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Francesca Magri,
Roberto Del Bo,
Maria Grazia D'Angelo,
Monica Sciacco,
Sandra Gandossini,
Alessandra Govoni,
Laura Napoli, Patrizia Ciscato,
Francesco Fortunato,
Erika Brighina,
Sara Bonato,
Andreina Bordoni,
Valeria Lucchini,
Stefania Corti,
Maurizio Moggio,
Nereo Bresolin,
Giacomo Pietro Comi
[show abstract]
[hide abstract]
ABSTRACT: Limb-girdle muscular dystrophy (LGMD) 2L, caused by mutations in the anoctamin 5 (ANO5) gene, is the third most common LGMD in Northern and Central Europe, where the c.191dupA mutation causes the majority of cases. We evaluated data from 228 Italian LGMD patients to determine the prevalence of LGMD2L and the c.191dupA mutation, and to describe the clinical, muscle biopsy, and magnetic resonance imaging findings in these patients. Forty-three patients who lacked molecular diagnosis were studied for ANO5 mutations, and four novel mutations were found in three probands. Only one proband carried the c.191dupA mutation, which was compound heterozygous with c.2516T>G. Two probands were homozygous for the c.1627dupA and c.397A>T mutations, respectively, while a fourth proband had a compound heterozygous status (c.220C>T and c.1609T>C). Therefore occurrence and molecular epidemiology of LGMD2L in this Italian cohort differed from those observed in other European countries. ANO5 mutations accounted for ∼2% of our sample. Affected patients exhibited benign progression with variable onset and an absence of cardiac and respiratory impairment; muscle biopsy generally showed mild signs, except when performed on the quadriceps muscles; MRI showed predominant involvement of the posterior thigh. Overall these common clinical, morphological and imaging findings could be useful in differential diagnosis.
Neuromuscular Disorders 06/2012; · 2.80 Impact Factor
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Francesca Magri,
Alessandra Govoni,
Maria Grazia D’Angelo,
Roberto Del Bo,
Serena Ghezzi,
Gandossini Sandra,
Anna Carla Turconi,
Monica Sciacco, Patrizia Ciscato,
Andreina Bordoni, [......],
Francesco Fortunato,
Valeria Lucchini,
Sara Bonato,
Costanza Lamperti,
Domenico Coviello,
Yvan Torrente,
Stefania Corti,
Maurizio Moggio,
Nereo Bresolin,
Giacomo Pietro Comi
[show abstract]
[hide abstract]
ABSTRACT: Duchenne and Becker muscular dystrophy (DMD and BMD, respectively) are allelic disorders with different clinical presentations
and severity determined by mutations in the gene DMD, which encodes the sarcolemmal protein dystrophin. Diagnosis is based on clinical aspects and muscle protein analysis, followed
by molecular confirmation. We revised the main aspects of the natural history of dystrophinopathies to define genotype–phenotype
correlations in large patient cohorts with extended follow-up. We also specifically explored subjects carrying nucleotide
substitutions in the DMD gene, a comparatively less investigated DMD/BMD subgroup. We studied 320 dystrophinopathic patients (205 DMD and 115 BMD),
defining muscular, cardiac, respiratory, and cognitive involvement. We also subdivided patients according to the kind of molecular
defect (deletions, duplications, nucleotide substitutions or other microrearrangements) and the mutation sites (proximal/distal
to exon 45), studying phenotype–genotype correlations for each group. In DMD, mutation type did not influence clinical evolution;
mutations located in distal regions (irrespective of their nature) are more likely to be associated with lower IQ levels (p=0.005). BMD carrying proximal deletions showed a higher degree of cardiac impairment than BMD with distal deletions (p=0.0046). In the BMD population, there was a strong correlation between the entity of muscle dystrophin deficiency and clinical
course (p=0.002). An accurate knowledge of natural history may help in the clinical management of patients. Furthermore, several
clinical trials are ongoing or are currently planned, some of which aim to target specific DMD mutations: a robust natural history is therefore essential to correctly design these experimental trials.
