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G.P.6 03 NT-proBNP is not associated with dilated cardiomyopathy in Becker and Duchenne muscular dystrophies
698
Abstracts / Neuromuscular Disorders 16 (2006) 644–726
phin observed through western blot analysis in these patients. As the presence of utrophin in untransformed human skin fibroblasts has been also found to be high, here we hypothesized if connective tissue replacing degenerated muscle fibers could also contribute to the higher quantity of utrophin observed in these dystrophic muscles. To test this hypothesis, we compared the expression of utrophin in muscle samples from three groups of patients with diverse degree of degeneration and connective tissue replacement parameters: (a) merosin-deficiency (b) DMD; (c) mildly degenerated LGMD muscles. Although muscle samples from the merosin-deficient patients were the most severely affected and with the highest quantity of connective tissue replacement, utrophin was present in less quantity, followed by the group of DMD. The group of LGMD patients presented the highest amount of utrophin associated to the lesser amount of connective tissue replacement. These results suggest that utrophin retention in the sarcolemma, as a marker of an arrest in muscle maturation, would be the most significant source of the protein in the dystrophic muscle. FAPESPCEPID, CNPq, and ABDIM.
bands compared with the control, more evident in the heart and less prominent in skeletal muscles, especially by NCL-Dys2 antibodies. Multiplex QF-PCR identified the same deletion in the DNA extracted respectively from skeletal and cardiac muscles with mutation proportion of 41% and 61%. Karyotyping showed a normal male pattern. One of his four asymptomatic daughters presented also an increased CPK level and was carrying the same deletion of exons 49–52 in the lymphocytes while it was not detected in his second daughter demonstrating that this mosaicism is not only somatic but also germinal. To our knowledge this is the first case of somatic and germinal mosaicism dystrophinopathic patient with an out-of-frame deletion of exons 49–52 in dystrophin gene with a predominant cardiac involvement. Our data demonstrate that IF study is better than WB analysis to detect a mosaicism pattern, multiplex QF-PCR technique remains the best quantitative technique to identify the coexistence of two dystrophin alleles in different tissues. doi:10.1016/j.nmd.2006.05.177
doi:10.1016/j.nmd.2006.05.176 G.P.6 03
G.P.6 02 Somatic and germinal mosaicism of an out-of-frame deletion in the dystrophin gene in an adult male patient with predominant dilated cardiomyopathy Y.H. Hu 1,4,5,*, F. Leturcq 2, D. He´ron 3, D. Loge´art 4, S. Llense 2, J. Chelly 2, D. Re´can 2, N.B. Romero 1, P. Lafoˆret 1, B. Eymard 1 1 Institut de Myologie, Groupe Hospitalier Pitie´-Salpeˆtrie`re, AP-HP, Paris, France; 2 Laboratoire de Biochimie et ge´ne´tique mole´culaire, Hoˆpital Cochin, AP-HP, Paris, France; 3 Service de ge´ne´tique me´dicale, Groupe Hospitalier Pitie´-Salpeˆtrie`re, AP-HP, Paris, France; 4 Service de Cardiologie, Hoˆpital Beaujon, Clichy, France; 5 Department of Neurology, RUI JIN Hospital, Shanghai, China Dystrophin gene mutations lead to different allelic disorders including Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) and X-linked dilated cardiomyopathy (XL-DCM). In all three entities, cardiac involvement is of variable severity. Somatic mosaicism predominating in the cardiac muscle has been reported in several female carriers of DMD. Few cases of somatic mosaicism in the skeletal muscle and/or in the cardiac muscle of DMD or BMD patients displaying principally skeletal muscle involvement have also been reported. we report a patient carrying an out-of-frame deletion in dystrophin gene, presenting a predominant DCM with a different proportion of mosaicisms in heart, skeletal muscle and lymphocytes. The patient developed DCM leading to heart transplantation at 41-year-old, associated with