Poster Presentations P3 S505

P3-185 CSF BIOMARKERS IN AUTOPSY-CONFIRMED

DEMENTIA WITH LEWY BODIES

Kazutomi Kanemaru, Akiko Kanemaru, Yasushi Nishina,

Tadashi Komiya, Shigeo Murayama, Tokyo Metropolitan Geriatric Hospi-tal, Tokyo, Japan. Contact e-mail: kazukane@tmghig.jp

Background: Previously, we demonstrated that the levels of CSF tau were

normal and the levels of CSF amyloid beta42 (Ab42) were decreased in de-

mentia with Lewy bodies (DLB). To confirm these results, we investigated

the levels of CSF biomarkers (tau, ptau-181 and Abeta42) in autopsy-con-

firmed DLB. Methods: We measured the levels of CSF biomarkers (tau,

ptau-181 and Abeta42) in 56 autopsy-confirmed dementia patients (18

females, 38 males, age 75.7 6 9.2 years). The subjects include 6 patients

with Alzheimer’s disease (AD), 10 patients with DLB, 9 patients with pro-

gressive supranuclear palsy (PSP), 3 patients with corticobasal degeneration

(CBD) and 3 patients with Creutzfeldt-Jakob disease (CJD). The antemortem

CSF samples were obtained by lumbar puncture after informed consent was

obtained. The levels of CSF tau, ptau-181 and Ab42 were measured by

ELISA (Innogenetics) according to the manufacturer’s protocol. Results:

The CSF levels of tau (pg/ml) were 648.2 6 397.1 (mean 6 SD) in the

AD group, 102.5 6 60.0 in the DLB group and 119.1 6 107.3 in the PSP

group. The CSF levels of ptau-181 (pg/ml) were 62.4 6 23.2 in the AD

group, 39.8 6 11.4 in the DLB group and 39.3 6 15.2 in the PSP group.

The CSF levels of Ab42 (pg/ml) were 236.5 6 115.3 in the AD group,

407.1 6 128.1 in the DLB group and 579.0 6 147.6 in the PSP group.

The CSF levels of tau and ptau were increased in the AD group, whereas,

the CSF levels of tau and ptau were normal in the DLB group. The CSF levels

of Ab42 showed no difference between the two groups. Conclusions: The

analysis of the CSF levels of tau or ptau may be useful in distinguishing

DLB patients from AD patients.

P3-186 EXPRESSION OF 8-OXOGUANINE DNA

GLYCOSYLASE 1 (OGG1) AND THE LEVEL OF P53

AND TNF-ALPHA PROTEINS IN PERIPHERAL

LYMPHOCYTES IN PATIENTS WITH

ALZHEIMER’S DISEASE

Jolanta Dorszewska1, Mateusz Dezor1, Jolanta Florczak2,

Wojciech Kozubski2, 1Laboratory of Neurobiology, Department of Neu-rology, Poznan University of Medical Sciences, Poznan, Poland; 2Chair and

Department of Neurology, Poznan University of Medical Sciences, Poznan,

Poland. Contact e-mail: dorszewskaj@yahoo.com

Background: Reactive oxygen species are highly reactive and may oxidize

especially nucleic acids (DNA). 8-Oxo-2’-deoxyguanosine (8-oxo2dG) is

one of the crucial lesions produced in DNA by oxygen radical-forming

agents. OGG1 is a main DNA repair enzyme that excises 8-oxo2dG from

DNA. It was postulated that decreased expression of OGG1 may lead to

higher background mutation frequency and could increase the DNA damages

risk. 8-Oxo2dG is known to induce GC-TA transversion type point muta-

tions, and this type of mutation is commonly observed in the tumor suppres-

sor p53 gene. Moreover, in mice OGG1 coupled with lower level of TNF-

alpha. Damage of genomic DNA may lead to the cell death by apoptosis

in results it causes degenerative disorders. P53 and TNF-alpha may induce

apoptotic process in the cells. Methods: The studies were conducted on

41 patients with AD, including 25 women and 16 men aging 34-84 years.

