Title: (R,S)-salbutamol plasma concentrations in severe asthma Authors: Jacobson, G. A. Chong, F. V. Wood-Baker, R. Auth Address: School of Pharmacy, University of Tasmania, GPO BOX 252-26, Hobart, Tasmania 7001, Australia. glenn.jacobson@utas.edu.au Pages: 235-8 Volume: 28 Number: 3 Keywords: Administration, Inhalation Adult Albuterol/*blood/chemistry/therapeutic use Asthma/blood/*drug therapy Bronchodilator Agents/*blood/chemistry/therapeutic use Dose-Response Relationship, Drug Female Humans Linear Models Male Pilot Projects Status Asthmaticus/*blood Stereoisomerism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12795782 Abstract: OBJECTIVE: Salbutamol is commonly delivered as a racemic mixture of pharmacologically active (R)-salbutamol and inactive (S)-salbutamol. This study investigated inactive (S)- and active (R)-salbutamol plasma levels and their relationship to dose in patients with severe asthma. METHODS: Basic demographics, racemic-salbutamol dose in the preceding 24 h, lung function tests at baseline and 1 h, and a 10 mL plasma sample were obtained from subjects presenting to the Department of Emergency Medicine with acute asthma. Plasma determinations were carried out using an LC-MS assay with solid phase extraction. RESULTS: All patients (n = 5) had detectable levels of drug in plasma with range of 0.9-7.7 and 4.7-27.4 ng/mL for (R)-salbutamol and (S)-salbutamol respectively. These were correlated to total racemic salbutamol dose. The range of the (S) : (R) ratio was 2.0-5.2, with (R)-salbutamol representing 16-33% of the total plasma concentration, which did not correlate with total salbutamol dose. CONCLUSION: Only a small fraction of total plasma salbutamol concentration was found to consist of active enantiomer in patients with an acute severe exacerbation of asthma actively undergoing treatment with racemic-salbutamol. As a result of the possible contribution of (S)-salbutamol to poor asthma control further enantioselective investigations are warranted in severe asthma. Year: 2003 Month: 6 Title: 134th ENMC International Workshop: Outcome Measures and Treatment of Spinal Muscular Atrophy, 11-13 February 2005, Naarden, The Netherlands Authors: Bertini, E. Burghes, A. Bushby, K. Estournet-Mathiaud, B. Finkel, R. S. Hughes, R. A. Iannaccone, S. T. Melki, J. Mercuri, E. Muntoni, F. Voit, T. Reitter, B. Swoboda, K. J. Tiziano, D. Tizzano, E. Topaloglu, H. Wirth, B. Zerres, K. Auth Address: Department of Laboratories, Unit of Molecular Medicine, Bambino Gesu' Children's Research Hospital, Piazza S. Onofrio 4, 00165 Rome, Italy. ebertini@tin.it Pages: 802-16 Volume: 15 Number: 11 Keywords: Animals Humans Muscular Atrophy, Spinal/classification/*diagnosis/*therapy Netherlands Research Support, Non-U.S. Gov't *Treatment Outcome Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16202598 Abstract: Year: 2005 Month: 11 Title: 157th ENMC International Workshop: patient registries for rare, inherited muscular disorders 25-27 January 2008 Naarden, The Netherlands Authors: Sarkozy, A. Bushby, K. Beroud, C. Lochmuller, H. Auth Address: Institute of Human Genetics, University of Newcastle, TREAT-NMD Office, Newcastle upon Tyne NE1 3BZ, UK. Pages: 997-1001 Volume: 18 Number: 12 Keywords: Data Collection/methods Genetic Screening/methods Humans Muscular Atrophy, Spinal/diagnosis/genetics/therapy Muscular Diseases/classification/congenital/*diagnosis Muscular Dystrophy, Duchenne/diagnosis/genetics/therapy Registries/*statistics & numerical data Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18948006 Abstract: Year: 2008 Month: 12 Title: 17-AAG, an Hsp90 inhibitor, ameliorates polyglutamine-mediated motor neuron degeneration Authors: Waza, M. Adachi, H. Katsuno, M. Minamiyama, M. Sang, C. Tanaka, F. Inukai, A. Doyu, M. Sobue, G. Auth Address: Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan. Pages: 1088-95 Volume: 11 Number: 10 Keywords: Animals Cell Line Cells, Cultured Dose-Response Relationship, Drug Gene Expression Regulation/drug effects HSP90 Heat-Shock Proteins/*antagonists & inhibitors/metabolism Male Mice Mice, Transgenic Motor Neurons/*drug effects/*pathology Muscular Atrophy, Spinal/drug therapy/*genetics/*pathology Mutation Peptides/*genetics Phenotype Receptors, Androgen/genetics/metabolism Research Support, Non-U.S. Gov't Rifabutin/*analogs & derivatives/pharmacology/therapeutic use Trinucleotide Repeat Expansion/genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16155577 Abstract: Heat-shock protein 90 (Hsp90) functions as part of a multichaperone complex that folds, activates and assembles its client proteins. Androgen receptor (AR), a pathogenic gene product in spinal and bulbar muscular atrophy (SBMA), is one of the Hsp90 client proteins. We examined the therapeutic effects of 17-allylamino-17-demethoxygeldanamycin (17-AAG), a potent Hsp90 inhibitor, and its ability to degrade polyglutamine-expanded mutant AR. Administration of 17-AAG markedly ameliorated motor impairments in the SBMA transgenic mouse model without detectable toxicity, by reducing amounts of monomeric and aggregated mutant AR. The mutant AR showed a higher affinity for Hsp90-p23 and preferentially formed an Hsp90 chaperone complex as compared to wild-type AR; mutant AR was preferentially degraded in the presence of 17-AAG in both cells and transgenic mice as compared to wild-type AR. 17-AAG also mildly induced Hsp70 and Hsp40. 17-AAG would thus provide a new therapeutic approach to SBMA and probably to other related neurodegenerative diseases. Year: 2005 Month: 10 Title: 17-DMAG ameliorates polyglutamine-mediated motor neuron degeneration through well-preserved proteasome function in an SBMA model mouse Authors: Tokui, K. Adachi, H. Waza, M. Katsuno, M. Minamiyama, M. Doi, H. Tanaka, K. Hamazaki, J. Murata, S. Tanaka, F. Sobue, G. Auth Address: Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan. Pages: 898-910 Volume: 18 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19066230 Abstract: The ubiquitin-proteasome system (UPS) is the principal protein degradation system that tags and targets short-lived proteins, as well as damaged or misfolded proteins, for destruction. In spinal and bulbar muscular atrophy (SBMA), the androgen receptor (AR), an Hsp90 client protein, is such a misfolded protein that tends to aggregate in neurons. Hsp90 inhibitors promote the degradation of Hsp90 client proteins via the UPS. In a transgenic mouse model of SBMA, we examined whether a functioning UPS is preserved, if it was capable of degrading polyglutamine-expanded mutant AR, and what might be the therapeutic effects of 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (17-DMAG), an oral Hsp90 inhibitor. Ubiquitin-proteasomal function was well preserved in SBMA mice and was even increased during advanced stages when the mice developed severe phenotypes. Administration of 17-DMAG markedly ameliorated motor impairments in SBMA mice without detectable toxicity and reduced amounts of monomeric and nuclear-accumulated mutant AR. Mutant AR was preferentially degraded in the presence of 17-DMAG in both SBMA cell and mouse models when compared with wild-type AR. 17-DMAG also significantly induced Hsp70 and Hsp40. Thus, 17-DMAG would exert a therapeutic effect on SBMA via preserved proteasome function. Year: 2009 Title: 2 Years' experience with inspiratory muscle training in patients with neuromuscular disorders Authors: Koessler, W. Wanke, T. Winkler, G. Nader, A. Toifl, K. Kurz, H. Zwick, H. Auth Address: Ludwig Boltzmann-Institute for Environmental Pulmonology and Pulmonary Department, City Hospital Lainz, Vienna, Austria. Kow@khl.pul.magwien.gv.at Pages: 765-9 Volume: 120 Number: 3 Keywords: Adolescent Adult *Breathing Exercises Forced Expiratory Volume Humans Muscular Atrophy, Spinal/*complications/physiopathology Muscular Dystrophy, Duchenne/*complications/physiopathology Respiratory Insufficiency/etiology/*rehabilitation Respiratory Muscles/*physiopathology Vital Capacity Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11555507 Abstract: PURPOSE: The aim of our study was to assess the long-term effects of specific inspiratory muscle training (IMT) in patients with neuromuscular disorders (NMDs) who have various degrees of respiratory impairment. Patients and methods: Twenty-seven patients with NMDs (Duchenne's muscular dystrophy, 18 patients; spinal muscular atrophy, 9 patients) underwent 24 months of IMT. Patients were divided into three groups according to their vital capacity (VC) values. VC was measured as the parameter for the respiratory system involvement of the disease. Maximal inspiratory pressure (PImax) was assessed as the parameter for respiratory muscle strength, and the results of the 12-s maximum voluntary ventilation test (12sMVV) were assessed as the parameter for respiratory muscle endurance. Pulmonary and inspiratory muscle function parameters were assessed 6 months before training, at the beginning of training, and then every 3 months. RESULTS: The PImax values improved in group A (VC, 27 to 50% predicted) from 51.45 to 87.00 cm H(2)O, in group B (VC, 51 to 70% predicted) from 59.38 to 94.4 cm H(2)O, and in group C (VC, 71 to 96% predicted) from 71.25 to 99.00 cm H(2)O. The 12sMVV values improved in group A from 52.69 to 69.50 L/min, in group B from 53.18 to 62.40 L/min, and in group C from 59.48 to 70.5 L/min. For all three groups, there was a significant improvement of PImax (p < 0.007) and 12sMVV (p < 0.015) until the 10th month when a plateau phase was reached with no decline in the following month until the end of training. CONCLUSION: With IMT, respiratory muscle function can be improved in the long term of up to 2 years. Year: 2001 Month: 9 Title: 2-Deoxy-D-glucose reduces epilepsy progression by NRSF-CtBP-dependent metabolic regulation of chromatin structure Authors: Garriga-Canut, M. Schoenike, B. Qazi, R. Bergendahl, K. Daley, T. J. Pfender, R. M. Morrison, J. F. Ockuly, J. Stafstrom, C. Sutula, T. Roopra, A. Auth Address: [1] Department of Neurology, Medical Science Center, Room 1715, University of Wisconsin-Madison, 1300 University Avenue, Madison, Wisconsin 53706, USA. [2] These authors contributed equally to this work. Pages: 1382-7 Volume: 9 Number: 11 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17041593 Abstract: Temporal lobe epilepsy is a common form of drug-resistant epilepsy that sometimes responds to dietary manipulation such as the 'ketogenic diet'. Here we have investigated the effects of the glycolytic inhibitor 2-deoxy-D-glucose (2DG) in the rat kindling model of temporal lobe epilepsy. We show that 2DG potently reduces the progression of kindling and blocks seizure-induced increases in the expression of brain-derived neurotrophic factor and its receptor, TrkB. This reduced expression is mediated by the transcription factor NRSF, which recruits the NADH-binding co-repressor CtBP to generate a repressive chromatin environment around the BDNF promoter. Our results show that 2DG has anticonvulsant and antiepileptic properties, suggesting that anti-glycolytic compounds may represent a new class of drugs for treating epilepsy. The metabolic regulation of neuronal genes by CtBP will open avenues of therapy for neurological disorders and cancer. Year: 2006 Month: 11 Title: 4th UK spinal muscular atrophy (SMA) researchers network meeting Authors: Parkinson, N. J. Auth Address: Department of Physiology, Anatomy and Genetics, University of Oxford, Henry Wellcome Centre for Gene Function, South Parks Road, Oxford OX1 3QX, United Kingdom. Pages: 349-53 Volume: 18 Number: 4 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18396402 Abstract: Year: 2008 Month: 4 Title: 59th ENMC International Workshop: Spinal Muscular Atrophies: recent progress and revised diagnostic criteria 17-19 April 1998, Soestduinen, The Netherlands Authors: Zerres, K. Davies, K. E. Auth Address: Institute for Human Genetics, Technical University, Aachen Germany. Pages: 272-8 Volume: 9 Number: 4 Keywords: Humans *Muscular Atrophy, Spinal/diagnosis/genetics/pathology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10399757 Abstract: Year: 1999 Month: 6 Title: A beginner's guide to NF-kappaB signaling pathways Authors: Delhalle, S. Blasius, R. Dicato, M. Diederich, M. Auth Address: Laboratoire de Biologie Moleculaire et Cellulaire du Cancer, Hopital Kirchberg, L-2540 Luxembourg, Luxembourg. Pages: 1-13 Volume: 1030 Number: Keywords: Animals Humans NF-kappa B/*metabolism Research Support, Non-U.S. Gov't *Signal Transduction Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15659775 Abstract: Nuclear factor kappaB (NF-kappaB) belongs to a family of heterodimeric transcription factors that play a key role in inflammatory and stress responses as well as in tumor cell resistance to apoptosis. These effects are due to the NF-kappaB-dependent transcription of many proinflammatory and antiapoptotic genes, whose products ensure various cell responses to environmental conditions. The signal transduction pathways leading to NF-kappaB activation are well characterized, and the different steps implicated in these pathways involve proteins that could constitute targets for NF-kappaB inhibition. Several inhibitors aiming to prevent NF-kappaB activity and thus the transcription of target genes are studied, and a few compounds seem particularly promising. We try here to summarize the advantages that can issue from various studies on NF-kappaB. Year: 2004 Month: 12 Title: A behavioural characterisation of the FVB/N mouse strain Authors: Pugh, P. L. Ahmed, S. F. Smith, M. I. Upton, N. Hunter, A. J. Auth Address: Neurology and GI CEDD, GlaxoSmithKline, Third Avenue, Harlow, Essex CM19 5AW, UK. Pippa_L_Pugh@gsk.com Pages: 283-9 Volume: 155 Number: 2 Keywords: Aggression/*physiology Animals Behavior, Animal/*physiology Behavioral Research/instrumentation/*methods Circadian Rhythm/genetics/*physiology Cognition/*physiology Comparative Study Female *Genetics, Behavioral Genotype Learning/*physiology Male Mice Mice, Inbred C57BL Mice, Transgenic Models, Animal Motor Activity/genetics/physiology Retina/physiopathology Species Specificity Visual Perception/genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15364488 Abstract: The use of transgenic models in scientific research has made an enormous contribution to our understanding of the causes and symptoms of many diseases, including neurodegenerative conditions such as Alzheimer's Disease (AD) and Parkinson's Disease (PD). In the creation of transgenic models of neurodegenerative disease, effects of the background strain of the animal on the resulting genotype must be taken into consideration. This is particularly true for behavioural studies in which the background strain of the mouse may mask the phenotype of the genetic manipulation. Here, the behaviour of two mouse strains used in transgenic models, FVB/N and C57BL6/J, were compared. Studies of circadian wheel activity, cognition and aggression revealed considerable phenotypic differences between strains. These data also indicate that the FVB/N strain is not appropriate as a background strain in the behavioural assessment of transgenic mouse models. Year: 2004 Title: A cell system with targeted disruption of the SMN gene: functional conservation of the SMN protein and dependence of Gemin2 on SMN Authors: Wang, J. Dreyfuss, G. Auth Address: Howard Hughes Medical Institute and Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia 19104, USA. Pages: 9599-605 Volume: 276 Number: 13 Keywords: Amino Acid Sequence Animals Blotting, Southern Blotting, Western Cell Death Cell Line Cell Survival Cells, Cultured Chickens Conserved Sequence Cyclic AMP Response Element-Binding Protein DNA, Complementary/metabolism Electrophoresis, Polyacrylamide Gel Exons Flow Cytometry Gene Library Humans Models, Genetic Molecular Sequence Data Muscular Atrophy, Spinal/metabolism *Mutagenesis, Site-Directed Nerve Tissue Proteins/chemistry/*genetics/*metabolism Plasmids Promoter Regions (Genetics) RNA-Binding Proteins Recombination, Genetic Research Support, U.S. Gov't, P.H.S. Tetracycline/metabolism Time Factors Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11121410 Abstract: The motor neuron degenerative disease spinal muscular atrophy is caused by reduced expression of the survival motor neuron (SMN) protein. Here we report a genetic system developed in the chicken pre-B cell line DT40, in which the endogenous SMN gene is disrupted by homologous recombination, and SMN protein is expressed from a chicken SMN cDNA under control of a tetracycline (tet)-repressible promoter. Addition of tet results in depletion of SMN protein and consequent cell death, which directly demonstrates that SMN is required for cell viability. The tet-induced lethality can be rescued by expression of human SMN, indicating that the function of SMN is highly conserved between the two species. Cells expressing low levels of SMN display slow growth proportional to the amount of SMN they contain. Interestingly, the level of the SMN-interacting protein Gemin2 decreases significantly following depletion of SMN, supporting the conclusion that SMN and Gemin2 form a stable complex in vivo. This system provides a powerful setting for studying the function of SMN in vivo and for screening for potential therapeutics for spinal muscular atrophy. Year: 2001 Title: A collaborative study on the natural history of childhood and juvenile onset proximal spinal muscular atrophy (type II and III SMA): 569 patients Authors: Zerres, K. Rudnik-Schoneborn, S. Forrest, E. Lusakowska, A. Borkowska, J. Hausmanowa-Petrusewicz, I. Auth Address: Institute for Human Genetics, University of Bonn, Germany. zerres@snphysio2.wilhelm.uni-bonn.de Pages: 67-72 Volume: 146 Number: 1 Keywords: Adolescent Adult Age of Onset Child Child, Preschool *Databases, Factual Disease Progression Female Germany Humans Infant Infant, Newborn Male Muscular Atrophy, Spinal/pathology/*physiopathology Poland Probability Research Support, Non-U.S. Gov't Spinal Muscular Atrophies of Childhood/pathology/*physiopathology Walking/physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9077498 Abstract: We analyzed clinical data of 569 patients in two combined series with childhood and juvenile proximal SMA. This cohort included only patients who had achieved the ability to sit unaided (type II and III SMA). The survival rate among 240 type II patients (who sat but never walked) was 98.5% at 5 years and 68.5% at 25 years. SMA III (n = 329) (those who walked and had symptoms before age 30 years) was subdivided into those with an onset before and after age 3 years (type IIIa, n = 195; SMA IIIb, n = 134). In patients with SMA III, life expectancy is not significantly less than a normal population. The probabilities of being able to walk at 10 years after onset was 70.3%, and at 40 years, 22.0% in SMA IIa. For SMA IIIb, 96.7% were walking 10 years after onset and 58.7% at 40 years. The subdivision of type III SMA was justified by the probability of being ambulatory depending on age at onset; the prognosis differed for those with onset before or after age 3 years. The data provide a reliable basis of the natural history of proximal SMA and support a classification system that is based primarily on age at onset and the achievement of motor milestones. Year: 1997 Title: A comprehensive interaction map of the human SMN-complex Authors: Otter, S. Grimmler, M. Neuenkirchen, N. Chari, A. Sickmann, A. Fischer, U. Auth Address: Biochemistry, Theodor Boveri Institute, University of Wuerzburg, Wuerzburg D-97074. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17178713 Abstract: Assembly of the Sm-class of U-rich small nuclear ribonucleoprotein particles (U snRNPs) is a process facilitated by the macromolecular SMN-complex. This entity promotes the binding of a set of factors, termed LSm/Sm proteins, onto snRNA to form the core structure of these particles. Nine factors, including the survival motor neuron (SMN) protein, the product of the spinal muscular atrophy (SMA) disease gene, Gemins 2-8 and unrip have been identified as the major components of the SMN-complex. So far however, only little is known about the architecture of this complex and the contribution of individual components to its function. Here, we present a comprehensive interaction map of all core components of the SMN-complex based upon in vivo and in vitro methods. Our studies reveal a modular composition of the SMN-complex with the three proteins SMN, Gemin8 and Gemin7 in its center. Onto this central building block the other components are bound via multiple interactions. Furthermore, by employing a novel assay, we were able to reconstitute the SMN-complex from individual components and confirm the interaction map. Interestingly, SMN protein carrying an SMA-causing mutation was severely impaired in formation of the SMN-complex. Finally, we show that the peripheral component Gemin5 contributes an essential activity to the SMN-complex, most likely the transfer of Sm proteins onto the U snRNA. Collectively, the data presented here provide a basis for the detailed mechanistic and structural analysis of the assembly machinery of U snRNPs. Year: 2006 Title: A CSF biomarker panel for identification of patients with amyotrophic lateral sclerosis Authors: Mitchell, R. M. Freeman, W. M. Randazzo, W. T. Stephens, H. E. Beard, J. L. Simmons, Z. Connor, J. R. Auth Address: Department of Neurosurgery, Pennsylvania State University College of Medicine/Milton S. Hershey Medical Center, Hershey, PA 17033-0850, USA. Pages: 14-9 Volume: 72 Number: 1 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18987350 Abstract: BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease with complicated pathogenesis that poses challenges with respect to diagnosis and monitoring of disease progression. OBJECTIVES: To identify a biomarker panel that elucidates ALS disease pathogenesis, distinguishes patients with ALS from neurologic disease controls, and correlates with ALS disease characteristics, and to determine the effect of HFE gene variants, a potential risk factor for sporadic ALS, on the biomarker profile. METHODS: We obtained CSF samples by lumbar puncture from 41 patients with ALS and 33 neurologic disease controls. All patients were genotyped for HFE polymorphisms. We performed a multiplex cytokine and growth factor analysis and immunoassays for iron-related analytes. Classification statistics were generated using a support vector machine algorithm. RESULTS: The groups of patients with ALS and neurologic disease controls were each associated with distinct profiles of biomarkers. Fourteen biomarkers differed between patients with ALS and the control group. The five proteins with the lowest p values differentiated patients with ALS from controls with 89.2% accuracy, 87.5% sensitivity, and 91.2% specificity. Expression of IL-8 was higher in those patients with lower levels of physical function. Expression of beta2-microglobulin was higher in subjects carrying an H63D HFE allele, while expression of several markers was higher in subjects carrying a C282Y HFE allele. CONCLUSIONS: A CSF inflammatory profile associated with amyotrophic lateral sclerosis (ALS) pathogenesis may distinguish patients with ALS from neurologic disease controls, and may serve as a biomarker panel to aid in the diagnosis of ALS pending further validation. Some of these biomarkers differ by HFE genotype. Year: 2009 Title: A defined long-term in vitro tissue engineered model of neuromuscular junctions Authors: Das, M. Rumsey, J. W. Bhargava, N. Stancescu, M. Hickman, J. J. Auth Address: Hybrid Systems Lab, NanoScience Technology Center, Suite 402, 12424 Research Parkway, University of Central Florida, Orlando, FL 32826, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20346499 Abstract: Neuromuscular junction (NMJ) formation, occurring between motoneurons and skeletal muscle, is a complex multistep process involving a variety of signaling molecules and pathways. In vitro motoneuron-muscle co-cultures are powerful tools to study the role of different growth factors, hormones and cellular structures involved in NMJ formation. In this study, a serum-free culture system utilizing defined temporal growth factor application and a non-biological substrate resulted in the formation of robust NMJs. The system resulted in long-term survival of the co-culture and selective expression of neonatal myosin heavy chain, a marker of myotube maturation. NMJ formation was verified by colocalization of dense clusters of acetylcholine receptors visualized using alpha-bungarotoxin and synaptophysin containing vesicles present in motoneuron axonal terminals. This model will find applications in basic NMJ research and tissue engineering applications such as bio-hybrid device development for limb prosthesis and regenerative medicine as well as for high-throughput drug and toxin screening applications. Title: A degron created by SMN2 exon 7 skipping is a principal contributor to spinal muscular atrophy severity Authors: Cho, S. Dreyfuss, G. Auth Address: Howard Hughes Medical Institute and Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA. Pages: 438-42 Volume: 24 Number: 5 Keywords: Amino Acid Sequence Cell Line Exons/*genetics Humans Molecular Sequence Data Muscular Atrophy, Spinal/genetics/metabolism/*pathology Mutation/genetics Protein Stability Sequence Alignment Severity of Illness Index Signal Transduction/genetics Survival of Motor Neuron 2 Protein/genetics/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20194437 Abstract: Spinal muscular atrophy (SMA) is caused by homozygous survival of motor neurons 1 (SMN1) gene deletions, leaving a duplicate gene, SMN2, as the sole source of SMN protein. However, most of the mRNA produced from SMN2 pre-mRNA is exon 7-skipped ( approximately 80%), resulting in a highly unstable and almost undetectable protein (SMNDelta7). We show that this splicing defect creates a potent degradation signal (degron; SMNDelta7-DEG) at SMNDelta7's C-terminal 15 amino acids. The S270A mutation inactivates SMNDelta7-DEG, generating a stable SMNDelta7 that rescues viability of SMN-deleted cells. These findings explain a key aspect of the SMA disease mechanism, and suggest new treatment approaches based on interference with SMNDelta7-DEG activity. Title: A direct interaction between the survival motor neuron protein and p53 and its relationship to spinal muscular atrophy Authors: Young, P. J. Day, P. M. Zhou, J. Androphy, E. J. Morris, G. E. Lorson, C. L. Auth Address: Department of Biology, Arizona State University, Tempe, Arizona 85287, USA. Pages: 2852-9 Volume: 277 Number: 4 Keywords: Apoptosis Biosensing Techniques Cell Line Cyclic AMP Response Element-Binding Protein Dimerization Exons Fibroblasts/metabolism Glutathione Transferase/metabolism Humans Immunohistochemistry Microscopy, Fluorescence Muscular Atrophy, Spinal/*genetics/*metabolism Mutation Mutation, Missense Nerve Tissue Proteins/*genetics/*metabolism Precipitin Tests Protein Binding Protein Structure, Tertiary RNA-Binding Proteins Recombinant Proteins/metabolism Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Time Factors Transfection Tumor Cells, Cultured Tumor Suppressor Protein p53/*metabolism/*physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11704667 file:///S:/Clinical%20Research%20Papers/Young,%20A%20direct%20interaction%20between%20the%20survival%20motor%20neuron%20protein%20and%20p53%20and%20its%20relationship%20to%20spinal%20muscular%20atrophy,%20J%20Bio%20Chem%202002.pdf Abstract: Mutations in the SMN1 (survival motor neuron 1) gene cause spinal muscular atrophy (SMA). We now show that SMN protein, the SMN1 gene product, interacts directly with the tumor suppressor protein, p53. Pathogenic missense mutations in SMN reduce both self-association and p53 binding by SMN, and the extent of the reductions correlate with disease severity. The inactive, truncated form of SMN produced by the SMN2 gene in SMA patients fails to bind p53 efficiently. SMN and p53 co-localize in nuclear Cajal bodies, but p53 redistributes to the nucleolus in fibroblasts from SMA patients. These results suggest a functional interaction between SMN and p53, and the potential for apoptosis when this interaction is impaired may explain motor neuron death in SMA. Year: 2002 Title: A direct role for FMRP in activity-dependent dendritic mRNA transport links filopodial-spine morphogenesis to fragile X syndrome Authors: Dictenberg, J. B. Swanger, S. A. Antar, L. N. Singer, R. H. Bassell, G. J. Auth Address: Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Dictenberg@genectr.hunter.cuny.edu Pages: 926-39 Volume: 14 Number: 6 Keywords: Animals Cells, Cultured Dendrites/drug effects/*metabolism Disease Models, Animal Fragile X Mental Retardation Protein/chemistry/genetics/*metabolism Fragile X Syndrome/*genetics/metabolism Green Fluorescent Proteins/metabolism Hippocampus/cytology In Situ Hybridization, Fluorescence Kinesin/antagonists & inhibitors Mice Mice, Knockout Microscopy, Video Models, Biological Protein Structure, Tertiary Pseudopodia/*metabolism/pathology *RNA Transport RNA, Messenger/*metabolism Sulfuric Acid Esters/pharmacology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18539120 Abstract: The function of local protein synthesis in synaptic plasticity and