an asymmetric right upper limb muscular weakness and moderately elevated CPK level. Histological, immunofluorescence (IF), Western blot (WB) studies on the deltoid and cardiac muscles, dystrophin gene analysis by multiplex QF-PCR technique using DNA extracted from lymphocytes, heart and skeletal muscle were performed. Genetic analysis firstly detected an out-of-frame deletion of exons 49–52 in the lymphocytes. In addition, the normal dystrophin gene was also observed, indicating mosaicism in the blood (80%/20%). On deltoid muscle, histological study showed a dystrophic pattern. IF revealed <1% negative muscle fibres just with NCL-Dys2 and specific anti-exon 47–48, 48–50 antibodies. On cardiac muscle, histological examinations showed mainly fibrous replacement of myocardiocytes. IF analysis highlighted a mosaic pattern with about 50% dystrophin negative fibres. WB revealed the discriminating amounts reduction of the dystrophin
NT- proBNP is not associated with dilated cardiomyopathy in Becker and Duchenne muscular dystrophies S.M. Schade van Westrum 1,*, L. Dekker 2, E. Endert 3, R.J. de Haan 4, A.A.M. Wilde 2, M. de Visser 1, A.J. van der Kooi 1 1 Academic Medical Center, University of Amsterdam, Department of Neurology, Amsterdam, The Netherlands; 2 Academic Medical Center, University of Amsterdam, Department of Cardiology, Amsterdam, The Netherlands; 3 Academical Medical Center, University of Amsterdam, Department of Clinical Chemistry, Laboratory of Endocrinology, Amsterdam, The Netherlands; 4 Academic Medical Center, University of Amsterdam, Departments of Clinical Epidemiology and Biostatistics, Amsterdam, The Netherlands N-terminal proBrain natriuretic peptide (NT-proBNP) is a stable protein produced by myocytes of mainly the left cardiac ventricle as a response to stretch. It is increasingly used as predictor for the presence of cardiac failure and as parameter of therapeutic effect. In Becker and Duchenne muscular dystrophies (BMD and DMD) dilated cardiomyopathy (DCM) with cardiac failure is frequently observed among patients and carriers. Although the curvilinear relationship between BNP and systolic dysfunction is known, the relationship between the NT-proBNP level and the presence of DCM in patients and carriers is not. To assess whether NT-Pro BNP can serve as predictor of dilated cardiomyopathy in DMD/BMD patients and carriers, we included 394 individuals (84 DMD (17%), 92 BMD (24%), 147 DMD-carriers (37%), and 81 BMD-carriers (21%)). All were interviewed, neurologically examined and the following tests were done. NT-proBNP in peripheral blood was measured (normal: <0.35 pmol/l). M-mode and 2D echocardiography was used to asses the presence of DCM, defined as an enlarged left ventricle corrected for age and height with a global left ventricle dysfunction or fractional shortening less than 28%. A v2 test was used for associations and ROC-analysis for sensitivity and specificity of NT-proBNP for DCM. In DMD patients mean age was 13.6 years (SD 6.8), 62% was wheelchair bound. The mean age of BMD patients was 30 years (SD 13), 73% had some functional weakness of the leg muscles. Of the DMD-carriers (mean age 44 years, SD 11.8) and of BMD-carriers (43.3 years, SD 12.4), 14% and 13%, respectively, had myalgia, cramps or weakness. Overall, clinical symptoms of cardiac failure were present in 39%. The mean NT-proBNP was 0.64 pmol/l (SD 0.64). 62 (17%) out of 365 individuals in whom echocardiography was feasible, had DCM. Nine patients with DCM had a normal NT-proBNP, none were treated for DCM. The association between an elevated NT-proBNP and DCM was not significant (p = 0.12). A ROC-analysis showed no NT-proBNP value where sen-
Abstracts / Neuromuscular Disorders 16 (2006) 644–726 sitivity and specificity of any significance was reached (area under ROC curve = 0,58). NT-proBNP is not associated with echocardiographic defined dilated cardiomyopathy in DMD/BMD patients and carriers. doi:10.1016/j.nmd.2006.05.178
