RESUMEN
BACKGROUND: Autosomal recessive limb-girdle muscular dystrophies (LGMD2) include a number of disorders with heterogeneous etiology that cause predominantly weakness and wasting of the shoulder and pelvic girdle muscles. In this study, we determined the frequency of LGMD subtypes within a cohort of Czech LGMD2 patients using mutational analysis of the CAPN3, FKRP, SGCA, and ANO5 genes. METHODS: PCR-sequencing analysis; sequence capture and targeted resequencing. RESULTS: Mutations of the CAPN3 gene are the most common cause of LGMD2, and mutations in this gene were identified in 71 patients in a set of 218 Czech probands with a suspicion of LGMD2. Totally, we detected 37 different mutations of which 12 have been described only in Czech LGMD2A patients. The mutation c.550delA is the most frequent among our LGMD2A probands and was detected in 47.1% of CAPN3 mutant alleles. The frequency of particular forms of LGMD2 was 32.6% for LGMD2A (71 probands), 4.1% for LGMD2I (9 probands), 2.8% for LGMD2D (6 probands), and 1.4% for LGMD2L (3 probands).Further, we present the first results of a new approach established in the Czech Republic for diagnosis of neuromuscular diseases: sequence capture and targeted resequencing. Using this approach, we identified patients with mutations in the DYSF and SGCB genes. CONCLUSIONS: We characterised a cohort of Czech LGMD2 patients on the basis of mutation analysis of genes associated with the most common forms of LGMD2 in the European population and subsequently compared the occurrence of particular forms of LGMD2 among countries on the basis of our results and published studies.
Asunto(s)
Calpaína/genética , Canales de Cloruro/genética , Proteínas Musculares/genética , Distrofia Muscular de Cinturas/genética , Proteínas/genética , Sarcoglicanos/genética , Anoctaminas , República Checa , Análisis Mutacional de ADN , Genotipo , Humanos , Pentosiltransferasa , Reacción en Cadena de la PolimerasaRESUMEN
Dystroglycanopathies are a clinically and genetically diverse group of recessively inherited conditions ranging from the most severe of the congenital muscular dystrophies, Walker-Warburg syndrome, to mild forms of adult-onset limb-girdle muscular dystrophy. Their hallmark is a reduction in the functional glycosylation of α-dystroglycan, which can be detected in muscle biopsies. An important part of this glycosylation is a unique O-mannosylation, essential for the interaction of α-dystroglycan with extracellular matrix proteins such as laminin-α2. Mutations in eight genes coding for proteins in the glycosylation pathway are responsible for â¼50% of dystroglycanopathy cases. Despite multiple efforts using traditional positional cloning, the causative genes for unsolved dystroglycanopathy cases have escaped discovery for several years. In a recent collaborative study, we discovered that loss-of-function recessive mutations in a novel gene, called isoprenoid synthase domain containing (ISPD), are a relatively common cause of Walker-Warburg syndrome. In this article, we report the involvement of the ISPD gene in milder dystroglycanopathy phenotypes ranging from congenital muscular dystrophy to limb-girdle muscular dystrophy and identified allelic ISPD variants in nine cases belonging to seven families. In two ambulant cases, there was evidence of structural brain involvement, whereas in seven, the clinical manifestation was restricted to a dystrophic skeletal muscle phenotype. Although the function of ISPD in mammals is not yet known, mutations in this gene clearly lead to a reduction in the functional glycosylation of α-dystroglycan, which not only causes the severe Walker-Warburg syndrome but is also a common cause of the milder forms of dystroglycanopathy.