its dysregulation in fragile X syndrome (FXS) is well studied, however the contribution of regulated mRNA transport to this function remains unclear. We report a function for the fragile X mental retardation protein (FMRP) in the rapid, activity-regulated transport of mRNAs important for synaptogenesis and plasticity. mRNAs were deficient in glutamatergic signaling-induced dendritic localization in neurons from Fmr1 KO mice, and single mRNA particle dynamics in live neurons revealed diminished kinesis. Motor-dependent translocation of FMRP and cognate mRNAs involved the C terminus of FMRP and kinesin light chain, and KO brain showed reduced kinesin-associated mRNAs. Acute suppression of FMRP and target mRNA transport in WT neurons resulted in altered filopodia-spine morphology that mimicked the FXS phenotype. These findings highlight a mechanism for stimulus-induced dendritic mRNA transport and link its impairment in a mouse model of FXS to altered developmental morphologic plasticity. Year: 2008 Month: 6 Title: A Drosophila melanogaster model of spinal muscular atrophy reveals a function for SMN in striated muscle Authors: Rajendra, T. K. Gonsalvez, G. B. Walker, M. P. Shpargel, K. B. Salz, H. K. Matera, A. G. Auth Address: Department of Genetics, Case Western Reserve University School of Medicine, Cleveland, OH 44106. Pages: 831-41 Volume: 176 Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17353360 Abstract: Mutations in human survival motor neurons 1 (SMN1) cause spinal muscular atrophy (SMA) and are associated with defects in assembly of small nuclear ribonucleoproteins (snRNPs) in vitro. However, the etiological link between snRNPs and SMA is unclear. We have developed a Drosophila melanogaster system to model SMA in vivo. Larval-lethal Smn-null mutations show no detectable snRNP reduction, making it unlikely that these animals die from global snRNP deprivation. Hypomorphic mutations in Smn reduce dSMN protein levels in the adult thorax, causing flightlessness and acute muscular atrophy. Mutant flight muscle motoneurons display pronounced axon routing and arborization defects. Moreover, Smn mutant myofibers fail to form thin filaments and phenocopy null mutations in Act88F, which is the flight muscle-specific actin isoform. In wild-type muscles, dSMN colocalizes with sarcomeric actin and forms a complex with alpha-actinin, the thin filament crosslinker. The sarcomeric localization of Smn is conserved in mouse myofibrils. These observations suggest a muscle-specific function for SMN and underline the importance of this tissue in modulating SMA severity. Year: 2007 Title: A Dutch guideline for the treatment of scoliosis in neuromuscular disorders Authors: Mullender, M. Blom, N. De Kleuver, M. Fock, J. Hitters, W. Horemans, A. Kalkman, C. Pruijs, J. Timmer, R. Titarsolej, P. Van Haasteren, N. Jager, M. V. Van Vught, A. Van Royen, B. Auth Address: Dept, Orthopaedic Surgery, Vrije Universiteit Medical Center (VUmc), Research Institute MOVE, Amsterdam, The Netherlands. m.mullender@vumc.nl. Pages: 14 Volume: 3 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18822133 Abstract: ABSTRACT: BACKGROUND: Children with neuromuscular disorders with a progressive muscle weakness such as Duchenne Muscular Dystrophy and Spinal Muscular Atrophy frequently develop a progressive scoliosis. A severe scoliosis compromises respiratory function and makes sitting more difficult. Spinal surgery is considered the primary treatment option for correcting severe scoliosis in neuromuscular disorders. Surgery in this population requires a multidisciplinary approach, careful planning, dedicated surgical procedures, and specialized after care. METHODS: The guideline is based on scientific evidence and expert opinions. A multidisciplinary working group representing experts from all relevant specialties performed the research. A literature search was conducted to collect scientific evidence in answer to specific questions posed by the working group. Literature was classified according to the level of evidence. RESULTS: For most aspects of the treatment scientific evidence is scarce and only low level cohort studies were found. Nevertheless, a high degree of consensus was reached about the management of patients with scoliosis in neuromuscular disorders. This was translated into a set of recommendations, which are now officially accepted as a general guideline in the Netherlands. CONCLUSION: In order to optimize the treatment for scoliosis in neuromuscular disorders a Dutch guideline has been composed. This evidence-based, multidisciplinary guideline addresses conservative treatment, the preoperative, perioperative, and postoperative care of scoliosis in neuromuscular disorders. Year: 2008 Month: 1 Title: A feasibility study for the newborn screening of spinal muscular atrophy Authors: Pyatt, R. E. Prior, T. W. Auth Address: Department of Pathology, Ohio State University, Columbus, Ohio 43210, USA. Pages: 428-37 Volume: 8 Number: 7 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16845275 Abstract: PURPOSE: The natural history of spinal muscular atrophy suggests that for maximum effect, therapeutics will need to be administered in the earliest phases of the disease. This will require the adoption of techniques for the genetic analysis of affected individuals at the newborn stage. Our objective was to examine the feasibility surrounding the newborn screening for spinal muscular atrophy. METHODS: We investigated the application of real-time polymerase chain reaction technology for newborn screening. A multiplex assay was designed to identify homozygous deletions in SMN1 exon 7 and validated using 266 samples with defined SMN1 and SMN2 copy numbers. Sensitivity and specificity were then evaluated as part of a newborn screening strategy using DNA from 153 blood spots. RESULTS: Real-time technology validation demonstrated correct exclusion of all normal and carrier samples, and identified the homozygous SMN1 exon 7 deletions in all 32 affected samples. In the series of blood spots, all 59 affected samples were correctly identified yielding an analytic sensitivity of 100%; 56 normal and 39 carrier samples were correctly excluded yielding an analytic specificity of 100% for this blood spot series. CONCLUSION: We demonstrate that effective molecular technology exists and that ethics may soon warrant the newborn screening of spinal muscular atrophy. Year: 2006 Month: 7 Title: A first step in viral gene therapy for muscular dystrophy Authors: Hirano, M. Auth Address: Columbia University Medical Center, 630 West 168th Street, P&S 4-423, New York, NY 10032, USA. mh29@columbia.edu Pages: 71-2 Volume: 10 Number: 2 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20425230 Abstract: Month: 3 Title: A flexible data analysis tool for chemical genetic screens Authors: Kelley, B. P. Lunn, M. R. Root, D. E. Flaherty, S. P. Martino, A. M. Stockwell, B. R. Auth Address: Department of Biological Sciences, Columbia University, Fairchild Center, MC 2406, 1212 Amsterdam Avenue, New York, NY 10027 USA. Pages: 1495-503 Volume: 11 Number: 11 Keywords: Cyclic AMP Response Element-Binding Protein Databases, Genetic *Drug Evaluation, Preclinical Flavonoids/pharmacology Genes, Reporter Humans Muscular Atrophy, Spinal/genetics Nerve Tissue Proteins/genetics/metabolism RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. *Software Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15556000 Abstract: High-throughput assays generate immense quantities of data that require sophisticated data analysis tools. We have created a freely available software tool, SLIMS (Small Laboratory Information Management System), for chemical genetics which facilitates the collection and analysis of large-scale chemical screening data. Compound structures, physical locations, and raw data can be loaded into SLIMS. Raw data from high-throughput assays are normalized using flexible analysis protocols, and systematic spatial errors are automatically identified and corrected. Various computational analyses are performed on tested compounds, and dilution-series data are processed using standard or user-defined algorithms. Finally, published literature associated with active compounds is automatically retrieved from Medline and processed to yield potential mechanisms of actions. SLIMS provides a framework for analyzing high-throughput assay data both as a laboratory information management system and as a platform for experimental analysis. Year: 2004 Month: 11 Title: A Handheld Electrical Impedance Myography probe for the assessment of neuromuscular disease Authors: Ogunnika, O. T. Scharfstein, M. Cooper, R. C. Ma, H. Dawson, J. L. Rutkove, S. B. Auth Address: Microsystems Technology Laboratory, Massachusetts Institute of Technology, Cambridge, 02139 USA. Pages: 3566-9 Volume: 1 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19163479 Abstract: Electrical Impedance Myography (EIM) is a non-invasive, painless clinical technique for the diagnosis and monitoring of a variety of neuromuscular diseases including amyotrophic lateral sclerosis and focal nerve injuries. It involves the application of a low-intensity alternating current to a muscle group and the measurement of the consequent surface voltage patterns. This paper presents a system for the rapid and accurate acquisition of data employing an interrogating signal composed of multiple tones with frequencies between 10 kHz and 4 MHz. The use of this composite signal makes possible measurement of impedance at multiple frequencies simultaneously. In addition, this system takes impedance measurements at multiple orientations with respect to the muscle fibers by means of an electronically reconfigurable electrode array and utilizes the linearity of muscle tissue to reduce the required measurement time. Testing of the EIM system on beef has established the capability of this system to rapidly detect the anisotropic conductive properties of muscle tissue at multiple frequencies. Year: 2008 Month: 1 Title: A high-throughput drug screen targeted to the 5'untranslated region of Alzheimer amyloid precursor protein mRNA Authors: Bandyopadhyay, S. Ni, J. Ruggiero, A. Walshe, K. Rogers, M. S. Chattopadhyay, N. Glicksman, M. A. Rogers, J. T. Auth Address: Neurochemistry Laboratory, Genetics and Aging Research Unit, Psychiatry Department, Massachusetts General Hospital, Boston, MA, USA, and School of Biological and Environmental Science, University College Dublin, Belfield, Ireland. Pages: 469-80 Volume: 11 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16928984 Abstract: The authors employed a novel approach to identify therapeutics effective in Alzheimer disease (AD). The 5'untranslated region (5'UTR) of the mRNA of AD amyloid precursor protein (APP) is a significant regulator of the levels of the APP holoprotein and amyloid beta (Abeta) peptide in the central nervous system. The authors generated stable neuroblastoma SH-SY5Y transfectants that express luciferase under the translational control of the 146-nucleotide APP mRNA 5'UTR and green fluorescent protein (GFP) driven by a viral internal ribosomal entry site. Using a high-throughput screen (HTS), they screened for the effect of 110,000 compounds obtained from the library of the Laboratory for Drug Discovery on Neurodegeneration (LDDN) on the APP mRNA 5'UTR-controlled translation of the luciferase reporter. This screening yielded several nontoxic specific inhibitors of APP mRNA 5'UTR-driven luciferase that had no effect on the GFP expression in the stable SH-SY5Y transfectants. Moreover, these compounds either did not inhibit or inhibited to a much lower extent the expression of the luciferase reporter regulated by a prion protein (PrP) mRNA 5'UTR, used as an alternative mRNA structure to counterscreen APP mRNA 5'UTR in stably transfected SH-SY5Y cell lines. The hits obtained from this robust, specific, and highly quantitative HTS will be characterized to identify agents that may be developed into useful future therapeutic agents to limit APP translation and Abeta production for AD. Year: 2006 Month: 8 Title: A homozygous mutation in the SCO2 gene causes a spinal muscular atrophy like presentation with stridor and respiratory insufficiency Authors: Pronicki, M. Kowalski, P. Piekutowska-Abramczuk, D. Taybert, J. Karkucinska-Wieckowska, A. Szymanska-Debinska, T. Karczmarewicz, E. Pajdowska, M. Migdal, M. Milewska-Bobula, B. Sykut-Cegielska, J. Popowska, E. Auth Address: Department of Pathology, Children's Memorial Health Institute, 04-730 Warsaw, Poland. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19879173 Abstract: Infants with deficiency of cytochrome c oxidase (COX) due to SCO2 mutations observed so far usually demonstrated early cardiomyopathy, encephalopathy and lactic acidosis. Milder spinal muscular atrophy-like (SMA-like) phenotype was also rarely reported. The aim is to present 18 Polish patients with SCO2 mutations. Molecular study revealed p.E140K mutation in all cases (on 32 alleles); p.Q53X mutation and novel p.M177T change were identified in single patients. In three families no second mutation was found. Thirteen p.E140K homozygotes presented in infancy with floppiness and remarkable stridor. Survival motor neuron (SMN) gene deletion was excluded. Mild to moderate lactic academia was found. Neurological involvement manifested as spasticity and psychomotor retardation. In some patients strabismus, ptosis and episodes of seizures were seen. During second half of the year chronic respiratory failure with artificial respiration dependency appeared in all homozygotes. Heart involvement was never present at the beginning. Rapidly progressive hypertrophic cardiomyopathy developed in several patients at the terminal stage. The stridor was constant and striking feature. Skeletal muscle biopsy was performed in 16 patients including 11 homozygotes. Four pathological patterns were discerned - from neurogenic muscle changes, including spinal muscular atrophy (SMA) to unspecific findings. Histochemical cytochrome c oxidase (COX) deficit was not a constant feature. Significant decrease in respiratory chain complex IV activity was detected in muscle homogenate by spectrophotometric method only in 7 out of 12 examined cases. CONCLUSIONS: 1/Mutations of SCO2 gene should be considered as a possible cause of neurogenic skeletal muscle features (including SMA-like) in infants with encephalomyopathy even in the absence of heart involvement and COX deficit; 2/Inspiratory stridor may be symptomatic of SCO2 gene mutation(s). Year: 2009 Title: A microarray configuration to quantify expression levels and relative abundance of splice variants Authors: Fehlbaum, P. Guihal, C. Bracco, L. Cochet, O. Auth Address: ExonHit Therapeutics, 63/65 boulevard Massena, 75013 Paris, France. Pages: e47 Volume: 33 Number: 5 Keywords: *Alternative Splicing Cell Line Exons Female Gene Expression Profiling/*methods Humans Introns Oligonucleotide Array Sequence Analysis/*methods Oligonucleotide Probes/chemistry Protein Isoforms/analysis/genetics Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15760843 Abstract: Over the past decade, alternative RNA splicing has raised a great interest appearing to be of high importance in the generation of expression diversity. This regulatory process plays a critical role in the normal development and its impact on the initiation and development of human disorders as well as on the pharmacological properties of drugs is increasingly being recognized. Only few studies describe specific alternative splicing expression profiling. Microarray strategies have been conceived to address alternative splicing events but with very few experimental data related to their abilities to provide true quantification values. We have developed a specific microarray configuration relying on a few, well optimized probes per splice event. Basically, five probes of 24mer are used to fully characterize a splice event. These probes are of two types, exon probes and junction probes, and are either specific to a splice event or not. The performances of such a 'splice array' were validated on synthetic model systems and on complex biological materials. The results indicate that DNA chips based on this design combining exon and junction derived probes enable the detection and, absolute and relative quantification of splice variants. In addition, this strategy is compatible with all the microarrays that use oligonucleotide probes. Year: 2005 Month: 1 Title: A modified Hammersmith functional motor scale for use in multi-center research on spinal muscular atrophy Authors: Krosschell, K. J. Maczulski, J. A. Crawford, T. O. Scott, C. Swoboda, K. J. Auth Address: Department of Physical Therapy and Human Movement Sciences, Feinberg School of Medicine, Northwestern University, Suite 1100, 645 North Michigan Avenue, Chicago, IL 60611, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16750368 Abstract: The Hammersmith functional motor scale for children with spinal muscular atrophy was modified to establish a standard measure of functional ability in children with non-ambulant spinal muscular atrophy types 2 and 3 in a longitudinal multi-center clinical trial. This study assessed the intra- and interrater reliability and the test-retest stability of a modified version of the scale. Both intra- and interrater reliability were established. Results indicate that the scale is reliable and stable over a 6 month period. Reliability was maintained when patient sample criteria were expanded to include children younger than 30 months and children with popliteal angles greater than 20 degrees . These data establish the modified Hammersmith functional motor scale for children with spinal muscular atrophy as a reliable instrument for use in multi-center treatment trials in non-ambulant spinal muscular atrophy children. Our data provides additional support for the use of original scale items in terms of ease of administration, usefulness and reliability, while incorporating modifications to optimize its use in a multi-center clinical research setting. Year: 2006 Title: A motor function for the DEAD-box RNA helicase, Gemin3, in Drosophila Authors: Cauchi, R. J. Davies, K. E. Liu, J. L. Auth Address: Department of Physiology, Medical Research Council Functional Genomics Unit, University of Oxford, Oxford, United Kingdom. Pages: e1000265 Volume: 4 Number: 11 Keywords: Animals DEAD Box Protein 20/genetics/*physiology DEAD-box RNA Helicases/genetics/*physiology Drosophila melanogaster/embryology/*enzymology/*genetics Humans Larva/metabolism Mice Motor Neurons/*enzymology Mutation Survival of Motor Neuron 1 Protein/metabolism Transgenes Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19023405 Abstract: The survival motor neuron (SMN) protein, the determining factor for spinal muscular atrophy (SMA), is complexed with a group of proteins in human cells. Gemin3 is the only RNA helicase in the SMN complex. Here, we report the identification of Drosophila melanogaster Gemin3 and investigate its function in vivo. Like in vertebrates, Gemin3 physically interacts with SMN in Drosophila. Loss of function of gemin3 results in lethality at larval and/or prepupal stages. Before they die, gemin3 mutant larvae exhibit declined mobility and expanded neuromuscular junctions. Expression of a dominant-negative transgene and knockdown of Gemin3 in mesoderm cause lethality. A less severe Gemin3 disruption in developing muscles leads to flightless adults and flight muscle degeneration. Our findings suggest that Drosophila Gemin3 is required for larval development and motor function. Year: 2008 Month: 11 Title: A motor function measure for neuromuscular diseases. Construction and validation study Authors: Berard, C. Payan, C. Hodgkinson, I. Fermanian, J. Auth Address: Department of Paediatric Rehabilitation, l'Escale, Centre Hospitalier Lyon-Sud, France. carole.berard@chu-lyon.fr Pages: 463-70 Volume: 15 Number: 7 Keywords: Adolescent Adult Child *Disability Evaluation Extremities/physiopathology Factor Analysis, Statistical Female Humans Male Middle Aged Motor Activity/*physiology Neuromuscular Diseases/classification/*physiopathology Pain Measurement Predictive Value of Tests Reproducibility of Results *Severity of Illness Index Time Factors Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16106528 Abstract: A new scale for motor function measurement has been developed for neuromuscular diseases. The validation study included 303 patients, aged 6-62 years. Seventy-two patients had Duchenne muscular dystrophy, 32 Becker muscular dystrophy, 30 limb-girdle muscular dystrophy, 39 facio-scapulo-humeral dystrophy, 29 myotonic dystrophy, 21 congenital myopathy, 10 congenital muscular dystrophy, 35 spinal muscular atrophy and 35 hereditary neuropathy. The scale comprised 32 items, in three dimensions: standing position and transfers, axial and proximal motor function, distal motor function. Agreement coefficients for inter-rater reliability were excellent (kappa=0.81-0.94) for nine items, good (kappa=0.61-0.80) for 20 items and moderate (kappa=0.51-0.60) for three items. High correlations were found between the total score and other scores: Vignos (r=0.91) and Brooke (r=0.85) grades, Functional Independence Measure (r=0.91), the global severity of disability evaluated with visual analog scales by physicians (r=0.88) and physiotherapists (r=0.91). This scale is reliable, does not require any special equipment and is well-accepted by patients. Its sensitivity to change is being assessed to permit its use in clinical trials of neuromuscular diseases. Year: 2005 Month: 7 Title: A mouse model for spinal muscular atrophy Authors: Hsieh-Li, H. M. Chang, J. G. Jong, Y. J. Wu, M. H. Wang, N. M. Tsai, C. H. Li, H. Auth Address: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan. Pages: 66-70 Volume: 24 Number: 1 Keywords: Animals Base Sequence Cells, Cultured Cyclic AMP Response Element-Binding Protein DNA Primers Disease Models, Animal Female Humans Male Mice Mice, Transgenic Muscular Atrophy, Spinal/*genetics/pathology Nerve Tissue Proteins/*genetics RNA-Binding Proteins Research Support, Non-U.S. Gov't Reverse Transcriptase Polymerase Chain Reaction Transgenes Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10615130 Abstract: The survival motor neuron gene is present in humans in a telomeric copy, SMN1, and several centromeric copies, SMN2. Homozygous mutation of SMN1 is associated with proximal spinal muscular atrophy (SMA), a severe motor neuron disease characterized by early childhood onset of progressive muscle weakness. To understand the functional role of SMN1 in SMA, we produced mouse lines deficient for mouse Smn and transgenic mouse lines that expressed human SMN2. Smn-/- mice died during the peri-implantation stage. In contrast, transgenic mice harbouring SMN2 in the Smn-/- background showed pathological changes in the spinal cord and skeletal muscles similar to those of SMA patients. The severity of the pathological changes in these mice correlated with the amount of SMN protein that contained the region encoded by exon 7. Our results demonstrate that SMN2 can partially compensate for lack of SMN1. The variable phenotypes of Smn-/-SMN2 mice reflect those seen in SMA patients, providing a mouse model for this disease. Year: 2000 Month: 1 Title: A mouse model of spinal and bulbar muscular atrophy Authors: McManamny, P. Chy, H.S. Finkelstein, D.I. Craythorn, R.G. Crack, P.J. Kola, I. Cheema, S.S. Horne, M.K. Wreford, N.G. O'Bryan, M.K. De Kretser, D.M. Morrison, J.R. Auth Address: Pages: 2103-2011 Volume: 11 Number: Keywords: No keywords found Related urls: Abstract: Year: 2002 Month: 1 Title: A multistep validation process of biomarkers for preclinical drug development Authors: Freeman, W. M. Bixler, G. V. Brucklacher, R. M. Lin, C. M. Patel, K. M. Vanguilder, H. D. Lanoue, K. F. Kimball, S. R. Barber, A. J. Antonetti, D. A. Gardner, T. W. Bronson, S. K. Auth Address: [1] Functional Genomics Core Facility, Penn State College of Medicine, Hershey, PA, USA [2] Department of Pharmacology, Penn State College of Medicine, Hershey, PA, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19997081 Abstract: Biomarkers that can be measured in preclinical models in a high-throughput, reproducible manner offer the potential to increase the speed and efficacy of drug development. Development of therapeutic agents for many conditions is hampered by the limited number of validated preclinical biomarkers available to gauge pharmacoefficacy and disease progression, but the validation process for preclinical biomarkers has received limited attention. This report defines a five-step preclinical biomarker validation process and applies the process to a case study of diabetic retinopathy. By showing that a gene expression panel is highly reproducible, coincides with disease manifestation, accurately classifies individual animals and identifies animals treated with a known therapeutic agent, a biomarker panel can be considered validated. This particular biomarker panel consisting of 14 genes (C1inh, C1s, Carhsp1, Chi3l1, Gat3, Gbp2, Hspb1, Icam1, Jak3, Kcne2, Lama5, Lgals3, Nppa, Timp1) can be used in diabetic retinopathy pharmacotherapeutic research, and the biomarker development process outlined here is applicable to drug development efforts for other diseases.The Pharmacogenomics Journal advance online publication, 8 December 2009; doi:10.1038/tpj.2009.60. Year: 2009 Title: A mutation in the small heat-shock protein HSPB1 leading to distal hereditary motor neuronopathy disrupts neurofilament assembly and the axonal transport of specific cellular cargoes Authors: Ackerley, S. James, P. A. Kalli, A. French, S. Davies, K. E. Talbot, K. Auth Address: Department of Human Anatomy and Genetics, South Parks Road, Oxford OX1 3QX, UK. Pages: 347-54 Volume: 15 Number: 2 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16368711 Abstract: Distal hereditary motor neuronopathies (dHMNs) are a clinically and genetically heterogeneous group of disorders in which motor neurons selectively undergo age-dependant degeneration. Mutations in the small heat-shock protein HSPB1 (HSP27) are responsible for one form of dHMN. In this study, we have analysed the effect of expressing a form of mutant HSPB1 in primary neuronal cells in culture. Mutant (P182L) but not wild-type HSPB1 led to the formation of insoluble intracellular aggregates and to the sequestration in the cytoplasm of selective cellular components, including neurofilament middle chain subunit (NF-M) and p150 dynactin. These findings suggest a possible pathogenic mechanism for HSPB1 whereby the mutation may lead to preferential motor neuron loss by disrupting selective components essential for axonal structure and transport. Year: 2006 Title: A mutation in the vesicle-trafficking protein VAPB causes late-onset spinal muscular atrophy and amyotrophic lateral sclerosis Authors: Nishimura, A. L. Mitne-Neto, M. Silva, H. C. Richieri-Costa, A. Middleton, S. Cascio, D. Kok, F. Oliveira, J. R. Gillingwater, T. Webb, J. Skehel, P. Zatz, M. Auth Address: Human Genome Research Center, Department of Biology, Biosciences Institute, Sao Paulo University, Sao Paulo, Brazil. Pages: 822-31 Volume: 75 Number: 5 Keywords: Adult Amino Acid Sequence Amyotrophic Lateral Sclerosis/*genetics Brazil Calcium-Binding Proteins/*genetics Cells, Cultured Chromosome Mapping Chromosomes, Human, Pair 20/*genetics DNA Primers Founder Effect *Gene Expression Green Fluorescent Proteins Humans Kv Channel-Interacting Proteins Middle Aged Models, Molecular Molecular Sequence Data Muscular Atrophy, Spinal/*genetics Mutation, Missense/genetics Pedigree Protein Structure, Tertiary Research Support, Non-U.S. Gov't Sequence Alignment Sequence Analysis, DNA Vesicular Transport Proteins Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15372378 Abstract: Motor neuron diseases (MNDs) are a group of neurodegenerative disorders with involvement of upper and/or lower motor neurons, such as amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), progressive bulbar palsy, and primary lateral sclerosis. Recently, we have mapped a new locus for an atypical form of ALS/MND (atypical amyotrophic lateral sclerosis [ALS8]) at 20q13.3 in a large white Brazilian family. Here, we report the finding of a novel missense mutation in the vesicle-associated membrane protein/synaptobrevin-associated membrane protein B (VAPB) gene in patients from this family. Subsequently, the same mutation was identified in patients from six additional kindreds but with different clinical courses, such as ALS8, late-onset SMA, and typical severe ALS with rapid progression. Although it was not possible to link all these families, haplotype analysis suggests a founder effect. Members of the vesicle-associated proteins are intracellular membrane proteins that can associate with microtubules and that have been shown to have a function in membrane transport. These data suggest that clinically variable MNDs may be caused by a dysfunction in intracellular membrane trafficking. Year: 2004 Month: 11 Title: A natural history study of late onset spinal muscular atrophy types 3b and 4 Authors: Piepers, S. van den Berg, L. H. Brugman, F. Scheffer, H. Ruiterkamp-Versteeg, M. van Engelen, B. G. Faber, C. G. de Visser, M. van