KeywordsDuchenne muscular dystrophy–Becker muscular dystrophy–Dystrophin gene sequencing–Protein analysis–Cardiac involvement in DMD and BMD–Respiratory involvement in DMD and BMD
Journal of Neurology 04/2012; 258(9):1610-1623. · 3.47 Impact Factor
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Dario Ronchi,
Monica Sciacco,
Andreina Bordoni,
Monika Raimondi,
Michela Ripolone,
Elisa Fassone,
Alessio Di Fonzo,
Mafalda Rizzuti, Patrizia Ciscato,
Alessandra Cosi,
Maura Servida,
Maurizio Moggio,
Stefania Corti,
Nereo Bresolin,
Giacomo P Comi
[show abstract]
[hide abstract]
ABSTRACT: Although mutations in mitochondrial tRNAs constitute the most common mtDNA defect, the presence of pathological variants in mitochondrial tRNA(Asn) is extremely rare. We were able to identify a novel mtDNA tRNA(Asn) gene pathogenic mutation associated with a myopathic phenotype and a previously unreported respiratory impairment. Our proband is an adult woman with ophthalmoparesis and respiratory impairment. Her muscle biopsy presented several cytochrome c oxidase-negative (COX-) fibres and signs of mitochondrial proliferation (ragged red fibres). Sequence analysis of the muscle-derived mtDNA revealed an m.5709T>C substitution, affecting mitochondrial tRNA(Asn) gene. Restriction-fragment length polymorphism analysis of the mutation in isolated muscle fibres showed that a threshold of at least 91.9% mutated mtDNA results in the COX deficiency phenotype. The new phenotype further increases the clinical spectrum of mitochondrial diseases caused by mutations in the tRNA(Asn) gene.
European journal of human genetics: EJHG 12/2011; 20(3):357-60. · 3.56 Impact Factor
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Dario Ronchi,
Elisa Fassone,
Andreina Bordoni,
Monica Sciacco,
Valeria Lucchini,
Alessio Di Fonzo,
Mafalda Rizzuti,
Irene Colombo,
Laura Napoli, Patrizia Ciscato,
Maurizio Moggio,
Alessandra Cosi,
Martina Collotta,
Stefania Corti,
Nereo Bresolin,
Giacomo P Comi
[show abstract]
[hide abstract]
ABSTRACT: Maintenance and replication of mitochondrial DNA require the concerted action of several factors encoded by nuclear genome. The mitochondrial helicase Twinkle is a key player of replisome machinery. Heterozygous mutations in its coding gene, PEO1, are associated with progressive external ophthalmoplegia (PEO) characterised by ptosis and ophthalmoparesis, with cytochrome c oxidase (COX)-deficient fibres, ragged-red fibres (RRF) and multiple mtDNA deletions in muscle. Here we describe clinical, histological and molecular features of two patients presenting with mitochondrial myopathy associated with PEO. PEO1 sequencing disclosed two novel mutations in exons 1 and 4 of the gene, respectively. Although mutations in PEO1 exon 1 have already been described, this is the first report of mutation occurring in exon 4.
