Absolute Quantification of Dystrophin Protein in Human Muscle Biopsies using Parallel Reaction Monitoring (PRM)

AbstractThe need for a reliable and accurate method to quantify dystrophin proteins in human skeletal muscle biopsies has become crucial in order to assess the efficacy of dystrophin replacement therapies in Duchenne muscular dystrophy as well as to gain insight into the relationship between dystrophin levels and disease severity in Becker's muscular dystrophy. Current methods to measure dystrophin such as western blot and immunofluorescence, while straightforward and simple, lack precision and sometimes specificity. Here we standardized a targeted mass spectrometry method to determine the absolute amount of dystrophin in ng/mg of muscle using full length 13C6 ‐Arg, 13C6,15N2‐Lys‐labeled dystrophin and parallel reaction monitoring (PRM). The method was found to be reproducible with a limit of quantification as low as 30 pg of dystrophin protein per mg of total muscle proteins. The method was then tested to measure levels of dystrophin in muscle biop sies from a healthy donor and from Duchenne and Becker's muscular dystrophy patients.
Source: Journal of Mass Spectrometry - Category: Chemistry Authors: Tags: SPECIAL ISSUE ‐ RESEARCH ARTICLE Source Type: research

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The objective of this study was to assess respiratory muscle quality and function in DMD using magnetic resonance imaging and to determine the relationship to clinical respiratory function.MethodsIndividuals with DMD (n = 36) and unaffected controls (n = 12) participated in this cross sectional magnetic resonance imaging study. Participants underwent dynamic imaging of the thorax to assess diaphragm and chest wall mobility and chemical shift-encoded imaging of the chest and abdomen to determine fatty infiltration of the accessory respiratory muscles. Additionally, clinical pulmonary function mea...
Source: Journal of Neurology - Category: Neurology Source Type: research
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Source: Neuropediatrics - Category: Neurology Authors: Tags: Short Communication Source Type: research
by Luca Madaro, Alessio Torcinaro, Marco De Bardi, Federica F. Contino, Mattia Pelizzola, Giuseppe R. Diaferia, Giulia Imeneo, Marina Bouch è, Pier Lorenzo Puri, Francesca De Santa Satellite cells (SCs) are muscle stem cells that remain quiescent during homeostasis and are activated in response to acute muscle damage or in chronic degenerative conditions such as Duchenne Muscular Dystrophy. The activity of SCs is supported by specialized cells which either reside in the musc le or are recruited in regenerating skeletal muscles, such as for instance macrophages (MΦs). By using a dystrophic mouse model of transie...
Source: PLoS Genetics - Category: Genetics & Stem Cells Authors: Source Type: research
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Source: Journal of Molecular and Cellular Cardiology - Category: Cytology Authors: Source Type: research
Duchenne muscular dystrophy (DMD) is a progressive neuromuscular disease resulting in severe respiratory derangements. As such, DMD patients are at a high risk of nocturnal hypoventilation, thereby requiring n...
Source: Respiratory Research - Category: Respiratory Medicine Authors: Tags: Research Source Type: research
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Source: Cell Stress and Chaperones - Category: Cytology Authors: Tags: Cell Stress Chaperones Source Type: research
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Source: Cochrane Database of Systematic Reviews - Category: General Medicine Authors: Tags: Cochrane Database Syst Rev Source Type: research
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Source: GEO: Gene Expression Omnibus - Category: Genetics & Stem Cells Tags: Expression profiling by high throughput sequencing Homo sapiens Source Type: research
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Source: Journal of the American Society of Echocardiography - Category: Cardiology Authors: Source Type: research
Conditions:   Duchenne Muscular Dystrophy;   Pompe Disease (Late-onset) Intervention:   Genetic: Laboratory Tests Sponsor:   Turkish Society of Pediatric Gastroenterology, Hepatology and Nutrition Recruiting
Source: ClinicalTrials.gov - Category: Research Source Type: clinical trials
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