der Pol, W. L. Wokke, J. H. Auth Address: Dept. of Neurology, Rudolf Magnus Institute of Neuroscience, University Medical Center Utrecht, HP G 02.320, 85500, 3508, GA Utrecht, The Netherlands. s.piepers-2@umcutrecht.nl Pages: 1400-4 Volume: 255 Number: 9 Keywords: Adolescent Age of Onset Child Disease Progression Fatigue/epidemiology Female Follow-Up Studies Gene Dosage *Genetic Predisposition to Disease Humans Male Muscle Weakness/epidemiology Muscular Atrophy, Spinal/classification/epidemiology/*genetics Netherlands/epidemiology Prospective Studies Quality of Life Respiratory Insufficiency/epidemiology SMN Complex Proteins/*genetics Survival of Motor Neuron 1 Protein/genetics Time Factors Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18575920 Abstract: BACKGROUND: Spinal muscular atrophy (SMA) is caused by a homozygous deletion of the survival motor neuron (SMN)1 gene. The nearly identical SMN2 gene plays a disease modifying role. SMA is classified into four different subtypes based on age of onset and clinical course (SMA types 1-4). The natural history of early onset SMA types 1-3a has been studied extensively. Late onset SMA is rare and disease course has not been studied in detail. OBJECTIVE: To perform a prospective study on the clinical course and the correlation with SMN2 copy numbers of late onset SMA. METHODS: Patients fulfilling the diagnostic criteria for late onset SMA (types 3b and 4) were included in the study. At inclusion and follow-up, muscle strength, respiratory function, functional status and quality of life were assessed. SMN2 copy number was determined in all patients. RESULTS: Twelve patients were identified and included. Six patients were siblings from one family, two patients were brothers from a second family and four patients were sporadic cases. All patients carried four copies of the SMN2 gene. Median age of disease onset was 22.2 years (10-37). Age of disease onset in patients from family one was lower as compared to the other patients. None of the outcome measures changed after a follow-up of 2.5 years. Five patients reported an increase in fatigue and muscle weakness. None of the patients showed symptoms of respiratory insufficiency. CONCLUSIONS: This study indicates that late onset SMA is not characterized by disease progression and that alternative or surrogate disease markers are required for the design of future trials. This study confirms the finding that SMN2 copy number is a SMA disease course modifier. Year: 2008 Month: 9 Title: A negative element in SMN2 exon 7 inhibits splicing in spinal muscular atrophy Authors: Kashima, T. Manley, J. L. Auth Address: Department of Biological Sciences, Columbia University, New York, New York 10027, USA. Pages: 460-3 Volume: 34 Number: 4 Keywords: Base Sequence Cyclic AMP Response Element-Binding Protein Enhancer Elements (Genetics) Exons Hela Cells Heterogeneous-Nuclear Ribonucleoprotein Group A-B/metabolism Humans Models, Genetic Muscular Atrophy, Spinal/*genetics/metabolism Nerve Tissue Proteins/*genetics RNA/genetics/metabolism RNA Interference RNA Splicing/*genetics RNA-Binding Proteins Research Support, U.S. Gov't, P.H.S. Silencer Elements, Transcriptional Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12833158 Abstract: Spinal muscular atrophy (SMA) is a relatively common neurodegenerative disease caused by homozygous loss of the survival motor neuron 1 (SMN1) gene. Humans possess a linked, nearly identical gene, SMN2, which produces a functional SMN protein but at levels insufficient to compensate for loss of SMN1 (refs. 1,2). A C/T transition at position +6 in exon 7 is all that differentiates the two genes, but this is sufficient to prevent efficient exon 7 splicing in SMN2 (refs. 2,3). Here we show that the C/T transition functions not to disrupt an exonic splicing enhancer (ESE) in SMN1 (ref. 4), as previously suggested, but rather to create an exonic splicing silencer (ESS) in SMN2. We show that this ESS functions as a binding site for a known repressor protein, hnRNP A1, which binds to SMN2 but not SMN1 exon 7 RNA. We establish the physiological importance of these results by using small interfering RNAs to reduce hnRNP A protein levels in living cells and show that this results in efficient SMN2 exon 7 splicing. Our findings not only define a new mechanism underlying the inefficient splicing of SMN2 exon 7 but also illustrate more generally the remarkable sensitivity and precision that characterizes control of mRNA splicing. Year: 2003 Month: 8 Title: A Negatively-Acting Bifunctional RNA Increases Survival Motor Neuron in vitro and in vivo Authors: Dickson, A. Osman, E. Lorson, C. Auth Address: University of Missouri, Molecular Microbiology and Immunology, COLUMBIA, Missouri, United States; amtpn7@missouri.edu. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18724800 Abstract: Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder and is the leading genetic cause of infant mortality. SMA is caused by the loss of survival motor neuron-1 (SMN1). In humans, a nearly identical copy gene is present called SMN2, but this gene cannot compensate for the loss of SMN1 due to a single silent nucleotide difference in SMN2 exon 7. This single nucleotide difference attenuates an exonic splice enhancer, resulting in the production of an alternatively spliced isoform lacking exon 7, which is essential for protein function. SMN2, however, is a critical disease modifier and is an outstanding target for therapeutic intervention since all SMA patients retain SMN2 and SMN2 maintains the same coding sequence as SMN1. Therefore, compounds or molecules that increase SMN2 exon 7 inclusion hold great promise for SMA therapeutics. Bifunctional RNAs have been previously utilized to increase SMN protein levels and derive their name from the presence of two domains: an antisense RNA sequence specific to the target RNA and an untethered RNA segment that serves as a binding platform for splicing factors. This study was designed to develop negatively-acting bifunctional RNAs that recruit hnRNPA1 to exon 8 and block the general splicing machinery from the exon 8. By blocking the downstream splice site, this could competitively favor the inclusion of SMN exon 7 and therefore increase full-length SMN production. Here we identify a bifunctional RNA that stimulated full-length SMN expression in a variety of cell-based assays including SMA patient fibroblasts. Importantly, this molecule was also able to induce SMN expression in a previously described mouse model of SMA and demonstrates a novel therapeutic approach for SMA as well as a variety of diseases caused by a defect in splicing. Year: 2008 Title: A new splice site mutation in the SMN1 gene causes discrepant results in SMN1 deletion screening approaches Authors: Eggermann, T. Eggermann, K. Elbracht, M. Zerres, K. Rudnik-Schoneborn, S. Auth Address: Institute of Human Genetics, Technical University of Aachen, Pauwelsstr. 30, D-52074 Aachen, Germany. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18155522 Abstract: In most patients with infantile spinal muscular atrophy (SMA) both exons 7 and 8 of the SMN1 gene are deleted, but the deletion may also be restricted to exon 7. We report on an SMA type I patient who was initially diagnosed to be homozygous for an exon 7 deletion only. However, multiplex ligation-dependent probe amplification (MLPA) analyses revealed a heterozygous deletion of exons 7 and 8 of the SMN1 gene. By sequencing a new subtle splice site mutation (IVS6-2A>G) was identified. This variant affects the target sequence of oligonucleotides of all applied tests in a way that it has contrary effects on the efficiencies of the different assays. The results have major impacts on genetic counselling and carrier detection of the patient's paternal relatives. Year: 2007 Title: A non-sequence-specific requirement for SMN protein activity: the role of aminoglycosides in inducing elevated SMN protein levels Authors: Wolstencroft, E. C. Mattis, V. Bajer, A. A. Young, P. J. Lorson, C. L. Auth Address: Department of Veterinary Pathobiology, University of Missouri, Columbia, MO 65211-7310, USA. Pages: 1199-210 Volume: 14 Number: 9 Keywords: Alanine/metabolism Amikacin/pharmacology Amino Acid Substitution Aminoglycosides/*metabolism/pharmacology Anti-Bacterial Agents/pharmacology Antibodies, Monoclonal/metabolism Blotting, Western Cells, Cultured Codon, Terminator Cyclic AMP Response Element-Binding Protein/*genetics/*metabolism Dose-Response Relationship, Drug Epithelial Cells/drug effects/metabolism Exons Fibroblasts/drug effects/metabolism Fluorescein-5-isothiocyanate Fluorescent Antibody Technique Fluorescent Dyes Gene Deletion Hela Cells Homozygote Humans Immunohistochemistry Indoles Kinetics Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/deficiency/*genetics/*metabolism RNA-Binding Proteins/*genetics/*metabolism Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Tissue Distribution Tobramycin/pharmacology Transcription, Genetic Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15790598 Abstract: Spinal muscular atrophy (SMA) is caused by homozygous loss of the survival motor neuron (SMN1) gene. In virtually all SMA patients, a nearly identical copy gene is present, SMN2. SMN2 cannot fully compensate for the loss of SMN1 because the majority of transcripts derived from SMN2 lack a critical exon (exon 7), resulting in a dysfunctional SMN protein. Therefore, the critical distinction between a functional and a dysfunctional SMN protein is the inclusion or the exclusion of the exon 7 encoded peptide. To determine the role of the 16 amino acids encoded by SMN exon 7, a panel of synthetic mutations were transiently expressed in SMA patient fibroblasts and HeLa cells. Consistent with previous reports, the protein encoded by SMN exons 1-6 was primarily restricted to the nucleus. However, a variety of heterologous sequences fused to the C-terminus of SMN exons 1-6 allowed mutant SMN proteins to properly distribute to the cytoplasm and to the nuclear gems. These data demonstrate that the SMN exon 7 sequence is not specifically required, rather this region functions as a non-specific 'tail' that facilitates proper localization. Therefore, a possible means to restore additional activity to the SMNDelta7 protein could be to induce a longer C-terminus by suppressing recognition of the native stop codon. To address this possibility, aminoglycosides were examined for their ability to restore detectable levels of SMN protein in SMA patient fibroblasts. Aminoglycosides can suppress the accurate identification of translation termination codons in eukaryotic cells. Consistent with this, treatment of SMA patient fibroblasts with tobramycin and amikacin resulted in a quantitative increase in SMN-positive gems and an overall increase in detectable SMN protein. Taken together, this work describes the role of the critical exon 7 region and identifies a possible alternative approach for therapeutic intervention. Year: 2005 Title: A novel association of the SMN protein with two major non-ribosomal nucleolar proteins and its implication in spinal muscular atrophy Authors: Lefebvre, S. Burlet, P. Viollet, L. Bertrandy, S. Huber, C. Belser, C. Munnich, A. Auth Address: INSERM U393, IRNEM Institute, Hopital Necker-Enfants Malades, 149 Rue de Sevres, 75743 Paris Cedex 15, France. lefebvre@necker.fr Pages: 1017-27 Volume: 11 Number: 9 Keywords: Animals Antineoplastic Agents/pharmacology COS Cells/metabolism Cell Nucleolus/physiology Cells, Cultured Cyclic AMP Response Element-Binding Protein Fibroblasts/metabolism Fluorescent Antibody Technique, Indirect Frameshift Mutation Humans Immunoblotting Microscopy, Confocal Mimosine/pharmacology Muscular Atrophy, Spinal/*metabolism Nerve Tissue Proteins/genetics/*metabolism Nocodazole/pharmacology Nuclear Proteins/*metabolism Nucleolus Organizer Region/metabolism Phosphoproteins/*metabolism Precipitin Tests RNA-Binding Proteins/*metabolism Recombinant Fusion Proteins/metabolism Research Support, Non-U.S. Gov't Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11978761 Abstract: Spinal muscular atrophy (SMA) is caused by the loss of functional survival motor neuron 1 (SMN1) protein. This ubiquitously expressed protein is a component of a novel complex immunodetected in both the cytoplasm and the nucleus, which is associated with complexes involved in mRNA splicing, ribosome biogenesis and transcription. Here, we study a mutant protein corresponding to the N-terminal half of the protein that is encoded by the SMA frameshift mutation SMN 472del5. We show by confocal microscopy that the resulting mutant protein exhibits various distribution patterns in different transiently transfected COS cells. The mutant distributes into the nucleoplasm and/or the nucleolus, whereas the normal SMN protein accumulates at discrete nucleocytoplasmic dot-like structures previously named gems/Cajal bodies. The cell population with the nucleolar distribution is enriched upon treatment with mimosine, a synchronizing drug in late G(1) phase. Co-immunoprecipitation studies carried out on nuclear extracts reveal that both the endogenous SMN and mutant proteins are associated with complexes containing two major non-ribosomal nucleolar proteins, namely nucleolin and protein B23, and that the association is mediated, by among other things, RNA moieties. Both the association of the SMN protein with nucleolin-containing complexes and the nucleolin/B23 complex are disrupted in fibroblasts derived from a type I SMA patient harboring a homozygous SMN1 gene deletion. These findings suggest that altered assembly and/or stability of ribonucleoprotein complexes may contribute to the pathophysiological processes in SMA. Year: 2002 Title: A novel cell immunoassay to measure survival of motor neurons protein in blood cells Authors: Kolb, S. J. Gubitz, A. K. Olszewski, R. F., Jr. Ottinger, E. Sumner, C. J. Fischbeck, K. H. Dreyfuss, G. Auth Address: Howard Hughes Medical Institute And Department of Biochemistry & Biophysics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6148, USA. skolb@mail.med.upenn.edu Pages: 6 Volume: 6 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16451734 Abstract: BACKGROUND: The motor neuron degenerative disease spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality and is caused by mutations in the survival of motor neurons (SMN) gene that reduce the expression levels of the SMN protein. A major goal of current therapeutic approaches is to increase SMN levels in SMA patients. The purpose of this study was to develop a reliable assay to measure SMN protein levels from peripheral blood samples. METHODS: We developed a novel cell immunoassay to quantitatively measure SMN levels from peripheral blood mononuclear cells (PBMCs) using a single anti-SMN antibody. RESULTS: SMN levels determined by the cell immunoassay are comparable to levels determined by Western blot, but in contrast, the immunoassay does not involve cell lysis, requires a small amount of patient material, and can be done on a large number of samples simultaneously. SMN levels from PBMCs are not influenced by cell type heterogeneity. CONCLUSION: SMN levels measured from total PBMCs provide an important snapshot of SMN protein expression, which should be a useful aid in SMA diagnosis, and a surrogate marker of efficacy of treatment in SMA clinical trials. Year: 2006 Month: 1 Title: A novel function for SMN, the spinal muscular atrophy disease gene product, in pre-mRNA splicing Authors: Pellizzoni, L. Kataoka, N. Charroux, B. Dreyfuss, G. Auth Address: Howard Hughes Medical Institute and Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia 19104-6148, USA. Pages: 615-24 Volume: 95 Number: 5 Keywords: Autoantigens/metabolism Cyclic AMP Response Element-Binding Protein Hela Cells Humans Microscopy, Fluorescence Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/genetics/*physiology Nuclear Proteins/metabolism Open Reading Frames Organelles/metabolism/ultrastructure Phosphoproteins/metabolism RNA Precursors/*genetics *RNA Splicing RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Ribonucleoproteins, Small Nuclear/metabolism Spliceosomes/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9845364 Abstract: Spinal muscular atrophy (SMA) is a common motor neuron degenerative disease that results from reduced levels of, or mutations in, the Survival of Motor Neurons (SMN) protein. SMN is found in the cytoplasm and the nucleus where it is concentrated in gems. SMN interacts with spliceosomal snRNP proteins and is critical for snRNP assembly in the cytoplasm. We show that a dominant-negative mutant SMN (SMNdeltaN27) causes a dramatic reorganization of snRNPs in the nucleus. Furthermore, SMNdeltaN27 inhibits pre-mRNA splicing in vitro, while wild-type SMN stimulates splicing. SMN mutants found in SMA patients cannot stimulate splicing. These findings demonstrate that SMN plays a crucial role in the generation of the pre-mRNA splicing machinery and thus in mRNA biogenesis, and they link the function of SMN in this pathway to SMA. Year: 1998 Title: A novel method for oral delivery of drug compounds to the neonatal SMNDelta7 mouse model of spinal muscular atrophy Authors: Butchbach, M. E. Edwards, J. D. Schussler, K. R. Burghes, A. H. Auth Address: Department of Molecular and Cellular Biochemistry, College of Medicine, Ohio State University, 363 Hamilton Hall, 1645 Neil Avenue, Columbus, OH 43210, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17161463 Abstract: Spinal muscular atrophy (SMA) is a devastating motor neuron disease that is one of the leading genetic causes of infant mortality. Currently, there is no cure for SMA. Mouse models that genetically and phenotypically resemble SMA have been generated and have the potential to be used for the discovery of novel therapeutics. Oral administration is a commonly used mode of drug delivery in humans as well as in rodents. Unfortunately, there is no method of drug delivery that can accurately and reliably deliver drug compounds orally to neonatal mice. In this report, we describe a novel method to orally administer compounds to neonatal SMA mice. Oral delivery to neonatal mice, usually starting at postnatal day 4 (PND04), is both rapid and safe to the pup. Oral delivery of two different commonly used vehicle formulations, distilled water and 2-hydroxypropyl-beta-cyclodextrin, does not affect the survival of SMA mice. After oral delivery for 3 days, 5-bromo-2'-deoxyuridine could be detected in the kidneys, brains and spinal cords of treated non-SMA as well as SMA neonatal pups. In conclusion, we have developed a method by which drugs can be safely and reliably administered orally to neural targets of neonatal mice. This approach offers a simple and rapid means by which potential therapeutics for SMA can be identified. Year: 2006 Title: A novel nuclear structure containing the survival of motor neurons protein Authors: Liu, Q. Dreyfuss, G. Auth Address: Howard Hughes Medical Institute, University of Pennsylvania School of Medicine, Philadelphia 19104-6148, USA. Pages: 3555-65 Volume: 15 Number: 14 Keywords: 3T3 Cells Animals Antibodies, Monoclonal Antibody Specificity Cell Line Cell Nucleus/metabolism/*ultrastructure Chromosomal Proteins, Non-Histone/metabolism Cyclic AMP Response Element-Binding Protein Fluorescent Antibody Technique, Indirect Hela Cells Heterogeneous-Nuclear Ribonucleoprotein U Heterogeneous-Nuclear Ribonucleoproteins Humans Mice Mice, Inbred A Nerve Tissue Proteins/*metabolism RNA Processing, Post-Transcriptional RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Ribonucleoproteins/metabolism Ribonucleoproteins, Small Nuclear/metabolism Spliceosomes/metabolism Xenopus Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8670859 Abstract: Spinal muscular atrophy (SMA) is a common, often fatal, autosomal recessive disease leading to progressive muscle wasting and paralysis as a result of degeneration of anterior horn cells of the spinal cord. A gene termed survival of motor neurons (SMN), at 5q13, has been identified as the determining gene of SMA (Lefebvre et al., 1995). The SMN gene is deleted in > 98% of SMA patients, but the function of the SMN protein is unknown. In searching for hnRNP-interacting proteins we found that SMN interacts with the RGG box region of hnRNP U, with itself, with fibrillarin and with several novel proteins. We have produced monoclonal antibodies to the SMN protein, and we report here on its striking cellular localization pattern. Immunolocalization studies using SMN monoclonal antibodies show several intense dots in HeLa cell nuclei. These structures are similar in number (2-6) and size (0.1-1.0 micron) to coiled bodies, and frequently are found near or associated with coiled bodies. We term these prominent nuclear structures gems, for Gemini of coiled bodies. Year: 1996 Title: A novel peroxisome proliferator-activated receptor (PPAR)gamma agonist, NIP-222, reduces urinary albumin excretion in streptozotocin-diabetic mice independent of PPARgamma activation Authors: Yotsumoto, T. Naitoh, T. Kanaki, T. Matsuda, M. Tsuruzoe, N. Auth Address: Biological Research Laboratories, Nissan Chemical Industries Ltd, Minamisaitama, Saitama, Japan. Pages: 1633-7 Volume: 52 Number: 12 Keywords: Albuminuria/complications/*drug therapy Angiotensin-Converting Enzyme Inhibitors/pharmacology Animals Blood Glucose/metabolism Blood Pressure/drug effects Body Weight/drug effects Captopril/pharmacology Chromans/pharmacology Diabetes Mellitus, Experimental/*complications/urine Diabetes Mellitus, Type 2/genetics/metabolism Diabetic Nephropathies/metabolism/pathology Genes, Reporter/genetics Hypoglycemic Agents/*pharmacology Kidney/drug effects/pathology Luciferases/genetics Male Mice Organ Size/drug effects Plasmids/genetics Receptors, Cytoplasmic and Nuclear/*agonists/*metabolism Thiazolidinediones/pharmacology Transcription Factors/*agonists/*metabolism Urodynamics/drug effects Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14669169 Abstract: NIP-222 is a novel peroxisome proliferator-activated receptor (PPAR)gamma agonist. This study provides evidence that NIP-222 decreases urinary albumin excretion (UAE) in diabetic mice independent of its PPARgamma activation. We compared the effect of NIP-222 and another PPARgamma agonist, troglitazone, on UAE, plasma glucose level, blood pressure, and creatinine clearance (C(cr)) in streptozotocin (STZ)-induced diabetic mice. Treatment for 3 weeks with NIP-222 (30 mg/kg) was associated with a significant decrease in UAE without any change in blood pressure, creatinine clearance, or plasma glucose level. In contrast, UAE did not decrease in mice treated with troglitazone (300 mg/kg). These results indicate that NIP-222 has PPARgamma independent effects on UAE in diabetic mice and suggest that this agent may have potential to minimize the development and progression of diabetic nephropathy. Year: 2003 Month: 12 Title: A novel, efficient, randomized selection trial comparing combinations of drug therapy for ALS Authors: Gordon, P. H. Cheung, Y. K. Levin, B. Andrews, H. Doorish, C. Macarthur, R. B. Montes, J. Bednarz, K. Florence, J. Rowin, J. Boylan, K. Mozaffar, T. Tandan, R. Mitsumoto, H. Kelvin, E. A. Chapin, J. Bedlack, R. Rivner, M. McCluskey, L. F. Pestronk, A. Graves, M. Sorenson, E. J. Barohn, R. J. Belsh, J. M. Lou, J. S. Levine, T. Saperstein, D. Miller, R. G. Scelsa, S. N. Auth Address: Department of Neurology, Columbia University, New York, USA. PHG8@columbia.edu Pages: 212-22 Volume: 9 Number: 4 Keywords: Adult Aged Aged, 80 and over Amyotrophic Lateral Sclerosis/*drug therapy Creatine/administration & dosage/*therapeutic use Double-Blind Method Drug Eruptions Drug Therapy, Combination Female Humans Male Middle Aged Minocycline/administration & dosage/adverse effects/*therapeutic use Neuroprotective Agents/administration & dosage/*therapeutic use Patient Selection Pyrazoles/administration & dosage/*therapeutic use Sulfonamides/administration & dosage/*therapeutic use Treatment Outcome Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18608093 Abstract: Combining agents with different mechanisms of action may be necessary for meaningful results in treating ALS. The combinations of minocycline-creatine and celecoxib-creatine have additive effects in the murine model. New trial designs are needed to efficiently screen the growing number of potential neuroprotective agents. Our objective was to assess two drug combinations in ALS using a novel phase II trial design. We conducted a randomized, double-blind selection trial in sequential pools of 60 patients. Participants received minocycline (100 mg)-creatine (10 g) twice daily or celecoxib (400 mg)-creatine (10 g) twice daily for six months. The primary objective was treatment selection based on which combination best slowed deterioration in the ALS Functional Rating Scale-Revised (ALSFRS-R); the trial could be stopped after one pool if the difference between the two arms was adequately large. At trial conclusion, each arm was compared to a historical control group in a futility analysis. Safety measures were also examined. After the first patient pool, the mean six-month decline in ALSFRS-R was 5.27 (SD=5.54) in the celecoxib-creatine group and 6.47 (SD=9.14) in the minocycline-creatine group. The corresponding decline was 5.82 (SD=6.77) in the historical controls. The difference between the two sample means exceeded the stopping criterion. The null hypothesis of superiority was not rejected in the futility analysis. Skin rash occurred more frequently in the celecoxib-creatine group. In conclusion, the celecoxib-creatine combination was selected as preferable to the minocycline-creatine combination for further evaluation. This phase II design was efficient, leading to treatment selection after just 60 patients, and can be used in other phase II trials to assess different agents. Year: 2008 Month: 8 Title: A phase 1 trial of riluzole in spinal muscular atrophy Authors: Russman, B. S. Iannaccone, S. T. Samaha, F. J. Auth Address: Oregon Health Sciences University and Shriners Hospital for Children, Portland, OR 97201, USA. brussman@shrinenet.org Pages: 1601-3 Volume: 60 Number: 11 Keywords: Age of Onset Child Child, Preschool Excitatory Amino Acid Antagonists/adverse effects/*therapeutic use Humans Infant Infant, Newborn Muscular Atrophy, Spinal/*drug therapy/mortality Research Support, Non-U.S. Gov't Riluzole/adverse effects/*therapeutic use Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14623733 Abstract: BACKGROUND: Severe spinal muscular atrophy (SMA) (Werdnig-Hoffmann disease, acute SMA, and SMA I) is a disease of the motor neuron characterized by onset before 6 months of age, failure ever to achieve sitting without support, and a life expectancy of 2 years or less. There is no known treatment for SMA, and, until recently, no therapeutic trials have been attempted. There is reason to believe that glutamate, an excitatory neurotransmitter, enhances programmed cell death of anterior horn cells. Riluzole, a glutamate inhibitor, has been shown to slow the rate of decline in patients with amyotrophic lateral sclerosis, another form of motor neuron disease. OBJECTIVES: To determine whether a glutamate inhibitor might be tolerated by infants with SMA and, furthermore, whether this medication could have a positive effect on life expectancy. DESIGN: Subjects with homozygous deletions of the survival motor neuron gene were recruited from pediatric neuromuscular clinics and randomized in a 2:1 ratio, 2 riluzole to 1 placebo. Neurologic examination was performed at the first visit by one of the investigators. Complete blood count, hepatic and renal screens, and urinalysis were performed at baseline, 2 weeks, 1 month, 2 months, 3 months, 6 months, and 9 months after drug or placebo was started. An electrocardiogram was done at baseline, 3 months, 6 months, and 12 months. Treatment was stopped after 9 months, and blood work was repeated at 12 months. Treatment was reinstituted at 1 year if requested by the parents. The enrollment goal was 30 patients; however, support from the pharmaceutical company was withdrawn when Rhone-Poulenc Rorer was taken over by Aventis. The investigational review boards of the participating centers approved the protocol and consent forms. RESULTS: Seven patients received riluzole and 3 received placebo medication. All 3 patients in the placebo group died (mean age, 9 months). Three of 7 who received active drug are still living at ages 513 years, 4 years, and 30 months. None of the 10 subjects experienced adverse effects or changes in laboratory test results. None showed any change in motor abilities. CONCLUSIONS: Riluzole appears to be safe in young children. This was a limited study with insufficient power to show a difference between the 2 groups. Because there is a suggestion of possible benefit in treated subjects, we recommend further study of riluzole in pediatric patients with SMA. Year: 2003 Month: 11 Title: A placebo-controlled trial of gabapentin in spinal muscular atrophy Authors: Miller, R. G. Moore, D. H. Dronsky, V. Bradley, W. Barohn, R. Bryan, W. Prior, T. W. Gelinas, D. F. Iannaccone, S. Kissel, J. Leshner, R. Mendell, J. Mendoza, M. Russman, B. Samaha, F. Smith, S. Auth Address: Department of Neurology, California Pacific Medical Center, Forbes Norris MSDA/ALS Center, 2324 Sacramento Street, #150, San Francisco, CA 94115, USA. rmiller@cooper.cpmc.org Pages: 127-31 Volume: 191 Number: 1-2 Keywords: Acetic Acids/*therapeutic use Adult *Amines Arm/physiopathology *Cyclohexanecarboxylic Acids Double-Blind Method Excitatory Amino Acid Antagonists/*therapeutic use Feasibility Studies Female Hand Strength Humans Male Muscle Contraction/drug effects Research Support, Non-U.S. Gov't Severity of Illness Index Sickness Impact Profile Spinal Muscular Atrophies of Childhood/*drug therapy/physiopathology Treatment Outcome United States Vital Capacity/drug effects *gamma-Aminobutyric Acid Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11677003 Abstract: OBJECTIVE: To evaluate the efficacy of gabapentin in increasing muscle strength of patients with spinal muscular atrophy (SMA). BACKGROUND: Preclinical data in experimental models of motor neuron disease suggest a neuroprotective effect of gabapentin. METHODS: Gabapentin (1200 mg), or placebo, was administered three times daily in a randomized, double-blind trial for 12 months. The primary outcome measure was the average percent change from baseline, based on the measurement of strength in four muscles (elbow flexion and hand grip bilaterally) for each patient. Drug efficacy was examined by comparing the percent change in strength for patients on drug vs. placebo. Secondary efficacy variables included: forced vital capacity (FVC), SMA functional rating scale (SMAFRS), and mini-Sickness Impact Profile (SIP). RESULTS: Eighty-four patients, with type II or III SMA, were enrolled at eight sites across the United States. There were no differences in baseline features. There was no difference between the placebo and drug groups in any outcome measure. CONCLUSIONS: This study demonstrates the feasibility of this trial design and provides data for the design of future clinical trials in SMA. Year: 2001 Title: A population-based study of genotypic and phenotypic variability in children with spinal muscular atrophy Authors: Arkblad, E. Tulinius, M. Kroksmark, A. K. Henricsson, M. Darin, N. Auth Address: Department of Clinical Genetics, Sahlgrenska University Hospital, Goteborg, Sweden. