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Poster P3:: Tuesday Posters
plement factor-H; r⫽0.34, p⬍0.03) with hippocampal NAA/mI across all
groups. Complement factor-H (CFH) also showed a significant negative
correlation with hippocampal mI/Cr⫹PCr (r⫽ -0.33, p⬍0.04) while alpha2-macroglobulin (A2M) showed a similar but non-significant correlation with hippocampal mI/Cr⫹PCr (r⫽ -0.3, p⬍0.07).When we examined
the AD group alone, we found a considerably stronger positive correlation
between A2M and NAA/mI (r⫽0.64, p⬍0.02), while CFH showed a
positive correlation tending to significance (r⫽0.55, p⬍0.06). Within the
AD group, there was a strong negative correlation between CFH (r⫽ -0.63,
p⬍0.02) and A2M (r⫽ -0.57, p⬍0.04) with mI/Cr⫹PCr. Conclusions:
Using a combined proteomic-neuroimaging strategy, we identified plasma
A2M and CFH as proteins showing significant correlations with hippocampal metabolite ratios known to reflect neurodegeneration in AD.
P3-094
DEFICIT OF EMOTIONAL MEMORY
CORRELATES WITH LEVELS OF THE KINASE
P70S6K IN LYMPHOCYTES OF PATIENTS WITH
ALZHEIMER DISEASE
Marie-Christine Perault-Pochat1, Marc Paccalin2, Fuad Al Khidir1,
Claudette Pluchon3, Stéphanie Pain-Barc1, Agnès Rioux-Bilan1,
Roger Gil3, Jacques Hugon3, 1Poitiers University Hospital - Clinical
Pharmacology, La Miletrie - BP577 - 86021 Poitiers, France; 2Poitiers
University Hospital - Department of Geriatrics, La Miletrie - BP577 86021 Poitiers, France; 3Poitiers University Hospital - Department of
Neurology, La Miletrie - BP577 - 86021 Poitiers, France. Contact
e-mail: m.c.perault-pochat@chu-poitiers.fr
Background: Alzheimer Disease (AD) is characterized by memory disturbances affecting explicit memory. The molecular process of learning
and memory seems to implicate protein synthesis and its control includes
the mTOR/p70S6k signaling pathway. These two processes are modified in
the brain of AD patients. We have previously shown that levels of the
kinase p70S6k in peripheral lymphocytes were correlated with memory
and cognitive deficits in patients with AD (Lafay et al J Neurochem 2005,
Paccalin et al Arch Int Med 2005). Emotional memory is also disturbed in
AD but seems to better resist than neutral explicit memory to the neurodegenerative process. Objective(s): The goal of the study was to assess the
difference between emotional and neutral explicit memories (emotional
delta) and to correlate these results with levels of the ratio between
phosphorylated and total p70S6k in lymphocytes of patients with AD.
Methods: Twenty eight AD patients were enrolled in this study. MMSE
scores ranged from 25 to 10. Explicit memory was assessed in all patients
with the Gröber and Buschke (GB) test. Total recall tests for emotional
memory (emotional words) and neutral memory (neutral words) were
carried out by patients and at the same time blood lymphocytes were
extracted and the ratio between phosphorylated and total p70S6k were
evaluated in the same patients using western blots. The Pearson correlation
test was used for statistical analysis. Results: The results showed:
1-MMSE scores were positively correlated with activated p70S6k levels
(p⬍.001), 2-GB test scores were correlated with p70S6klevels (p⬍.01),
3-Emotional deltas were also positively correlated with the ratio between
phosphorylated and total p70S6k (p⬍.05). Conclusions: Our results demonstrate that levels of peripheral and activated p70S6k are also a good
marker for the evolution of emotional memory in AD as shown for the
explicit memory, language and working memory.
P3-095
HIGH FREQUENCIES OF THYROID DISEASE IN
MIXED MEMORY CLINIC PATIENTS
Anne-Mette Hejl1, Kieu Phung1, Birgitte B. Andersen1,
Ulla Feldt-Rasmussen2, Gunhild Waldemar1, 1Memory Disorders
Research Group, Copenhagen University Hospital, Denmark; 2Dept. of
Endocrinology, Copenhagen University Hospital, Denmark. Contact
e-mail: hejl@inet.uni2.dk
Background: Memory impairment may be the first sign of Alzheimer’s
disease, but not every case represents an irreversible degenerative dementia
disorder and some reversible conditions may cause or mimic dementia.