Journal of the neurological sciences 06/2011; 308(1-2):173-6. · 2.32 Impact Factor
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Francesca Magri,
Alessandra Govoni,
Maria Grazia D'Angelo,
Roberto Del Bo,
Serena Ghezzi,
Gandossini Sandra,
Anna Carla Turconi,
Monica Sciacco, Patrizia Ciscato,
Andreina Bordoni, [......],
Francesco Fortunato,
Valeria Lucchini,
Sara Bonato,
Costanza Lamperti,
Domenico Coviello,
Yvan Torrente,
Stefania Corti,
Maurizio Moggio,
Nereo Bresolin,
Giacomo Pietro Comi
[show abstract]
[hide abstract]
ABSTRACT: Duchenne and Becker muscular dystrophy (DMD and BMD, respectively) are allelic disorders with different clinical presentations and severity determined by mutations in the gene DMD, which encodes the sarcolemmal protein dystrophin. Diagnosis is based on clinical aspects and muscle protein analysis, followed by molecular confirmation. We revised the main aspects of the natural history of dystrophinopathies to define genotype-phenotype correlations in large patient cohorts with extended follow-up. We also specifically explored subjects carrying nucleotide substitutions in the DMD gene, a comparatively less investigated DMD/BMD subgroup. We studied 320 dystrophinopathic patients (205 DMD and 115 BMD), defining muscular, cardiac, respiratory, and cognitive involvement. We also subdivided patients according to the kind of molecular defect (deletions, duplications, nucleotide substitutions or other microrearrangements) and the mutation sites (proximal/distal to exon 45), studying phenotype-genotype correlations for each group. In DMD, mutation type did not influence clinical evolution; mutations located in distal regions (irrespective of their nature) are more likely to be associated with lower IQ levels (p = 0.005). BMD carrying proximal deletions showed a higher degree of cardiac impairment than BMD with distal deletions (p = 0.0046). In the BMD population, there was a strong correlation between the entity of muscle dystrophin deficiency and clinical course (p = 0.002). An accurate knowledge of natural history may help in the clinical management of patients. Furthermore, several clinical trials are ongoing or are currently planned, some of which aim to target specific DMD mutations: a robust natural history is therefore essential to correctly design these experimental trials.
Journal of Neurology 03/2011; 258(9):1610-23. · 3.47 Impact Factor
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Francesca Magri,
Roberto Del Bo,
Maria G D'Angelo,
Alessandra Govoni,
Serena Ghezzi,
Sandra Gandossini,
Monica Sciacco, Patrizia Ciscato,
Andreina Bordoni,
Silvana Tedeschi,
Francesco Fortunato,
Valeria Lucchini,
Matteo Cereda,
Stefania Corti,
Maurizio Moggio,
Nereo Bresolin,
Giacomo P Comi
[show abstract]
[hide abstract]
ABSTRACT: Duchenne and Becker Muscular dystrophies (DMD/BMD) are allelic disorders caused by mutations in the dystrophin gene, which encodes a sarcolemmal protein responsible for muscle integrity. Deletions and duplications account for approximately 75% of mutations in DMD and 85% in BMD. The implementation of techniques allowing complete gene sequencing has focused attention on small point mutations and other mechanisms underlying complex rearrangements.
We selected 47 patients (41 families; 35 DMD, 6 BMD) without deletions and duplications in DMD gene (excluded by multiplex ligation-dependent probe amplification and multiplex polymerase chain reaction analysis). This cohort was investigated by systematic direct sequence analysis to study sequence variation. We focused our attention on rare mutational events which were further studied through transcript analysis.
We identified 40 different nucleotide alterations in DMD gene and their clinical correlates; altogether, 16 mutations were novel. DMD probands carried 9 microinsertions/microdeletions, 19 nonsense mutations, and 7 splice-site mutations. BMD patients carried 2 nonsense mutations, 2 splice-site mutations, 1 missense substitution, and 1 single base insertion. The most frequent stop codon was TGA (n=10 patients), followed by TAG (n=7) and TAA (n=4). We also analyzed the molecular mechanisms of five rare mutational events. They are two frame-shifting mutations in the DMD gene 3'end in BMD and three novel splicing defects: IVS42: c.6118-3C>A, which causes a leaky splice-site; c.9560A>G, which determines a cryptic splice-site activation and c.9564-426 T>G, which creates pseudoexon retention within IVS65.
The analysis of our patients' sample, carrying point mutations or complex rearrangements in DMD gene, contributes to the knowledge on phenotypic correlations in dystrophinopatic patients and can provide a better understanding of pre-mRNA maturation defects and dystrophin functional domains. These data can have a prognostic relevance and can be useful in directing new therapeutic approaches, which rely on a precise definition of the genetic defects as well as their molecular consequences.