Asunto(s)
Distrofias Musculares/congénito , Distrofias Musculares/genética , Mutación , Nucleotidiltransferasas/genética , Adolescente , Niño , Preescolar , Distroglicanos/genética , Distroglicanos/metabolismo , Femenino , Glicosilación , Humanos , Imagen por Resonancia Magnética , Masculino , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Distrofias Musculares/metabolismo , Distrofias Musculares/patología , Distrofia Muscular de Cinturas/genética , Distrofia Muscular de Cinturas/metabolismo , Distrofia Muscular de Cinturas/patología , Adulto JovenRESUMEN
Hereditary sensory and autonomic neuropathy type I (HSAN-I) is an axonal peripheral neuropathy associated with progressive distal sensory loss and severe ulcerations. Mutations in the first subunit of the enzyme serine palmitoyltransferase (SPT) have been associated with HSAN-I. The SPT enzyme catalyzes the first and rate-limiting step in the de novo sphingolipid synthesis pathway. However, different studies suggest the implication of other genes in the pathology of HSAN-I. Therefore, we screened the two other known subunits of SPT, SPTLC2 and SPTLC3, in a cohort of 78 HSAN patients. No mutations were found in SPTLC3, but we identified three heterozygous missense mutations in the SPTLC2 subunit of SPT in four families presenting with a typical HSAN-I phenotype. We demonstrate that these mutations result in a partial to complete loss of SPT activity in vitro and in vivo. Moreover, they cause the accumulation of the atypical and neurotoxic sphingoid metabolite 1-deoxy-sphinganine. Our findings extend the genetic heterogeneity in HSAN-I and enlarge the group of HSAN neuropathies associated with SPT defects. We further show that HSAN-I is consistently associated with an increased formation of the neurotoxic 1-deoxysphinganine, suggesting a common pathomechanism for HSAN-I.
Asunto(s)
Neuropatías Hereditarias Sensoriales y Autónomas/genética , Serina C-Palmitoiltransferasa/genética , Secuencia de Bases , Línea Celular , Clonación Molecular , Estudios de Cohortes , Análisis Mutacional de ADN , Cartilla de ADN/genética , Prueba de Complementación Genética , Humanos , Repeticiones de Microsatélite/genética , Datos de Secuencia Molecular , Mutación Missense/genética , Serina C-Palmitoiltransferasa/metabolismoRESUMEN
Toxoplasmosis is a rare opportunistic protozoal infection, which may occur in patients after hematopoietic stem cell transplantation. This disease originates almost exclusively from reactivation of latent infection in seropositive recipients. We present a case report of one patient with diagnosis of acute myeloid leukemia undergoing two allogeneic stem cell transplantations at two years interval. The second transplantation was complicated by the development of the toxoplasmic encephalitis in early posttransplant course. The initial neurological symptoms included diplopia caused by the paresis of right side motor branches of the 3rd and 6th cranial nerves due to a compressive lesion in basal ganglia. Patient suddenly deteriorated after an epileptic seizure followed by a loss of consciousness, bilateral ptosis and right side mydriasis. Prolonged sopor and bilateral mydriasis appeared because of the further lesion progression in basal ganglia and compression of the 3rd cranial nerve. After targeted therapy of Toxoplasma gondii the patient's clinical status improved and she regained consciousness. Unfortunately, examination of bone marrow later revealed the relapse of leukemia. We compared risk factors of the latent reactivation of infection in immunocompromised patients with published data. It is of interest that the toxoplasmosis of the brain developed in this patient after the second transplantation.
Asunto(s)
Trasplante de Células Madre/efectos adversos , Toxoplasmosis Cerebral/etiología , Adulto , Femenino , Humanos , Huésped Inmunocomprometido , Leucemia Mieloide Aguda/terapiaRESUMEN
Duchenne muscular dystrophy (DMD) affects 1:3500-5000 newborn boys and manifests with progressive skeletal muscle wasting, respiratory failure and eventual heart failure. Symptoms show different onset from patients' childhood to the second decade of age. We reprogrammed fibroblasts from two independent DMD patients with a complete loss of dystrophin expression, carrying deletions of exons 45-50 and 48-50. The resulting hiPSCs show expression of pluripotency markers (NANOG, OCT4, SSEA4), differentiation capacity into all three germ layers, normal karyotype, genetic identity to the originating parental fibroblasts and the patient-specific dystrophin mutation.