Objective: To investigate the prevalence of thyroid disease in a prospective memory clinic cohort of younger and elderly patients with cognitive
symptoms. Methods: The study included the primary analysis of 2,296
consecutive patients referred during a period of 86 months to a university
hospital multidisciplinary memory clinic based in neurology. All patients
were referred for diagnostic evaluation and treatment of cognitive symptoms. As part of the diagnostic evaluation laboratory screening tests were
performed in all patients. Results: The mean age of the patients was 67
years (range 17-98) and the frequency of women was 54%. Abnormal
thyroid stimulating hormone (TSH) was found in 9.8% of all patients; 4.2%
had increased level of TSH suggesting hypothyroidism and 5.6% had
reduced TSH indicating hyperthyroidism. Conclusions: We found a high
frequency of TSH abnormalities in a mixed cohort of memory clinic
patients. Hyperthyroidism as well as hypothyroidism may contribute to
cognitive impairment as co-morbidity in dementia, or may represent the
main cause of mild subjective cognitive symptoms.
P3-096
KINASES MTOR AND PKR CONTROLLING
TRANSLATION IN ALZHEIMER’S DISEASE
POTENTIAL BIOMARKERS?
Marc Paccalin, Stephanie Pain Barc, Claire Paquet, Claudette Pluchon,
Agnes Roiux Bilan, Roger Gil, Jacques Hugon, University of Poitiers,
Poitiers, France. Contact e-mail: j.hugon@chu-poitiers.fr
Background: The initiation phase of protein translation includes the kinases mTOR and p70S6k and modulates memory, cell growth and proliferation. The pathway involving the kinase PKR and the eukaryotic initiation factor 2␣ (eIF2␣) decreases protein synthesis and is associated with
apoptosis. Both pathways are altered in the brains of patients with Alzheimer’s disease (AD). We recently reported significant alterations of the
expression and activation of p70S6k in lymphocytes of AD patients that
were correlated with memory and cognitive test scores Objective(s): In
this report we analyze the levels of the kinases mTOR, PKR and eIF2␣ in
lymphocytes of AD patients and control individuals and describe the
correlations between these levels and cognition tests in patients. Methods:
Diagnosis of AD was made in 37 patients (25F/12M; 77.8 ⫾ 1.1 years)
according to the NINCDS-ADRDA criteria. AD patients and age-matched
controls (n⫽33) were included after informed consent. Blood lymphocytes
were isolated and protein determined with a protein assay kit. The levels of
the activated kinases were measured by western blotting using the ratio
phosphorylated form / total form after quantification. Cognitive tests included MMSE, the explicit memory test: Gröber and Bushke (GB) and the
digit span tests (DST) for immediate and working memories. Results: The
results show that mTOR levels were significantly decreased (p ⬍ 0.01) in
lymphocytes of AD patients compared with controls. The mTOR ratios had
a positive correlation with MMSE scores (p ⬍ 0.05), GB scores (p ⬍ 0.05)
and reverse DST (p ⬍ 0.005). PKR levels were significantly increased (p ⬍
0.005) in lymphocytes of AD patients. The PKR ratios had a negative
correlation with MMSE scores (p ⬍ 0.05) and GB scores (p ⬍ 0.01).
EIF2␣ levels were significantly increased (p ⬍ 0.05) in lymphocytes of AD
patients and were negatively correlated with MMSE scores (p ⬍ 0.05), GB
scores (p ⬍ 0.01) and DST (p ⬍ 0.05). Conclusions: In conclusion, these
results suggest that the levels of kinases controlling translation in lymphocytes of AD patients are correlated to the cognitive decline and could be
helpful to analyze the response to new drugs in patients.