BMC Medical Genetics 03/2011; 12:37. · 2.33 Impact Factor
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Dario Ronchi,
Roberta Virgilio,
Andreina Bordoni,
Elisa Fassone,
Monica Sciacco, Patrizia Ciscato,
Maurizio Moggio,
Alessandra Govoni,
Stefania Corti,
Nereo Bresolin,
Giacomo P Comi
[show abstract]
[hide abstract]
ABSTRACT: Mitochondrial disorders are often associated with mutations in mitochondrial tRNA. Independent observation of the same molecular defect in unrelated subjects is a generally required proof of pathogenicity. A sporadic case of chronic external ophthalmoplegia (cPEO) with ragged red fibres (RRFs) has been previously related to an m.12316G>A substitution in tRNA(Leu(CUN)). Sequencing muscle-derived mtDNA, we found the m.12316G>A substitution in an adult woman with mitochondrial myopathy and respiratory impairment. Her muscle biopsy presented several cytochrome c oxidase-negative (COX-) fibres, and RRFs as signs of mitochondrial proliferation. Restriction-fragment length polymorphism (RFLP) analysis of the mutation in isolated muscle fibres showed a threshold of at least 60% of mutated mtDNA to determine a COX deficiency phenotype. This second report of the m.12316G>A mutation in a sporadic patient consolidates its pathogenic nature and provides further elements for genetic counselling.
Journal of the neurological sciences 02/2010; 292(1-2):107-10. · 2.32 Impact Factor
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Michela Guglieri,
Francesca Magri,
Maria Grazia D'Angelo,
Alessandro Prelle,
Lucia Morandi,
Carmelo Rodolico,
Rachele Cagliani,
Marina Mora,
Francesco Fortunato,
Andreina Bordoni, [......],
Mohammed Aguennouz, Patrizia Ciscato,
Claudia Di Blasi,
Alessandra Ruggieri,
Isabella Moroni,
Anna Turconi,
Antonio Toscano,
Maurizio Moggio,
Nereo Bresolin,
Giacomo P Comi
[show abstract]
[hide abstract]
ABSTRACT: Limb girdle muscular dystrophies (LGMD) are characterized by genetic and clinical heterogeneity: seven autosomal dominant and 12 autosomal recessive loci have so far been identified. Aims of this study were to evaluate the relative proportion of the different types of LGMD in 181 predominantly Italian LGMD patients (representing 155 independent families), to describe the clinical pattern of the different forms, and to identify possible correlations between genotype, phenotype, and protein expression levels, as prognostic factors. Based on protein data, the majority of probands (n=72) presented calpain-3 deficiency; other defects were as follows: dysferlin (n=31), sarcoglycans (n=32), alpha-dystroglycan (n=4), and caveolin-3 (n=2). Genetic analysis identified 111 different mutations, including 47 novel ones. LGMD relative frequency was as follows: LGMD1C (caveolin-3) 1.3%; LGMD2A (calpain-3) 28.4%; LGMD2B (dysferlin) 18.7%; LGMD2C (gamma-sarcoglycan) 4.5%; LGMD2D (alpha-sarcoglycan) 8.4%; LGMD2E (beta-sarcoglycan) 4.5%; LGMD2F (delta-sarcoglycan) 0.7%; LGMD2I (Fukutin-related protein) 6.4%; and undetermined 27.1%. Compared to Northern European populations, Italian patients are less likely to be affected with LGMD2I. The order of decreasing clinical severity was: sarcoglycanopathy, calpainopathy, dysferlinopathy, and caveolinopathy. LGMD2I patients showed both infantile noncongenital and mild late-onset presentations. Age at disease onset correlated with variability of genotype and protein levels in LGMD2B. Truncating mutations determined earlier onset than missense substitutions (20+/-5.1 years vs. 36.7+/-11.1 years; P=0.0037). Similarly, dysferlin absence was associated with an earlier onset when compared to partial deficiency (20.2+/-standard deviation [SD] 5.2 years vs. 28.4+/-SD 11.2 years; P=0.014).