G.P.6 04 Autoantibodies to myocardium are elevated at high rate in patients with muscular dystrophy T. Matsumura 1,*, T. Yoshio 2, T. Okazaki 3, T. Saito 1, H. Fujimura 1, S. Shinno 1 1 Department of Neurology, National Hospital Organization Toneyama National Hospital, Toyonaka, Japan; 2 Division of Rheumatology and Clinical Immunology, School of Medicine, Jichi Medical University, Tochigi, Japan; 3 21st century COE formation, Department of Medical Chemistry, Graduate School of Medicine, Kyoto University, Kyoto, Japan Cardiac dysfunction is one of the most serious complications for muscular dystrophies. However, its severity is not solely determined by genetic mutation. Even affected siblings in a pedigree frequently present quite different course in cardiac involvements. It seems that there are some modifier factors for cardiac degeneration of muscular dystrophy. Immune factor is one of the major causes of idiopathic cardiomyopathies and autoimmune antibodies to myocardium are frequently detected in these patients. Thus, the immune system may also play some role in cardiac involvements of muscular dystrophy. To reveal this question, we measured anti-b 1 adrenoreceptor antibody (ARAb) and anti-cardiac troponin I antibody (TnIAb) in 134 patients with muscular dystrophy consisting of 108 Duchenne muscular dystrophy patients, 15 myotonic dystrophy patients and 11 other patients. Other serological indexes including brain natriuretic peptide (BNP) and noradrenalin (NA) were examined simultaneously. Echocardiogram and Holter electrocardiogram were also studied. The titers were abnormally elevated in 31.5% in ARAb and 36.0% in TnIAb. These ratios were elevated to 81.3% and 80% in patients with symptomatic cardiac failures. ARAb was weakly correlated to left ventricular ejection fraction, BNP, NA and severity of premature ventricular contractions (Lown grade). We could examine ARAb before and after expression of cardiac failure in four patients and all these patients showed increase of ARAb. Although cardiac function of patients receiving beta blocker (carvedilol) was lower than patients having angiotensin converting enzyme inhibitor alone, the titer of ARAb was rather low. As for TnIAb, we could not find obvious correlation to any cardiac indexes. In conclusion, autoantibodies for myocardium exist in muscular dystrophy in certain ratio as is the case with cardiomyopathies. It is quite likely that immune response has certain influence on cardiac dysfunction even in patients with muscular dystrophy, although we must accumulate more data to elucidate the impact. doi:10.1016/j.nmd.2006.05.179
G.P.6 05 Revising the cardiac phenotype of Duchenne muscular dystrophy L.W. Markham 1,*, A. Barone 1, K. Kinnett 1, R. Spicer 1, B. Wong 2, D.W. Benson 1, L. Cripe 1 1 Division of Cardiology, Cincinnati Children’s Hospital Medical Center, University of Cincinnati, Cincinnati, OH, USA; 2 Division of Neurology, Cincinnati Children’s Hospital Medical Center, University of Cincinnati, Cincinnati, OH, USA
699
Duchenne muscular dystrophy (DMD) is a progressive skeletal and cardiac myopathy with a reported classic electrocardiogram (ECG) consisting of sinus tachycardia along with additional ECG abnormalities. Inappropriate sinus tachycardia has been postulated as precipitating the development of ventricular dysfunction. The objectives of this study were to determine the prevalence of sinus tachycardia and other ECG abnormalities in association with ventricular dysfunction. We hypothesized that the DMD ECG profile would change with age and the degree of ventricular dysfunction. From 1991 to 2006, 105 DMD boys had 503 ECGs and 322 echocardiograms performed. Subjects were divided into five groups based on age of initial cardiac evaluation – A, 0–5 years (n = 17); B, 6–10 years (n = 34); C, 11–15 years (n = 24); D, 16–20 years (n = 19); and E, >20 years (n = 11). Left ventricular (LV) dysfunction was defined as a shortening fraction <28%. By group, mean age in years at evaluation was: A, 4.3; B, 