Asunto(s)
Línea Celular/citología , Células Madre Pluripotentes Inducidas/citología , Distrofia Muscular de Duchenne/fisiopatología , Adolescente , Diferenciación Celular , Línea Celular/metabolismo , Niño , Distrofina/genética , Distrofina/metabolismo , Exones , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Masculino , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/metabolismo , Factor 3 de Transcripción de Unión a Octámeros/genética , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , Eliminación de SecuenciaRESUMEN
Spinal muscular atrophy (SMA) is caused by homozygous deletion of the SMN1 gene in approximately 96% of cases. Four percent of SMA patients have a combination of the deletion or conversion on one allele and an intragenic mutation on the second one. We performed analysis of point mutations in a set of our patients with suspicion of SMA and without homozygous deletion of the SMN1 gene. A quantitative test determining SMN1 copy number (using real-time PCR and/or MLPA analysis) was performed in 301 patients and only 1 SMN1 copy was detected in 14 of them. When these 14 patients were screened for the presence of point mutations we identified 6 mutations, p.Y272C (in three patients) and p.T274I, p.I33IfsX6, and p.A188S (each in one case). The mutations p.I33IfsX6 and p.A188S were found in two SMAI patients and were not detected previously. Further, evaluation of the relationship between mutation type, copy number of the SMN2 gene and clinical findings was performed. Among our SMA patients with a SMN1 homozygous deletion, we found a family with two patients: the son with SMAII possesses 3 SMN2 copies and the nearly asymptomatic father has a homozygous deletion of SMN1 exon 7 and carries 4 SMN2 copies. Generally, our results illustrate that an increased SMN2 gene copy number is associated with a milder SMA phenotype.
Asunto(s)
Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Dosificación de Gen , Atrofia Muscular Espinal/genética , Proteínas del Tejido Nervioso/genética , Mutación Puntual/genética , Proteínas de Unión al ARN/genética , Adolescente , Adulto , Preescolar , ADN/genética , Exones/genética , Eliminación de Gen , Homocigoto , Humanos , Técnicas de Amplificación de Ácido Nucleico , Fenotipo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas del Complejo SMN , Proteína 1 para la Supervivencia de la Neurona Motora , Proteína 2 para la Supervivencia de la Neurona MotoraRESUMEN
Limb girdle muscular dystrophy type 2A (LGMD2A) is caused by single or small nucleotide changes widespread along the CAPN3 gene, which encodes the muscle-specific proteolytic enzyme calpain-3. About 356 unique allelic variants of CAPN3 have been identified to date. We performed analysis of the CAPN3 gene in LGMD2A patients at both the mRNA level using reverse transcription-PCR, and at the DNA level using PCR and denaturing high performance liquid chromatography. In four patients, we detected homozygous occurrence of a missense mutation or an in-frame deletion at the mRNA level although the DNA was heterozygous for this mutation in conjunction with a frame-shift mutation. The relationship observed in 12 patients between the quantity of CAPN3 mRNA, determined using real-time PCR, and the genotype leads us to propose that CAPN3 mRNAs which contain frame-shift mutations are degraded by nonsense-mediated mRNA decay. Our results illustrate the importance of DNA analysis for reliable establishment of mutation status, and provide a new insight into the process of mRNA decay in cells of LGMD2A patients.
Asunto(s)
Calpaína/genética , Codón sin Sentido/genética , Proteínas Musculares/genética , Distrofia Muscular de Cinturas/genética , ARN Mensajero/genética , Adolescente , Adulto , Western Blotting , Niño , Preescolar , ADN/biosíntesis , ADN/genética , Femenino , Mutación del Sistema de Lectura/genética , Eliminación de Gen , Genotipo , Humanos , Lactante , Intrones/genética , Masculino , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
We report a sporadic case of congenital muscular dystrophy (CMD) in a 13-year-old girl with early manifestation of muscle weakness and hypotonia, severe contractures, bulbar syndrome, progressive external ophtalmoplegia, and white matter changes on magnetic resonance imaging (MRI) of the brain, but no mental defect. Serum creatine kinase (CK) level was normal. Muscle biopsy revealed a dystrophic picture with a prominent inflammatory infiltrate mimicking inflammatory myopathy-typical histological findings in CMD. Immunostaining showed normal expression of merosin, alpha and beta-dystroglycans. Mutation analyses of calpain3, dysferlin, and SEPN1 genes were negative. An electron microscopy revealed the accumulation of abnormally enlarged mitochondria located under the sarcolemma. Measurement of respiratory chain enzyme activities did not reveal any biochemical defect and mitochondrial genetic studies, including sequencing of the entire mitochondrial genome, were unremarkable. Phenotypic presentation of our patient is very unusual and differs considerably from other CMD variants.