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19154529 Abstract: Aims: To describe the occurrence of spinal muscular atrophy (SMA) in childhood; to evaluate if any of the genes in the SMA region on chromosome 5q13 correlates with disease severity; to make genotype-phenotype correlations; to evaluate the variability of different disease alleles in carriers and the sensitivity of multiplex ligation-dependent probe amplification (MLPA) for detecting carriers. Methods: In a population-based study from Western Sweden MLPA was used to determine the copy-numbers of several genes in the SMA region (SMN1, SMN2, BIRC1, GTF2H2 and SERF1A) in SMA-patients and their parents. Results: We estimated the incidence of SMN1-related SMA in childhood at 1 in 11 800 live births and confirmed the relationship between the number of SMN2 copies and the severity of disease. No other direct relationships were found. All but one of the analysed parents were confirmed as carriers by MLPA analysis. A total of at least 30 different disease alleles were identified and no specific disease allele represented more than 15% of the total. Conclusion: The childhood incidence of SMA in the Swedish population is around 1 in 12 000 live births and it is unlikely that there is any founder effect involved in SMA in western Sweden. Year: 2009 Title: A predominantly nuclear protein affecting cytoplasmic localization of beta-actin mRNA in fibroblasts and neurons Authors: Gu, W. Pan, F. Zhang, H. Bassell, G. J. Singer, R. H. Auth Address: Department of Anatomy, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Pages: 41-51 Volume: 156 Number: 1 Keywords: Actins/*genetics Amino Acid Sequence Animals Blotting, Western Brain/cytology/metabolism Chick Embryo Chickens/genetics Chromatography, Affinity Cloning, Molecular Cytoplasm/*genetics/*metabolism DNA, Complementary/genetics Electrophoretic Mobility Shift Assay Fibroblasts/*metabolism Gene Expression Regulation, Developmental Humans In Situ Hybridization Molecular Sequence Data Mutation/genetics Neurons/*metabolism Nuclear Proteins/chemistry/genetics/isolation & purification/*metabolism Protein Binding Protein Structure, Tertiary RNA, Messenger/genetics/*metabolism RNA-Binding Proteins/chemistry/genetics/isolation & purification/*metabolism Research Support, U.S. Gov't, P.H.S. *Trans-Activators Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11781334 Abstract: The localization of beta-actin mRNA to the leading lamellae of chicken fibroblasts and neurite growth cones of developing neurons requires a 54-nt localization signal (the zipcode) within the 3' untranslated region. In this study we have identified and isolated five proteins binding to the zipcode. One of these we previously identified as zipcode binding protein (ZBP)1, a 4-KH domain protein. A second is now investigated in detail: a 92-kD protein, ZBP2, that is especially abundant in extracts from embryonic brain. We show that ZBP2 is a homologue of the human hnRNP protein, KSRP, that appears to mediate pre-mRNA splicing. However, ZBP2 has a 47-amino acid (aa) sequence not present in KSRP. Various portions of ZBP2 fused to GFP indicate that the protein most likely shuttles between the nucleus and the cytoplasm, and that the 47-aa insert promotes the nuclear localization. Expression of a truncated ZBP2 inhibits the localization of beta-actin mRNA in both fibroblast and neurons. These data suggest that ZBP2, although predominantly a nuclear protein, has a role in the cytoplasmic localization of beta-actin mRNA. Year: 2002 Title: A preliminary report on spinal muscular atrophy lymphoblastoid cell lines: Are they an appropriate tool for drug screening? Authors: Dayangac-Erden, D. Topaloglu, H. Erdem-Yurter, H. Auth Address: Department of Medical Biology, Hacettepe University, Faculty of Medicine, Ankara, Turkey, didayan@hacettepe.edu.tr. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18345520 Abstract: INTRODUCTION: Spinal muscular atrophy (SMA) is a neurodegenerative disease of the motor neurons that results in progressive muscle weakness. It is also the leading hereditary cause of infant mortality. Homozygous loss of the survival motor neuron (SMN1) gene causes SMA, and the number of copies of the SMN2 gene modulates the severity of the disease. Increasing the expression of the SMN2 gene by pharmacological agents is one of the therapeutic approaches currently being implemented. METHODS: In this preliminary study, we investigated the effect of phenylbutyrate, a histone deacetylase (HDAC) inhibitor, on SMN2 expression in two SMA type III Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines to understand the suitability of lymphoblastoid cell lines in drug screening. These cell lines are regarded as a good source as they can easily be established from the peripheral leucocytes of patients. Quantitative analysis of SMN2 mRNA was performed on established cell lines treated with various concentrations of phenylbutyrate and for a range of incubation periods using real-time polymerase chain reaction. Western blot analysis was used to determine SMN protein levels. RESULTS: Real-time polymerase chain reaction and Western blot analysis demonstrated that the levels of SMN2 full-length (fl-SMN2) transcripts and protein were not increased in phenylbutyrate-treated cell lines compared to non-treated controls. CONCLUSION: These results suggest that EBV-transformed lymphoblastoid cell lines are not suitable for studying the effect of certain HDAC inhibitors on SMN2 gene expression. Year: 2008 Title: A presynaptic giant ankyrin stabilizes the NMJ through regulation of presynaptic microtubules and transsynaptic cell adhesion Authors: Pielage, J. Cheng, L. Fetter, R. D. Carlton, P. M. Sedat, J. W. Davis, G. W. Auth Address: Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158-2822, USA. Pages: 195-209 Volume: 58 Number: 2 Keywords: Amino Acid Motifs/physiology Animals Animals, Genetically Modified Ankyrins/*genetics Cell Adhesion Molecules, Neuronal/metabolism Drosophila Drosophila Proteins/*genetics Gene Expression Regulation/genetics Horseradish Peroxidase/metabolism Larva Luminescent Proteins/genetics/metabolism Microscopy, Electron, Transmission/methods Microtubules/*metabolism Mutation/genetics Neuromuscular Junction/*physiology/ultrastructure Presynaptic Terminals/*metabolism/ultrastructure Synapsins/metabolism Synaptic Transmission/genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18439405 Abstract: In a forward genetic screen for mutations that destabilize the neuromuscular junction, we identified a novel long isoform of Drosophila ankyrin2 (ank2-L). We demonstrate that loss of presynaptic Ank2-L not only causes synapse disassembly and retraction but also disrupts neuronal excitability and NMJ morphology. We provide genetic evidence that ank2-L is necessary to generate the membrane constrictions that normally separate individual synaptic boutons and is necessary to achieve the normal spacing of subsynaptic protein domains, including the normal organization of synaptic cell adhesion molecules. Mechanistically, synapse organization is correlated with a lattice-like organization of Ank2-L, visualized using extended high-resolution structured-illumination microscopy. The stabilizing functions of Ank2-L can be mapped to the extended C-terminal domain that we demonstrate can directly bind and organize synaptic microtubules. We propose that a presynaptic Ank2-L lattice links synaptic membrane proteins and spectrin to the underlying microtubule cytoskeleton to organize and stabilize the presynaptic terminal. Year: 2008 Title: A randomized study of alglucosidase alfa in late-onset Pompe's disease Authors: van der Ploeg, A. T. Clemens, P. R. Corzo, D. Escolar, D. M. Florence, J. Groeneveld, G. J. Herson, S. Kishnani, P. S. Laforet, P. Lake, S. L. Lange, D. J. Leshner, R. T. Mayhew, J. E. Morgan, C. Nozaki, K. Park, D. J. Pestronk, A. Rosenbloom, B. Skrinar, A. van Capelle, C. I. van der Beek, N. A. Wasserstein, M. Zivkovic, S. A. Auth Address: Department of Pediatrics, Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, Rotterdam, The Netherlands. a.vanderploeg@erasmusmc.nl Pages: 1396-406 Volume: 362 Number: 15 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20393176 Abstract: BACKGROUND: Pompe's disease is a metabolic myopathy caused by a deficiency of acid alpha glucosidase (GAA), an enzyme that degrades lysosomal glycogen. Late-onset Pompe's disease is characterized by progressive muscle weakness and loss of respiratory function, leading to early death. We conducted a randomized, placebo-controlled trial of alglucosidase alfa, a recombinant human GAA, for the treatment of late-onset Pompe's disease. METHODS: Ninety patients who were 8 years of age or older, ambulatory, and free of invasive ventilation were randomly assigned to receive biweekly intravenous alglucosidase alfa (20 mg per kilogram of body weight) or placebo for 78 weeks at eight centers in the United States and Europe. The two primary end points were distance walked during a 6-minute walk test and percentage of predicted forced vital capacity (FVC). RESULTS: At 78 weeks, the estimated mean changes from baseline in the primary end points favored alglucosidase alfa (an increase of 28.1+/-13.1 m on the 6-minute walk test and an absolute increase of 3.4+/-1.2 percentage points in FVC; P=0.03 and P=0.006, respectively). Similar proportions of patients in the two groups had adverse events, serious adverse events, and infusion-associated reactions; events that occurred only in patients who received the active study drug included anaphylactic reactions and infusion-associated reactions of urticaria, flushing, hyperhidrosis, chest discomfort, vomiting, and increased blood pressure (each of which occurred in 5 to 8% of the patients). CONCLUSIONS: In this study population, treatment with alglucosidase alfa was associated with improved walking distance and stabilization of pulmonary function over an 18-month period. (ClinicalTrials.gov number, NCT00158600.) Title: A randomized, double-blind, futility clinical trial of creatine and minocycline in early Parkinson disease Authors: Auth Address: Pages: 664-71 Volume: 66 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16481597 Abstract: BACKGROUND: Creatine and minocycline were prioritized for testing in Phase II clinical trials based on a systematic evaluation of potentially disease modifying compounds for Parkinson disease (PD). OBJECTIVE: To test whether creatine and minocycline alter the course of early PD relative to a predetermined futility threshold for progression of PD in a randomized, double-blind, Phase II futility clinical trial. Agents that do not perform better than the futility threshold are rejected as futile and are not considered for further study. METHODS: Participants had a diagnosis of PD within 5 years, but did not require medications for the management of symptoms. The primary outcome was the change in the total Unified Parkinson's Disease Rating Scale (UPDRS) score from baseline to either the time when there was sufficient disability to warrant symptomatic therapy for PD or 12 months, whichever came first. Subjects were randomized 1:1:1 to receive creatine 10 g/day, minocycline 200 mg/day, or matching placebo. The futility threshold was set as a 30% reduction in UPDRS progression based on the placebo/tocopherol arm of the Deprenyl And Tocopherol Antioxidative Therapy Of Parkinsonism (DATATOP) trial. p values < or = 0.1 indicate futility. RESULTS: Two hundred subjects were randomized to the three groups. Neither creatine (p = 0.96) nor minocycline (p = 0.66) could be rejected as futile based on the DATATOP futility threshold. The rate of progression for the calibration placebo group fell outside the 95% CI for the DATATOP historical control. In a sensitivity analysis, based on the threshold derived from the calibration placebo group, again neither drug could be rejected as futile. Tolerability was 91% in the creatine group and 77% in the minocycline group. Common adverse events included upper respiratory symptoms (26%), joint pain (19%), and nausea (17%). CONCLUSIONS: Both creatine and minocycline should be considered for definitive Phase III trials to determine if they alter the long term progression of Parkinson disease (PD). Additional factors must be weighed before selecting agents for Phase III trials, including safety, tolerability, activity, cost, and availability of these two agents in comparison with other agents currently in development for PD. Year: 2006 Title: A rare SMN2 variant in a previously unrecognized composite splicing regulatory element induces exon 7 inclusion and reduces the clinical severity of spinal muscular atrophy Authors: Vezain, M. Saugier-Veber, P. Goina, E. Touraine, R. Manel, V. Toutain, A. Fehrenbach, S. Frebourg, T. Pagani, F. Tosi, M. Martins, A. Auth Address: Inserm U614, IFRMP, Institute for Biomedical Research, Faculty of Medicine, University of Rouen, 76183 Rouen, France. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19953646 Abstract: Spinal muscular atrophy (SMA) is a common neuromuscular disorder caused by homozygous inactivation of the SMN1 (Survival Motor Neuron 1) gene. The disease severity is mainly influenced by the copy number of SMN2, a nearly identical gene from which only low amounts of full-length mRNA are produced. This correlation is not absolute, suggesting the existence of yet unknown factors modulating disease progression. We identified and characterized the rare variant c.859G>C (p.Gly287Arg) in exon 7 in both SMN2 copies of a male patient affected with type III SMA, a milder form of the disease rarely associated with only two SMN2 copies. We demonstrated in vivo, in this patient and in a second unrelated patient, and ex vivo, using SMN splicing assays, that the variant induces inclusion of exon 7 into SMN2 mRNA. Moreover, we show that the c.859G>C variation is located in a composite splicing regulatory element in the centre of exon 7. The variation does not affect binding of HTra2a nor creates a novel SF2/ASF enhancer, but disrupts an hnRNP A1 binding site. The natural occurrence of enhanced inclusion of SMN2 exon 7 in milder SMA cases supports the current therapeutic strategies based on splicing modulation in SMA patients. (c) 2009 Wiley-Liss, Inc. Year: 2009 Title: A retrospective, multinational, multicenter study on the natural history of infantile-onset Pompe disease Authors: Kishnani, P. S. Hwu, W. L. Mandel, H. Nicolino, M. Yong, F. Corzo, D. Auth Address: Division of Medical Genetics, Department of Pediatrics, Duke University Medical Center, Durham, NC 27710, USA. kishn001@mc.duke.edu Pages: 671-676 Volume: 148 Number: 5 Keywords: Cohort Studies Disease Progression Europe/epidemiology Female Glycogen Storage Disease Type II/*complications/*epidemiology/therapy Humans Infant Infant, Newborn Israel/epidemiology Male North America/epidemiology Respiration, Artificial Retrospective Studies Survival Rate Taiwan/epidemiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16737883 Abstract: OBJECTIVE: To characterize the natural progression of infantile-onset Pompe disease. STUDY DESIGN: Retrospective chart reviews of 168 patients with documented acid alpha-glucosidase deficiency and symptom onset by 12 months of age; Kaplan-Meier analysis of total and ventilator-free survival time; Cox proportional hazards regression modeling of mortality risk factors. RESULTS: The median age at symptom onset was 2.0 months (range 0 to 12 months), 4.7 months at diagnosis (range: prenatal to 4.2 months), 5.9 months at first ventilator support (range 0.1 to 31.1 months), and 8.7 months at death (range 0.3 to 73.4 months). Survival rates at 12 months of age were 25.7% overall and 16.9% ventilator-free; at 18 months 12.3% and 6.7%. Cardiomegaly (92%), hypotonia (88%), cardiomyopathy (88%), respiratory distress (78%), muscle weakness (63%), feeding difficulties (57%), and failure to thrive (53%) appeared after a median age of approximately 4.0 months. Multiple covariate analysis confirmed that early symptom onset increased risk of early death. CONCLUSION: Despite frequent therapeutic interventions, infantile-onset Pompe disease remains lethal. Year: 2006 Month: 5 Title: A role for Caenorhabditis elegans in understanding the function and interactions of human disease genes Authors: Culetto, E. Sattelle, D. B. Auth Address: MRC Functional Genetics Unit, Department of Human Anatomy and Genetics, University of Oxford, UK. Pages: 869-77 Volume: 9 Number: 6 Keywords: Amino Acid Sequence Animals Caenorhabditis elegans/*genetics Disease Models, Animal Humans *Ligases Molecular Sequence Data Muscular Atrophy, Spinal/genetics Muscular Dystrophies Polycystic Kidney Diseases/genetics Proteins/chemistry/genetics Research Support, Non-U.S. Gov't Sequence Homology, Amino Acid *Ubiquitin-Protein Ligases Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10767309 Abstract: A growing number of medical research teams have begun to explore the experimental advantages of using a genetic animal model, the nematode worm Caenorhabditis elegans, with a view to enhancing our understanding of genes underlying human congenital disorders. In this study, we have compared sequences of positionally cloned human disease genes with the C.elegans database of predicted genes. Drawing on examples from spinal muscular atrophy, polycystic kidney disease, muscular dystrophy and Alzheimer's disease, we illustrate how data from C.elegans can yield new insights into the function and interactions of human disease genes. Year: 2000 Title: A role for complexes of survival of motor neurons (SMN) protein with gemins and profilin in neurite-like cytoplasmic extensions of cultured nerve cells Authors: Sharma, A. Lambrechts, A. Hao le, T. Le, T. T. Sewry, C. A. Ampe, C. Burghes, A. H. Morris, G. E. Auth Address: Biochemistry Group, N. E. Wales Institute, Mold Road, Wrexham LL11 2AW, UK. Pages: 185-97 Volume: 309 Number: 1 Keywords: Amino Acid Sequence Animals Cyclic AMP Response Element-Binding Protein/genetics/*metabolism Mice Mice, Inbred BALB C Molecular Sequence Data Motor Neurons/*metabolism/pathology/ultrastructure Muscular Atrophy, Spinal/metabolism/pathology Neoplasm Proteins/metabolism Nerve Tissue Proteins/genetics/*metabolism Neurites/*metabolism/ultrastructure Nuclear Proteins/*metabolism PC12 Cells Profilins/*metabolism RNA-Binding Proteins/genetics/*metabolism Rabbits Rats Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15975577 Abstract: Spinal muscular atrophy (SMA) is caused by reduced levels of SMN (survival of motor neurons protein) and consequent loss of motor neurons. SMN is involved in snRNP transport and nuclear RNA splicing, but axonal transport of SMN has also been shown to occur in motor neurons. SMN also binds to the small actin-binding protein, profilin. We now show that SMN and profilin II co-localise in the cytoplasm of differentiating rat PC12 cells and in neurite-like extensions, especially at their growth cones. Many components of known SMN complexes were also found in these extensions, including gemin2 (SIP-1), gemin6, gemin7 and unrip (unr-interacting protein). Coilin p80 and Sm core protein immunoreactivity, however, were seen only in the nucleus. SMN is known to associate with beta-actin mRNA and specific hnRNPs in axons and in neurite extensions of cultured nerve cells, and SMN also stimulates neurite outgrowth in cultures. Our results are therefore consistent with SMN complexes, rather than SMN alone, being involved in the transport of actin mRNPs along the axon as in the transport of snRNPs into the nucleus by similar SMN complexes. Antisense knockdown of profilin I and II isoforms inhibited neurite outgrowth of PC12 cells and caused accumulation of SMN and its associated proteins in cytoplasmic aggregates. BIAcore studies demonstrated a high affinity interaction of SMN with profilin IIa, the isoform present in developing neurons. Pathogenic missense mutations in SMN, or deletion of exons 5 and 7, prevented this interaction. The interaction is functional in that SMN can modulate actin polymerisation in vitro by reducing the inhibitory effect of profilin IIa. This suggests that reduced SMN in SMA might cause axonal pathfinding defects by disturbing the normal regulation of microfilament growth by profilins. Year: 2005 Title: A sensitive assay for measuring SMN mRNA levels in peripheral blood and in muscle samples of patients affected with spinal muscular atrophy Authors: Vezain, M. Saugier-Veber, P. Melki, J. Toutain, A. Bieth, E. Husson, M. Pedespan, J. M. Viollet, L. Penisson-Besnier, I. Fehrenbach, S. Bou, J. Frebourg, T. Tosi, M. Auth Address: 1Department of Genetics, Rouen University Hospital and INSERM U614, Institute for Biomedical Research, University of Rouen, Rouen, France. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17609673 Abstract: Different therapeutic strategies are currently evaluated in spinal muscular atrophy (SMA) that are aimed at increasing full-length (FL) mRNA levels produced from the SMN2 gene. Assays measuring SMN mRNA levels are needed. We have developed a sensitive, comparative assay based on multiplex fluorescent reverse-transcription polymerase chain reaction (RT-PCR) that can measure, in the same reaction, the levels of SMN mRNA with and without exon 7 sequences as well as those of total SMN mRNA. This assay allows to calculate directly the ratios of FL SMN mRNA to SMN mRNA without exon 7 (Delta7). We have used this assay to compare the levels of SMN transcripts in the blood of 75 unrelated normal subjects and of 48 SMA patients, and in muscle samples of 8 SMA patients. The SMN1 and the SMN2 genes produced very similar levels of total mRNA. Levels of transcripts lacking exon 7 were linearly dependent on the number of SMN2 copies, both in SMA patients and in controls. In patients, FL mRNA levels correlated with SMN2 copy number. A significant but weaker inverse correlation was also observed between FL or Delta7 mRNA levels and disease severity, but patients with three SMN2 copies and different SMA types displayed similar mRNA levels. A significantly higher FL to Delta7 ratio was measured in blood cells than in skeletal muscle (0.80+/-0.18 versus 0.47+/-0.11). This assay can be used as a sensitive biomarker for monitoring the effects of various drugs in forthcoming clinical trials of SMA.European Journal of Human Genetics advance online publication, 4 July 2007; doi:10.1038/sj.ejhg.5201885. Year: 2007 Title: A short antisense oligonucleotide masking a unique intronic motif prevents skipping of a critical exon in spinal muscular atrophy Authors: Singh, N. N. Shishimorova, M. Cao, L. C. Gangwani, L. Singh, R. N. Auth Address: Department of Biomedical Sciences, Iowa State University, Ames, IA, USA. Pages: Volume: 6 Number: 3 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19430205 Abstract: Spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality. Most SMA cases are associated with the low levels of SMN owing to deletion of Survival Motor Neuron 1 (SMN1). SMN2, a nearly identical copy of SMN1, fails to compensate for the loss of SMN1 due to predominant skipping of exon 7. Hence, correction of aberrant splicing of SMN2 exon 7 holds the potential for cure of SMA. Here we report an 8-mer antisense oligonucleotide (ASO) to have a profound stimulatory response on correction of aberrant splicing of SMN2 exon 7 by binding to a unique GC-rich sequence located within intron 7 of SMN2. We confirm that the splicing-switching ability of this short ASO comes with a high degree of specificity and reduced off-target effect compared to larger ASOs targeting the same sequence. We further demonstrate that a single low nanomolar dose of this 8-mer ASO substantially increases the levels of SMN and a host of factors including Gemin 2, Gemin 8, ZPR1, hnRNP Q and Tra2-beta1 known to be down-regulated in SMA. Our findings underscore the advantages and unmatched potential of very short ASOs in splicing modulation in vivo. Year: 2009 Title: A simple multiplex real-time PCR methodology for the SMN1 gene copy number quantification Authors: Passon, N. Pozzo, F. Molinis, C. Bregant, E. Gellera, C. Damante, G. Lonigro, R. I. Auth Address: Dipartimento di Scienze e Tecnologie Biomediche, Universita degli Studi, Udine, Italy. Pages: 37-42 Volume: 13 Number: 1 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19309272 Abstract: Spinal muscular atrophy (SMA) is an autosomal recessive disease caused, in about 95% of SMA cases, by homozygous deletion of the survival motor neuron 1 (SMN1) gene or its conversion to the highly homologous SMN2 gene. The molecular diagnosis of SMA is usually carried out by a PCR-Restriction fragment length polymorphism (RFLP) approach. However, this approach is not useful for identification of healthy deletion carriers. TaqMan technology is one of the most reliable and widely adopted techniques for the SMN1 copy number evaluation. However, several limitations of this technique have been described. Particularly, DNA extraction methods and accurate template quantification have been shown to be critical for reliable results. In this work, we set up a reliable, highly reproducible, and easy-to-perform TaqMan technology-based protocol to obtain the SMN1 gene copy number assessment. We demonstrate that PCR amplification of both target gene and reference gene in the same reaction mix, instead of separated mixes, greatly reduces reported criticisms of simplex TaqMan technology. The multiplex real-time PCR we describe allows interlaboratory samples and data exchange, without the need to equalize the DNA isolation technique. Further, the protocol described below requires fewer replica tests than the simplex methodology does, leading to reduced overall cost for the diagnostic assay. Year: 2009 Month: 2 Title: A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays Authors: Zhang, J. H. Chung, T. D. Oldenburg, K. R. Auth Address: DuPont Pharmaceuticals Research Laboratories, Leads Discovery, DuPont Pharmaceuticals Company, Wilmington, Delaware. Pages: 67-73 Volume: 4 Number: 2 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10838414 Abstract: The ability to identify active compounds ((3)hits(2)) from large chemical libraries accurately and rapidly has been the ultimate goal in developing high-throughput screening (HTS) assays. The ability to identify hits from a particular HTS assay depends largely on the suitability or quality of the assay used in the screening. The criteria or parameters for evaluating the (3)suitability(2) of an HTS assay for hit identification are not well defined and hence it still remains difficult to compare the quality of assays directly. In this report, a screening window coefficient, called (3)Z-factor,(2) is defined. This coefficient is reflective of both the assay signal dynamic range and the data variation associated with the signal measurements, and therefore is suitable for assay quality assessment. The Z-factor is a dimensionless, simple statistical characteristic for each HTS assay. The Z-factor provides a