S401 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, S402 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 A␤42 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 (A␤40) and 42 (A␤42), 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 A␤40 and A␤42, 1-4 years after the initial measurement. In the longi-