Asunto(s)
Encéfalo/patología , Creatina Quinasa/sangre , Distrofia Miotónica , Neuroglía/patología , Oftalmoplejía/etiología , Adolescente , Femenino , Humanos , Imagen por Resonancia Magnética/métodos , Distrofia Miotónica/sangre , Distrofia Miotónica/complicaciones , Distrofia Miotónica/patologíaRESUMEN
A new multi-pin probe head was installed and tested at the COMPASS tokamak. The probe head consists of several ball-pen and Langmuir probes in similar geometric configurations, which enables fast, simultaneous measurements of the radial and poloidal electric field components from differences of plasma or floating potentials using appropriately positioned ball-pen or Langmuir probes, respectively. The fast measurements of the radial electric field are compared with other methods of measuring selected frequency components of the radial electric field. The radial profiles of the Reynolds stress calculated from correlations of electric field fluctuations obtained by either probe type are compared along with their spectral composition. Lower Reynolds stress values are found for Langmuir probes in comparison to ball-pen probes due to negative contributions from higher frequency fluctuations possibly associated with electron temperature fluctuations.
RESUMEN
BACKGROUND: Neuromuscular weakness in paediatric patients with sepsis and multiple organ dysfunction is increasingly reported. However, many aspects of neuromuscular involvement in critically ill children are not completely understood. As more patients survive the critical illness, an understanding of the long-term outcomes of this condition is needed. AIMS: To describe clinical and electrophysiological features and evaluate the long-term outcomes in critically ill paediatric patients with neuromuscular complications. METHODS: A case series of five critically ill children was observed prospectively for a 1-month period. Selected clinical and laboratory parameters were evaluated. Electrophysiological studies were performed during the first week and then 1 month later in order to detect signs of critical illness polyneuromyopathy (CIPM). Patients with neuromuscular involvement completed a 1-year follow-up. RESULTS: Electrophysiological abnormalities were detected in two patients. Flaccid quadriplegia was a clinical presentation. Both children had electromyographic evidence of chronic partial denervation at follow-up, findings indicative of a preceding axonal neuropathy. Marked but incomplete recovery within 1 year after the onset of the disease occured in both patients. With a mild residual functional handicap the health-related quality of life was not significantly impaired (Barthel Index > 80). CONCLUSIONS: In both our patients with CIPM, the long-term clinical outcome is markedly better than we expected when electromyography in the 1-year follow-up demonstrated a persistent severe chronic partial denervation. These findings can have important implications for the management and rehabilitation of paediatric intensive care survivors.
