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0227 Lack of association between a functional polymorphism in the tyrosine hydroxylase gene and Parkinson's disease
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Monday, November 7, 2005
adverse drug effect are very poorly understood, as are the factors involved in determining individual susceptibility. This study aimed to develop a m o u s e model of AED-induced bone loss, and to identify different genetic strains that are sensitive and resistant to tiffs effect. Method: Seven different inbred mice strains (in - 40 per strain, n -- 10 per diet) were placed on a diet mixed with 0, 2, 4 or 6 g/kg valproate (VPA) for 8 weeks. Total BMC, fat mass and lean mass were assessed using dual energy x-ray absorptiometry (DXA). B M C was corrected for total body weight and total lean mass to account for differences in animal size. Results: Strain 1 was identified as being sensitive to VPA-induced bone disease, showing significant differences (195% CI) of 10.4% (12.7%18.2%) and 8.4 % (0.1%-16.2%), respectively, in weight-adjusted B M C compared with control mice whilst on the 2 a n d 6 g/kg V P A diets (p < 0.05). Strain 2 was identified as a strain resistant to the effects of V P A on B M C at all doses. Two other VPA-sensitive and resistant strains have also been identified. Conclusions: This study has produced, for the first time, a n animal model of A E D associated bone loss, and identified differential effects in mice of differing genetic, backgrounds. This should provide an invaluable tool to allow tire identification of genes and the mechanisnrs that mrderlie these effects. Tiffs approach will facilitate the design of therapeutic strategies for tire prevention and treatnrent of AEDassociated bone disease.
0226 The Role of Polymorphis~rns of Genes - Regulators of Programmed Cell Death in Formation of Brain Infarction and Severity of Atherothrombotie Ischemic Stroke (IS) Skvortsova, V ~, Shetova, 1-a, Koltsova, E ~, Slominsky, p2, Tupit sina, T 2, Limborska, S2. ZDepartment of Fundamental and Clinical Neurology,
Russian State Medical University, Moscow; Russian Federation; 2Institute of Molecular Generics, Moscow; Russian Federation Purpose: The aim was to study the role of PARP-1, A I F and p53 polymorphisnls in formation of the brain infarction and IS severity. Methods: Rsa-1- PARP-1, Bcl-1 A I F and Banr HI p53 diallelic polymorphisnts were studied in 96 patients with atherothrombotic IS. M R I with m o r p h o m e t r y was applied on days 1, 3, 7 and 21 for calculation of brain infarction volume (BIV). Presence of restriction site for all polymorphisms marked as (+), absence - as (-). CSF P A R level was assessed on days 1 and 3 using ELISA. Results: A close association between Rsa-I PARP-1 and Banr HI p53 polymorphisnts and BIV was revealed. The predominance of the bigsize infarctions (i>90 cm 3) was found in patients with ( - / - ) Rsa-I PARP-1 genotype vs. carriers o f (+) Rsa-I allele on all the days (p < 0.02-0.007), and in carriers o f ( + ) Barn HI p53 allele in comparison with ( / ) Banr HI genotype on days 3 and 7 ( P < 0.001). Carriage of ( / ) Rsa-I PARP-1 genotype and (+)Barn HI p53 allele was also connected with worse neurological status (p < 0.001). Tire associations of PARP-1 and p53 gene polymorphisms with BIV and stroke severity were independent on location of artery occlusion (p < 0.002). The correlation between CSF P A R level on day 1 and both BIV (p -0.03; r -- 0.67) and clinical score (p -- 0.04; r -0.65) on day 3 was revealed. Higher P A R level occurred in carriers o f (+) Barn HI p53 allele (~v _ 0.03). Discussion: The results confirm a substantial contribution o f programmed cell death in both formation of brain infarction and severity of IS.
0227 Lack of association between a Functional Polymorphism in the Tyrosine Hydroxylase Gene and Parkinson's Disease Stevens, J~, Sutherland, G a, Lee, L a, Double, K a, Rowe, D 2, Halliday, G 1. z Prince of Wales Medical Research Institute and the
Poster Abstracts University of New South Wa[es, Sydney, Australia; ~Deparmwnt of Neurology, Royal North Shore Hospital, Sydney, Australia Background: Tyrosine hydroxylase (TH) catalyses tire rate-limiting step in dopanffne production and is therefore a potential candidate gene for tire aetiology of Parkinson's disease (PD). A functional polymorphic microsatellite, H U M T H 0 1 , is found in intron 1 of the T H gene, although a single study suggests no disease association and mutations in the T H gene have not been found in families with PD. We have evaluated a larger Australian cohort to determine whether a genetic influence on T H function associates with PD. Methods: Genomic D N A was extracted from whole blood of PD patients (in - 135) and age a n d sex matched controls (in - 126) recruited through the Royal N o r t h Shore Hospital movement disorders clinic. The H U M T H 0 1 tetranucleotide repeat region was anrplified using the polymerase chain reaction a n d the products electrophoresed on a 5% denaturing polyacrylanffde gel with a Corbett Gelscan 2000. Allelic and genotypic frequencies of cases and controls were compared using Chi-square analyses. Results: Five H U M T H 0 1 alleles were identified and designated a, b, c, d a n d e (relative sizes: 310, 314, 318, 322 and 326 bp respectively). There were no sigtffficant differences in either allelic (X~ - 1.11, p > 0.9) or genotypic frequencies ()~2 _ 12.81, p > 0.7) between the PD cases and controls. Conehisions: Our results suggest that there is no association between the H U M T H 0 1 functional polymorphism and PD, consistent with previous findings. 0228 Genetic variation of alphaB-crystallin (CRYAB) and CSF alptlaB-erystallin in patients with ulultiple sclerosis Stoevring B ~, Frederiksen J~, Vang 0 3, C.hristiansen M 1..
