Donepezil treatment of Alzheimer's disease patients in California clinical practice: One year follow-up

Z?4. Degenerative and neurological disorders modifications to insolubile trophic neurites.

microscopic

size inclusions

in dys-

Materials and Methods: We analyzed insoluble protein mass from parietal cortex of autopsy specimen of patients with AD. Proteins were fragmented by trypsin and peptides were resolved by HPLC. After subtraction of the elution profile of peptides present in non-demented contols, peaks specific to diseased samples were analyzed by microsequencing and MALDI-TOF mass spectrometry. Results and Discussion: Sequence and mass analysis of ADspecific peptides indicates that NFT-z is rendered insoluble by covalent cross-linking oft protein chains via (poly)ubiquitin and HSP 27 protein or, to lesser extent alpha-synuclein. Our results indicate a pivotal role for transglutaminase-mediated isopeptide cross-linking of ubiquitinylated t. Conclusions: The formation of pathological insoluble inclusions inside neurons is a consequence of transglutaminase activity. Besides polyglutamine repeat diseases, our results propose an pathogenic role of transglutaminase-mediated protein crosslinking in Alzheimer’s Dementia as well.

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neuropathological methods. The genomic DNA from HCs was extracted from the peripheral blood according to the standard method. In the PiD cases, DNA was extracted from paraffin embedded brain tissues by the method of Jackson et al. (1990) and amplified by PCR. The amplified product was digested by Mb01 restriction enzyme. After electrophoresis on 8% polyacrylamide gels, DNA fragments were visualized by UV illumination. Results: Tbe homozygous Glu 298 genotype was underrepresented in PiD population (26.2%) compared with HC (43.2%). We did not find any homozygous Asp/Asp in the HC and the PiD groups. In the presence of apoE ~4 allele, the homozygous Glu 298 genotype was also underrepresented in PiD (18.8%) compared with the HC (28.6%) population. Comparing the Glu/Glu genotype between the genders, it was found, that the presence of this genotype was more frequent in women in both groups (PiD women: 30%, men: 16.7%, HC women: 48.2%, men: 37.5%). Conclusion: From these results we can conclude, that there is no association between the apoE ~4 allele and the NOS3 Glu/Glu genotype and it is not a risk factor for PiD patients in the Hungarian population. This work was supported by Hungarian Health Scientific Board (ETT 01817/2000) grant.

References [l] Nemes, Z., Marekov L.N., F&is L. and Steinert, E M. (1999) A novel function for transghnaminase 1: attachment of long-chain omegahydroxyceramides to involucrin by ester bond formation. Proc Nat1 Acad Sci U S A, 96,8402-8407. [2] Nemes, Z., F&is, L, Keszthelyi, A, Egerhiizi, A. and Degrell, I. M. (2001) w(y-glutamyl)lysine in cerebrospinal fluid marks Aizheimer type and vascular dementia. Neurobiol. Aging 22, in the press. [3] Nemes, Z., Mbdi, A., Marekov, L N., Piacentini, M., Steinert, P M. and F&is, L. (2001) Analysis of protein transglutamylation in apoptosis. Meth. Cell Biol. 66, 11 l-134.

m(

Nitric oxide synthase gene polymorphism in Pick’s disease in the Hungarian population

A. Juhbz, A. Rimanoczy, J. Kahnan, K. Majdnyi’, Z. Jar&a. University of Szeged, Department of Psychiatry, Szeged; ‘Department of Neuropathologx National Institute of Psychiav and Neurology, Budapest, Hungary

References [l] Clark L., Wilhelmsen KC.: The genetics of Pick complex and adult onset dementia. In: Kertt%z A.! and Munoz D.: Pick’s Disease and Pick Complex. New York, Wiley-Lrss Inc., 1998; pp. 269-280. [2] K&ran J., Juh&z A., Majt&nyi K., et al.: Apolipoprotein E polymorphism in Pick’s disease and in Huntington’s disease. Neurobiol. Aging 2000; 21: 555-558. [3] Marsden PA., Heng HH., Scherer SW., et al.: Structure and chromosomal localization of the human constitutive endothelial nitric oxide syntbase gene. J. Biol. Chem. 1993; 268: 1747817488.