Asunto(s)
Enfermedades Musculares/terapia , Enfermedades del Sistema Nervioso Periférico/terapia , Adolescente , Niño , Cuidados Críticos , Enfermedad Crítica , Estimulación Eléctrica , Electromiografía , Electrofisiología , Femenino , Estudios de Seguimiento , Humanos , Masculino , Músculo Esquelético/fisiopatología , Enfermedades Musculares/fisiopatología , Examen Neurológico , Enfermedades del Sistema Nervioso Periférico/fisiopatología , Pronóstico , Estudios Prospectivos , Cuadriplejía/etiología , Respiración Artificial , Resultado del TratamientoRESUMEN
BACKGROUND: Publication of comprehensive clinical care guidelines for Duchenne muscular dystrophy (DMD) in 2010 was a milestone for DMD patient management. Our CARE-NMD survey investigates the neuromuscular, medical, and psychosocial care of DMD patients in Europe, and compares it to the guidelines. METHODS: A cross-sectional survey of 1677 patients contacted via the TREAT-NMD patient registries was conducted using self-report questionnaires in seven European countries. RESULTS: Survey respondents were 861 children and 201 adults. Data describe a European DMD population with mean age of 13.0 years (range 0.8-46.2) of whom 53% had lost ambulation (at 10.3 years of age, median). Corticosteroid medication raised the median age for ambulatory loss from 10.1 years in patients never medicated to 11.4 years in patients who received steroids (pâ<â0.0001). The majority of patients reported receiving care in line with guidelines, although we identified significant differences between countries and important shortcomings in prevention and treatment. Summarised, 35% of patients aged≥ nine years received no corticosteroid medication, 24% of all patients received no regular physiotherapy, echocardiograms were not performed regularly in 22% of patients, pulmonary function was not regularly assessed in 71% of non-ambulatory patients. Patients with regular follow-up by neuromuscular specialists were more likely to receive care according to guidelines, were better satisfied, and experienced shorter unplanned hospitalization periods.
Asunto(s)
Corticoesteroides/uso terapéutico , Adhesión a Directriz , Distrofia Muscular de Duchenne/terapia , Modalidades de Fisioterapia/estadística & datos numéricos , Pautas de la Práctica en Medicina , Sistema de Registros , Adolescente , Adulto , Factores de Edad , Niño , Preescolar , Estudios Transversales , Ecocardiografía/estadística & datos numéricos , Europa (Continente) , Humanos , Lactante , Masculino , Persona de Mediana Edad , Distrofia Muscular de Duchenne/fisiopatología , Guías de Práctica Clínica como Asunto , Pruebas de Función Respiratoria/estadística & datos numéricos , Nivel de Atención , Encuestas y Cuestionarios , Adulto JovenRESUMEN
Calpain3 (CAPN3, p94) is a muscle-specific nonlysosomal cysteine proteinase. Loss of proteolytic function or change of other properties of this enzyme (such as stability or ability to interact with other muscular proteins) is manifested as limb girdle muscular dystrophy type 2A (LGMD2A, calpainopathy). These pathological changes in properties of calpain3 are caused by mutations in the calpain3 gene. The fact that the human gene for calpain3 is quite long led us to analyse its coding sequence by reverse transcription-PCR followed by sequence analysis. This study reports nine mutations that we found by analysing mRNA of seven unrelated LGMD patients in the Czech Republic. Three of these mutations were novel, not described on the Leiden muscular dystrophy pages so far. Further, we observed a reduction of dysferlin in muscle membrane in five of our seven LGMD2A patients by immunohistochemical analysis of muscle sections.
Asunto(s)
Calpaína/genética , Isoenzimas/genética , Proteínas Musculares/genética , Distrofia Muscular de Cinturas/genética , Mutación , Fenotipo , Adolescente , Adulto , Anciano , Arginina/genética , Western Blotting/métodos , Calpaína/metabolismo , Niño , República Checa , Análisis Mutacional de ADN/métodos , Disferlina , Exones/genética , Femenino , Humanos , Inmunohistoquímica/métodos , Isoenzimas/metabolismo , Masculino , Proteínas de la Membrana/metabolismo , Persona de Mediana Edad , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Distrofia Muscular de Cinturas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Triptófano/genéticaRESUMEN
OBJECTIVE: To evaluate the spectrum and time profile of electrophysiological parameters in the detection of neuromuscular involvement in critically ill patients and establish their correlation with biopsy findings. DESIGN: Prospective clinical and neurophysiological study. SETTING: One general and one neurological intensive care unit in a university hospital. PATIENTS: Forty-six critically ill patients with failure of at least two organ systems were enrolled and completed the 1-month follow up. INTERVENTIONS: Detailed clinical and electrophysiological evaluation including direct muscle stimulation was performed in all cases on entry and at the end of the follow-up. Muscle biopsy was performed in 11, and sural nerve biopsy in 5, cases. MEASUREMENTS AND RESULTS: Electrophysiological signs of new or progressing neuromuscular involvement at the end of the first month were detected in 26 patients (56%) and could be classified into three groups: "pure motor syndrome" (12 cases), combined motor syndrome and sensory polyneuropathy (13 cases) and isolated sensory polyneuropathy (1 case). Direct muscle stimulation showed decreased muscle membrane excitability in 11 of these abnormal cases. Muscle biopsy disclosed various myopathic abnormalities in all 11 cases examined with motor syndrome, in 7 of them in association with denervation/re-innervation changes. CONCLUSIONS: Electrophysiological and histological examinations showed significant overlapping of several pathogenic components of neuromuscular involvement in critically ill patients, namely decreased muscle excitability, myopathy, axonal motor neuropathy and sensory neuropathy. The characterisation of the electrophysiological components of a complex polyneuromyopathy is preferred to the strict categorisation of abnormalities into critical illness myopathy and polyneuropathy.