useful tool for comparison and evaluation of the quality of assays, and can be utilized in assay optimization and validation. Year: 1999 Month: 1 Title: A single nucleotide difference that alters splicing patterns distinguishes the SMA gene SMN1 from the copy gene SMN2 Authors: Monani, U. R. Lorson, C. L. Parsons, D. W. Prior, T. W. Androphy, E. J. Burghes, A. H. McPherson, J. D. Auth Address: Department of Neurology, The Ohio State University, Columbus 43210, USA. Pages: 1177-83 Volume: 8 Number: 7 Keywords: *Alternative Splicing Cyclic AMP Response Element-Binding Protein Genotype Humans Molecular Sequence Data Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/*genetics Point Mutation RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sequence Analysis, DNA Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10369862 Abstract: Spinal muscular atrophy (SMA) is a recessive disorder characterized by loss of motor neurons in the spinal cord. It is caused by mutations in the telomeric survival motor neuron 1 ( SMN1 ) gene. Alterations within an almost identical copy gene, the centromeric survival motor neuron 2 ( SMN2 ) gene produce no known phenotypic effect. The exons of the two genes differ by just two nucleotides, neither of which alters the encoded amino acids. At the genomic level, only five nucleotides that differentiate the two genes from one another have been reported. The entire genomic sequence of the two genes has not been determined. Thus, differences which might explain why SMN1 is the SMA gene are not readily apparent. In this study, we have completely sequenced and compared genomic clones containing the SMN genes. The two genes show striking similarity, with the homology being unprecedented between two different yet functional genes. The only critical difference in an approximately 32 kb region between the two SMN genes is the C->T base change 6 bp inside exon 7. This alteration but not other variations in the SMN genes affects the splicing pattern of the genes. The majority of the transcript from the SMN1 locus is full length, whereas the majority of the transcript produced by the SMN2 locus lacks exon 7. We suggest that the exon 7 nucleotide change affects the activity of an exon splice enhancer. In SMA patients, the loss of SMN1 but the presence of SMN2 results in low levels of full-length SMN transcript and therefore low SMN protein levels which causes SMA. Year: 1999 Month: 7 Title: A SMN missense mutation complements SMN2 restoring snRNPs and rescuing SMA mice Authors: Workman, E. Saieva, L. Carrel, T. L. Crawford, T. O. Liu, D. Lutz, C. Beattie, C. E. Pellizzoni, L. Burghes, A. H. Auth Address: Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH, 43210 USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19329542 Abstract: Spinal muscular atrophy (SMA) is an autosomal recessive neurodegenerative disease. Loss of the survival motor neuron (SMN1) gene, in the presence of the SMN2 gene causes SMA. SMN functions in snRNP assembly in all cell types, however, it is unclear how this function results in specifically motor neuron cell death. Lack of endogenous mouse SMN (Smn) in mice results in embryonic lethality. Introduction of 2 copies of human SMN2 results in a mouse with severe SMA, while 1 copy of SMN2 is insufficient to overcome embryonic lethality. We show that SMN(A111G), an allele capable of snRNP assembly, can rescue mice that lack Smn and contain either one or two copies of SMN2 (SMA mice). The correction of SMA in these animals was directly correlated with snRNP assembly activity in spinal cord, as was correction of snRNA levels. These data support snRNP assembly as being the critical function affected in SMA and suggests that the levels of snRNPs are critical to motor neurons. Furthermore, SMN(A111G) cannot rescue Smn-/- mice without SMN2 suggesting that both SMN(A111G) and SMN from SMN2 undergo intragenic complementation in vivo to function in heteromeric complexes that have greater function than either allele alone. The oligomer composed of limiting full-length SMN and SMN(A111G) has substantial snRNP assembly activity. Also, the SMN(A2G) and SMN(A111G) alleles in vivo did not complement each other leading to the possibility that these mutations could affect the same function. Year: 2009 Title: A SMNDelta7 read-through product confers functionality to the SMNDelta7 protein Authors: Mattis, V. B. Bowerman, M. Kothary, R. Lorson, C. L. Auth Address: Department of Veterinary Pathobiology, Life Sciences Center, Room 404, University of Missouri, Columbia, MO 65211, United States. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18601974 Abstract: Spinal muscular atrophy (SMA) affects about 1 in every 6000 children born and is the leading genetic cause of infant death. SMA is a recessive disorder caused by the mutation or deletion of Survival Motor Neuron-1 (SMN1). SMN2, a nearly identical copy gene, has the potential to encode the same protein as SMN1 and is retained in all SMA patients. The majority of SMN2-derived transcripts are alternatively spliced and therefore encode a truncated isoform lacking exon 7 (SMNDelta7), which is a defective protein because it is unstable, has a reduced ability to self-associate and is unable to efficiently function in SMN cellular activities. However, we have shown that the SMN C-terminus functions non-specifically, since heterologous sequences can compensate for the exon 7 sequence. Several classes of compounds identified in SMN-inducing high throughput screens have been proposed to function through a read-through mechanism; however, a functional analysis of the SMNDelta7 read-through product has not been performed. In this report, the SMNDelta7 read-through product is characterized and compared to the SMNDelta7 protein. In a series of in vitro and cell based assays, SMNDelta7 read-through product is shown to increase protein stability, promote neurite outgrowths in SMN deficient neurons, and significantly elevate SMN-dependent UsnRNP assembly in extracts from SMA patient fibroblasts. Collectively, these results demonstrate that SMNDelta7 read-through product is more active than the SMNDelta7 protein and suggest that SMA therapeutics that specifically induce SMNDelta7 read-through may provide an alternative platform for drug discovery. Year: 2008 Title: A spatio-temporal analysis of motoneuron survival, axonal regeneration and neurotrophic factor expression after lumbar ventral root avulsion and implantation Authors: Eggers, R. Tannemaat, M. R. Ehlert, E. M. Verhaagen, J. Auth Address: Laboratory for Neuroregeneration, Netherlands Institute for Neuroscience, An Institute of the Royal Academy of Arts and Sciences, Meibergdreef 47, 1105 BA, Amsterdam, The Netherlands. r.eggers@nin.knaw.nl Pages: 207-20 Volume: 223 Number: 1 Keywords: Animals Cell Survival/physiology Choline O-Acetyltransferase/metabolism Disease Models, Animal Enzyme-Linked Immunosorbent Assay/methods Female Gene Expression Regulation/*physiology Hindlimb/pathology Lumbosacral Plexus/pathology/physiopathology Motor Neurons/*pathology Muscle, Skeletal/physiopathology Nerve Growth Factors/genetics/*metabolism Nerve Regeneration/*physiology *Radiculopathy/pathology/physiopathology/surgery Rats Rats, Wistar Replantation/*methods Sciatic Nerve/metabolism/pathology Time Factors Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19646436 Abstract: Reimplantation of avulsed rat lumbar spinal ventral roots results in poor recovery of function of the denervated hind limb muscles. In contrast, reimplantation of cervical or sacral ventral roots is a successful repair strategy that results in a significant degree of regeneration. A possible explanation for this difference could be that following lumbar root avulsion, axons have to travel longer distances towards their target muscles, resulting in prolonged denervation of the distal nerve and a diminished capacity to support regeneration. Here we present a detailed spatio-temporal analysis of motoneuron survival, axonal regeneration and neurotrophic factor expression following unilateral avulsion and implantation of lumbar ventral roots L3, L4, and L5. Reimplantation prolongs the survival of motoneurons up to one month post-lesion. The first regenerating motor axons entered the reimplanted ventral roots during the first week and large numbers of fibers gradually enter the lumbar plexus between 2 and 4 weeks, indicating that axons enter the reimplanted roots and plexus over an extended period of time. However, motor axon counts show that relatively few axons reach the distal sciatic nerve in the 16 week post-lesion period. The observed initial increase and subsequent decline in expression of glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor correlate with the apparent spatio-temporal decline in the regenerative capacity of motor axons, indicating that the distal nerve is losing its capacity to support regenerating motor axons following prolonged denervation. These findings have important implications for future strategies to promote long-distance regeneration through distal, chronically denervated peripheral nerves. Month: 5 Title: A splicing component adapted to gene silencing Authors: Roca, X. Krainer, A. R. Auth Address: Pages: 250-1 Volume: 27 Number: 3 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19270672 Abstract: Year: 2009 Month: 3 Title: A survival motor neuron:tetanus toxin fragment C fusion protein for the targeted delivery of SMN protein to neurons Authors: Francis, J. W. Figueiredo, D. vanderSpek, J. C. Ayala, L. M. Kim, Y. S. Remington, M. P. Young, P. J. Lorson, C. L. Ikebe, S. Fishman, P. S. Brown, R. H., Jr. Auth Address: Cecil B. Day Laboratory for Neuromuscular Research, Massachusetts General Hospital, Building 114, 16th Street, Room 3003, Charlestown, MA 02129, USA. jwfrancis@partners.org Pages: 84-96 Volume: 995 Number: 1 Keywords: Animals Animals, Newborn Antibodies/immunology Cell Membrane/drug effects/immunology/metabolism Cells, Cultured Cyclic AMP Response Element-Binding Protein Cytotoxins/genetics/immunology/pharmacology Diphtheria Toxin/genetics/immunology Dose-Response Relationship, Drug Endocytosis/immunology Immunohistochemistry Muscular Atrophy, Spinal/*drug therapy/genetics/metabolism Nerve Tissue Proteins/*genetics/immunology Neurons/drug effects/metabolism Peptide Fragments/*genetics/immunology Protein Binding/immunology Protein Structure, Tertiary/physiology Protein Transport/drug effects/immunology RNA-Binding Proteins Rats Recombinant Fusion Proteins/genetics/immunology/*pharmacology Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Tetanus Toxin/*genetics/immunology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14644474 Abstract: Spinal muscular atrophy (SMA) is a degenerative disorder of spinal motor neurons caused by homozygous mutations in the survival motor neuron (SMN1) gene. Because increased tissue levels of human SMN protein (hSMN) in transgenic mice reduce the motor neuron loss caused by murine SMN knockout, we engineered a recombinant SMN fusion protein to deliver exogenous hSMN to the cytosolic compartment of motor neurons. The fusion protein, SDT, is comprised of hSMN linked to the catalytic and transmembrane domains of diphtheria toxin (DTx) followed by fragment C of tetanus toxin (TTC). Following overexpression in Escherichia coli, SDT possessed a subunit molecular weight of approximately 130 kDa as revealed by both SDS-PAGE and immunoblot analyses with anti-SMN, anti-DTx, and anti-TTC antibodies. Like wild-type SMN, purified SDT showed specific binding in vitro to an RG peptide derived from Ewing's sarcoma protein. The fusion protein also bound to cultured primary neurons in amounts similar to those achieved by TTC. Unlike the case with TTC, however, immunolabeling of SDT-treated neurons with anti-TTC and anti-SMN antibodies showed staining restricted to the cell surface. Results from cytotoxicity studies in which the DTx catalytic domain of SDT was used as a reporter protein for internalization and membrane translocation activity suggest that the SMN moiety of the fusion protein is interfering with one or both of these processes. While these studies indicate that SDT may not be useful for SMA therapy, the use of the TTC:DTx fusion construct to deliver other passenger proteins to the neuronal cytosol should not be ruled out. Year: 2004 Title: A transgene carrying an A2G missense mutation in the SMN gene modulates phenotypic severity in mice with severe (type I) spinal muscular atrophy Authors: Monani, U. R. Pastore, M. T. Gavrilina, T. O. Jablonka, S. Le, T. T. Andreassi, C. DiCocco, J. M. Lorson, C. Androphy, E. J. Sendtner, M. Podell, M. Burghes, A. H. Auth Address: Department of Neurology, Ohio State University, Columbus, OH 43210, USA. monani.2@osu.edu Pages: 41-52 Volume: 160 Number: 1 Keywords: Animals Axons/metabolism Blotting, Southern Blotting, Western Cyclic AMP Response Element-Binding Protein DNA, Complementary/metabolism Dose-Response Relationship, Drug Electromyography Electrophysiology Genotype Glutathione Transferase/metabolism Homozygote Immunohistochemistry Mice Mice, Knockout Mice, Transgenic Models, Biological Models, Genetic Motor Neurons/pathology Muscles/cytology/metabolism/pathology Muscular Atrophy, Spinal/*genetics Mutation *Mutation, Missense Nerve Tissue Proteins/*genetics Phenotype Protein Binding RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Reverse Transcriptase Polymerase Chain Reaction Time Factors Tissue Distribution *Transgenes Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12515823 Abstract: 5q spinal muscular atrophy (SMA) is a common autosomal recessive disorder in humans and the leading genetic cause of infantile death. Patients lack a functional survival of motor neurons (SMN1) gene, but carry one or more copies of the highly homologous SMN2 gene. A homozygous knockout of the single murine Smn gene is embryonic lethal. Here we report that in the absence of the SMN2 gene, a mutant SMN A2G transgene is unable to rescue the embryonic lethality. In its presence, the A2G transgene delays the onset of motor neuron loss, resulting in mice with mild SMA. We suggest that only in the presence of low levels of full-length SMN is the A2G transgene able to form partially functional higher order SMN complexes essential for its functions. Mild SMA mice exhibit motor neuron degeneration, muscle atrophy, and abnormal EMGs. Animals homozygous for the mutant transgene are less severely affected than heterozygotes. This demonstrates the importance of SMN levels in SMA even if the protein is expressed from a mutant allele. Our mild SMA mice will be useful in (a) determining the effect of missense mutations in vivo and in motor neurons and (b) testing potential therapies in SMA. Year: 2003 Title: A two-site ELISA can quantify upregulation of SMN protein by drugs for spinal muscular atrophy Authors: Thi Man, N. Humphrey, E. Lam, L. T. Fuller, H. R. Lynch, T. A. Sewry, C. A. Goodwin, P. R. Mackenzie, A. E. Morris, G. E. Auth Address: From the Wolfson Centre for Inherited Neuromuscular Disease (N.t.M., E.H., L.T.L., H.R.F., T.A.L., C.A.S., G.E.M.), Oswestry, UK; Hallmark Analytical Ventures (P.R.G.), Saltney, Flintshire, UK; Children's Hospital of Eastern Ontario Research Institute (A.E.M.), Ottawa, Canada; and Institute for Science and Technology in Medicine (G.E.M.), Keele University, UK. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18633133 Abstract: OBJECTIVES: Spinal muscular atrophy (SMA) is an autosomal recessive disorder characterized by loss of lower motor neurons during early or postnatal development. Severity is variable and is inversely related to the levels of survival of motor neurons (SMN) protein. The aim of this study was to produce a two-site ELISA capable of measuring both the low, basal levels of SMN protein in cell cultures from patients with severe SMA and small increases in these levels after treatment of cells with drugs. METHODS: A monoclonal antibody against recombinant SMN, MANSMA1, was selected for capture of SMN onto microtiter plates. A selected rabbit antiserum against refolded recombinant SMN was used for detection of the captured SMN. RESULTS: The ratio of SMN levels in control fibroblasts to levels in SMA fibroblasts was greater than 3.0, consistent with Western blot data. The limit of detection was 0.13 ng/mL and SMN could be measured in human NT-2 neuronal precursor cells grown in 96-well culture plates (3 x 10(4) cells per well). Increases in SMN levels of 50% were demonstrable by ELISA after 24 hours treatment of 10(5) SMA fibroblasts with valproate or phenylbutyrate. CONCLUSION: A rapid and specific two-site, 96-well ELISA assay, available in kit format, can now quantify the effects of drugs on survival of motor neurons protein levels in cell cultures. Year: 2008 Title: A two-stage design for a phase II clinical trial of coenzyme Q10 in ALS Authors: Levy, G. Kaufmann, P. Buchsbaum, R. Montes, J. Barsdorf, A. Arbing, R. Battista, V. Zhou, X. Mitsumoto, H. Levin, B. Thompson, J. L. Auth Address: Department of Biostatistics, Mailman School of Public Health, Columbia University, New York, NY 10032, USA. GL227@columbia.edu Pages: 660-3 Volume: 66 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16534103 Abstract: BACKGROUND: The combination of a small pool of patients at any given time with the availability of many potential neuroprotective agents to be tested in ALS requires efficient phase II trial designs. OBJECTIVE: To describe the design of the Clinical Trial of High Dose Coenzyme Q10 (CoQ10) in ALS (QALS study)--a phase II, randomized, placebo-controlled, double-blind, multicenter clinical trial. METHODS: The study design features two stages. The first stage (dose selection) identifies which of two doses of CoQ10 (1800 mg or 2700 mg) is preferred using a selection procedure rather than a formal hypothesis test. The second stage (early efficacy test) compares the preferred dose of CoQ10 against placebo using a non-superiority or futility design. Data from patients assigned to the preferred dose of CoQ10 in the first stage are also used in the second stage. The primary outcome measure is the decline in Amyotrophic Lateral Sclerosis Functional Rating Scale-revised (ALSFRSr) score from baseline to 9 months. RESULTS: The total sample size required is 185 patients, as compared to a much larger sample size estimated to be necessary using a conventional superiority design (total: 852 patients). The authors report a bias correction made necessary by the inclusion of patient data from the first stage in the second stage. CONCLUSIONS: Several features of the Clinical Trial of High Dose Coenzyme Q10 in ALS study design promote efficiency. These features may be beneficial in phase II trials in amyotrophic lateral sclerosis and other fields. Year: 2006 Title: Aberrant E2F activation by polyglutamine expansion of androgen receptor in SBMA neurotoxicity Authors: Suzuki, E. Zhao, Y. Ito, S. Sawatsubashi, S. Murata, T. Furutani, T. Shirode, Y. Yamagata, K. Tanabe, M. Kimura, S. Ueda, T. Fujiyama, S. Lim, J. Matsukawa, H. Kouzmenko, A. P. Aigaki, T. Tabata, T. Takeyama, K. Kato, S. Auth Address: Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan. Pages: 3818-22 Volume: 106 Number: 10 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19237573 Abstract: Spinal and bulbar muscular atrophy (SBMA) is a neurodegenerative disorder caused by a polyglutamine repeat (polyQ) expansion within the human androgen receptor (AR). Unlike other neurodegenerative diseases caused by abnormal polyQ expansion, the onset of SBMA depends on androgen binding to mutant human polyQ-AR proteins. This is also observed in Drosophila eyes ectopically expressing the polyQ-AR mutants. We have genetically screened mediators of androgen-induced neurodegeneration caused by polyQ-AR mutants in Drosophila eyes. We identified Rbf (Retinoblastoma-family protein), the Drosophila homologue of human Rb (Retinoblastoma protein), as a neuroprotective factor. Androgen-dependent association of Rbf or Rb with AR was remarkably potentiated by aberrant polyQ expansion. Such potentiated Rb association appeared to attenuate recruitment of histone deacetyltransferase 1 (HDAC1), a corepressor of E2F function. Either overexpression of Rbf or E2F deficiency in fly eyes reduced the neurotoxicity of the polyQ-AR mutants. Induction of E2F function by polyQ-AR-bound androgen was suppressed by Rb in human neuroblastoma cells. We conclude that abnormal expansion of polyQ may potentiate innate androgen-dependent association of AR with Rb. This appears to lead to androgen-dependent onset of SBMA through aberrant E2F transactivation caused by suppressed histone deacetylation. Year: 2009 Title: Aberrant RNA processing events in neurological disorders Authors: Anthony, K. Gallo, J. M. Auth Address: MRC Centre for Neurodegeneration Research, King's College London, Institute of Psychiatry, Department of Clinical Neuroscience, PO37, De Crespigny Park, London, SE5 8AF, United Kingdom. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20226177 Abstract: The importance of aberrant RNA processing in neurodegeneration is becoming increasingly clear; a recent example being the identification of the splicing factor TDP-43 as the major component of inclusions characteristic of a number of neurodegenerative conditions including amyotrophic lateral sclerosis (ALS). Due to the enormous diversity generated by alternative splicing and its importance in the nervous system, it is no surprise that defective alternative splicing in disease has been particularly well documented. However, in addition to splicing, other RNA processing events such as RNA editing, polyadenylation and mRNA stability are also disrupted in some neurological disorders. For instance: the editing efficiency of specific ionotropic receptors is reduced in ALS affecting ion permeability and the function of RNA-processing proteins is affected by their sequestration to trinucleotide repeat expanded mRNAs in several disorders. Due to the extensive coupling between RNA processing events and the multifunctionality of the RNA processing factors that regulate them it is important to consider RNA processing as a whole. Here we review RNA processing events, their extensive coupling to one another and detail the associations of RNA processing including, but not exclusively, alternative splicing with neurodegeneration. Title: Abnormal fatty acid metabolism in childhood spinal muscular atrophy Authors: Crawford, T. O. Sladky, J. T. Hurko, O. Besner-Johnston, A. Kelley, R. I. Auth Address: Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Pages: 337-43 Volume: 45 Number: 3 Keywords: Age Distribution Child Child, Preschool Fatty Acids/*metabolism Humans Infant Lauric Acids/metabolism Muscular Atrophy, Spinal/*metabolism Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10072048 Abstract: Our previous experience with abnormal fatty acid metabolism in several children with spinal muscular atrophy (SMA) prompted evaluation of fatty acid metabolism in a larger cohort. Thirty-three infants with severe infantile SMA were shown to have a significantly increased ratio of dodecanoic to tetradecanoic acid in plasma compared with normal infants and 6 infants affected with equally debilitating, non-SMA denervating disorders. Seventeen children with milder forms of SMA had normal fatty acid profiles. In addition, all 5 infants with severe SMA evaluated in a fasting state developed a distinctive and marked dicarboxylic aciduria, including saturated, unsaturated, and 3-hydroxy forms, comparable in severity with the dicarboxylic aciduria of children with primary defects of mitochondrial fatty acid beta-oxidation. Nine children with chronic SMA and 23 control patients did not develop an abnormal dicarboxylic aciduria during fasting. No known disorder of fatty acid metabolism explains all of the abnormalities we find in SMA. Our data suggest, however, that the abnormalities are not a consequence of SMA-related immobility, systemic illness, muscle denervation, or muscle atrophy. These abnormalities in fatty acid metabolism may be caused by changes in cellular physiology related to the molecular defects of the SMA-pathogenic survival motor neuron gene or neighboring genes. Year: 1999 Month: 3 Title: Abnormal Mitochondrial Dynamics and Neurodegenerative Diseases Authors: Su, B. Wang, X. Zheng, L. Perry, G. Smith, M. A. Zhu, X. Auth Address: Department of Pathology, Case Western Reserve University, Cleveland, Ohio 44106, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19799998 Abstract: Mitochondrial dysfunction is a prominent feature of various neurodegenerative diseases. A deeper understanding of the remarkably dynamic nature of mitochondria, characterized by a delicate balance of fission and fusion, has helped to fertilize a recent wave of new studies demonstrating abnormal mitochondrial dynamics in neurodegenerative diseases. This review highlights mitochondrial dysfunction and abnormal mitochondrial dynamics in Alzheimer disease, Parkinson disease, amyotrophic lateral sclerosis, and Huntington disease and discusses how these abnormal mitochondrial dynamics may contribute to mitochondrial and neuronal dysfunction. We propose that abnormal mitochondrial dynamics represents a key common pathway that mediates or amplifies mitochondrial dysfunction and neuronal dysfunction during the course of neurodegeneration. Year: 2009 Title: Abnormal motoneuron migration, differentiation, and axon outgrowth in spinal muscular atrophy Authors: Simic, G. Mladinov, M. Seso Simic, D. Jovanov Milosevic, N. Islam, A. Pajtak, A. Barisic, N. Sertic, J. Lucassen, P. J. Hof, P. R. Kruslin, B. Auth Address: Department of Neuroscience, School of Medicine, Croatian Institute for Brain Research, Medical School Zagreb, Zagreb University, Salata 12, 10000 Zagreb, Croatia. gsimic@hiim.hr Pages: 313-26 Volume: 115 Number: 3 Keywords: Axons/metabolism/*pathology Brain/metabolism/pathology Cell Differentiation/*physiology Cell Movement/*physiology Humans Immunohistochemistry In Situ Nick-End Labeling Motor Neurons/metabolism/*pathology Muscle, Skeletal/metabolism/pathology Spinal Cord/metabolism/pathology Spinal Muscular Atrophies of Childhood/metabolism/*pathology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18075747 Abstract: The role of heterotopic (migratory) motoneurons (HMN) in the pathogenesis of spinal muscular atrophy (SMA) is still controversial. We examined the occurrence and amount of HMN in spinal cord tissue from eight children with SMA (six with SMA-I and two with SMA-II). All affected subjects were carrying a homozygous deletion of exon 7 in the SMN1 gene. Unlike controls, virtually free from HMN, all SMA subjects showed a significant number of HMN at all levels of the spinal cord. Heterotopic neurons were hyperchromatic, located mostly in the ventral white matter and had no axon or dendrites. More than half of the HMN were very undifferentiated, as judged from their lack of immunoreactivity for NeuN and MAP2 proteins. Small numbers of more differentiated heterotopic neurons were also found in the dorsal and lateral white matter region. As confirmed by ultrastructural analysis, in situ end labeling (ISEL) and CD68 immunoreactivity, HMN in the ventral outflow were found to have no synapses, to activate microglial cells, and to eventually die by necrosis. An unbiased quantitative analysis showed a significant negative correlation between age of SMA subjects (a reflection of the clinical severity) and the number of HMN. Subjects who died at older ages had increased number of GFAP-positive astrocytes. Complementing our previous report on motoneuron apoptosis within the ventral horns in SMA, we now propose that abnormal migration, differentiation, and lack of axonal outgrowth may induce motoneuron apoptosis predominantly during early stages, whereas a slower necrosis-like cell death of displaced motoneurons which "escaped" apoptosis characterizes later stages of SMA. Year: 2008 Month: 3 Title: Abnormal motor phenotype in the SMNDelta7 mouse model of spinal muscular atrophy Authors: Butchbach, M. E. Edwards, J. D. Burghes, A. H. Auth Address: Department of Molecular and Cellular Biochemistry, College of Medicine, The Ohio State University, Columbus, OH, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17561409 Abstract: Spinal muscular atrophy (SMA) is a recessive motor neuron disease that affects motor neurons in the anterior horn of the spinal cord. SMA results from the reduction of SMN (survival motor neuron) protein. Even though SMN is ubiquitously expressed, motor neurons are more sensitive to the reduction in SMN than other cell types. We have previously generated mouse models of SMA with varying degrees of clinical severity. So as to more clearly understand the pathogenesis of motor neuron degeneration in SMA, we have characterized the phenotype of the SMNDelta7 SMA mouse which normally lives for 13.6+/-0.7 days. These mice are smaller than their non-SMA littermates and begin to lose body mass at 10.4+/-0.4 days. SMNDelta7 SMA mice exhibit impaired responses to surface righting, negative geotaxis and cliff aversion but not to tactile stimulation. Spontaneous motor activity and grip strength are also significantly impaired in SMNDelta7 SMA mice. In summary, we