1Department of Clinical Biochemistry, Statens Serum Institut, Copenhagen, Denmark *[email protected]; 2Tile MS clinic, Department of Neurology, University of Copenhagen, Glostrup Hospital, Glostrup, Denmark; 3Department of Life Sciences and Chemistry, Roskilde University, Roskilde, Denmark Background: e~B-crystallin is a molecular chaperone a n d potential myelin antigen, up-regulated in the earlier stages of Multiple Sclerosis (MS) lesions. In the ~B-crystallin gene (CRYAB), single nucleofide polymovphisms (SNPs) have been associated with disease susceptibility ( - 6 5 2 A > G ) and a rapidly progressive clinical course (-650c > 6 ) . Method: CRYAB was genetically screened in MS patients (96 R R M S , 96 SPMS, 82 PPMS) a n d 96 controls. GenomJc D N A was extracted and the coding and untranslated regions were anrplified by PCR. Subsequently, the products were analysed by SSCP technique followed by D N A sequencing of aberrant conformers. In addition, an anti G, 650C> G and - 6 5 2 A > G), coding region (1162G > A) and intron (2398T > G). The -249C > G genotype distribution was significant different between groups (Zz, p - 0.01), caused by differences between R R M S and controls (Z2, p -- 0.025) and SPMS and controls (Z2, p - 0.05). In addition, a sigtffficant difference was observed in the 249C> G allele distribution (X2, p - 0.04), caused by a difference between SPMS and controls (Xa, p - 0.01). Likewise, a significant difference in 2398T > G genotype distribution was observed (Xa, p -- 0.04), caused by difference between R R M S and controls (iXz, p -- 0.05) a n d between SPMS and PPMS (X2, p -- 0.05). Conelasion: ~B-crystallin is present in the CSF and m a y be involved in tire pathogenesis of MS.
Monday, November 7, 2005
adverse drug effect are very poorly understood, as are the factors involved in determining individual susceptibility. This study aimed to develop a m o u s e model of AED-induced bone loss, and to identify different genetic strains that are sensitive and resistant to tiffs effect. Method: Seven different inbred mice strains (in - 40 per strain, n -- 10 per diet) were placed on a diet mixed with 0, 2, 4 or 6 g/kg valproate (VPA) for 8 weeks. Total BMC, fat mass and lean mass were assessed using dual energy x-ray absorptiometry (DXA). B M C was corrected for total body weight and total lean mass to account for differences in animal size. Results: Strain 1 was identified as being sensitive to VPA-induced bone disease, showing significant differences (195% CI) of 10.4% (12.7%18.2%) and 8.4 % (0.1%-16.2%), respectively, in weight-adjusted B M C compared with control mice whilst on the 2 a n d 6 g/kg V P A diets (p < 0.05). Strain 2 was identified as a strain resistant to the effects of V P A on B M C at all doses. Two other VPA-sensitive and resistant strains have also been identified. Conclusions: This study has produced, for the first time, a n animal model of A E D associated bone loss, and identified differential effects in mice of differing genetic, backgrounds. This should provide an invaluable tool to allow tire identification of genes and the mechanisnrs that mrderlie these effects. Tiffs approach will facilitate the design of therapeutic strategies for tire prevention and treatnrent of AEDassociated bone disease.