Asunto(s)
Electromiografía , Polineuropatías/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Humanos , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Insuficiencia Multiorgánica/complicaciones , Insuficiencia Multiorgánica/diagnóstico , Músculo Esquelético/patología , Enfermedades Musculares/complicaciones , Enfermedades Musculares/diagnóstico , Polineuropatías/complicaciones , Polineuropatías/patología , Estudios Prospectivos , Nervio Sural/patologíaRESUMEN
BACKGROUND: Spinal muscular atrophy is a rare hereditary neuromuscular disorder (with a prevalence of 1 per 30,000) that greatly debilitates patients and, in most cases, shortens their life expectancy. Although there is no causal therapy, improvements in symptomatic therapy have extended patients' life expectancy and increased their quality of life. Unfortunately, the advancements in care vary from country to country. To improve the care for children with spinal muscular atrophy in the Czech Republic, we created a survey to obtain the baseline information about their quality of life and compared the data with equivalent data from the United States. METHODS: We used the Pediatric Quality of Life Inventory 3.0 Neuromuscular Measurement Model, which is a health-related quality of life questionnaire specific to children with neuromuscular disorders. The survey was conducted on 35 children with genetically proven spinal muscular atrophy and their parents. RESULTS: Compared with the US data, the Czech data generally show a lower quality of life, mainly in the family resources part. The greatest score was achieved in the section about communication. Altogether, the parents' scores are lower than those of the children. CONCLUSION: In the Czech Republic, patients with spinal muscular atrophy and, especially their parents, have a significantly lower quality of life compared with US patients, mostly because of economic factors and a lack of social support. Our results reveal areas toward which improvement should be directed. The need for family support through social care as well as civic, patient, or organizational support is accentuated.
Asunto(s)
Atrofia Muscular Espinal/psicología , Calidad de Vida , Adolescente , Niño , Preescolar , República Checa , Femenino , Humanos , Masculino , Atrofia Muscular Espinal/economía , Padres/psicología , Apoyo Social , Encuestas y Cuestionarios , Estados UnidosRESUMEN
Myotonia congenita (MC) is a genetic disease caused by mutations in the skeletal muscle chloride channel gene (CLCN1) encoding the skeletal muscle chloride channel (ClC-1). Mutations of CLCN1 result in either autosomal dominant MC (Thomsen disease) or autosomal recessive MC (Becker disease). The ClC-1 protein is a homodimer with a separate ion pore within each monomer. Mutations causing recessive myotonia most likely affect properties of only the mutant monomer in the heterodimer, leaving the wild type monomer unaffected, while mutations causing dominant myotonia affect properties of both subunits in the heterodimer. Our study addresses two points: 1) molecular genetic diagnostics of MC by analysis of the CLCN1 gene and 2) structural analysis of mutations in the homology model of the human dimeric ClC-1 protein. In the first part, 34 different types of CLCN1 mutations were identified in 51 MC probands (14 mutations were new). In the second part, on the basis of the homology model we identified the amino acids which forming the dimer interface and those which form the Cl(-) ion pathway. In the literature, we searched for mutations of these amino acids for which functional analyses were performed to assess the correlation between localisation of a mutation and occurrence of a dominant-negative effect (corresponding to dominant MC). This revealed that both types of mutations, with and without a dominant-negative effect, are localised at the dimer interface while solely mutations without a dominant-negative effect occur inside the chloride channel. This work is complemented by structural analysis of the homology model which provides elucidation of the effects of mutations, including a description of impacts of newly detected missense mutations.