The control groups included 51 individuals, 20 women and 31 men aging

22-83 years. The level of 8-oxo2dG was determined using HPLC/EC/UV

technique and the level of OGG1 and p53, TNF-alpha proteins was deter-

mined with Western Blot method. Results: We were observed increase of

the level of 8-oxo2dG after 60 years of age (insignificant) and in AD patients

(p < 0.05) as compared to the controls. Simultaneously, the levels of OGG1

and TNF-alpha proteins were decreased in individuals after 60 years of age

(OGG1, p < 0.01; TNF-alpha, p < 0.05) and in AD patients (OGG1, p <

0.001; TNF-alpha, p < 0.05) as compared to the controls when the level

of p53 protein was increased in individuals after 60 years as well as in AD

patients (Mann-Whitney test, p < 0.05) as compared to the controls. How-

ever, in patient with mild dementia (in MMSE scale) were observed the low-

est level of 8-oxo2-dG and OGG1 and the highest levels of p53 and TNF-

alpha proteins, when in the patients with moderate dementia (in MMSE

scale) were observed the highest level of OGG1 (p < 0.05, as compared to

the patient with mild dementia) and the lowest level of TNF-alpha. Conclu-

sions: It is possible that OGG1 and p53 and TNF-alpha proteins are involved

in pathogenesis of AD by repair of oxidative DNA damage and apoptosis.

P3-187 COMPLEXES OF 17BETA-HYDROXYSTEROID

DEHYDROGENASE TYPE 10 AND OF AMYLOID-

BETA PEPTIDES IN CEREBROSPINAL FLUID OF

PATIENTS WITH ALZHEIMER’S DISEASE AND OF

NEUROINFLAMMATORY CONTROLS

Daniela Rıpova1, Zdena Kristofikova1, Jan Rı�cny1, Katerina Hegnerova2,

Marketa Bockova2, Ales Bartos3, Linda �Cechova4, Jirı Homola2, 1Prague

Psychiatric Center, Prague, Czech Republic; 2Institute of Photonics and

Electronics, Prague, Czech Republic; 3Prague Psychiatric Center andCharles University, Third Faculty of Medicine, Prague, Czech Republic;4Charles University, Third Faculty of Medicine, Prague, Czech Republic.

Contact e-mail: ripova@pcp.lf3.cuni.cz

Background: It has been suggested that the multifunctional mitochondrial

enzyme 17beta-hydroxysteroid dehydrogenase type 10 could play a role in

the development of Alzheimer disease (AD) via its high-affinity binding to

amyloid beta peptides or its overexpression and that total levels of enzyme

or its bound forms e.g. in cerebrospinal fluid could be used as biomarkers

of this type of dementia. However, our earlier experiments indicated that

the complexes of enzyme with amyloid beta peptides could be a more spe-

cific biomarker of AD than the total enzyme levels. Methods: In the study,

we have evaluated 40 samples of cerebrospinal fluid of people with AD and

of neuroinflammatory/non-neuroinflammatory controls by means of ELISA

and surface plasmon resonance (SPR) biosensor. The results were compared

with those obtained for triplet (amyloid beta peptide 1-42, protein tau and

phospho-tau). Results: Our experiments suggest that: i) total enzyme levels

estimated by ELISA are significantly elevated in people with AD when com-

pared to non-neuroinflammatory controls (but not to neuroinflammatory

ones), ii) levels of complexes estimated by ELISA are increased in neuroin-

flammatory controls/people with AD when compared to non-neuroinflam-

matory controls, iii) levels of amyloid beta peptide 1-42 are decreased in

neuroinflammatory controls/people with AD when compared to non-neuro-

inflammatory controls, iv) response of SPR biosensor with immobilized an-

tibody against amyloid beta peptides to cerebrospinal fluid is increased in

neuroinflammatory when compared to non-neuroinflammatory controls,

and finally iv) levels of tau/phospho-tau are increased in people with AD

when compared to both control groups. Results of correlation analysis indi-

cated i) positive correlations between enzyme and its complexes/tau/phos-

pho-tau estimated by ELISA, ii) positive correlation between complexes

estimated by ELISA and response of SPR biosensor with immobilized anti-

body against amyloid beta peptides to cerebrospinal fluid, iii) negative cor-

relation between amyloid beta 1-42 and tau, and finally iv) positive

correlation between tau and phospho-tau. Conclusions: Enhanced levels

of the complexes of enzyme with amyloid beta peptides in cerebrospinal fluid

appear to sensitively reflect neuroinflammation mechanisms. Nor enzyme

nor its complexes are therefore the fully specific biomarkers of AD, but

both can be compared with the triplet.