Human Mutation 03/2008; 29(2):258-66. · 5.69 Impact Factor
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Costanza Lamperti,
Rachele Cagliani, Patrizia Ciscato,
Isabella Moroni,
Maurizio Viri,
Antonio Romeo,
Gigliola Fagiolari,
Alessandro Prelle,
Giacomo Pietro Comi,
Nereo Bresolin,
Maurizio Moggio
[show abstract]
[hide abstract]
ABSTRACT: Congenital muscular dystrophies (CMD) are autosomal recessive infantile disorders characterized by dystrophic changes at muscle biopsy and contractures. Central nervous system (CNS) abnormalities associated with mental retardation are often present. We describe a patient affected with muscle weakness, psychomotor developmental delay and normal brain MRI. Muscle biopsy showed complete absence of the alpha-dystroglycan (DG) glycosylated epitope and preservation of alpha-dystroglycan (alpha-DG) protein core. The analysis of FKRP, LARGE, POMT1 and POMGnT1 genes did not show any pathogenic mutations, suggesting that at least another gene may account for CMD with secondary glycosylated alpha-DG deficiency.
Journal of the Neurological Sciences 05/2006; 243(1-2):47-51. · 2.35 Impact Factor
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Davide Gabellini,
Giuseppe D'Antona,
Maurizio Moggio,
Alessandro Prelle,
Chiara Zecca,
Raffaella Adami,
Barbara Angeletti, Patrizia Ciscato,
Maria Antonietta Pellegrino,
Roberto Bottinelli,
Michael R Green,
Rossella Tupler
[show abstract]
[hide abstract]
ABSTRACT: Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder that is not due to a classical mutation within a protein-coding gene. Instead, almost all FSHD patients carry deletions of an integral number of tandem 3.3-kilobase repeat units, termed D4Z4, located on chromosome 4q35 (ref. 3). D4Z4 contains a transcriptional silencer whose deletion leads to inappropriate overexpression in FSHD skeletal muscle of 4q35 genes located upstream of D4Z4 (ref. 4). To identify the gene responsible for FSHD pathogenesis, we generated transgenic mice selectively overexpressing in skeletal muscle the 4q35 genes FRG1, FRG2 or ANT1. We find that FRG1 transgenic mice develop a muscular dystrophy with features characteristic of the human disease; by contrast, FRG2 and ANT1 transgenic mice seem normal. FRG1 is a nuclear protein and several lines of evidence suggest it is involved in pre-messenger RNA splicing. We find that in muscle of FRG1 transgenic mice and FSHD patients, specific pre-mRNAs undergo aberrant alternative splicing. Collectively, our results suggest that FSHD results from inappropriate overexpression of FRG1 in skeletal muscle, which leads to abnormal alternative splicing of specific pre-mRNAs.
Nature 03/2006; 439(7079):973-7. · 36.28 Impact Factor
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Monica Sciacco,
Alessandro Prelle,
Gigliola Fagiolari,
Andreina Bordoni,
Marco Crimi,
Alessio Di Fonzo, Patrizia Ciscato,
Costanza Lamperti,
Elisabetta D'Adda,
Stefano Jann,
Nereo Bresolin,
Giacomo P Comi,
Maurizio Moggio
[show abstract]
[hide abstract]
ABSTRACT: A 45-year-old male patient had an episode of acute renal failure with myoglobinuria, myalgias, weakness, and markedly increased serum CK levels. Similar episodes had occurred in the past. Carnitine palmitoyl-transferase II (CPT II) deficiency was documented both biochemically and genetically. Interestingly, muscle biopsy also showed some ragged red fibers (RRF) and complete mitochondrial DNA (mtDNA) sequence disclosed a homoplasmic T3394C point mutation. This mutation is described in Leber's hereditary optic neuropathy (LHON) or in patients with diabetes mellitus.