Ip.4.0041

Doneperil treatment of Alzheimer’s disease patients in California clinical practice: One year follow-up

Introduction: Pick’s disease (PiD) is a rare neurodegenerative

J. Tinklenberg, L. Newkirk, J. Thompson, L. Ross, H. Davies, R. Shakoori, J. Taylor, R. O’Hara, H. Kraemer, J. Yesavage, The Alzheimer’s Disease Research Centers of California. VA Palo Alto Health Care System and Stanford Universi& USA

disorder in which focal atrophy involving the frontal and temporal lobes is present. The clinical syndrome is characterized by a deterioration of the intellect and slowly processing changes in the character, language, memory, and social functioning, with occasional extrapyramidal symptoms. The etiology of this dementia is unknown, but genetic (1) and inflammatory factors have been implicated. Apolipoprotein E (apoE) gene is considered as a susceptibility locus for Alzheimer’s dementia and vascular dementia. Moreover, the apoE ~4 allele was suggested as a risk factor for PiD also (2). Other candidate gene in dementia is the endothelial nitric oxide synthase (NOS3) gene, which is located on chromosome 7q35 (3). We hypothesized that polymorphic variants in the NOS3 gene might influence the pathology and the degree of the genetic risk for PiD. We compared the frequency of common structural polymorphism, Glu/Asp in codon 298 (exon 7) of NOS3 in PiD and healthy control (HC) populations. Subjects and Methods: A total number of 93 subjects was enrolled in this study. Fifty-one control individuals were healthy volunteers without family history of dementia. Forty-two PiD cases were selected from the tissue bank at the Department of Neuropathology, National Institute of Psychiatry and Neurology, Budapest. The clinical diagnosis of PiD was verified by

Statement of Purpose: The available evidence indicates that donepezil and other cholinesterase inhibitors do not directly affect the neurodegeneration of the Alzheimer disease process. Any positive effects of these drugs on the cognition and behavior of Alzheimer patients usually do not persist more than a few weeks after drug discontinuation. Therefore, patients must continue cholinesterase treatment to maintain benefits. The purpose of this one-year follow-up study was to identify clinically useful predictors of those patients who continue donepezil treatment compared to those who stop. Methods: The baseline diagnostic assessments of the 163 patients were conducted using the same standardized protocol at 6 Alzheimer’s Research Centers of California (ARCC). All patients fulfilled NINCDS-ADRDA criteria for probable or possible Alzheimer’s disease with or without other CNS diagnoses. Each of these patients was currently taking donepezil prescribed by their physician according to his/her usual clinical criteria. Donepezil treatment status was determined at the standard one-year ARCC follow-up visit. Results and Statistical Assessments: Eighty-one of the original 163 patients were taking donepezil at the one-year follow-up.

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p4. Degenerative and neurological disorders

Twenty-five patients had stopped taking donepezil. Forty were lost to follow-up and 17 had died. A logistical regression analysis using baseline predictors as covariates indicated that only the initial Mini-Mental State Exam scores were a significant predictor (‘p < 0.03). Higher baseline scores predicted continued donepezil treatment. Patient age, age at symptom onset, gender, and years of education were not useful predictors. Conclusion: About l/4* of our California AD patients stopped taking donepezil before our one year follow-up. In another routine clinical practice follow-up study in Leeds, England, 38 out of 113 AD patients (34%) prescribed donepezil at initial assessment stopped donepezil treatment before or at their 3 month visit (1). In the California sample, patients who had lower initial Mini-mental State Exam scores were more likely to stop medication.

the ARCI, were all greater for methylphenidate 40 mg than those measured after placebo. Scores for atomoxetine were not different from placebo for any of these measures (stimulant subscales). Scores for non-pleasurable subscales of the VAS and ARC1 were significantly higher with atomoxetine 90 mg than with placebo. Conclusions: The profile associated with atomoxetine differed from that typically observed with stimulants, and significant scores on non-pleasurable sub-scales suggest atomoxetine is unlikely to have significant abuse liability. References

VI American Psychiatric Association. (1994) Diagnostic and Statistical Manual of Mental Disorders: DSM It: Fourth Edition. 4th ed. Washington, DC: American Psychiatric Association.