Asunto(s)
Canales de Cloruro/química , Canales de Cloruro/genética , Músculo Esquelético/metabolismo , Mutación , Miotonía Congénita/genética , Adolescente , Adulto , Canales de Cloruro/metabolismo , República Checa , Femenino , Humanos , Masculino , Modelos Moleculares , Mutación Missense , Miotonía Congénita/diagnóstico , Fenotipo , Conformación Proteica , Multimerización de Proteína , Adulto JovenRESUMEN
Effective planning of clinical trials requires an appropriate number of patients who fulfil given inclusion criteria. In the case of so called "orphan" diseases, such as Duchenne and Becker muscular dystrophy (DMD/BMD), the number of suitable patients within one country is usually limited. We developed a detailed registry of Czech and Slovak DMD/BMD patients which may contribute to cooperation on the European level. The registry uses internet and database technologies with a multilevel architecture. Patients may view their own data. As of May 2008, 163 patients have been registered in the database. The registry provides a detailed phenotypic and genotypic description of patients. The main purpose of such a registry is the time-effective recruitment of eligible patients for a clinical trial or therapy and may allow the anticipation of possible future effects of appropriate therapy on individual patients. The importance of the DMD/BMD patient registries has recently also been rising with new clinical trials focused on mutation-specific approaches. Other outputs include assessment of epidemiology, phenotype and genotype relationships, or standards of care.
Asunto(s)
Bases de Datos como Asunto/organización & administración , Distrofia Muscular de Duchenne/epidemiología , Sistema de Registros/estadística & datos numéricos , Niño , Ensayos Clínicos como Asunto/métodos , Ensayos Clínicos como Asunto/tendencias , República Checa/epidemiología , Análisis Mutacional de ADN/métodos , Análisis Mutacional de ADN/tendencias , Recolección de Datos/métodos , Bases de Datos como Asunto/tendencias , Femenino , Tamización de Portadores Genéticos/métodos , Predisposición Genética a la Enfermedad/genética , Pruebas Genéticas/métodos , Pruebas Genéticas/tendencias , Humanos , Masculino , Proteínas Musculares/genética , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/terapia , Mutación/genética , Selección de Paciente , Eslovaquia/epidemiologíaRESUMEN
OBJECTIVE: To evaluate the safety and efficacy of pregabalin in the management of chemotherapy-induced neuropathic pain in patients with childhood solid tumors and leukaemia. MATERIALS AND METHODS: In an open-label study, 30 children (11 boys and 19 girls; mean age 13.5 years) who were treated for solid tumors and leukaemia, and developed a painful peripheral neuropathy, were medicated with pregabalin in the daily dose of 150-300 mg for 8 weeks. RESULTS: Twenty-eight patients completed the 8-week follow-up. A significant and long-lasting pain relief was noted in 86% of these patients. Median VAS score decreased by 59% at the 8th week from baseline. Adverse effects were infrequent and transient. CONCLUSION: The treatment with pregabalin resulted in a significant improvement in pain symptoms. The use of pregabalin in children is off-label so far. However, this drug seems to be a safe and effective remedy, which could significantly broaden the therapeutic spectrum in paediatric oncological patients suffering from neuropathic pain.
