GSS were also transmissible to a
But the transmission of features of Alzheimer's pathology is a different
matter, given
the much greater frequency of this disease and raises the unanswered
question whether
some cases are the result of a transmissible prion. The only tenable public
line will
be that "more research is required" before that hypothesis could be
evaluated. The
possibility on a transmissible prion remains open. In the meantime MRC needs
carefully to consider the range and sequence of studies needed to follow
through from
the preliminary observations in these two cases. Not a particularly
message, but until we know more about the causation of Alzheimer's disease
the total
reassurance is not practical.

Room 509
Richmond House
Pager No: 081-884 3344
Callsign: DOH 832



also, see the increase of Alzheimer's from 1981 to 1986

Proof Mad Cow Is The Same
As Alzheimer's And CJD ???

How Many Of Them Are Really Mad Cow/vCJD/TSEs ???

How Can Government Claims Of Just 'One In A Million' Be Accurate
When CJD Is Not A Reportable Disease? And When The Elderly Do
Not Get Routinely Autopsied??

By Terry Singletary, Sr

Note - This extensive, powerful assemblage of science was first posted on
1-24-3. The
following data is even more important today. -ed


Regarding Alzheimer's disease

(note the substantial increase on a yearly basistss)


The pathogenesis of these diseases was compared to Alzheimer's disease at a
molecular level...


And NONE of this is relevant to BSE?

There is also the matter whether the spectrum of ''prion disease'' is wider
than that recognized at present.

Human BSE


These are not relevant to any possible human hazard from BSE nor to the much
more common dementia, Alzheimers.



From: TSS
Subject: CJD or Alzheimer's, THE PA STUDY...full text
Date: May 7, 2001 at 10:24 am PST

Diagnosis of dementia: Clinicopathologic correlations

Francois Boller, MD, PhD; Oscar L. Lopez, MD; and John Moossy, MD

Article abstract--Based on 54 demented patients consecutively autopsied at
the University of Pittsburgh, we studied the accuracy of clinicians in
predicting the pathologic diagnosis. Thirty-nine patients (72.2%) had
Alzheimer's disease, while 15 (27.7%) had other CNS diseases (four
multi-infarct dementia; three Creutzfeldt-Jakob disease; two thalamic and
subcortical gliosis; three Parkinson's disease; one progressive supranuclear
palsy; one Huntington's disease; and one unclassified). Two neurologists
independently reviewed the clinical records of each patient without
knowledge of the patient's identity or clinical or pathologic diagnoses;
each clinician reached a clinical diagnosis based on criteria derived from
those of the NINCDS/ADRDA. In 34 (63 %) cases both clinicians were correct,
in nine (17%) one was correct, and in 11 (20%) neither was correct. These
results show that in patients with a clinical diagnosis of dementia, the
etiology cannot be accurately predicted during life.

NEUROLOGY 1989;39:76-79

Several recent papers and reports have addressed the problem of improving




Subject: Re: Hello Dr. Manuelidis Date: Fri, 22 Dec 2000 17:47:09 -0500
From: laura manuelidis Reply-To: Organization:
Yale Medical School To: "Terry S. Singeltary Sr."

References: <> <>
<> <>
<> <>

Dear Terry,

One of our papers (in Alzheimer's Disease Related Disord. 3:100-109, 1989)
in text cites 6 of 46 (13%) of clinical AD as CJD. There may be a later
paper from another lab showing the same higher than expected incidence but I
can't put my hands on it right now. We also have a lot of papers from 1985
on stating that there are likely many silent (non-clinical) CJD infections,
i.e. much greater than the "tip of the iceberg" of long standing end-stage
cases with clinical symptoms. Hope this helps.

best wishes for the new year laura manuelidis

"Terry S. Singeltary Sr." wrote: Hello again Dr. Manuelidis, could you
please help me locate the 2 studies that were done on CJD where it showed
that up to 13% of the people diagnosed as having Alzheimer's actually had
CJD. trying to find reference... thank you, > Terry S. Singeltary Sr.





full text;

Proof Mad Cow Is The Same As Alzheimer's And CJD
As Alzheimer's And CJD How Many Of Them Are Really Mad Cow/vCJD/TSEs ??? ...
I have posted some data below on CJD and Alzheimer's that you may find
interest ...


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