Journal of the Neurological Sciences 01/2006; 239(1):21-4. · 2.35 Impact Factor
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Simona Zanotti,
Tiziana Negri,
Cristina Cappelletti,
Pia Bernasconi,
Eleonora Canioni,
Claudia Di Blasi,
Elena Pegoraro,
Corrado Angelini, Patrizia Ciscato,
Alessandro Prelle,
Renato Mantegazza,
Lucia Morandi,
Marina Mora
[show abstract]
[hide abstract]
ABSTRACT: Biglycan and decorin are small extracellular proteoglycans that interact with cytokines, whose activity they may modulate, and with matrix proteins, particularly collagens. To better understand their role in muscle fibrosis, we investigated expression of decorin and biglycan transcripts and protein in muscle of several forms of muscular dystrophy, and also expression of perlecan, an extracellular proteoglycan unrelated to collagen deposition. In Duchenne muscular dystrophy (DMD) and LAMA2-mutated congenital muscular dystrophy (MDC1A) we also quantitated transcript levels of the profibrotic cytokine TGF-beta1. We examined muscle biopsies from nine DMD patients, aged 2-8 years; 14 BMD (Becker muscular dystrophy) patients (nine aged 1-5 years; five aged 30-37 years); four MDC1A patients (aged 2-7 years); six dysferlin-deficient patients (aged 19-53 years) with mutation ascertained in two, and normal expression of proteins related to limb girdle muscular dystrophies in the others; 10 sarcoglycan-deficient patients: seven with alpha-sarcoglycan mutation, two with beta-sarcoglycan mutation and one with gamma-sarcoglycan mutation (five aged 8-15 years; five aged 26-43 years); and nine children (aged 1-6 years) and 12 adults (aged 16-61 years) suspected of neuromuscular disease, but who had normal muscle on biopsy. Biglycan mRNA levels varied in DMD and MDC1A depending on the quantitation method, but were upregulated in BMD, sarcoglycanopathies and dysferlinopathy. Decorin mRNA was significantly downregulated in DMD and MDC1A, whereas TGF-beta1 was significantly upregulated. Decorin mRNA was normal in paediatric BMD, but upregulated in adult BMD, sarcoglycanopathies and dysferlinopathy. Perlecan transcript levels were similar to those of age-matched controls in all disease groups. By immunohistochemistry, decorin and biglycan were mainly localized in muscle connective tissue; their presence increased in relation to increased fibrosis in all dystrophic muscle. By visual inspection, decorin bands on immunoblot did not differ from those of age-matched controls in all patient groups. However, when the intensity of the bands was quantitated against vimentin and normalized against sarcomeric actin, in DMD and MDC1A the ratio of band intensities was significantly lower than in age-matched controls. Variations in the transcript and protein levels of these proteoglycans in different muscular dystrophies probably reflect the variable disruption of extracellular matrix organization that occurs in these diseases. The significantly lowered decorin levels in DMD and MDC1A may be related to the increased TGF-beta1 levels, suggesting a therapeutic role of decorin in these severe dystrophies.