have demonstrated an impairment of neonatal motor responses in SMNDelta7 SMA mice. This phenotype characterization could be used to assess the effectiveness of potential therapies for SMA. Year: 2007 Title: Abnormal serum concentrations of proteins in Parkinson's disease Authors: Goldknopf, I. L. Bryson, J. K. Strelets, I. Quintero, S. Sheta, E. A. Mosqueda, M. Park, H. R. Appel, S. H. Shill, H. Sabbagh, M. Chase, B. Kaldjian, E. Markopoulou, K. Auth Address: Power3 Medical Products, Inc., The Woodlands, TX 77381, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19723509 Abstract: Blood serum was used to identify protein biomarkers for diagnosis of Parkinson's disease (PD) using analytically validated quantitative 2D-gel electrophoresis, and single variable and multivariate statistics. Using banked samples from a first medical center, we identified 57 specific protein spot biomarkers with disease-specific abnormal levels in serum of patients with PD, Alzheimer's disease, amyotrophic lateral sclerosis and similar neurodegenerative conditions (337 samples), when compared to age-matched normal controls (132 samples). To further assess their clinical usefulness in Parkinson's disease, we obtained prospective newly drawn blood serum samples from a second (56 PD, 30 controls) and third (6 PD, 48 controls) medical center. The protein concentrations of the 57 biomarkers were assessed by 2D-gel electrophoresis. Stepwise linear discriminant analysis selected a combination of 21 of the 57 as optimal to distinguish PD patients from controls. When applied to the samples from the second site, the 21 proteins had sensitivity of 93.3% (52 of 56 PD correctly classified), specificity of 92.9% (28 of 30 controls correctly classified); 15 of 15 patients with mild, 28 of 30 with moderate to severe symptoms, and all of the 6 PD patients from the third site were correctly classified. Eleven of the 21 proteins showed statistically significant abnormal concentrations in patients with mild symptoms, and 14 in patients with moderate-severe symptoms. The protein identities reflect the heterogeneity of Parkinson's disease, and thus may provide the capability of monitoring the blood for a diverse range of PD pathophysiological mechanisms: cellular degeneration, oxidative stress, inflammation, and transport. Year: 2009 Title: Abnormal SMN1 gene copy number is a susceptibility factor for amyotrophic lateral sclerosis Authors: Corcia, P. Mayeux-Portas, V. Khoris, J. de Toffol, B. Autret, A. Muh, J. P. Camu, W. Andres, C. Auth Address: INSERM U316, Tours, France. Pages: 243-6 Volume: 51 Number: 2 Keywords: Amyotrophic Lateral Sclerosis/epidemiology/*genetics Cyclic AMP Response Element-Binding Protein Gene Dosage Genetic Predisposition to Disease Humans Nerve Tissue Proteins/*genetics RNA-Binding Proteins Risk Factors Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11835381 Abstract: The etiology of amyotrophic lateral sclerosis remains unknown in the majority of cases. Homozygous SMN1 (survival motor neuron) gene deletion causes spinal muscular atrophy, and SMN2 gene deletions are possible risk factors in lower motor neuron disease. We studied SMN1 and SMN2 genes copy numbers in 167 amyotrophic lateral sclerosis patients and in 167 matched controls. We noted that 16% of amyotrophic lateral sclerosis patients had an abnormal copy number of the SMN1 gene (1 or 3 copies), compared with 4% of controls. An abnormal SMN1 gene locus may be a susceptibility factor for amyotrophic lateral sclerosis. Year: 2002 Month: 2 Title: Abolishing Bax-Dependent Apoptosis Shows Beneficial Effects on Spinal Muscular Atrophy Model Mice Authors: Tsai, M. S. Chiu, Y. T. Wang, S. H. Hsieh-Li, H. M. Lian, W. C. Li, H. Auth Address: Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16564230 Abstract: Spinal muscular atrophy (SMA) is the most common genetic motoneuron degenerative disorder, but the mechanism(s) of motoneuron degeneration is unclear. We previously generated SMA model mice, which genotypically and phenotypically mimicked human SMA patients, by a combination of knockout and transgenic techniques. Here, we used these SMA model mice to decipher the apoptotic mechanism(s) involved in SMA motoneuron degeneration. We found a significant increase in proapoptotic Bax expression in the spinal cords of SMA mice in comparison with their wild-type littermates. After crossing SMA mice with Bax knockout mice, we produced in vivo evidence indicating that Bax protein plays an important role in the degeneration of SMA spinal motoneurons. Progeny Bax-deficient SMA mice showed milder disease severity, longer life spans, and significant increases in spinal motoneuron densities compared to SMA littermates with wild-type Bax genes. Our results strongly suggest that suppression of Bax-involved apoptosis has the potential for amelioration of SMA. Year: 2006 Title: Abolishing Trp53-dependent apoptosis does not benefit spinal muscular atrophy model mice Authors: Tsai, M. S. Chiu, Y. T. Wang, S. H. Hsieh-Li, H. M. Li, H. Auth Address: [1] 1Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan [2] 2Department of Bioindustry Technology, Da-Yeh University, Da-Tsuen, Chang-Hua, Taiwan. Pages: 372-5 Volume: 14 Number: 3 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16391561 Abstract: Spinal muscular atrophy (SMA) is the most common genetic motoneuron degenerative disorder, but the mechanism(s) of motoneuron death is unclear. Previously, a direct interaction between tumor-suppressive TP53 protein and the SMA determinant gene product, survival motor neuron protein, was identified and therefore it has been suggested that a mechanism of TP53-dependent apoptosis plays an important role in motoneuron degeneration in SMA. We used our SMA model mice, generated by a combination of knockout and transgenic techniques, to decipher the role of TP53 protein in the motoneuron degeneration in SMA. We detected a significant increase of Trp53 expression in the spinal cord of SMA-like mice compared to their normal littermates. After crossing SMA-like mice with Trp53 knockout mice, the progeny Trp53-deficient SMA-like mice did not show milder disease severity or longer lifespan compared to SMA littermates with wild-type Trp53 genes. Our studies provide in vivo evidence indicating that Trp53-dependent apoptosis does not play a crucial role in motoneuron degeneration in SMA-like mice.European Journal of Human Genetics (2006) 14, 372-375. doi:10.1038/sj.ejhg.5201556; published online 4 January 2006. Year: 2006 Month: 3 Title: AChR channel conversion and AChR-adjusted neuronal survival during embryonic development Authors: Yampolsky, P. Gensler, S. McArdle, J. Witzemann, V. Auth Address: Max-Planck-Institute for Medical Research, Department of Cell Physiology, Jahnstrasse 29, D-69120 Heidelberg, Germany. Pages: 634-45 Volume: 37 Number: 3 Keywords: Acetylcholinesterase/metabolism Animals Animals, Newborn Autoradiography Bungarotoxins/metabolism Cell Survival Embryo, Mammalian *Embryonic Development Gene Expression Regulation, Developmental/*physiology Humans Luminescent Proteins/genetics Mice Mice, Inbred C57BL Mice, Transgenic Motor Neurons/cytology/*physiology Muscle, Smooth/embryology/metabolism *Neuromuscular Junction/embryology/growth & development/metabolism Protein Binding/drug effects Protein Transport/genetics Receptors, Cholinergic/*physiology Receptors, Nicotinic/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18226545 Abstract: We generated knock-in mice that express GFP-labeled embryonic-type acetylcholine receptors (AChR) to follow postsynaptic differentiation and innervation during embryonic development and to visualize the postnatally occurring channel conversion from embryonic- to adult-type AChR. The dynamics of AChRgamma/AChRepsilon conversion at the neuromuscular junction indicates that muscle-specific programs of receptor subunit gene transcription control AChR replacement. While conversion proceeds from peripheral to central regions for individual endplates, it does not occur simultaneously for all endplates. Although GFP-labeled receptors were expressed at reduced levels, the localization of postsynaptic sites and synapse formation was not noticeably altered. However, these alterations correlated with a striking reduction of motoneuron programmed cell death, transient increase of neurite growth and axon branching. This animal model refines the view on reciprocal neuromuscular signaling and suggests that motoneuron survival and axon branching are directly regulated by AChR function to enable optimal innervation and timing of neurally evoked muscle contraction. Year: 2008 Month: 3 Title: Aclarubicin treatment restores SMN levels to cells derived from type I spinal muscular atrophy patients Authors: Andreassi, C. Jarecki, J. Zhou, J. Coovert, D. D. Monani, U. R. Chen, X. Whitney, M. Pollok, B. Zhang, M. Androphy, E. Burghes, A. H. Auth Address: Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, OH 43210, USA. Pages: 2841-9 Volume: 10 Number: 24 Keywords: Aclarubicin/*pharmacology Alternative Splicing/drug effects Animals Blotting, Western Cell Line Cyclic AMP Response Element-Binding Protein Dose-Response Relationship, Drug Drug Evaluation, Preclinical Exons Feasibility Studies Fibroblasts Humans Immunohistochemistry Mice Motor Neurons/drug effects/metabolism Nerve Tissue Proteins/genetics/*physiology RNA, Messenger/metabolism RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Reverse Transcriptase Polymerase Chain Reaction Spinal Muscular Atrophies of Childhood/*drug therapy/genetics Transcription, Genetic/drug effects Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11734549 Abstract: Proximal spinal muscular atrophy (SMA) is a common motor neuron disorder caused by mutation of the telomeric survival of motor neuron gene SMN1. The centromeric survival of motor neuron SMN2 gene is retained in all SMA patients but does not produce sufficient SMN protein to prevent the development of clinical symptoms. The SMN1 and SMN2 genes differ functionally by a single nucleotide change. This change affects the efficiency with which exon 7 is incorporated into the mRNA transcript. Thus, SMN2 produces less full-length mRNA and protein than SMN1. We have screened a library of compounds in order to identify ones that can alter the splicing pattern of the SMN2 gene. Here, we report that the compound aclarubicin increases the retention of exon 7 into the SMN2 transcript. We show that aclarubicin effectively induces incorporation of exon 7 into SMN2 transcripts from the endogenous gene in type I SMA fibroblasts as well as into transcripts from a SMN2 minigene in the motor neuron cell line NSC34. In type I fibroblasts, treatment resulted in an increase in SMN protein and gems to normal levels. Our results suggest that alteration of splicing pattern represents a new approach to modification of gene expression in disease treatment and demonstrate the feasibility of high throughput screens to detect compounds that affect the splicing pattern of a gene. Year: 2001 Title: ACOG Committee Opinion No. 432: Spinal Muscular Atrophy Authors: Auth Address: Pages: 1194-6 Volume: 113 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19384151 Abstract: Year: 2009 Month: 5 Title: Activation of RNA metabolism-related genes in mouse but not human tissues deficient in SMN Authors: Olaso, R. Joshi, V. Fernandez, J. Roblot, N. Courageot, S. Bonnefont, J. P. Melki, J. Auth Address: Molecular Neurogenetics Laboratory, Institut National de la Sante et de la Recherche Medicale (INSERM), E-223, University of Evry, Genopole, Evry, France. Pages: 97-104 Volume: 24 Number: 2 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16118268 Abstract: Mutations of the survival of motor neuron gene (SMN1) are responsible for spinal muscular atrophies (SMA), a frequent recessive autosomal motor neuron disease. SMN is involved in various processes including RNA metabolism. However, the molecular pathway linking marked deficiency of SMN to SMA phenotype remains unclear. Homozygous deletion of murine Smn exon 7 directed to neurons or skeletal muscle causes severe motor axonal or myofiber degeneration, respectively. With the use of cDNA microarrays, expression profiles of 8,400 genes were analyzed in skeletal muscle and spinal cord of muscular and neuronal mutants, respectively, and compared with age-matched controls. A high proportion of genes (20 of 429, 5%) was involved in pre-mRNA splicing, ribosomal RNA processing, or RNA decay, and 18 of them were upregulated in mutant tissues. By analyzing other neuromuscular disorders, we showed that most of them (14 of 18) were specific to the SMN defect. Quantitative PCR analysis of these transcripts showed that gene activation was an early adaptive response to the lack but not reduced amount of full-length SMN in mouse mutant tissues. In human SMA tissues, activation of this program was not observed, which could be ascribed to the reduction but not the absence of full-length SMN. Year: 2006 Title: Active transport of the survival motor neuron protein and the role of exon-7 in cytoplasmic localization Authors: Zhang, H. L. Pan, F. Hong, D. Shenoy, S. M. Singer, R. H. Bassell, G. J. Auth Address: Department of Neuroscience, Rose F. Kennedy Center for Mental Retardation, Albert Einstein College of Medicine, Bronx, New York 10461, USA. Pages: 6627-37 Volume: 23 Number: 16 Keywords: Animals Cell Nucleus/metabolism Cells, Cultured Chick Embryo Cyclic AMP Response Element-Binding Protein Cytoplasm/*metabolism Cytoplasmic Granules/metabolism Exons/physiology Fibroblasts/cytology/metabolism Genes, Reporter Growth Cones/metabolism Humans Microfilaments/physiology Microtubules/physiology Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/genetics/*metabolism Neurites/metabolism Neurons/cytology/*metabolism Protein Transport/physiology RNA-Binding Proteins Rats Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sequence Deletion Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12878704 Abstract: Spinal muscular atrophy (SMA) is a neurodegenerative disease caused by deletion and/or mutation of the survival motor neuron protein Gene (SMN1) that results in the expression of a truncated protein lacking the C terminal exon-7. Whereas SMN has been shown to be an important component of diverse ribonucleoprotein (RNP) complexes, its function in neurons is unknown. We hypothesize that the active transport of SMN may be important for neurite outgrowth and that disruption of exon-7 could impair its normal intracellular trafficking. SMN was localized in granules that were associated with cytoskeletal filament systems and distributed throughout neurites and growth cones. Live cell imaging of enhanced green fluorescent protein (EGFP)-SMN granules revealed rapid, bidirectional and cytoskeletal-dependent movements. Exon-7 was necessary for localization of SMN into the cytoplasm but was not sufficient for granule formation and transport. A cytoplasmic targeting signal within exon-7 was identified that could completely redistribute the nuclear protein D-box binding factor 1 into the cytoplasm. Neurons transfected with SMN lacking exon-7 had significantly shorter neurites, a defect that could be rescued by redirecting the exon-7 deletion mutant into neurites by a targeting sequence from growth-associated protein-43. These findings provide the first demonstration of cytoskeletal-based active transport of SMN in neuronal processes and the function of exon-7 in cytoplasmic localization. Such observations provide motivation to investigate possible transport defects or inefficiency of SMN associated RNPs in motor neuron axons in SMA. Year: 2003 Title: Activity-dependent release of precursor nerve growth factor, conversion to mature nerve growth factor, and its degradation by a protease cascade Authors: Bruno, M. A. Cuello, A. C. Auth Address: Departments of Pharmacology and Therapeutics, Anatomy and Cell Biology, and Neurology and Neurosurgery, McGill University, Montreal, QC, Canada H3G 1Y6. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16618925 Abstract: In this report, we provide direct demonstration that the neurotrophin nerve growth factor (NGF) is released in the extracellular space in an activity-dependent manner in its precursor form (proNGF) and that it is in this compartment that its maturation and degradation takes place because of the coordinated release and the action of proenzymes and enzyme regulators. This converting protease cascade and its endogenous regulators (including tissue plasminogen activator, plasminogen, neuroserpin, precursor matrix metalloproteinase 9, and tissue inhibitor metalloproteinase 1) are colocalized in neurons of the cerebral cortex and released upon neuronal stimulation. We also provide evidence that this mechanism operates in in vivo conditions, as the CNS application of inhibitors of converting and degrading enzymes lead to dramatic alterations in the tissue levels of either precursor NGF or mature NGF. Pathological alterations of this cascade in the CNS might cause or contribute to a lack of proper neuronal trophic support in conditions such as cerebral ischemia, seizure and Alzheimer's disease or, conversely, to excessive local production of neurotrophins as reported in inflammatory arthritis pain. Year: 2006 Title: Acute and chronic effects of neurotrophic factors BDNF and GDNF on responses mediated by thermo-sensitive TRP channels in cultured rat dorsal root ganglion neurons Authors: Ciobanu, C. Reid, G. Babes, A. Auth Address: Department of Animal Physiology and Biophysics, Faculty of Biology, University of Bucharest, Splaiul Independentei 91-95, 050095 Bucharest, Romania. Pages: 54-67 Volume: 1284 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19524560 Abstract: Neurotrophic factors (NTFs), beside regulating neuronal survival in the central and peripheral nervous system, are also involved in the modulation of neuronal function in the adult animal. Both brain-derived neurotrophic factor (BDNF) and glial cell-derived neurotrophic factor (GDNF) levels are altered in pathological pain states, and exogenous BDNF and GDNF have multiple effects on pain behavior, depending on the animal model (i.e. inflammatory vs. neuropathic). Thermally gated TRP channels TRPM8, TRPA1 and TRPV1 play a significant role in pain signaling and their pattern and level of expression as well as their biophysical properties are altered in chronic pain states. Our aim was to investigate the effect of long-term and acute exposure to BDNF and GDNF on the functional expression of these thermoTRP channels in cultured rat dorsal root ganglion (DRG) neurons. We found that while BDNF treatment primarily increased the fraction of capsaicin-sensitive (TRPV1-expressing) neurons, GDNF exposure led to an increase in the allyl isothiocyanate (AITC)-responding (TRPA1-expressing) population. Moreover, BDNF treatment increased the amplitude of the response to both AITC and capsaicin. Acute treatment with both NTFs leads to a reduction in the magnitude of tachyphylaxis to noxious stimuli (heat and AITC). Overall, our data provides evidence for a role of BDNF and GDNF in regulating the pattern of expression and level of activity of the transducer channels TRPA1 and TRPV1, leading to enhanced neuronal sensitivity to painful stimuli and increased co-expression of thermoTRP channels. Year: 2009 Title: Adenosine receptor A2A-R contributes to motoneuron survival by transactivating the tyrosine kinase receptor TrkB Authors: Wiese, S. Jablonka, S. Holtmann, B. Orel, N. Rajagopal, R. Chao, M. V. Sendtner, M. Auth Address: Institute for Clinical Neurobiology, University of Wurzburg, Josef Schneider Strasse 11, D-97080 Wurzburg, Germany. Pages: 17210-5 Volume: 104 Number: 43 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17940030 Abstract: Neurotrophins are potent survival factors for developing and injured neurons. However, they are not being used to treat neurodegenerative diseases because of difficulties in administration and numerous side effects that have been encountered in previous clinical trials. Their biological activities use Trk (tropomyosin-related kinase) transmembrane tyrosine kinases. Therefore, one alternative approach is to use transactivation pathways such as adenosine 2A receptor agonists, which can activate Trk receptor signaling independent of neurotrophin binding. However, the relevance in vivo and applicability of these transactivation events during neurodegenerative and injury conditions have never been extensively studied. Here we demonstrate that motoneuron survival after facial nerve lesioning is significantly enhanced by transactivation of Trk receptor tyrosine kinases by adenosine agonists. Moreover, survival of motoneurons directly required the activation of the BDNF receptor TrkB and an increase in Akt (AKT8 virus oncogene cellular homolog) activity. The ability of small molecules to activate a trophic response by using Trk signaling provides a unique mechanism to promote survival signals in motoneurons and suggests new strategies for using transactivation in neurodegenerative diseases. Year: 2007 Title: Adenoviral gene transfer to spinal-cord neurons: intrathecal vs. intraparenchymal administration Authors: Mannes, A. J. Caudle, R. M. O'Connell, B. C. Iadarola, M. J. Auth Address: NAB/NIDR/NIH, Bethesda, MD 20892, USA. Pages: 1-6 Volume: 793 Number: 1-2 Keywords: Adenoviridae/*genetics Animals Cytomegalovirus/genetics Gene Expression/drug effects *Gene Transfer Techniques Immunohistochemistry Injections, Spinal Male Neurons/drug effects/*metabolism Promoter Regions, Genetic/genetics Rats Rats, Sprague-Dawley Spinal Cord/*cytology/drug effects beta-Galactosidase/diagnostic use/genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9630472 Abstract: The spinal cord is the site of many chronic, debilitating, neurological disorders that may be amenable to gene therapy. The present study, using quantitative and anatomical methods, examines the ability of replication deficient adenovirus to transfer a transcription cassette composed of the cytomegalovirus promoter driving the expression of the LacZ reporter gene (AdCMVbetagal) to spinal-cord neurons. Rats were microinjected with AdCMVbetagal into the spinal-cord parenchyma or subarachnoid space and sacrificed between 1 and 60 days post-infusion. The spinal cord was assayed for beta-galactosidase (beta-gal) activity fluorometrically (MUG). Intraparenchymal injection resulted in significant beta-gal activity at day 1, which peaked at day 7, and decreased at day 14 (21-, 57- and 9.8-fold of control respectively). The spatial distribution of beta-gal activity on day 7 was confined to the 1-cm section containing the injection site but was detected 2 cm caudal to this section by day 14. Histochemical staining and immunocytochemistry revealed a prominent reaction product in neurons, particularly motor neurons, and glia within the ventral grey matter bilaterally. Intrathecal viral injections showed comparatively modest, yet significant increases in beta-gal activity throughout the spinal cord with the greatest activity (170% control) closest to the catheter tip. This study demonstrates that AdCMVbetagal injected into the ventral spinal cord results in extensive in vivo neuronal gene transfer with beta-gal activity reaching a peak by day 7 and remaining detectable at 60 days. Intrathecal viral injections result in greater spatial distribution but a comparatively lower level of expression. Year: 1998 Title: Advances in signaling in vertebrate regeneration as a prelude to regenerative medicine Authors: Stoick-Cooper, C. L. Moon, R. T. Weidinger, G. Auth Address: Department of Pharmacology, Howard Hughes Medical Institute, and Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, Washington 98195, USA. Pages: 1292-315 Volume: 21 Number: 11 Keywords: Animals Humans Muscle, Skeletal/*physiology Regeneration/*physiology *Regenerative Medicine *Signal Transduction Vertebrates/physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17545465 Abstract: While all animals have evolved strategies to respond to injury and disease, their ability to functionally recover from loss of or damage to organs or appendages varies widely damage to skeletal muscle, but, unlike amphibians and fish, they fail to regenerate heart, lens, retina, or appendages. The relatively young field of regenerative medicine strives to develop therapies aimed at improving regenerative processes in humans and is predicated on >40 years of success with bone marrow transplants. Further progress will be accelerated by implementing knowledge about the molecular mechanisms that regulate regenerative processes in model organisms that naturally possess the ability to regenerate organs and/or appendages. In this review we summarize the current knowledge about the signaling pathways that regulate regeneration of amphibian and fish appendages, fish heart, and mammalian liver and skeletal muscle. While the cellular mechanisms and the cell types involved in regeneration of these systems vary widely, it is evident that shared signals are involved in tissue regeneration. Signals provided by the immune system appear to act as triggers of many regenerative processes. Subsequently, pathways that are best known for their importance in regulating embryonic development, in particular fibroblast growth factor (FGF) and Wnt/beta-catenin signaling (as well as others), are required for progenitor cell formation or activation and for cell proliferation and specification leading to tissue regrowth. Experimental activation of these pathways or interference with signals that inhibit regenerative processes can augment or even trigger regeneration in certain contexts. Year: 2007 Title: Advances in SMA research: review of gene deletions Authors: Morrison, K. E. Auth Address: Department of Clinical Neurology, University of Oxford, UK. Pages: 397-408 Volume: 6 Number: 6 Keywords: Chromosome Aberrations/genetics Chromosome Disorders Cloning, Molecular *Gene Deletion Genetic Diseases, Inborn/genetics Humans Motor Neurons/physiology Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/genetics Neuronal Apoptosis-Inhibitory Protein Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9027847 Abstract: The term spinal muscular atrophy (SMA) is used to encompass a group of inherited disorders in which the striking pathological feature is loss of the cell bodies of alpha motor neurons in the anterior horn cell of the spinal cord and, in some cases, of the bulbar motor nuclei. Although the pathological features of these disorders have been well characterized, the nature of the primary underlying biochemical abnormality remains to be determined. In the 1990s genetic linkage was established for the childhood onset recessive forms of SMA (types I, II and III) to markers mapping to the chromosomal region 5q11.2-13.3. Physical maps of the region were then constructed, several candidate genes isolated and in 1995 deletions in two genes, the survival motor neuron (SMN) gene and the neuronal apoptosis inhibitory protein (NAIP) gene, were identified in significant numbers of patients. Already the impact of the characterization of these deletions is being seen in clinical practice in terms of aiding diagnosis in symptomatic cases and in prenatal diagnosis. As discussed in this review however, several questions remain unresolved. It is unclear whether deletions in one or both of these genes, or indeed in other, as yet unidentified, genes are important in generating the SMA phenotype. The function of the protein product of the SMN gene is unknown. The NAIP gene encodes a protein which inhibits apoptosis in a mammalian cell line: is it disruption of this function which is relevant in SMA? What underlies the variation in disease severity evident both between and within families? Resolution of such issues is of crucial importance if the identification of these deleted gene sequences is to lead to the development of rational therapies for motor neuron diseases. Year: 1996 Month: 12 Title: AEG3482 is an antiapoptotic compound that inhibits Jun kinase activity and cell death through induced expression of heat shock protein 70 Authors: Salehi, A. H. Morris, S. J. Ho, W. C. Dickson, K. M. Doucet, G. Milutinovic, S. Durkin, J. Gillard, J. W. Barker, P. A. Auth Address: Centre for Neuronal Survival, Montreal Neurological Institute, McGill University, Quebec, Canada. Pages: 213-23 Volume: 13 Number: 2 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16492569 Abstract: We describe a group of small-molecule inhibitors of Jun kinase (JNK)-dependent apoptosis. AEG3482, the parental compound, was identified in a screening effort designed to detect compounds that reduce apoptosis of neonatal sympathetic neurons after NGF withdrawal. We show that AEG3482 blocks apoptosis induced by the p75 neurotrophin receptor (p75NTR) or its cytosolic interactor, NRAGE, and demonstrate that AEG3482 blocks proapoptotic JNK activity. We show that AEG3482 induces production of heat shock protein 70 (HSP70), an endogenous inhibitor of JNK, and establish that HSP70 accumulation is required for the AEG3482-induced JNK blockade. We show that AEG3482 binds HSP90 and induces HSF1-dependent HSP70 mRNA expression and find that AEG3482 facilitates HSP70 production while retaining