8.5; C, 13.5; D, 18.5; and E, 23.1. Sinus tachycardia (A, 0%; B, 12%; C, 30%; D, 48%; and E, 55%) increased significantly with age (p < 0.01). The percent with LV dysfunction (A, 5%; B, 15%; C, 33%; D, 53%; and E, 64%) also increased significantly with age (p < 0.01). Sinus tachycardia occurred in association with LV dysfunction (p < 0.001). Sinus tachycardia has a sensitivity of 84%, specificity of 95%, predictive value positive of 87% and predictive value negative of 93% for LV dysfunction. Additional ECG abnormalities were frequently noted in later stages of cardiac disease. In DMD, sinus tachycardia increases with age, rarely occurs prior to LV dysfunction, and is temporally related to LV dysfunction. The ‘‘classic’’ ECG findings of DMD as reported in the literature are only seen late in the cardiac disease process. Additional ECG abnormalities become more prevalent with age and correlate with the degree of ventricular dysfunction. doi:10.1016/j.nmd.2006.05.180
G.P.6 06 Patterns of dystrophin gene deletion in Egyptian Duchenne/Becker muscular dystrophy patients R.L. El Sherif *, N. Fahmy, M.A. Etribi Neurology Department, Ain Shams University, Cairo, Egypt Large variations in the proportion of intragenic deletion in the dystrophin gene have been observed in different populations. Although dystrophin gene deletion was extensively studied all over the world, only few studies were done on Egyptian population and there was no account on the dystrophin gene duplication. In this study, we present our results on the pattern of deletion of the dystrophin gene together with the usage of quantitative PCR as a method for duplication analysis within the dystrophin gene in Egyptian patients. Forty-one Duchenne/ Becker muscular dystrophy patients were included in this study. The diagnosis was based on detailed clinical assessment, serum CK level, neurophysiological study and muscle biopsy for histopathological analysis. DNA was extracted from 10 ml peripheral blood according to basic protocol, and multiplex polymerase chain reaction for dystrophin gene using both Chamberlaine and Beggs sets of primers amplifying 18 exons covering the two main dystrophin gene hot spots. In addition primers from Abbs set were used when it was necessary to check the exon borders. DNA from cases with no detectable deletion were analysed for dystrophin gene duplication using Quantitative PCR technique. Furthermore, all cases with no deletion or duplication were subjected for immunohistochemical study using dystrophin antibodies to confirm DMDnBMD diagnosis. In our results five patients were excluded after the immunohistochemical study (where the dystrophin was intact), 22 cases of the 36 showed detectable deletion (61%), 16 patients had deletion within the gene
Abstracts / Neuromuscular Disorders 16 (2006) 644–726
phin observed through western blot analysis in these patients. As the presence of utrophin in untransformed human skin fibroblasts has been also found to be high, here we hypothesized if connective tissue replacing degenerated muscle fibers could also contribute to the higher quantity of utrophin observed in these dystrophic muscles. To test this hypothesis, we compared the expression of utrophin in muscle samples from three groups of patients with diverse degree of degeneration and connective tissue replacement parameters: (a) merosin-deficiency (b) DMD; (c) mildly degenerated LGMD muscles. Although muscle samples from the merosin-deficient patients were the most severely affected and with the highest quantity of connective tissue replacement, utrophin was present in less quantity, followed by the group of DMD. The group of LGMD patients presented the highest amount of utrophin associated to the lesser amount of connective tissue replacement. These results suggest that utrophin retention in the sarcolemma, as a marker of an arrest in muscle maturation, would be the most significant source of the protein in the dystrophic muscle. FAPESPCEPID, CNPq, and ABDIM.