P3-188 NONINVASIVE OPTICAL IMAGING OF RETINAL

AMYLOID PLAQUES IN MICE MODEL OF

ALZHEIMER’S DISEASE: EARLY DETECTION AND

TREATMENT ASSESSMENT

Yosef Koronyo1, Maya Koronyo-Hamaoui1, Alexander V. Ljubimov1,

Carol A. Miller2, MinHee K. Ko1, Keith L. Black1, Michal Schwartz3,

Daniel L. Farkas1, 1Cedars-Sinai Medical Center, Los Angeles, CA, USA;

Poster Presentations P3S506

2University of Southern California, Los Angeles, CA, USA; 3The Weizmann

Institute of Science, Rehovot, Israel. Contact e-mail: koronyoy@cshs.org

Background: A definite diagnosis of Alzheimer’s disease (AD) is deter-

mined following brain autopsy by detection of hallmark pathologies of

the disease: an extracellular aggregates of amyloid-b (Ab) peptides, termed

as Ab plaques, and an intracellular neurofibrillary tangles. Presently, non-

invasive monitoring of Ab plaques in the brains of living patients is, how-

ever, inadequate due to restricted resolution and specificity. We, thus,

hypothesized that the eyes can serve as a window to the brain for direct

and noninvasive imaging of AD. Methods: To this end, we used

APPswe/PS1dE9 double-transgenic AD mice, which faithfully represent

some of the key pathologies found in the human disease. We investigated

presence and formation of amyloid plaques in retinas from these AD mice,

as well as estimated the correlation between retinal amyloid plaques and

the plaque pathology in the brain. Lastly, we evaluated responsiveness of

retinal plaques to an immune-based therapy and the ability to image

them in live mice. Results: Here, we detected Ab plaques in retinal sam-

ples of AD mice, and found that Ab plaques appear in the retina signifi-

cantly earlier than in the brain. Retinal plaques accumulated during

disease progression, and in good correlation with brain pathology. More-

over, retinal Ab plaque number and size were significantly reduced follow-

ing an immune-based therapy, which we found to be effective in reducing

plaques in the brain. Systemic administration of a natural compound that

binds and labels Ab plaques, allowed in vivo noninvasive visualization

of retinal Ab plaques in live AD mice, with high resolution and specificity.

Conclusions: These studies establish the potential of direct retinal Ab pla-

que imaging in live subjects as a promising tool for early AD diagnosis,

prognosis and assessment of therapies.

P3-189 ASSOCIATION BETWEEN GENE EXPRESSION OF

PERIPHERAL BLOOD OXIDATIVE STRESS

MARKERS AND ALZHEIMER’S DISEASE: THE

NUN STUDY

Jon D. Wilson, Bharat Thyragarajan, Myron D. Gross, University of Minne-

sota, Fairview Hospital, Minneapolis, MN, USA. Contact e-mail: wils1355@

umn.edu

Background: There is increasing evidence that oxidative stress plays an im-

portant role in the pathogenesis of several neurodegenerative disorders,

including Alzheimer’s disease. Circulating concentrations of both SOD1

and HMOX1 protein have been associated with oxidative stress in neurons.

We performed a case control study of 22 participants with clinical dementia

and 22 controls within the Nun Study Cohort to evaluate the association be-

tween Copper-Zinc Superoxide Dismutase (SOD1) and Heme Oxygenase 1

(HMOX1) expression in peripheral blood and clinical dementia. Methods:

The Nun Study cohort was begun with 678 sisters who were at least 75 years

of age at enrollment. We randomly selected twenty-two individuals with

severe clinical dementia, and twenty-two normal individuals from the Nun

Study for evaluation. RNA was extracted with the RNeasy method from pe-

ripheral blood samples collected in PaxGENE tubes. The mRNA was used to

assess relative expression of SOD1 and HMOX1 using Taqman gene expres-

sion assays (Applied Biosystems Inc.). The 18s ribosomal RNA was used as

a housekeeping gene to standardize gene expression levels across all sam-

ples. Relative gene expression (copy fold change) was calculated using the

Pfaffl method. Results: Expression of SOD1 was lower in cases as compared

to controls (mean 6 SD: 0.68 6 0.26 vs. 0.77 6 0.15 respectively; p¼ 0.13).

In contrast, expression of HMOX1 was higher in cases as compared to con-

trols (mean 6 SD: 1.00 6 0.23 vs. 0.84 6 0.23 respectively; p ¼ 0.17).

Assay variability, based on 8 blinded duplicates, was 5.8% for both SOD1

and HMOX1 genes. Conclusions: This pilot study demonstrates the feasibil-

ity of utilizing expression levels of SOD1 and HMOX1 in peripheral blood

and testing as risk factors for Alzheimer’s disease. Though the results were

not statistically significant in this study with limited sample size, the direction

of effect observed is consistent with previous studies that have shown asso-

ciations between HMOX1, and SOD1 protein concentrations and Alzheim-

er’s disease. Future studies with larger samples will be needed to fully

evaluate oxidative stress markers in peripheral blood as risk factors for

Alzheimer’s disease.