0226 The Role of Polymorphis~rns of Genes - Regulators of Programmed Cell Death in Formation of Brain Infarction and Severity of Atherothrombotie Ischemic Stroke (IS) Skvortsova, V ~, Shetova, 1-a, Koltsova, E ~, Slominsky, p2, Tupit sina, T 2, Limborska, S2. ZDepartment of Fundamental and Clinical Neurology,
Russian State Medical University, Moscow; Russian Federation; 2Institute of Molecular Generics, Moscow; Russian Federation Purpose: The aim was to study the role of PARP-1, A I F and p53 polymorphisnls in formation of the brain infarction and IS severity. Methods: Rsa-1- PARP-1, Bcl-1 A I F and Banr HI p53 diallelic polymorphisnts were studied in 96 patients with atherothrombotic IS. M R I with m o r p h o m e t r y was applied on days 1, 3, 7 and 21 for calculation of brain infarction volume (BIV). Presence of restriction site for all polymorphisms marked as (+), absence - as (-). CSF P A R level was assessed on days 1 and 3 using ELISA. Results: A close association between Rsa-I PARP-1 and Banr HI p53 polymorphisnts and BIV was revealed. The predominance of the bigsize infarctions (i>90 cm 3) was found in patients with ( - / - ) Rsa-I PARP-1 genotype vs. carriers o f (+) Rsa-I allele on all the days (p < 0.02-0.007), and in carriers o f ( + ) Barn HI p53 allele in comparison with ( / ) Banr HI genotype on days 3 and 7 ( P < 0.001). Carriage of ( / ) Rsa-I PARP-1 genotype and (+)Barn HI p53 allele was also connected with worse neurological status (p < 0.001). Tire associations of PARP-1 and p53 gene polymorphisms with BIV and stroke severity were independent on location of artery occlusion (p < 0.002). The correlation between CSF P A R level on day 1 and both BIV (p -0.03; r -- 0.67) and clinical score (p -- 0.04; r -0.65) on day 3 was revealed. Higher P A R level occurred in carriers o f (+) Barn HI p53 allele (~v _ 0.03). Discussion: The results confirm a substantial contribution o f programmed cell death in both formation of brain infarction and severity of IS.
0227 Lack of association between a Functional Polymorphism in the Tyrosine Hydroxylase Gene and Parkinson's Disease Stevens, J~, Sutherland, G a, Lee, L a, Double, K a, Rowe, D 2, Halliday, G 1. z Prince of Wales Medical Research Institute and the
Poster Abstracts University of New South Wa[es, Sydney, Australia; ~Deparmwnt of Neurology, Royal North Shore Hospital, Sydney, Australia Background: Tyrosine hydroxylase (TH) catalyses tire rate-limiting step in dopanffne production and is therefore a potential candidate gene for tire aetiology of Parkinson's disease (PD). A functional polymorphic microsatellite, H U M T H 0 1 , is found in intron 1 of the T H gene, although a single study suggests no disease association and mutations in the T H gene have not been found in families with PD. We have evaluated a larger Australian cohort to determine whether a genetic influence on T H function associates with PD. Methods: Genomic D N A was extracted from whole blood of PD patients (in - 135) and age a n d sex matched controls (in - 126) recruited through the Royal N o r t h Shore Hospital movement disorders clinic. The H U M T H 0 1 tetranucleotide repeat region was anrplified using the polymerase chain reaction a n d the products electrophoresed on a 5% denaturing polyacrylanffde gel with a Corbett Gelscan 2000. Allelic and genotypic frequencies of cases and controls were compared using Chi-square analyses. Results: Five H U M T H 0 1 alleles were identified and designated a, b, c, d a n d e (relative sizes: 310, 314, 318, 322 and 326 bp respectively). There were no sigtffficant differences in either allelic (X~ - 1.11, p > 0.9) or genotypic frequencies ()~2 _ 12.81, p > 0.7) between the PD cases and controls. Conehisions: Our results suggest that there is no association between the H U M T H 0 1 functional polymorphism and PD, consistent with previous findings. 0228 Genetic variation of alphaB-crystallin (CRYAB) and CSF alptlaB-erystallin in patients with ulultiple sclerosis Stoevring B ~, Frederiksen J~, Vang 0 3, C.hristiansen M 1..
1Department of Clinical Biochemistry, Statens Serum Institut, Copenhagen, Denmark *[email protected]; 2Tile MS clinic, Department of Neurology, University of Copenhagen, Glostrup Hospital, Glostrup, Denmark; 3Department of Life Sciences and Chemistry, Roskilde University, Roskilde, Denmark Background: e~B-crystallin is a molecular chaperone a n d potential myelin antigen, up-regulated in the earlier stages of Multiple Sclerosis (MS) lesions. In the ~B-crystallin gene (CRYAB), single nucleofide polymovphisms (SNPs) have been associated with disease susceptibility ( - 6 5 2 A > G ) and a rapidly progressive clinical course (-650c > 6 ) . Method: CRYAB was genetically screened in MS patients (96 R R M S , 96 SPMS, 82 PPMS) a n d 96 controls. GenomJc D N A was extracted and the coding and untranslated regions were anrplified by PCR. Subsequently, the products were analysed by SSCP technique followed by D N A sequencing of aberrant conformers. In addition, an anti