P3-097
CLASSIFYING ALZHEIMER’S DISEASE
THROUGH MULTIGENE EXPRESSION
SIGNATURE PROFILING
Catherine L. Clelland1,2, Steven H. Ferris3, Laura L. Read2,3,
James D. Clelland2,3, Jaclyn Cocchiola2,1, Robyn Waters3, Thet Oo3,
Alok Vedvyas3, 1Mount Sinai School of Medicine, New York City, NY,
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Poster P3:: Tuesday Posters
USA; 2The Nathan Kline Institute, Orangeburg, NY, USA; 3NYU School
of Medicine, New York City, NY, USA. Contact e-mail:
Catherine.Clelland@mssm.edu
Background: The NINCDS-ADRDA and DSM-IV criteria are currently
widely used for diagnosis of probable AD. These clinical criteria have a
number of limitations, including lack of specificity and sensitivity in the
diagnosis, and have an error rate of ⬃10% even in academic research
centers. Furthermore, diagnosis based on cognitive function can only be
made post-symptomatically, when medications that may inhibit AD development or delay its progression will likely be less effective. The imaging
and biological marker diagnostic methods currently under development
have additional drawbacks in terms of their need for highly specialized
equipment, and specificity and sensitivity respectively, and thus may not be
useful for early screening. There is however, growing evidence of specific
peripheral responses in AD, e.g. Tcell CD45 isoforms, HO1 and HSP70
RNA levels, increased circulating and in-vitro produced cytokines (CD4,
CD25, CD28 antigen), decreased CD7, CD8 and increased T-lymphocyte
IL-6 receptor. Objectives: We hypothesize that gene expression changes in
a surrogate tissue (leukocytes), induced by the Alzheimer’s disease (AD)
process as a primary or secondary event, can be utilized to form an overall
biological classifier of AD. Our hypothesis is based on literature describing
individual genes as peripheral markers of AD, plus positive data that we
have obtained in a preliminary study. We are now extending this research,
via funding from the NIA. Methods: The Aging and Dementia Research
Center provides access to a large elderly population, who are evaluated
there. The Core population of ⬎2,250 people includes patients with AD,
vascular dementia, Mild Cognitive Impairment, and nondemented controls.
For our study, 15 ml bloods are collected, leukocyte RNA isolated, and
samples processed for hybridization to Affymetrix microarrays containing
⬎40,000 genes. Results: We have been encouraged with our results to
date, suggesting that leukocyte gene expression can be employed for the
classification between subjects with AD and age-matched controls. Conclusions: We believe the larger numbers (n⫽60) being recruited will
increase the significance of these results and that the findings of our larger
study may be of significance for improving clinical AD diagnosis.
P3-098
PROTEOMIC ANALYSIS OF AMYLOID BETA
BINDING PROTEINS IN HUMAN PLASMA
Pattie S. Green, J. Jacob Kulstad, Helen Chea, Suzanne Craft,
University of Washington, Seattle, WA, USA. Contact e-mail:
psgreen@u.washington.edu
Background: Amyloid- peptide (A) circulates in plasma and interacts
extensively with plasma proteins. Several plasma proteins have been identified which can interact with A in purified form; however, a paucity of
studies examine A-protein binding relationships in the intact plasma
milieu. Published studies suggest that plasma sequestration of A by
protein interaction can alter brain levels of A. A-protein interactions
could also interfere with removal of A from the plasma. Objective(s):
The present study uses affinity-purification with A followed by mass
spectrometric identification of proteins to identify endogenous A binding
proteins in human plasma. Methods: The affinity purified proteins were
separated by denaturing electrophoresis and identified after in-gel trypsin
digests by peptide map fingerprint analysis using matrix-assisted laser
desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS)
and subsequently verified using tandem analysis (MALDI-TOF/TOF MS)
of the digested samples. All identified proteins were identified from three
different plasma samples. Results and Conclusions: A binding proteins
identified in this study include proteins involved in coagulation processes
such as fibrinogen , fibrinogen ␥, coagulation factor V and vitronectin.