Brain 12/2005; 128(Pt 11):2546-55. · 9.46 Impact Factor
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Rachele Cagliani,
Francesca Magri,
Antonio Toscano,
Luciano Merlini,
Francesco Fortunato,
Costanza Lamperti,
Carmelo Rodolico,
Alessandro Prelle,
Manuela Sironi,
Mohammed Aguennouz, Patrizia Ciscato,
Antonino Uncini,
Maurizio Moggio,
Nereo Bresolin,
Giacomo P Comi
[show abstract]
[hide abstract]
ABSTRACT: Mutations in the DYSF gene underlie two main muscle diseases: Limb Girdle Muscular Dystrophy (LGMD) 2B and Miyoshi myopathy (MM). Dysferlin is involved in muscle membrane-repair and is thought to interact with other dysferlin molecules and annexins A1 and A2 at the sarcolemma. We performed genotype/phenotype correlations in a large cohort of dysferlinopathic patients and explored the possible role of annexins as modifier factors in LGMD-2B and MM. In particular, clinical examination, expression of sarcolemmal proteins and genetic analysis were performed on 27 dysferlinopathic subjects. Expression of A1 and A2 annexins was investigated in LGMD-2B/MM subjects and in patients with other muscle disorders. We identified 24 different DYSF mutations, 10 of them being novel. We observed no clear correlation between mutation type and clinical phenotype, but MM patients were found to display muscle symptoms significantly earlier in life than LGMD subjects. Remarkably, dysferlinopathic patients and subjects suffering from other muscular disorders expressed higher levels of both annexins compared to controls; a significant correlation was observed between annexin expression levels and clinical severity scores. Also, annexin amounts paralleled the degree of muscle histopathologic changes. In conclusion, our data indicate that the pathogenesis of different inherited and acquired muscle disorders involves annexin overexpression, probably because these proteins actively participate in the plasmalemma repair process. The positive correlation between annexin A1 and A2 and clinical severity, as well as muscle histopathology, suggests that their level may be a prognostic indicator of disease.
Human Mutation 10/2005; 26(3):283. · 5.69 Impact Factor
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[show abstract]
[hide abstract]
ABSTRACT: Myotonic dystrophy is a multisystemic disorder, due to a CTG triplet expansion at the 3'UTR of the DM1 gene encoding for myotonic dystrophy protein kinase. Recent studies indicate that decreased DMPK levels could account for part of the symptoms suggesting a role of this protein in skeletal muscle differentiation. To investigate this aspect, polyclonal antibodies were raised against two peptides of the catalytic domain and against the human full-length DMPK (DMFL). In western blots, anti-hDMFL antibody was able to detect low amounts of purified human recombinant protein and recognized the splicing isoforms in heart and stomach of overexpressing mice. In human muscle extracts, this antibody specifically recognized a protein of apparent molecular weight of 85 kDa and it specifically stained neuromuscular junctions in skeletal muscle sections. In contrast, both anti-peptide antibodies demonstrated low specificity for either denatured or native DMPK, suggesting that these two epitopes are probably cryptic sites. Using anti-hDMFL, the expression and localization of DMPK was studied in human skeletal muscle cells (SkMC). Western blot analysis indicated that the antibody recognizes a main protein of apparent MW of 75 kDa, which appears to be expressed during differentiation into myotubes. Immunolocalization showed low levels of DMPK in the cytoplasm of undifferentiated cells; during differentiation the staining became more intense and was localized to the terminal part of the cells, suggesting that DMPK might have a role in cell elongation and fusion.
Cell Biology International 10/2005; 29(9):742-53. · 1.48 Impact Factor
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[show abstract]
[hide abstract]
ABSTRACT: Duchenne muscular dystrophy (DMD) and murine X-linked muscular dystrophy (mdx) are genetically homologous and both characterized by absence of dystrophin. The function of this protein is not defined nor is the pathogenesis of the severe muscle necrosis and progressive weakness found in DMD but not in mdx. Recently we found that anionic phospholipid (AP) calcium binding sites are lacking at the muscle cell surface in DMD and we correlated these data with dystrophin deficiency and muscle necrosis. In order to verify the role of AP lack in the pathogenesis of muscle necrosis in DMD we studied the ultrastructural localization of these Ca++ receptors in mdx muscle membrane showing that they are normally represented as they are in control mouse and normal human muscle. The absence of AP in DMD compared with a normal distribution in mdx suggests that these calcium binding site alterations play an important and specific role in muscle fiber necrosis. © 1994 John & Sons, Inc.