bands compared with the control, more evident in the heart and less prominent in skeletal muscles, especially by NCL-Dys2 antibodies. Multiplex QF-PCR identified the same deletion in the DNA extracted respectively from skeletal and cardiac muscles with mutation proportion of 41% and 61%. Karyotyping showed a normal male pattern. One of his four asymptomatic daughters presented also an increased CPK level and was carrying the same deletion of exons 49–52 in the lymphocytes while it was not detected in his second daughter demonstrating that this mosaicism is not only somatic but also germinal. To our knowledge this is the first case of somatic and germinal mosaicism dystrophinopathic patient with an out-of-frame deletion of exons 49–52 in dystrophin gene with a predominant cardiac involvement. Our data demonstrate that IF study is better than WB analysis to detect a mosaicism pattern, multiplex QF-PCR technique remains the best quantitative technique to identify the coexistence of two dystrophin alleles in different tissues. doi:10.1016/j.nmd.2006.05.177
doi:10.1016/j.nmd.2006.05.176 G.P.6 03
G.P.6 02 Somatic and germinal mosaicism of an out-of-frame deletion in the dystrophin gene in an adult male patient with predominant dilated cardiomyopathy Y.H. Hu 1,4,5,*, F. Leturcq 2, D. He´ron 3, D. Loge´art 4, S. Llense 2, J. Chelly 2, D. Re´can 2, N.B. Romero 1, P. Lafoˆret 1, B. Eymard 1 1 Institut de Myologie, Groupe Hospitalier Pitie´-Salpeˆtrie`re, AP-HP, Paris, France; 2 Laboratoire de Biochimie et ge´ne´tique mole´culaire, Hoˆpital Cochin, AP-HP, Paris, France; 3 Service de ge´ne´tique me´dicale, Groupe Hospitalier Pitie´-Salpeˆtrie`re, AP-HP, Paris, France; 4 Service de Cardiologie, Hoˆpital Beaujon, Clichy, France; 5 Department of Neurology, RUI JIN Hospital, Shanghai, China Dystrophin gene mutations lead to different allelic disorders including Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) and X-linked dilated cardiomyopathy (XL-DCM). In all three entities, cardiac involvement is of variable severity. Somatic mosaicism predominating in the cardiac muscle has been reported in several female carriers of DMD. Few cases of somatic mosaicism in the skeletal muscle and/or in the cardiac muscle of DMD or BMD patients displaying principally skeletal muscle involvement have also been reported. we report a patient carrying an out-of-frame deletion in dystrophin gene, presenting a predominant DCM with a different proportion of mosaicisms in heart, skeletal muscle and lymphocytes. The patient developed DCM leading to heart transplantation at 41-year-old, associated with an asymmetric right upper limb muscular weakness and moderately elevated CPK level. Histological, immunofluorescence (IF), Western blot (WB) studies on the deltoid and cardiac muscles, dystrophin gene analysis by multiplex QF-PCR technique using DNA extracted from lymphocytes, heart and skeletal muscle were performed. Genetic analysis firstly detected an out-of-frame deletion of exons 49–52 in the lymphocytes. In addition, the normal dystrophin gene was also observed, indicating mosaicism in the blood (80%/20%). On deltoid muscle, histological study showed a dystrophic pattern. IF revealed <1% negative muscle fibres just with NCL-Dys2 and specific anti-exon 47–48, 48–50 antibodies. On cardiac muscle, histological examinations showed mainly fibrous replacement of myocardiocytes. IF analysis highlighted a mosaic pattern with about 50% dystrophin negative fibres. WB revealed the discriminating amounts reduction of the dystrophin
NT- proBNP is not associated with dilated cardiomyopathy in Becker and Duchenne muscular dystrophies S.M. Schade van Westrum 1,*, L. Dekker 2, E. Endert 3, R.J. de Haan 4, A.A.M. Wilde 2, M. de Visser 1, A.J. van der Kooi 1 1 Academic Medical Center, University of Amsterdam, Department of Neurology, Amsterdam, The Netherlands; 2 Academic Medical Center, University of Amsterdam, Department of Cardiology, Amsterdam, The Netherlands; 3 Academical Medical Center, University of Amsterdam, Department of Clinical Chemistry, Laboratory of Endocrinology, Amsterdam, The Netherlands; 4 Academic Medical Center, University of Amsterdam, Departments of Clinical Epidemiology and Biostatistics, Amsterdam, The Netherlands N-terminal proBrain natriuretic peptide (NT-proBNP) is a stable protein produced by myocytes of mainly the left cardiac ventricle as a response to stretch. It is increasingly used as predictor for the