P3-190 A NOVEL DIAGNOSTIC AND PROGNOSTIC FLUID

BIOMARKER FOR EARLY ALZHEIMER’S

DISEASE: YKL-40

Rebecca Craig-Schapiro1, Richard J. Perrin1, Catherine M. Roe2,

Elizabeth A. Grant2, Deborah Carter2, Nigel J. Cairns2, Mark A. Mintun2,

Elaine R. Peskind3, Ge Li3, Douglas R. Galasko4, Christopher M. Clark5,

Joseph F. Quinn6, Gina D’Angelo2, James P. Malone2, R. Reid Townsend2,

John C. Morris2, Anne M. Fagan2, David M. Holtzman2, 1Washington

University in St. Louis, St. Louis, MO, USA; 2Washington University in St.

Louis, St. Louis, MO, USA; 3University of Washington, Seattle, WA, USA;4University of California San Diego, San Diego, CA, USA; 5University ofPennsylvania, Philadelphia, PA, USA; 6Oregon Health Science University,

Portland, OR, USA. Contact e-mail: rebeccajcs@hotmail.com

Background: Clinicopathological studies suggest that Alzheimer’s dis-

ease (AD) pathology begins w10-20 years before the resulting cognitive

impairment draws medical attention. Currently, AD diagnosis relies upon

clinical assessment, without a definitive confirmatory laboratory test.

Biomarkers that can detect AD pathology in its early stages and predict

dementia onset and progression would, therefore, be invaluable for pa-

tient care and efficient clinical trial design. Methods: To discover such

AD biomarkers, we used two-dimensional difference gel electrophoresis

and tandem mass spectrometry to identify cerebrospinal fluid (CSF) pro-

teins that are increased or decreased in subjects with early AD relative to

age-matched cognitively normal controls. YKL-40 (chitinase 3-like 1),

a secreted 40-kDa glycoprotein of poorly understood function, was found

to be significantly more abundant in AD CSF. To confirm this finding, we

measured YKL-40 concentrations in the original cohort of CSF samples

by enzyme-linked immunosorbent assay (ELISA). Results: Mean CSF

YKL-40 levels were significantly higher in subjects with mild AD (N

¼ 23, Clinical Dementia Rating [CDR] 1) relative to age-matched con-

trols (N ¼ 24, CDR 0). Subsequent evaluation of YKL-40 by ELISA

in a larger, independent set of CSF samples (N ¼ 292) confirmed that

YKL-40 is more abundant in the CSF of those with very mild and

mild AD-type dementia. Importantly, CSF YKL-40 levels also predicted

risk of developing dementia (conversion from CDR 0 to CDR>0) and of

dementia progression from CDR 0.5 to 1; subjects with high CSF YKL-

40 values (�327 ng/mL, corresponding to the upper tertile) converted

and progressed faster than subjects with lower CSF YKL-40 values. In

plasma, mean levels of YKL-40, evaluated in 237 subjects of the larger

cohort, were significantly higher in the CDR>0 groups vs. the CDR

0 group. Plasma and CSF YKL-40 levels were correlated, with levels

roughly 5-fold higher in CSF than in plasma. In an immunohistological

study of human AD brain tissue, we detected YKL-40 within astrocytes

in the vicinity of a subset of amyloid plaques, implicating YKL-40 as part

of a neuroinflammatory response to Ab deposition. Conclusions: To-

gether, these data suggest that YKL-40, as a putative indicator of neuro-

inflammation, has potential utility as a diagnostic and prognostic

biomarker for the earliest stages of AD.

P3-191 GLYCOSYLATION OF SERUM PROTEIN AS A NEW

DIAGNOSTIC MARKER FOR ALZHEIMER’S

DISEASE

Miyako Taniguchi1, Minoru Kouduki1, Yuka Okayama2, Mai Okazaki1,

Daiki Jimbo1, Masashi Inoue1, Katsuya Urakami1, 1Tottori University, Yonago,

Japan; 2Tottori University Hospital, Yonago, Japan. Contact e-mail: miyako@med.tottori-u.ac.jp

Background: We previously discovered that a few glycoproteins in the ce-

rebrospinal fluid of Alzheimer’s disease patients (AD) had less lectin-binding

activities. Especially one of them showed high diagnostic sensitivity and

specificity for the AD against other non-AD groups even tauopathies. In

this study, we examined the some serum glycoproteins as a new diagnostic

marker for AD. Methods: We investigated serum samples obtained from