Asunto(s)
Analgésicos/uso terapéutico , Antineoplásicos/efectos adversos , Neuralgia/inducido químicamente , Neuralgia/tratamiento farmacológico , Ácido gamma-Aminobutírico/análogos & derivados , Adolescente , Antineoplásicos/uso terapéutico , Niño , Femenino , Estudios de Seguimiento , Humanos , Masculino , Neoplasias/tratamiento farmacológico , Dimensión del Dolor , Pregabalina , Ácido gamma-Aminobutírico/uso terapéuticoRESUMEN
Duchenne and Becker muscular dystrophies (DMD/BMD) are associated with mutations in the DMD gene. We determined the mutation status of 47 patients with dystrophinopathy without deletion or duplication in the DMD gene by screening performed by reverse transcription-PCR, protein truncation test, and DNA sequencing. We describe three patients with a mutation creating a premature termination codon (p.E55X, p.E1110X, and p.S3497PfsX2) but with a mild phenotype, which present three different ways of rescuing the DMD phenotype. In one patient we detected the insertion of a repetitive sequence AluYa5 in intron 56, which led to skipping of exon 57. Further, using quantitative analysis of DMD mRNA carrying various mutated alleles, we examine levels of mRNA degradation due to nonsense mediated mRNA decay. The quantity of dystrophin mRNA is different depending on the presence of a mutation leading to a premature termination codon, and position of the analysed mRNA region with respect to its 5' end or 3' end. Average relative amounts of DMD mRNAs carrying a premature termination codon is 48% and 17%, when using primers amplifying the 5' and 3' cDNA regions, respectively.
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Distrofina/genética , Distrofia Muscular de Duchenne/genética , Fenotipo , Mutación Puntual , Secuencia de Bases , República Checa , Análisis Mutacional de ADN/métodos , Distrofina/metabolismo , Exones , Femenino , Humanos , Masculino , Datos de Secuencia Molecular , ARN Mensajero/metabolismoAsunto(s)
Enfermedades de los Caballos/patología , Músculo Esquelético/patología , Distrofia Miotónica/veterinaria , Animales , Biopsia con Aguja/veterinaria , Electromiografía/veterinaria , Femenino , Enfermedades de los Caballos/diagnóstico , Enfermedades de los Caballos/etiología , Caballos , Microscopía Electrónica/veterinaria , Músculo Esquelético/ultraestructura , Distrofia Miotónica/diagnóstico , Distrofia Miotónica/etiología , Distrofia Miotónica/patologíaRESUMEN
Limb-girdle muscular dystrophy type 2A (LGMD2A) is an autosomal-recessive disorder characterized by selective atrophy and progressive weakness of proximal girdle muscles. LGMD2A, the most prevalent form of LGMD, is caused by mutations in the CAPN3 gene that encodes the skeletal muscle-specific member of the calpain family, calpain-3 (p 94). We examined the histopathologic and molecular pathologic findings in 14 Czech LGMD2A patients. Analysis of the CAPN3 gene was performed at the mRNA level, using reverse transcription-polymerase chain reaction (RT-PCR) and sequencing, and/or DNA level, using PCR and denaturing high-performance liquid chromatography (DHPLC). Our results confirm that mutation 550 delA is the most frequent CAPN3 defect in Czech LGMD2A patients (9 alleles of 28). Furthermore, we established that, in a patient with the 550 delA/R490W genotype, mRNA carrying frameshift mutation 550 delA was not detected, probably due to its degradation by nonsense-mediated mRNA decay. In muscle biopsies of two LGMD2A patients, a neurogenic pattern simulating a neurogenic lesion was observed. Immunoblot analysis revealed the deficiency of p 94 in all genetically confirmed cases of LGMD2A, and secondary dysferlin deficiency was demonstrated on muscle membranes in 6 patients using immunofluorescence. Thus, we find a combination of DNA and mRNA mutational analysis to be useful in the diagnosis of LGMD2A. Moreover, our study expands the spectrum of calpainopathies to cases that simulate a neurogenic lesion in muscle biopsies, and the knowledge of possible secondary deficiencies of muscular proteins also contributes to a diagnosis of LGMD2A.