presence of cardiac failure and as parameter of therapeutic effect. In Becker and Duchenne muscular dystrophies (BMD and DMD) dilated cardiomyopathy (DCM) with cardiac failure is frequently observed among patients and carriers. Although the curvilinear relationship between BNP and systolic dysfunction is known, the relationship between the NT-proBNP level and the presence of DCM in patients and carriers is not. To assess whether NT-Pro BNP can serve as predictor of dilated cardiomyopathy in DMD/BMD patients and carriers, we included 394 individuals (84 DMD (17%), 92 BMD (24%), 147 DMD-carriers (37%), and 81 BMD-carriers (21%)). All were interviewed, neurologically examined and the following tests were done. NT-proBNP in peripheral blood was measured (normal: <0.35 pmol/l). M-mode and 2D echocardiography was used to asses the presence of DCM, defined as an enlarged left ventricle corrected for age and height with a global left ventricle dysfunction or fractional shortening less than 28%. A v2 test was used for associations and ROC-analysis for sensitivity and specificity of NT-proBNP for DCM. In DMD patients mean age was 13.6 years (SD 6.8), 62% was wheelchair bound. The mean age of BMD patients was 30 years (SD 13), 73% had some functional weakness of the leg muscles. Of the DMD-carriers (mean age 44 years, SD 11.8) and of BMD-carriers (43.3 years, SD 12.4), 14% and 13%, respectively, had myalgia, cramps or weakness. Overall, clinical symptoms of cardiac failure were present in 39%. The mean NT-proBNP was 0.64 pmol/l (SD 0.64). 62 (17%) out of 365 individuals in whom echocardiography was feasible, had DCM. Nine patients with DCM had a normal NT-proBNP, none were treated for DCM. The association between an elevated NT-proBNP and DCM was not significant (p = 0.12). A ROC-analysis showed no NT-proBNP value where sen-
Abstracts / Neuromuscular Disorders 16 (2006) 644–726 sitivity and specificity of any significance was reached (area under ROC curve = 0,58). NT-proBNP is not associated with echocardiographic defined dilated cardiomyopathy in DMD/BMD patients and carriers. doi:10.1016/j.nmd.2006.05.178
G.P.6 04 Autoantibodies to myocardium are elevated at high rate in patients with muscular dystrophy T. Matsumura 1,*, T. Yoshio 2, T. Okazaki 3, T. Saito 1, H. Fujimura 1, S. Shinno 1 1 Department of Neurology, National Hospital Organization Toneyama National Hospital, Toyonaka, Japan; 2 Division of Rheumatology and Clinical Immunology, School of Medicine, Jichi Medical University, Tochigi, Japan; 3 21st century COE formation, Department of Medical Chemistry, Graduate School of Medicine, Kyoto University, Kyoto, Japan Cardiac dysfunction is one of the most serious complications for muscular dystrophies. However, its severity is not solely determined by genetic mutation. Even affected siblings in a pedigree frequently present quite different course in cardiac involvements. It seems that there are some modifier factors for cardiac degeneration of muscular dystrophy. Immune factor is one of the major causes of idiopathic cardiomyopathies and autoimmune antibodies to myocardium are frequently detected in these patients. Thus, the immune system may also play some role in cardiac involvements of muscular dystrophy. To reveal this question, we measured anti-b 1 adrenoreceptor antibody (ARAb) and anti-cardiac troponin I antibody (TnIAb) in 134 patients with muscular dystrophy consisting of 108 Duchenne muscular dystrophy patients, 15 myotonic dystrophy patients and 11 other patients. Other serological indexes including brain natriuretic peptide (BNP) and noradrenalin (NA) were examined simultaneously. Echocardiogram and Holter electrocardiogram were also studied. The titers were abnormally elevated in 31.5% in ARAb and 36.0% in TnIAb. These ratios were elevated to 81.3% and 80% in patients with symptomatic cardiac failures. ARAb was weakly correlated to left ventricular ejection fraction, BNP, NA and severity of premature ventricular contractions (Lown grade). We could examine ARAb before and after expression of cardiac failure in four patients and all these patients showed increase of ARAb. Although cardiac function of patients receiving beta blocker (carvedilol) was lower than patients having angiotensin converting enzyme inhibitor alone, the titer of ARAb was rather low. As for TnIAb, we could not find obvious correlation to any cardiac indexes. In conclusion, autoantibodies for myocardium exist in muscular dystrophy in certain ratio as is the case with cardiomyopathies. It is quite likely that immune response has certain influence on cardiac dysfunction even in patients with muscular dystrophy, although we must accumulate more data to elucidate the impact. doi:10.1016/j.nmd.2006.05.179