Several lipoprotein-associated proteins were also identified suggesting lipoprotein association of A. These proteins include apolipoprotein (apo)
A-I, apoC-II, apoE, apoJ, and serum paraoxonase. The pattern of apolipoproteins identified suggests that the A is interacting predominately with
high density lipoprotein (HDL). Interestingly, apoE and apoJ were en-
riched relative to apo A-I suggesting that the A may interact with a subset
of lipoprotein particles. Additional proteins identified by A binding include the protease inhibitor ␣-2-macroglobulin, the thyroid binding protein
transthyretin, and complement C3. The nature of these A-protein interactions has implications in both A clearance and A equilibrium across
the blood-brain barrier.
P3-099
ANALYSIS OF SERUM PATIENTS WITH
ALZHEIMER’S DISEASE TOWARDS THE LEVEL
OF ADVANCED GLYCATION END-PRODUCTS
Jerzy W. Leszek, University of Wroclaw, Wroclaw, Poland. Contact
e-mail: jleszek@psych.am.wroc.pl
Background: The presence of advanced glycation end-products (AGEs) in
deposits formed in brain of Alzheimer disease (AD) patients have been
previously histopathologically documented, suggesting the relation of
AGE with degeneration of central nervous system. Objectives: In the
current study the content of AGEs was examined with an immunochemical
assay in the sera of AD patients, in the frame of search for a biochemical
marker of disease. Methods: Subjects with AD and vascular dementia
(VD) were included in the study (n⫽30, age 68-75 yrs). The results were
compared to the healthy control groups: older people (n⫽25, age 45-85
yrs), young people (n⫽45, age range 21-39 yrs). The ELISA inhibition test
for the determination of AGEs is based on a rabbit anti-AGE, affinity
purified antibody and a model AGE-myoglobin antigen, where serum
sample treated with proteinase K is used as an inhibitor. For the measurement of immune complexes and anti-AGE antibodies the corresponding
ELISA tests have been applied. Results: The AGEs level in VD group
(49.5 U AGE) was higher than in AD patients (46.1 UAGE). The level of
total AGEs in the sera of AD patients was significantly lower than in the
controls(50/51.6 UAGE).These relations was not observed regarding to the
immune complexes and IgG anti-AGE antibody levels in AD(70.2 UIC/
0.027 U IgG) and VD(83UIC/0.034 UIgG)patients,because the levels of
these parameters were similar to the controls(76.2 UIC/0.042 UIgG). The
level of IgM was significantly higher in AD patients(0.025 UIgM) than in
the control group(0.02 UIgM). Conclusion: The work revealed the lower
level of circulating serum advanced glycation end-products in patients with
AD, in relation to healthy controls.
P3-100
LONGITUDINAL CHANGES IN PLASMA A42
POSITIVELY CORRELATE WITH PROGRESSION
OF MILD COGNITIVE IMPAIRMENT
Michael C. Irizarry1, Alona Muzikansky1, Hang Lee1, Susan Raju1,
Teodoro Bottiglieri2, Bradley T. Hyman1, Marilyn S. Albert3,
Blacker Deborah1, 1Massachusetts General Hospital, Boston, MA, USA;
2
Baylor University Medical Center, Dallas, TX, USA; 3Johns Hopkins
University, Baltimore, MD, USA. Contact e-mail:
mirizarry@partners.org
Background: Plasma levels of amyloid -protein (A) and vascular risk
(total homocysteine, tHcy; high sensitivity C-reactive protein, hsCRP) are
implicated as risk factors for development of cognitive impairment and
Alzheimer’s disease (AD). Objective(s): To determine whether plasma
levels of A, tHcy, and hsCRP are associated with risk of progression from
normal cognition (Clinical Dementia Rating, CDR 0) to mild cognitive
impairment (MCI, defined here as CDR 0.5) or conversion from MCI to
AD. Methods: A ending at amino acid 40 (A40) and 42 (A42), tHcy,
and hsCRP were measured in nondemented individuals participating in a
longitudinal study of preclinical predictors of AD (MGH Memory and
Aging Study). A measures were repeated 1-4 years later. A semi-structured CDR interview was performed annually on all study participants to
quantify progression, and incident dementia was diagnosed by standardized
criteria. Results: The study cohort consisted of 68 individuals with CDR 0
and 153 person-years of follow-up, and 167 individuals with CDR 0.5 and
472 person-years of follow-up. 232 individuals had a repeat measurement
of A40 and A42, 1-4 years after the initial measurement. In the longi-