PI Biederman J, Spencer TJ. (1999) Attention-deficitiweractivitv &&4DHD)

References [l] Cameron, I., Curran, S., Newton, F!, Petty, D., Wattis, .I., 2000. Use of Donepezil for the treatment of mild-moderate Alzheimer’s disease:

An audit of the assessment and treatment of patients in routine clinical practice, Int J Geriatr Psych, 15, 887-891.

asa noradrenekgic disorder. Biol Psych&y

disor46 (91 1234-

t31 Murphy K, Barkby RA. (1996) Prevalence of DSM-IV symptoms of ADHD in adult licensed drivers: Implications Attention Disord 1 (3), 147-161.

for clinical diagnosis. J

IP.4.006( Atomoxetine and methylphenidate treatment Jp.4.005)

Subjective responses to LY139603 (atomoxetine) and methylphenidate

H.F. Laws’, S.H. Heil*, W.K. Bickel*, ST. Higgins*, D.E. Faries’, G. Badge?. ‘Eli Lilly and Company, Indianapolis, IN; ‘University of Vermont, Burlington, m, USA

Statement of Purpose: The onset of Attention-DeficitHyperactivity Disorder (ADHD) occurs early in childhood that affects 3% to 5% of school age children (1). Its pathophysiology appears to involve alterations in dopaminergic and noradrenergic pathways associated with control of attention and impulsivity (2) resulting in impairment of academic and social functioning. The extent of the burden associated with this disorder is compounded, as ADHD is also associated with familial and social dysfunction (3). Symptoms of this disorder have been shown to respond favorably to several pharmacological interventions, most notably stimulants such as methylphenidate and d-amphetamine. However, some patients fail to respond to stimulants or are unable to tolerate them. Additionally, many parents, caregivers, and physicians are concerned by the fact that these medications are considered controlled substances. For all of these reasons, there has been considerable interest in developing new treatments for ADHD. LY 139603 (atomoxetine, formerly known as tomoxetine) is an investigational non-stimulant compound being studied in children with ADHD. Atomoxetine enhances noradrenergic function through highly selective blockade of the pre-synaptic norepinephrine transporter. Atomoxetine has low afIinities for other neuronal transporters and for norepinephrine, dopamine and serotonin receptor sites. This mechanism of action is expected to have a different profile from that of the stimulants with respect to patient perception of drug effects. Methods: Subjects were healthy volunteers who had used recreational drugs previously. Each condition atomoxetine 20 mg, 45 mg, and 90 mg; methylphenidate 20 mg, and 40 mg; and placebo) was assessed on a separate day using a 7-item visual analog scale (VAS), the Addiction Research Center Inventory (ARCI) short form, and Adjective Rating Scale (ARS) questionnaires in an order-randomized, double-blind crossover study design. Results: Mean scores on the stimulant sub-scales of the VAS and ARCI, as well as the amphetamine and euphoria sub-scales of

in children with ADHD: A prospective, randomized, open-label trial R.W. Dittmann' , D.S. Fouche’, C. Kratochvi13, J.H. Heiligenstein4, T. SpenceI.5, J. Biedern&. ‘Eli Lilly and Company, Germany; ‘Eli Lilly and Company-Erl Wood, Surrey, UK; ‘University of Nebmsk-a Medical Center; Omaha, NE; 4Eli Lilly and Company, Indianapolis, IN; ‘Massachusetts General Hospital, Boston, MA, USA

Statement of Purpose: Attention-Deficit/Hyperactivity Disorder (ADHD) is a common disorder of childhood that affects 3% to 5% of school age children in the United States (1). The pathophysiology is not well understood, however it appears to involve alterations in dopaminergic and noradrenergic pathways associated with attention and control of impulsivity (2) resulting in impairment of academic and social functioning. Supportive of the above hypothesis is that ADHD symptoms have shown a favorable response to pharmacological interventions, in particular stimulants such as methylphenidate and d-amphetamine. However, some patients fail to respond to stimulants or are unable to tolerate them. Additionally, many parents, caregivers, and physicians are concerned with using stimulants due to their status as controlled substances. The underlying reason for DEA control is the fear for potential abuse. Additionally, there is also a concern of the potential impact of long-term use of stimulants on development. For these reasons, there has been considerable interest in developing new treatments for ADHD. The purpose of this study was to assess the comparability of atomoxetine, an investigational, nonstimulant therapy for ADHD and methylphenidate. Methods: Children with ADHD were randomized under openlabel conditions to either atomoxetine or methylphenidate therapy for an approximately lo-week period. Gutcomes were assessed using the Attention-Deficit/Hyperactivity Disorder Rating Scale-IVParent Version:Investigator Administered and Scored (ADHDRSIV-Parem:Inv). Results: A total of 228 patients were randomized to treatment (atomoxetine N = 184, methylphenidate N = 44). Both drugs were associated with marked improvement in inattentive and hyperactive/impulsive symptom clusters, and thus there were no statistically significant differences between treatment groups on