G.P.6 05 Revising the cardiac phenotype of Duchenne muscular dystrophy L.W. Markham 1,*, A. Barone 1, K. Kinnett 1, R. Spicer 1, B. Wong 2, D.W. Benson 1, L. Cripe 1 1 Division of Cardiology, Cincinnati Children’s Hospital Medical Center, University of Cincinnati, Cincinnati, OH, USA; 2 Division of Neurology, Cincinnati Children’s Hospital Medical Center, University of Cincinnati, Cincinnati, OH, USA
699
Duchenne muscular dystrophy (DMD) is a progressive skeletal and cardiac myopathy with a reported classic electrocardiogram (ECG) consisting of sinus tachycardia along with additional ECG abnormalities. Inappropriate sinus tachycardia has been postulated as precipitating the development of ventricular dysfunction. The objectives of this study were to determine the prevalence of sinus tachycardia and other ECG abnormalities in association with ventricular dysfunction. We hypothesized that the DMD ECG profile would change with age and the degree of ventricular dysfunction. From 1991 to 2006, 105 DMD boys had 503 ECGs and 322 echocardiograms performed. Subjects were divided into five groups based on age of initial cardiac evaluation – A, 0–5 years (n = 17); B, 6–10 years (n = 34); C, 11–15 years (n = 24); D, 16–20 years (n = 19); and E, >20 years (n = 11). Left ventricular (LV) dysfunction was defined as a shortening fraction <28%. By group, mean age in years at evaluation was: A, 4.3; B, 8.5; C, 13.5; D, 18.5; and E, 23.1. Sinus tachycardia (A, 0%; B, 12%; C, 30%; D, 48%; and E, 55%) increased significantly with age (p < 0.01). The percent with LV dysfunction (A, 5%; B, 15%; C, 33%; D, 53%; and E, 64%) also increased significantly with age (p < 0.01). Sinus tachycardia occurred in association with LV dysfunction (p < 0.001). Sinus tachycardia has a sensitivity of 84%, specificity of 95%, predictive value positive of 87% and predictive value negative of 93% for LV dysfunction. Additional ECG abnormalities were frequently noted in later stages of cardiac disease. In DMD, sinus tachycardia increases with age, rarely occurs prior to LV dysfunction, and is temporally related to LV dysfunction. The ‘‘classic’’ ECG findings of DMD as reported in the literature are only seen late in the cardiac disease process. Additional ECG abnormalities become more prevalent with age and correlate with the degree of ventricular dysfunction. doi:10.1016/j.nmd.2006.05.180
G.P.6 06 Patterns of dystrophin gene deletion in Egyptian Duchenne/Becker muscular dystrophy patients R.L. El Sherif *, N. Fahmy, M.A. Etribi Neurology Department, Ain Shams University, Cairo, Egypt Large variations in the proportion of intragenic deletion in the dystrophin gene have been observed in different populations. Although dystrophin gene deletion was extensively studied all over the world, only few studies were done on Egyptian population and there was no account on the dystrophin gene duplication. In this study, we present our results on the pattern of deletion of the dystrophin gene together with the usage of quantitative PCR as a method for duplication analysis within the dystrophin gene in Egyptian patients. Forty-one Duchenne/ Becker muscular dystrophy patients were included in this study. The diagnosis was based on detailed clinical assessment, serum CK level, neurophysiological study and muscle biopsy for histopathological analysis. DNA was extracted from 10 ml peripheral blood according to basic protocol, and multiplex polymerase chain reaction for dystrophin gene using both Chamberlaine and Beggs sets of primers amplifying 18 exons covering the two main dystrophin gene hot spots. In addition primers from Abbs set were used when it was necessary to check the exon borders. DNA from cases with no detectable deletion were analysed for dystrophin gene duplication using Quantitative PCR technique. Furthermore, all cases with no deletion or duplication were subjected for immunohistochemical study using dystrophin antibodies to confirm DMDnBMD diagnosis. In our results five patients were excluded after the immunohistochemical study (where the dystrophin was intact), 22 cases of the 36 showed detectable deletion (61%), 16 patients had deletion within the gene