Muscle & Nerve 10/2004; 17(5):485 - 488. · 2.37 Impact Factor
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Alessandro Prelle,
Lucia Tancredi,
Monica Sciacco,
Luca Chiveri,
Giacomo P Comi,
Alessandro Battistel,
Paola Bazzi,
Filippo Martinelli Boneschi,
Vincenzo Bagnardi, Patrizia Ciscato,
Andreina Bordoni,
Franco Fortunato,
Sandra Strazzer,
Nereo Bresolin,
Guglielmo Scarlato,
Maurizio Moggio
[show abstract]
[hide abstract]
ABSTRACT: A retrospective evaluation of asymptomatic subjects with persistent elevation of serum creatine kinase (CK) levels (hyperCKemia) was made in order to verify the presence of subclinical myopathy or idiopathic hyperCKemia and to define the most appropriate diagnostic pathway. Persistently increased serum CK levels are occasionally encountered in healthy individuals. In 1980 Rowland coined for them the term idiopathic hyperCKemia. Despite the increase of scientific knowledge, several healthy subjects with hyperCKemia still represent a problem for the clinician. We made a retrospective evaluation of 114 asymptomatic or minimally symptomatic individuals with incidentally detected persistent hyperCKemia. They underwent neurological examination and laboratory/instrumental evaluation. Skeletal muscle biopsy was performed and thoroughly investigated. Biochemical and genetic investigations were added in selected cases. Logistic regression analysis was applied. We diagnosed a neuromuscular disorder in 21 patients (18.4%), and found, by muscle biopsy and/or EMG, pathological but not conclusive findings in 57 subjects (50%). The statistic correlation between elevated serum CK levels and the probability of making a diagnosis changed according to the age of the patient. Conclusions: Muscle biopsy is the basic tool for screening asymptomatic subjects with hyperCKemia. It allowed us to make a diagnosis of disease in 18.4% of patients, and to detect skeletal muscle abnormalities in 38.6% of the subjects. Interestingly, 31.6% of individuals had completely normal muscle findings. These best fit the "diagnosis" of idiopathic hyperCKemia.
Journal of Neurology 04/2002; 249(3):305-11. · 3.47 Impact Factor
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Monica Sciacco,
Alessandro Prelle,
Giacomo P. Comi,
Laura Napoli,
Alessandro Battistel,
Nereo Bresolin,
Lucia Tancredi,
Costanza Lamperti,
Adreina Bordoni,
Gigliola Fagiolari, [......],
Luca Chiveri,
Maria Paola Perini,
Francesco Fortunato,
Laura Adobbati,
Stefano Messina,
Antonio Toscano,
Filippo Martinelli-Boneschi,
Alex Papadimitriou,
Guglielmo Scarlato,
Maurizio Moggio
[show abstract]
[hide abstract]
ABSTRACT: Mitochondrial disorders are human genetic diseases with extremely variable clinical and genetic features. To better define
them, we made a genotype-phenotype correlation in a series of 207 affected patients, and we examined most of them with six
laboratory examinations (serum CK and basal lactate levels, EMG, cardiac and EEG studies, neuroradiology). We found that,
depending on the genetic abnormality, hyperckemia occurs most often with either chronic progressive external ophthalmoplegia
(CPEO) and ptosis or with limb weakness. Myopathic EMGs are more common than limb weakness, except in patients with A8344G
mutations. Peripheral neuropathy, when present, is always axonal. About 80 % of patients with A3243G and A8344G mutations
have high basal lactate levels, whereas pure CPEO is never associated with increased lactate levels. Cardiac abnormalities
mostly consist of conduction defectsAbnormalities on CT or MRI of the brain are relatively common in A3243G mutations independently
of the clinical phenotype. Patients with multiple mtDNA deletions are somehow “protected” against the development of abnormalities
with any of the tests. We conclude that, despite the phenotypic heterogeneity of mitochondrial disorders, correlation of clinical
features and laboratory findings may give the clinician important clues to the genetic defect, allowing earlier diagnosis
and counselling.
Journal of Neurology 08/2001; 248(9):778-788. · 3.47 Impact Factor