HSP90 chaperone activity. These studies establish that AEG3482 inhibits JNK activation and apoptosis by a mechanism involving induced expression of HSP proteins. Year: 2006 Month: 2 Title: Aerobic conditioning: an effective therapy in McArdle's disease Authors: Haller, R. G. Wyrick, P. Taivassalo, T. Vissing, J. Auth Address: Neuromuscular Center, Institute for Exercise and Environmental Medicine of Presbyterian Hospital, Dallas, TX 75231, USA. ronald.haller@utsouthwestern.edu Pages: 922-8 Volume: 59 Number: 6 Keywords: 3-Hydroxyacyl CoA Dehydrogenases/metabolism Adult Cardiac Output/physiology Citrate (si)-Synthase/metabolism Creatine Kinase/blood *Exercise Therapy Female Glucose/metabolism Glycogen Storage Disease Type V/*therapy Humans Male Middle Aged Oxygen Consumption/physiology Physical Fitness/*physiology Quadriceps Muscle/enzymology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16718692 Abstract: OBJECTIVE: Susceptibility to exertional cramps and rhabdomyolysis in myophosphorylase deficiency (McArdle's disease [MD]) may lead patients to shun exercise. However, physical inactivity may worsen exercise intolerance by further reducing the limited oxidative capacity caused by blocked glycogenolysis. We investigated whether aerobic conditioning can safely improve exercise capacity in MD. METHODS: Eight MD patients (4 men and 4 women; age range, 33-61 years) pedalled a cycle ergometer for 30 to 40 minutes a day, 4 days a week, for 14 weeks, at an intensity corresponding to 60 to 70% of maximal heart rate. We monitored serum creatine kinase levels; changes in peak cycle work, oxygen uptake, and cardiac output; presence and magnitude of a spontaneous and glucose-induced second wind; and citrate synthase and beta-hydroxyacyl coenzyme A dehydrogenase enzyme activities in quadriceps muscle. RESULTS: The prescribed exercise program increased average work capacity (36%), oxygen uptake (14%), cardiac output (15%), and citrate synthase and beta-hydroxyacyl coenzyme A dehydrogenase enzyme levels (80 and 62%, respectively) without causing pain or cramping or increasing serum creatine kinase. A spontaneous and glucose-induced second wind was present and was of similar magnitude in each patient before and after training. INTERPRETATION: Moderate aerobic exercise is an effective means of improving exercise capacity in MD by increasing circulatory delivery and mitochondrial metabolism of bloodborne fuels. Year: 2006 Month: 6 Title: Aerobic training improves exercise performance in facioscapulohumeral muscular dystrophy Authors: Olsen, D. B. Orngreen, M. C. Vissing, J. Auth Address: Department of Neurology and the Copenhagen Muscle Research Center, National University Hospital Rigshospitalet, Copenhagen, Denmark. d.benee@mfi.ku.dk Pages: 1064-6 Volume: 64 Number: 6 Keywords: Adolescent Adult Age Factors Creatine Kinase/blood Energy Metabolism/physiology Exercise/*physiology Exercise Therapy/*methods Exercise Tolerance/physiology Female Humans Male Middle Aged Muscle Fibers, Skeletal/metabolism Muscle, Skeletal/blood supply/*physiology/physiopathology Muscular Dystrophy, Facioscapulohumeral/*therapy Oxygen Consumption/physiology Physical Fitness/*physiology Questionnaires Regional Blood Flow/physiology Sex Factors Treatment Outcome Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15781829 Abstract: Exercise programs have been shown to increase strength and endurance in patients with myopathic disorders. The authors investigated the effect of aerobic training in patients with facioscapulohumeral dystrophy (FSHD). Twelve weeks of low-intense aerobic exercise improved maximal oxygen uptake and workload with no signs of muscle damage. The authors conclude that aerobic training is a safe method to increase exercise performance in patients with FSHD. Year: 2005 Title: Aerobic training in patients with myotonic dystrophy type 1 Authors: Orngreen, M. C. Olsen, D. B. Vissing, J. Auth Address: Copenhagen Muscle Research Center and the Department of Neurology, National University Hospital, Rigshospitalet, Copenhagen, Denmark. rh10679@rh.dk Pages: 754-7 Volume: 57 Number: 5 Keywords: Activities of Daily Living Adult Anaerobic Threshold/physiology Cell Size Creatine Kinase/metabolism *Exercise Exercise Test *Exercise Therapy Female Humans Male Middle Aged Muscle Fibers, Skeletal/pathology Muscle, Skeletal/pathology/physiopathology Myotonic Dystrophy/pathology/physiopathology/*therapy Oxygen Consumption/physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15852373 Abstract: The effect of 12 weeks of aerobic training on a cycle ergometer was studied in 12 patients with myotonic dystrophy. Efficacy was evaluated by cycle testing and muscle morphology before and after training. Patients increased their maximal oxygen uptake by 14%, the maximal workload by 11%, muscle fiber diameter increased significantly, and creatine kinase did not increase with training. The study indicates that aerobic training is safe and can improve fitness effectively in patients with myotonic dystrophy. Year: 2005 Month: 5 Title: Aerobic training is safe and improves exercise capacity in patients with mitochondrial myopathy Authors: Jeppesen, T. D. Schwartz, M. Olsen, D. B. Wibrand, F. Krag, T. Duno, M. Hauerslev, S. Vissing, J. Auth Address: Department of Neurology, Neuromuscular Research Unit, The Copenhagen Muscle Research Centre, Rigshospitalet, Copenhagen, Denmark. dysgaard@rh.dk Pages: 3402-12 Volume: 129 Number: Pt 12 Keywords: Adult Aerobiosis/physiology Creatine Kinase/blood DNA, Mitochondrial/genetics Exercise Therapy/*methods Female Gene Deletion Heart Rate/physiology Humans Lactates/blood Male Middle Aged Mitochondria/enzymology Mitochondrial Myopathies/genetics/physiopathology/*therapy Muscle, Skeletal/enzymology/pathology/physiopathology Oxygen Consumption/physiology Point Mutation/genetics Quality of Life Treatment Outcome Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16815877 Abstract: Exercise intolerance is a prominent symptom in patients with mitochondrial myopathy (MM), but it is still unsettled whether exercise training is safe and beneficial for patients with MM. To address this, we studied the effect of 12 weeks cycle training on exercise capacity, quality of life and underlying molecular and cellular events in five patients with single large-scale deletions, one with a microdeletion and 14 with point mutations of mitochondrial DNA (mtDNA), and 13 healthy subjects. Each training session lasted 30 min, and was performed at an intensity of 70% of VO2max (maximal oxygen uptake). Each subject performed 50 training sessions in 12 weeks. All subjects were evaluated before and after training, and 13 MM patients were studied after 8 weeks of deconditioning. Evaluation included VO2max and mutation load and mtDNA quantity, mitochondrial enzymatic activity, and number of centrally nucleated, apoptotic, ragged red and cytochrome oxidase (COX)-negative fibres in muscle biopsies from the quadriceps muscle. After 12 weeks of training, VO2max and muscle citrate synthase increased in MM (26 and 67%) and healthy (17 and 65%) subjects, while mtDNA quantity in muscle only increased in the MM patients (81%). In the MM patients, training did not change mtDNA mutation load in muscle, mitochondrial enzyme complex activities, muscle morphology and plasma creatine kinase. After deconditioning, VO2max and citrate synthase activity returned to values before training, while muscle mtDNA mutation load decreased. These findings show that aerobic training efficiently improves oxidative capacity in MM patients. Based on unchanged levels of mutant load in muscle, morphological findings on muscle biopsy and plasma creatine kinase levels during training, the treatment appears to be safe. Regular, supervised aerobic exercise is therefore recommended in MM patients with the studied mutations. Year: 2006 Month: 12 Title: Akt/mTOR pathway is a crucial regulator of skeletal muscle hypertrophy and can prevent muscle atrophy in vivo Authors: Bodine, S. C. Stitt, T. N. Gonzalez, M. Kline, W. O. Stover, G. L. Bauerlein, R. Zlotchenko, E. Scrimgeour, A. Lawrence, J. C. Glass, D. J. Yancopoulos, G. D. Auth Address: Regeneron Pharmaceuticals, Inc. 777 Old Saw Mill River Road, Tarrytown, New York 10591-6707, USA. sue.bodine@regeneron.com Pages: 1014-9 Volume: 3 Number: 11 Keywords: Animals Calcineurin/metabolism Cardiomegaly/metabolism Cyclosporine/pharmacology Enzyme Inhibitors/pharmacology Female Muscle, Skeletal/*metabolism Muscular Atrophy/*metabolism Protein Kinases/*metabolism *Protein-Serine-Threonine Kinases Proto-Oncogene Proteins/*metabolism Proto-Oncogene Proteins c-akt Rats Rats, Sprague-Dawley Ribosomal Protein S6 Kinases/metabolism *Signal Transduction Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11715023 Abstract: Skeletal muscles adapt to changes in their workload by regulating fibre size by unknown mechanisms. The roles of two signalling pathways implicated in muscle hypertrophy on the basis of findings in vitro, Akt/mTOR (mammalian target of rapamycin) and calcineurin/NFAT (nuclear factor of activated T cells), were investigated in several models of skeletal muscle hypertrophy and atrophy in vivo. The Akt/mTOR pathway was upregulated during hypertrophy and downregulated during muscle atrophy. Furthermore, rapamycin, a selective blocker of mTOR, blocked hypertrophy in all models tested, without causing atrophy in control muscles. In contrast, the calcineurin pathway was not activated during hypertrophy in vivo, and inhibitors of calcineurin, cyclosporin A and FK506 did not blunt hypertrophy. Finally, genetic activation of the Akt/mTOR pathway was sufficient to cause hypertrophy and prevent atrophy in vivo, whereas genetic blockade of this pathway blocked hypertrophy in vivo. We conclude that the activation of the Akt/mTOR pathway and its downstream targets, p70S6K and PHAS-1/4E-BP1, is requisitely involved in regulating skeletal muscle fibre size, and that activation of the Akt/mTOR pathway can oppose muscle atrophy induced by disuse. Year: 2001 Month: 11 Title: Allelic variants of the canine heavy neurofilament (NFH) subunit and extensive phosphorylation in dogs with motor neuron disease Authors: Green, S. L. Westendorf, J. M. Jaffe, H. Pant, H. C. Cork, L. C. Ostrander, E. A. Vignaux, F. Ferrell, J. E., Jr. Auth Address: Department of Comparative Medicine, Stanford University, Stanford, CA 94305, USA. Pages: 33-50 Volume: 132 Number: 1 Keywords: *Alleles Amino Acid Sequence Animals Base Sequence Chromatography, High Pressure Liquid/veterinary Chromosome Mapping/veterinary Cloning, Molecular Dog Diseases/*genetics/metabolism/pathology Dogs Humans Mice Molecular Sequence Data Muscular Atrophy, Spinal/genetics/metabolism/pathology/*veterinary Neurofilament Proteins/chemistry/*genetics/metabolism Phosphorylation Polymorphism, Restriction Fragment Length Research Support, U.S. Gov't, P.H.S. Sequence Analysis, DNA/veterinary Spectrometry, Mass, Electrospray Ionization/veterinary Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15629478 Abstract: Aberrant accumulation of extensively phosphorylated heavy (high molecular weight) neurofilament (NFH) and neurodegeneration are features of hereditary canine spinal muscular atrophy (HCSMA), an animal model of human motor neuron disease. In this study, the canine NFH gene was mapped, cloned, and sequenced, and electrospray/mass spectrometry was used to evaluate the phosphorylation state of NFH protein from normal dogs and dogs with HCSMA. The canine NFH gene was localized to a region on canine chromosome 26 that corresponds to human NFH on chromosome 22q. The predicted length of the canine NFH protein is 1135 amino acids, and it shares an 80.3% identity with human NFH and >74.6% with murine NFH proteins. Direct sequencing of NFH cDNA from HCSMA dogs revealed no mutations, although cDNA sequence and restriction fragment length polymorphism (RFLP) analysis indicates that there are at least three canine NFH alleles, differing in the position and number (61 or 62) of Lys-Ser-Proline (KSP) motifs. The two longest alleles (L1 and L2), each with 62 KSP repeats, contain an additional 24-base insert and were observed in both normal and HCSMA dogs. However, the shorter allele (the C allele), with 61 KSP sites and lacking the 24-base insertion, was absent in dogs with HCSMA. Mass spectrometry data indicated that almost all of the NFH KSP phosphorylation sites were occupied. No new or extra sites were identified in native NFH purified from the HCSMA dogs. The predominance of the two longest NFH alleles and the additional KSP phosphorylation sites they confer probably account for the presence of extensively phosphorylated NFs detected immunohistochemically in dogs with HCSMA. Year: 2005 Month: 1 Title: Alterations in the ankyrin domain of TRPV4 cause congenital distal SMA, scapuloperoneal SMA and HMSN2C Authors: Auer-Grumbach, M. Olschewski, A. Papic, L. Kremer, H. McEntagart, M. E. Uhrig, S. Fischer, C. Frohlich, E. Balint, Z. Tang, B. Strohmaier, H. Lochmuller, H. Schlotter-Weigel, B. Senderek, J. Krebs, A. Dick, K. J. Petty, R. Longman, C. Anderson, N. E. Padberg, G. W. Schelhaas, H. J. van Ravenswaaij-Arts, C. M. Pieber, T. R. Crosby, A. H. Guelly, C. Auth Address: Institute of Human Genetics, Medical University of Graz, Austria. michaela.auergrumbach@medunigraz.at Pages: 160-4 Volume: 42 Number: 2 Keywords: Amino Acid Substitution/genetics *Ankyrin Repeat Calcium/metabolism Hela Cells Hereditary Sensory and Motor Neuropathy/complications/*genetics/physiopathology Humans Immunohistochemistry Intracellular Space/metabolism Ion Channel Gating Models, Molecular Molecular Sequence Data Muscular Atrophy, Spinal/complications/*congenital/*genetics/physiopathology Mutant Proteins/metabolism Mutation/*genetics Osmosis TRPV Cation Channels/*chemistry/*genetics Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20037588 Abstract: Spinal muscular atrophies (SMA, also known as hereditary motor neuropathies) and hereditary motor and sensory neuropathies (HMSN) are clinically and genetically heterogeneous disorders of the peripheral nervous system. Here we report that mutations in the TRPV4 gene cause congenital distal SMA, scapuloperoneal SMA, HMSN 2C. We identified three missense substitutions (R269H, R315W and R316C) affecting the intracellular N-terminal ankyrin domain of the TRPV4 ion channel in five families. Expression of mutant TRPV4 constructs in cells from the HeLa line revealed diminished surface localization of mutant proteins. In addition, TRPV4-regulated Ca(2+) influx was substantially reduced even after stimulation with 4alphaPDD, a TRPV4 channel-specific agonist, and with hypo-osmotic solution. In summary, we describe a new hereditary channelopathy caused by mutations in TRPV4 and present evidence that the resulting substitutions in the N-terminal ankyrin domain affect channel maturation, leading to reduced surface expression of functional TRPV4 channels. Month: 2 Title: Altered intracellular Ca2+ homeostasis in nerve terminals of severe spinal muscular atrophy mice Authors: Ruiz, R. Casanas, J. J. Torres-Benito, L. Cano, R. Tabares, L. Auth Address: Department of Medical Physiology and Biophysics, School of Medicine, University of Seville, 41009 Seville, Spain. Pages: 849-57 Volume: 30 Number: 3 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20089893 Abstract: Low levels of survival motor neuron (SMN) protein result in spinal muscular atrophy (SMA), a severe genetic disease characterized by motor impairment and premature lethality. Although SMN is a ubiquitous protein, motor neurons are much more vulnerable to low levels of SMN than other cells. To gain insight into the pathogenesis of SMA, we have compared synaptic function of motor terminals in wild-type and severe SMA mice at different ages and in two proximal muscles. Our results show that mutant muscle fibers fire normal action potentials and that multi-innervated terminals are functional. By studying the characteristics of the three main components of synaptic transmission in nerve terminals (spontaneous, evoked, and asynchronous release), we found that the kinetics of the postsynaptic potentials are slowed and evoked neurotransmitter release is decreased by approximately 55%. In addition, asynchronous release is increased approximately 300%, indicating an anomalous augmentation of intraterminal bulk Ca(2+) during repetitive stimulation. Together, these results show that the reduction of SMN affects synaptic maturation, evoked release, and regulation of intraterminal Ca(2+) levels. Title: Altered RNA splicing contributes to skeletal muscle pathology in Kennedy disease knock-in mice Authors: Yu, Z. Wang, A. M. Robins, D. M. Lieberman, A. P. Auth Address: Department of Pathology, The University of Michigan Medical School, Ann Arbor, MI 48109, USA. Pages: 500-7 Volume: 2 Number: 9-10 Keywords: Animals Bulbo-Spinal Atrophy, X-Linked/*genetics/*pathology Chloride Channels/genetics/metabolism Exons/genetics *Gene Knock-In Techniques Glutamine/genetics Ligands Male Mice Mice, Mutant Strains Muscle Denervation Muscle, Skeletal/innervation/*pathology RNA Splicing/*genetics RNA, Messenger/genetics/metabolism RNA-Binding Proteins/genetics/metabolism Receptors, Androgen/metabolism Trinucleotide Repeat Expansion/genetics Troponin T/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19692580 Abstract: Here, we used a mouse model of Kennedy disease, a degenerative disorder caused by an expanded CAG repeat in the androgen receptor (AR) gene, to explore pathways leading to cellular dysfunction. We demonstrate that male mice containing a targeted Ar allele with 113 CAG repeats (AR113Q mice) exhibit hormone- and glutamine length-dependent missplicing of Clcn1 RNA in skeletal muscle. Changes in RNA splicing are associated with increased expression of the RNA-binding protein CUGBP1. Furthermore, we show that skeletal muscle denervation in the absence of a repeat expansion leads to increased CUGBP1 expression. However, this induction of CUGBP1 is not sufficient to alter Clcn1 RNA splicing, indicating that changes mediated by both denervation and AR113Q toxicity contribute to altered RNA processing. To test this notion directly, we exogenously expressed the AR in vitro and observed hormone-dependent changes in the splicing of pre-mRNAs from a human cardiac troponin T minigene. These effects were notably similar to changes mediated by RNA with expanded CUG tracts, but not CAG tracts, highlighting unanticipated similarities between CAG and CUG repeat diseases. The expanded glutamine AR also altered hormone-dependent splicing of a calcitonin/calcitonin gene-related peptide minigene, suggesting that toxicity of the mutant protein additionally affects RNA processing pathways that are distinct from those regulated by CUGBP1. Our studies demonstrate the occurrence of hormone-dependent alterations in RNA splicing in Kennedy disease models, and they indicate that these changes are mediated by both the cell-autonomous effects of the expanded glutamine AR protein and by alterations in skeletal muscle that are secondary to denervation. Year: 2009 Title: Altered synaptic plasticity in a mouse model of fragile X mental retardation Authors: Huber, K. M. Gallagher, S. M. Warren, S. T. Bear, M. F. Auth Address: Department of Neuroscience, Howard Hughes Medical Institute, Brown University, Providence, RI 02912, USA. Pages: 7746-50 Volume: 99 Number: 11 Keywords: Animals Disease Models, Animal Fragile X Mental Retardation Protein Fragile X Syndrome/genetics/*physiopathology Humans Mental Retardation/genetics/physiopathology Mice Mice, Knockout Models, Neurological Mutation Nerve Tissue Proteins/*genetics Neuronal Plasticity/*physiology Protein Biosynthesis RNA, Messenger/genetics *RNA-Binding Proteins Synapses/*physiology Transcription, Genetic Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12032354 Abstract: Fragile X syndrome, the most common inherited form of human mental retardation, is caused by mutations of the Fmr1 gene that encodes the fragile X mental retardation protein (FMRP). Biochemical evidence indicates that FMRP binds a subset of mRNAs and acts as a regulator of translation. However, the consequences of FMRP loss on neuronal function in mammals remain unknown. Here we show that a form of protein synthesis-dependent synaptic plasticity, long-term depression triggered by activation of metabotropic glutamate receptors, is selectively enhanced in the hippocampus of mutant mice lacking FMRP. This finding indicates that FMRP plays an important functional role in regulating activity-dependent synaptic plasticity in the brain and suggests new therapeutic approaches for fragile X syndrome. Year: 2002 Title: Alternative splicing events are a late feature of pathology in a mouse model of spinal muscular atrophy Authors: Baumer, D. Lee, S. Nicholson, G. Davies, J. L. Parkinson, N. J. Murray, L. M. Gillingwater, T. H. Ansorge, O. Davies, K. E. Talbot, K. Auth Address: MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom. Pages: e1000773 Volume: 5 Number: 12 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20019802 Abstract: Spinal muscular atrophy is a severe motor neuron disease caused by inactivating mutations in the SMN1 gene leading to reduced levels of full-length functional SMN protein. SMN is a critical mediator of spliceosomal protein assembly, and complete loss or drastic reduction in protein leads to loss of cell viability. However, the reason for selective motor neuron degeneration when SMN is reduced to levels which are tolerated by all other cell types is not currently understood. Widespread splicing abnormalities have recently been reported at end-stage in a mouse model of SMA, leading to the proposition that disruption of efficient splicing is the primary mechanism of motor neuron death. However, it remains unclear whether splicing abnormalities are present during early stages of the disease, which would be a requirement for a direct role in disease pathogenesis. We performed exon-array analysis of RNA from SMN deficient mouse spinal cord at 3 time points, pre-symptomatic (P1), early symptomatic (P7), and late-symptomatic (P13). Compared to littermate control mice, SMA mice showed a time-dependent increase in the number of exons showing differential expression, with minimal differences between genotypes at P1 and P7, but substantial variation in late-symptomatic (P13) mice. Gene ontology analysis revealed differences in pathways associated with neuronal development as well as cellular injury. Validation of selected targets by RT-PCR confirmed the array findings and was in keeping with a shift between physiologically occurring mRNA isoforms. We conclude that the majority of splicing changes occur late in SMA and may represent a secondary effect of cell injury, though we cannot rule out significant early changes in a small number of transcripts crucial to motor neuron survival. Year: 2009 Month: 12 Title: AMPK and PPARdelta agonists are exercise mimetics Authors: Narkar, V. A. Downes, M. Yu, R. T. Embler, E. Wang, Y. X. Banayo, E. Mihaylova, M. M. Nelson, M. C. Zou, Y. Juguilon, H. Kang, H. Shaw, R. J. Evans, R. M. Auth Address: Gene Expression Laboratory, Salk Institute, La Jolla, CA 92037, USA. Pages: 405-15 Volume: 134 Number: 3 Keywords: AMP-Activated Protein Kinases Administration, Oral Aminoimidazole Carboxamide/administration & dosage/*analogs & derivatives/pharmacology Animals Biomimetics Male Mice Mice, Inbred C57BL Multienzyme Complexes/*metabolism Muscle, Skeletal/*metabolism PPAR delta/*agonists Physical Conditioning, Animal Physical Endurance/*drug effects Protein-Serine-Threonine Kinases/*metabolism Ribonucleotides/administration & dosage/*pharmacology Thiazoles/*pharmacology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18674809 Abstract: The benefits of endurance exercise on general health make it desirable to identify orally active agents that would mimic or potentiate the effects of exercise to treat metabolic diseases. Although certain natural compounds, such as reseveratrol, have endurance-enhancing activities, their exact metabolic targets remain elusive. We therefore tested the effect of pathway-specific drugs on endurance capacities of mice in a treadmill running test. We found that PPARbeta/delta agonist and exercise training synergistically increase oxidative myofibers and running endurance in adult mice. Because training activates AMPK and PGC1alpha, we then tested whether the orally active AMPK agonist AICAR might be sufficient to overcome the exercise requirement. Unexpectedly, even in sedentary mice, 4 weeks of AICAR treatment alone induced metabolic genes and enhanced running endurance by 44%. These results demonstrate that AMPK-PPARdelta pathway can be targeted by orally active drugs to enhance training adaptation or even to increase endurance without exercise. Year: 2008 Title: An 11 base pair duplication in exon 6 of the SMN gene produces a type I spinal muscular atrophy (SMA) phenotype: further evidence for SMN as the primary SMA-determining gene Authors: Parsons, D. W. McAndrew, P. E. Monani, U. R. Mendell, J. R. Burghes, A. H. Prior, T. W. Auth Address: Department of Pathology, College of Biological Sciences, Ohio State University College of Medicine, Columbus 43210, USA. Pages: 1727-32 Volume: 5 Number: 11 Keywords: Alleles Cyclic AMP Response Element-Binding Protein Exons/*genetics Female Frameshift Mutation/*genetics Gene Dosage Humans Infant Male Nerve Tissue Proteins/*genetics Phenotype Polymorphism, Single-Stranded Conformational RNA, Messenger/genetics RNA-Binding Proteins Research Support, Non-U.S. Gov't Spinal Muscular Atrophies of Childhood/*genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8922999 Abstract: The gene for autosomal recessive spinal muscular atrophy (SMA) has been mapped to 5q12 in a region that contains repeated markers and genes. Three cDNAs that detect deletions in SMA patients have been reported. One of these, the survival motor neuron (SMN) cDNA, is encoded by two genes (SMNT and SMNC) which are distinguished by base changes in exons 7 and 8. Exon 7 of the SMNT gene is not detectable in approximately 95% of SMA cases, due either to deletion or sequence conversion. There is limited information on the mutations in SMA patients that have detectable SMNT, these are critical for confirmation of SMNT as the SMA gene. Using SSCP analysis of the SMN exons we screened our SMA patients that possess at least one intact SMNT allele for mutations in SMNT. We identified one type I SMA patient with an 11 bp duplication in exon 6 which causes a frameshift and premature termination of the deduced SMNT protein. Dosage and SSCP analysis of SMNT in this family indicated that the father contributed a SMNT-deleted allele to the affected child whereas the mother passed on the 11 bp exon 6 duplication SMNT allele. Analysis of RNA by RT-PCR conclusively demonstrated that the 11 bp duplication is associated with the SMNT locus and not SMNC. This mutation provides strong support for SMN as the SMA-determining gene and indicates that disruption of SMNT on its own is sufficient to produce a severe type I SMA phenotype. Year: 1996 Month: 11 Title: An analysis of disease severity based on SMN2 copy number in adults with spinal muscular atrophy Authors: Elsheikh, B. Prior, T. Zhang, X. Miller, R. Kolb, S. J. Moore, D. Bradley, W. Barohn, R. Bryan, W. Gelinas, D. Iannaccone, S. Leshner, R. Mendell, J. R. Mendoza, M. Russman, B. Smith, S. King, W. Kissel, J. T. Auth Address: Department of Neurology, Ohio State University, 421 Means Hall, 1654 Upham Drive, Columbus Ohio 43210, USA. Pages: 652-656 Volume: 40 Number: 4 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19760790 Abstract: To evaluate the effect of SMN2 copy number on disease severity in spinal muscular atrophy (SMA), we stratified 45 adult SMA patients based on SMN2 copy number (3 vs. 4 copies). Patients with 3 copies had an earlier age of onset and lower spinal muscular atrophy functional rating scale (SMAFRS) scores and were more likely to be non-ambulatory. There was, however, no difference between the groups in quantitative muscle strength or pulmonary function testing. Functional scale may be a more discriminating outcome measure for SMA clinical trials. Muscle Nerve, 2009. Year: 2009 Title: An antisense microwalk reveals critical role of an intronic position linked to a unique long-distance interaction in pre-mRNA splicing Authors: Singh, N. N. Hollinger, K. Bhattacharya, D. Singh, R. N. Auth Address: Department of Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa 50011, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20413618 Abstract: Here we report a novel finding of an antisense oligonucleotide (ASO) microwalk in which we examined the position-specific role of intronic residues downstream from the 5' splice site (5' ss) of SMN2 exon 7, skipping of which is associated with spinal muscular atrophy (SMA), a leading genetic cause of infant mortality. Our results revealed the inhibitory role of a cytosine residue at the 10th intronic position ((10)C), which is neither conserved nor associated with any known splicing motif. Significance of (10)C emerged from the splicing pattern of SMN2 exon 7 in presence of a 14-mer ASO (L14) that sequestered two adjacent hnRNP A1 motifs downstream from (10)C and yet promoted SMN2 exon 7 skipping. Another 14-mer ASO (F14) that sequestered both, (10)C and adjacent hnRNP A1 motifs, led to a strong stimulation of SMN2 exon 7 inclusion. The inhibitory role of (10)C was found to be tightly linked to its unpaired status and specific positioning immediately upstream of a RNA:RNA helix formed between the targeting ASO and its intronic target. Employing a heterologous context as well as changed contexts of SMN2 intron 7, we show that the inhibitory effect of unpaired (10)C is dependent upon a long-distance interaction involving downstream intronic sequences. Our report furnishes one of the rare examples in which an ASO-based approach could be applied to unravel the critical role of an intronic position that may not belong to a linear motif and yet play significant role through long-distance interactions. Title: An assembly chaperone collaborates with the SMN complex to generate spliceosomal SnRNPs Authors: Chari, A. Golas, M. M. Klingenhager, M. Neuenkirchen, N. Sander, B. Englbrecht, C. Sickmann, A. Stark, H. Fischer, U. Auth Address: Department of Biochemistry, Biocenter, University of Wurzburg, Am Hubland, D-97074 Wurzburg, Germany. Pages: 497-509 Volume: 135 Number: 3 Keywords: Hela Cells Humans Models, Biological Molecular Chaperones/metabolism Nerve Tissue Proteins/metabolism Protein Methyltransferases/*chemistry/metabolism RNA/metabolism RNA-Binding Proteins/metabolism Ribonucleoproteins, Small Nuclear/chemistry/*metabolism Survival of Motor Neuron 1 Protein/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18984161 Abstract: Spliceosomal small nuclear ribonucleoproteins (snRNPs) are essential components of the nuclear pre-mRNA processing machinery. A hallmark of these particles is a ring-shaped core domain generated by the binding of Sm proteins onto snRNA. PRMT5 and SMN complexes mediate the formation of the core domain in vivo. Here, we have elucidated the mechanism of this reaction by both biochemical and structural studies. We show that pICln, a component of the PRMT5 complex, induces the formation of an otherwise unstable higher-order Sm protein unit. In this state, the Sm proteins are kinetically trapped, preventing their association with snRNA. The SMN complex subsequently binds to these Sm protein units, dissociates pICln, and catalyzes ring closure on snRNA. Our data identify pICln as an assembly chaperone and the SMN complex as a catalyst of spliceosomal snRNP formation. The mode of action of this combined chaperone/catalyst system is reminiscent of the mechanism employed by DNA clamp loaders. Year: 2008 Title: An autopsy case of spinal muscular atrophy type III (Kugelberg-Welander disease) Authors: Kuru, S. Sakai, M. Konagaya, M. Yoshida, M. Hashizume, Y. Saito, K. Auth Address: Department of Neurology, Suzuka National Hospital, Suzuka-shi, Mie, Japan. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18410269 Abstract: We report an autopsy case of a 67-year-old man clinicogenetically diagnosed as having spinal muscular atrophy (SMA) type III (Kugelberg-Welander disease), showing slowly progressive muscle wasting and weakness of the extremities. His brother showed similar manifestations. Autopsy revealed neuronal loss and severe gliosis in the anterior horns of the spinal cord, a marked neurogenic change of skeletal muscles and mild degeneration of cardiomyocytes. Chromatolytic change was seen in the anterior horn, but not in the Clarke's and thalamic nuclei. The anterior spinal roots were atrophic, and there was loss of myelinated fibers with abundant glial bundles. In addition, degeneration was also observed in the posterior column and dentate nucleus. The pathological features were essentially similar to those of SMA I. Chronic change was prominent while acute change was mild in degree, corresponding to a very long clinical course. Year: 2008 Title: An essential role for beta-actin mRNA localization and translation in Ca(2+)-dependent growth cone guidance Authors: Yao, J. Sasaki, Y. Wen, Z. Bassell, G. J. Zheng, J. Q. Auth Address: Department of Neuroscience and Cell Biology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854, USA. Pages: 1265-73 Volume: 9 Number: 10 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16980965 Abstract: Axon pathfinding requires directional responses of growth cones to extracellular cues, which have been shown to involve local synthesis of protein. The identity and functions of the locally produced proteins remain, however, unclear. Here we report that Ca(2+)-dependent bidirectional turning of Xenopus laevis growth cones requires localized distribution and translation of beta-actin messenger RNA. Both beta-actin mRNA and its zipcode-binding protein, ZBP1, are localized at the growth cone and become asymmetrically distributed upon local exposure to brain-derived neurotrophic factor (BDNF). Inhibition of protein synthesis or antisense interference with beta-actin mRNA-ZBP1 binding abolishes both Ca(2+)-mediated attraction and repulsion. In addition, attraction involves a local increase in beta-actin, whereas repulsion is accompanied by a local decrease in beta-actin; thus, both produce a synthesis- and ZBP1 binding-dependent beta-actin asymmetry but with opposite polarities. Together with a similar asymmetry in Src activity during bidirectional responses, our findings indicate that Ca(2+)-dependent spatial regulation of beta-actin synthesis through Src contributes to the directional motility of growth cones during guidance. Year: 2006 Month: 10 Title: An essential SMN interacting protein (SIP1) is not involved in the phenotypic variability of spinal muscular atrophy (SMA) Authors: Helmken, C. Wetter, A. Rudnik-Schoneborn, S. Liehr, T. Zerres, K. Wirth, B. Auth Address: Institute of Human Genetics, Bonn, Germany. Pages: 493-9 Volume: 8 Number: 7 Keywords: Alternative Splicing Child, Preschool Chromosomes, Human, Pair 14/genetics DNA/analysis/blood DNA Mutational Analysis DNA Primers/chemistry Exons Genetic Screening Genotype Humans In Situ Hybridization, Fluorescence Infant Introns Molecular Sequence Data Motor Neurons/metabolism/pathology Mutation Nerve Tissue Proteins/*genetics *Phenotype RNA, Messenger/analysis Research Support, Non-U.S. Gov't Reverse Transcriptase Polymerase Chain Reaction Spinal Muscular Atrophies of Childhood/*genetics/pathology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10909848 Abstract: The survival motor neuron (SMN) protein and the SMN interacting protein 1 (SIP1) are part of a 300 kD protein complex with a crucial role in snRNP biogenesis and pre-mRNA splicing. Both proteins are colocalised in nuclear structures called gems and in the cytoplasm. Approximately 96% of patients with autosomal recessive spinal muscular atrophy (SMA) show mutations in the SMN1 gene, while about 4% fail to show any mutation, despite a typical SMA phenotype. Additionally, sibs with identical 5q13 homologs and homozygous absence of SMN1 can show variable phenotypes which suggest that SMA is modified by other, yet unknown factors. Since both genes, SMN1 and SIP1, belong to the same pathway and are part of the same protein complex, it is obvious to ask whether mutations within SIP1 are responsible for both the phenotypic variability and the appearance of non-SMN mutated SMA patients. First, we identified the chromosomal location of SIP1 and assigned it to chromosomal region 14q13-q21 by fluorescence in situ hybridisation. No SMA related disorder has yet been assigned to this chromosomal region. Next, we determined the exon-intron structure of the SIP1 gene which encompasses 10 exons and identified five transcription isoforms. We sequenced either RT-PCR products or genomic DNA covering the complete coding region from 23 typical SMA patients who had failed to show any SMN1 mutation. No mutation and no polymorphism was found within SIP1. Additionally, we sequenced RT-PCR products or genomic fragments of the entire SIP1 coding region from 26 sibs of 11 SMA families with identical genotypes (delta7SMN/delta7SMN or delta7SMN/other mutation) but different phenotypes and again no mutation was found. Finally, we performed quantitative analysis of RT-PCR products from the same 26 sibs. No difference in expression level of the five isoforms among phenotypically variable sibs was observed. Based on these data, we suggest that neither the phenotypic variability nor the 5q-unlinked SMA are caused by mutations within SIP1. Year: 2000 Month: 7 Title: An exon skipping-associated nonsense mutation in the dystrophin gene uncovers a complex interplay between multiple antagonistic splicing elements Authors: Disset, A. Bourgeois, C. F. Benmalek, N. Claustres, M. Stevenin, J. Tuffery-Giraud, S. Auth Address: Laboratoire de Genetique Moleculaire, Institut Universitaire de Recherche Clinique (IURC), CHU Montpellier F34000, France. Pages: 999-1013 Volume: 15 Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16461336 Abstract: A nonsense mutation c.4250T>A (p.Leu1417X) in the dystrophin gene of a patient with an intermediate phenotype of muscular dystrophy induces partial in-frame skipping of exon 31. On the basis of UV cross-linking assays and pull-down analysis, we present evidence that the skipping of this exon is because of the creation of an exonic splicing silencer, which acts as a highly specific binding site (UAGACA) for a known repressor protein, hnRNP A1. Recombinant hnRNP A1 represses exon inclusion both in vitro and in vivo upon transient transfection of C2C12 cells with Duchenne muscular dystrophy (DMD) minigenes carrying the c.4250T>A mutation. Furthermore, we identified a downstream splicing enhancer in the central region of exon 31. This region functions as a Tra2beta-dependent exonic splicing enhancer (ESE) in vitro when inserted into a heterologous splicing reporter, and deletion of the ESE showed that incorporation of exon 31 depends on the Tra2beta-dependent enhancer both in the wild-type and mutant context. We conclude that dystrophin exon 31 contains juxtaposed sequence motifs that collaborate to regulate exon usage. This is the first elucidation of the molecular mechanism leading to exon skipping in the dystrophin gene and allowing the occurrence of a milder phenotype than the expected DMD phenotype. The knowledge of which cis-acting sequence within an exon is important for its definition will be essential for the alternative gene therapy approaches based on modulation of splicing to bypass DMD-causing mutations in the endogenous dystrophin gene. Year: 2006 Title: An exonic enhancer is required for inclusion of an essential exon in the SMA-determining gene SMN Authors: Lorson, C. L. Androphy, E. J. Auth Address: Department of Dermatology, New England Medical Center and Tufts University School of Medicine, Boston, MA 02111, USA. Pages: 259-65 Volume: 9 Number: 2 Keywords: Alternative Splicing/genetics Base Sequence Cyclic AMP Response Element-Binding Protein Enhancer Elements (Genetics)/*genetics Exons/*genetics Humans Molecular Sequence Data Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/*genetics RNA-Binding Proteins Research Support, Non-U.S. Gov't Tumor Cells, Cultured Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10607836 Abstract: The survival motor neuron genes, SMN1 and SMN2, encode identical proteins; however, only homo- zygous loss of SMN1 correlates with the development of spinal muscular atrophy (SMA). We have previously shown that a single non-polymorphic nucleotide difference in SMN exon 7 dramatically affects SMN mRNA processing. SMN1 primarily produces a full-length RNA whereas SMN2 expresses dramatically reduced full-length RNA and abundant levels of an aberrantly spliced transcript lacking exon 7. The importance of proper exon 7 processing has been underscored by the identification of several mutations within splice sites adjacent to exon 7. Here we show that an AG-rich exonic splice enhancer (ESE) in the center of SMN exon 7 is required for inclusion of exon 7. This region functioned as an ESE in a heterologous context, supporting efficient in vitro splicing of the Drosophila double-sex gene. Finally, the protein encoded by the exon-skipping event, Delta7, was less stable than full-length SMN, providing additional evidence of why SMN2 fails to compensate for the loss of SMN1 and leads to the development of SMA. Year: 2000 Title: An extended inhibitory context causes skipping of exon 7 of SMN2 in spinal muscular atrophy Authors: Singh, N. N. Androphy, E. J. Singh, R. N. Auth Address: Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01605-2324, USA. Pages: 381-8 Volume: 315 Number: 2 Keywords: Amino Acid Motifs Base Sequence Binding Sites Cells, Cultured Codon, Terminator Cyclic AMP Response Element-Binding Protein Enhancer Elements (Genetics) Exons Gene Silencing Heterogeneous-Nuclear Ribonucleoprotein Group A-B/*genetics Heterogeneous-Nuclear Ribonucleoproteins/genetics Humans Models, Genetic Molecular Sequence Data Muscular Atrophy, Spinal/*genetics Mutation Nerve Tissue Proteins/*genetics Plasmids/metabolism RNA/chemistry RNA Splicing RNA, Messenger/metabolism RNA-Binding Proteins Research Support, U.S. Gov't, P.H.S. Reverse Transcriptase Polymerase Chain Reaction Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14766219 Abstract: SMN1 and SMN2 represent the two nearly identical copies of the survival of motor neuron gene in humans. The most frequent cause of spinal muscular atrophy (SMA) is loss of SMN1 accompanied by the inability of SMN2 to compensate due to an inhibitory mutation at position 6 in exon 7 (C6U) that causes exon 7 exclusion. How this single exonic nucleotide regulates exon 7 recognition has been of major interest. Based on score matrices and in vitro assays, abrogation of an exonic splicing enhancer (ESE) associated with SF2/ASF has been considered as the cause of exon 7 exclusion. However, a recent report supports the creation of an exonic splicing silencer (ESS) associated with hnRNP A1 as the determining factor for exon 7 exclusion. Here we show that C6U strengthens an inhibitory context that covers a larger sequence than the hnRNP A1 binding site. The inhibitory context can also be strengthened by the addition of a G residue at the first position of exon 7 in SMN1, promoting exon 7 skipping despite the presence of SF2/ASF binding site. Through in vivo selection and a series of mutations we demonstrate that the strengthening of the extended inhibitory context at the 5' end of exon 7 is exercised through overlapping sequence motifs that collaborate to regulate exon usage. Year: 2004 Title: An extensive network of coupling among gene expression machines Authors: Maniatis, T. Reed, R. Auth Address: Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA. maniatis@mcb.harvard.edu Pages: 499-506 Volume: 416 Number: 6880 Keywords: *Gene Expression Models, Genetic Poly A/metabolism RNA Caps RNA Polymerase II/metabolism RNA Precursors/metabolism RNA Splicing RNA, Messenger/metabolism Transcription, Genetic Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11932736 Abstract: Gene expression in eukaryotes requires several multi-component cellular machines. Each machine carries out a separate step in the gene expression pathway, which includes transcription, several pre-messenger RNA processing steps and the export of mature mRNA to the cytoplasm. Recent studies lead to the view that, in contrast to a simple linear assembly line, a complex and extensively coupled network has evolved to coordinate the activities of the gene expression machines. The extensive coupling is consistent with a model in which the machines are tethered to each other to form 'gene expression factories' that maximize the efficiency and specificity of each step in gene expression. Year: 2002 Title: An in vivo reporter system for measuring increased inclusion of exon 7 in SMN2 mRNA: potential therapy of SMA Authors: Zhang, M. L. Lorson, C. L. Androphy, E. J. Zhou, J. Auth Address: Department of Dermatology, New England Medical Center and Tufts University School of Medicine, Boston, MA 02111, USA. Pages: 1532-8 Volume: 8 Number: 20 Keywords: Artificial Gene Fusion/methods Cell Line Cyclic AMP Response Element-Binding Protein Enzyme Inhibitors/pharmacology Exons Gene Expression Gene Therapy/*methods Green Fluorescent Proteins Humans Luciferases/genetics Luminescent Proteins/genetics Motor Neurons/*metabolism Muscular Atrophy, Spinal/genetics/metabolism/*therapy Nerve Tissue Proteins/*genetics/metabolism Phosphoric Monoester Hydrolases/antagonists & inhibitors Phosphorylation RNA Splicing/*drug effects RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Reverse Transcriptase Polymerase Chain Reaction Transfection/methods Vanadates/*pharmacology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11704813 Abstract: Spinal muscular atrophy (SMA) is a degenerative motor neuron disorder resulting from homozygous loss of the SMN1 gene. SMN2, a nearly identical copy gene, is preserved in SMA patients. A single nucleotide difference between SMN1 and SMN2 causes exon 7 skipping in the majority of SMN2 mRNA. Gene therapy through modulation of SMN2 gene transcription in SMA patients may be possible. We constructed a series of SMN mini-genes comprised of SMN exon 6 to exon 8 sequences fused to green fluorescence protein (GFP) or luciferase reporters, to monitor SMN exon 7 splicing. These reporters recapitulated the splicing patterns of the endogenous SMN gene in stable cell lines. The SMN1-luciferase reporter was approximately 3.5-fold more active than SMN2-luciferase and SMN1-GFP intensities were visually distinguishable from SMN2-GFP. We have screened chemical inducers and inhibitors of kinase pathways using stable SMN-reporter lines and found that the phosphatase inhibitor sodium vanadate specifically stimulated exon 7 inclusion within SMN2 mRNAs. This is the first compound identified that can stimulate exon 7 inclusion into transcripts derived from the endogenous SMN2 gene. These results demonstrate that this system can be utilized to identify small molecules that regulate the splicing of SMN exon 7. Year: 2001 Month: 10 Title: An integrated motion capture system for evaluation of neuromuscular disease patients Authors: Gamarnik, V. Pan, S. Malke, J. Chiu, C. Koo, B. Montes, J. Yeager, K. Marra, J. Dunaway, S. Montgomery, M. Strauss, N. De Vivo, D. C. Kaufmann, P. Morrison, B. Konofagou, E. Auth Address: Biomedical Engineering Department, Columbia University, New York, NY 10027 USA. Pages: 218-21 Volume: 1 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19964732 Abstract: There currently exist a variety of methods for evaluating movement in patients suffering from neuromuscular diseases (NMD). These tests are primarily performed in the clinical setting and evaluated by highly trained individuals, rather than evaluating patient in their natural environments (i.e., home or school). Currently available automated motion capture modalities offer a highly accurate means of assessing general motion, but are also limited to a highly controlled setting. Recent advances in MEMS technology have introduced the possibility of robust motion capture in uncontrolled environments, while minimizing user interference with self-initiated motion, especially in weaker subjects. The goal of this study is to design and evaluate a MEMS-sensor-based system for motion capture in the NMD patient population. The highly interdisciplinary effort has led to significant progress toward the implementation of a new device, which is accurate, clinically relevant, and highly affordable. Year: 2009 Month: 1 Title: An intronic element contributes to splicing repression in spinal muscular atrophy Authors: Kashima, T. Rao, N. Manley, J. L. Auth Address: Department of Biological Sciences, Columbia University, New York, NY 10027. Pages: 3426-31 Volume: 104 Number: 9 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17307868 Abstract: The neurodegenerative disease spinal muscular atrophy is caused by mutation of the survival motor neuron 1 (SMN1) gene. SMN2 is a nearly identical copy of SMN1 that is unable to prevent disease, because most SMN2 transcripts lack exon 7 and thus produce a nonfunctional protein. A key cause of inefficient SMN2 exon 7 splicing is a single nucleotide difference between SMN1 and SMN2 within exon 7. We previously provided evidence that this base change suppresses exon 7 splicing by creating an inhibitory element, a heterogeneous nuclear ribonucleoprotein (hnRNP) A1-dependent exonic splicing silencer. We now find that another rare nucleotide difference between SMN1 and SMN2, in intron 7, potentially creates a second SMN2-specific hnRNP A1 binding site. Remarkably, this single base change does indeed create a high-affinity hnRNP A1 binding site, and base substitutions that disrupt it restore exon 7 inclusion in vivo and prevent hnRNP A1 binding in vitro. We propose that interactions between hnRNP A1 molecules bound to the exonic and intronic sites cooperate to exclude exon 7 and discuss the significance of this exclusion with respect to SMN expression and splicing control more generally. Year: 2007 Title: An SMA Project Report: Neural Cell-Based Assays Derived from Human Embryonic Stem Cells Authors: Wilson, P. G. Cherry, J. J. Schwamberger, S. Adams, A. M. Zhou, J. Shin, S. Stice, S. L. Auth Address: Regenerative Bioscience Center, University of Georgia, Athens, GA 20602. Pages: 1027-42 Volume: 16 Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18078382 Abstract: Human embryonic stem (ES) cells are promising resources for developing new treatments for neurodegenerative diseases. Spinal muscular atrophy (SMA) is one of the leading causes of childhood paralysis and infant mortality. SMA is caused by inactivation of the survival motor neuron-1 (SMN1) gene. The nearly identical SMN2 gene contains a silent polymorphism that disrupts splicing and as a result cannot compensate for loss of SMN1. The SMA Project was established by the National Institute of Neurological Disorders and Stroke (NINDS) as a pilot effort to establish a fully transparent coalition between academics, industry, and government to create a centralized network of shared resources and information to identify and test new SMA therapeutics. As one of the funded projects, the work described here tested the feasibility of generating a SMA cell-based assay using neural lineages derived from human ES cells approved for National Institutes of Health (NIH)-funded research. Minigene cassettes were constructed, employing firefly luciferase or green fluorescent protein (GFP) as reporters for splicing efficiency of SMN1 and/ or SMN2 under the control of the SMN1, SMN2, or cytomegalovirus (CMV) promoters. Transient transfection of proliferating neuroprogenitors in a 96-well format with plasmid DNA or adenoviral vectors showed differential levels that correlated with the splicing minigene and the promoter used; luciferase activities with SMN1 splicing minigenes were higher than SMN2, and the CMV promoter generated higher levels of activity than the SMN1 and SMN2 promoters. Our results indicate that human ES cell-derived neuroprogenitors provide a promising new primary cell source for assays of new therapeutics for neurodegenerative diseases. Year: 2007 Month: 12 Title: An update of the mutation spectrum of the survival motor neuron gene (SMN1) in autosomal recessive spinal muscular atrophy (SMA) Authors: Wirth, B. Auth Address: Institute of Human Genetics, Bonn, Germany. bwirth@uni-bonn.de Pages: 228-37 Volume: 15 Number: 3 Keywords: Amino Acid Sequence Cyclic AMP Response Element-Binding Protein *Genes, Recessive Humans Molecular Sequence Data Muscular Atrophy, Spinal/*genetics Mutation Nerve Tissue Proteins/*genetics RNA-Binding Proteins Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10679938 Abstract: Spinal muscular atrophy (SMA) is characterized by degeneration of motor neurons in the spinal cord, causing progressive weakness of the limbs and trunk, followed by muscle atrophy. SMA is one of the most frequent autosomal recessive diseases, with a carrier frequency of 1 in 50 and the most common genetic cause of childhood mortality. The phenotype is extremely variable, and patients have been classified in type I-III SMA based on age at onset and clinical course. All three types of SMA are caused by mutations in the survival motor neuron gene (SMN1). There are two almost identical copies, SMN1 and SMN2, present on chromosome 5q13. Only homozygous absence of SMN1 is responsible for SMA, while homozygous absence of SMN2, found in about 5% of controls, has no clinical phenotype. Ninety-six percent of SMA patients display mutations in SMN1, while 4% are unlinked to 5q13. Of the 5q13-linked SMA patients, 96.4% show homozygous absence of SMN1 exons 7 and 8 or exon 7 only, whereas 3. 6% present a compound heterozygosity with a subtle mutation on one chromosome and a deletion/gene conversion on the other chromosome. Among the 23 different subtle mutations described so far, the Y272C missense mutation is the most frequent one, at 20%. Given this uniform mutation spectrum, direct molecular genetic testing is an easy and rapid analysis for most of the SMA patients. Direct testing of heterozygotes, while not trivial, is compromised by the presence of two SMN1 copies per chromosome in about 4% of individuals. The number of SMN2 copies modulates the SMA phenotype. Nevertheless, it should not be used for prediction of severity of the SMA. Year: 2000 Month: 1 Title: An ~140-kb deletion associated with feline spinal muscular atrophy implies an essential LIX1 function for motor neuron survival Authors: Fyfe, J. C. Menotti-Raymond, M. David, V. A. Brichta, L. Schaffer, A. A. Agarwala, R. Murphy, W. J. Wedemeyer, W. J. Gregory, B. L. Buzzell, B. G. Drummond, M. C. Wirth, B. O'Brien S, J. Auth Address: Laboratory of Comparative Medical Genetics, Department of Microbiology & Molecular Genetics, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan 48824, USA; Pages: 1084-90 Volume: 16 Number: 9 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16899656 Abstract: The leading genetic cause of infant mortality is spinal muscular atrophy (SMA), a clinically and genetically heterogeneous group of disorders. Previously we described a domestic cat model of autosomal recessive, juvenile-onset SMA similar to human SMA type III. Here we report results of a whole-genome scan for linkage in the feline SMA pedigree using recently developed species-specific and comparative mapping resources. We identified a novel SMA gene candidate, LIX1, in an ~140-kb deletion on feline chromosome A1q in a region of conserved synteny to human chromosome 5q15. Though LIX1 function is unknown, the predicted secondary structure is compatible with a role in RNA metabolism. LIX1 expression is largely restricted to the central nervous system, primarily in spinal motor neurons, thus offering explanation of the tissue restriction of pathology in feline SMA. An exon sequence screen of 25 human SMA cases, not otherwise explicable by mutations at the SMN1 locus, failed to identify comparable LIX1 mutations. Nonetheless, a LIX1-associated etiology in feline SMA implicates a previously undetected mechanism of motor neuron maintenance and mandates consideration of LIX1 as a candidate gene in human SMA when SMN1 mutations are not found. Year: 2006 Month: 9 Title: Analysis of Neuroprotective Effects of Valproic Acid on Primary Motor Neurons in Monoculture or Co-cultures with Astrocytes or Schwann Cells Authors: Ragancokova, D. Jahn, K. Kotsiari, A. Schlesinger, F. Haastert, K. Stangel, M. Petri, S. Krampfl, K. Auth Address: Department of Neurology and Clinical Neurophysiology, Hannover Medical School, OE 7210, Carl-Neuberg-Strasse 1, 30625, Hannover, Germany, ragancokova.daniela@mh-hannover.de. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19330445 Abstract: Chronic dysregulation of the intracellular Ca(2+) homeostasis (excitotoxicity) is thought to contribute to the development of motor neuron diseases. Valproic acid (VPA) is widely used as an antiepileptic drug and acts mainly by inhibition of sodium channels and by enhancing the level of the inhibitory neurotransmitter gamma-aminobutyric acid. Neuroprotective capacities of VPA are supposed to arise also from the inhibition of histone deacetylases. We investigated the viability of highly purified rat embryonic motor neurons cultured on glial feeder layers, composed of either astrocytes or Schwann cells, or in the absence of glia, monoculture in presence of VPA and/or kainate (KA) using immunocytochemistry and calcium imaging. A significant effect of the culture and co-culture conditions on the viability of motor neurons in our in vitro model of excitotoxicity was detected. The neuroprotective effect of VPA on primary embryonic motor neuron cultures was not proven. A functional interaction between VPA and KA occurred during the first 10 days in culture. Year: 2009 Title: Androgen-dependent pathology demonstrates myopathic contribution to the Kennedy disease phenotype in a mouse knock-in model Authors: Yu, Z. Dadgar, N. Albertelli, M. Gruis, K. Jordan, C. Robins, D. M. Lieberman, A. P. Auth Address: Department of Pathology, Department of Human Genetics, and Department of Neurology, University of Michigan Medical School, Ann Arbor, Michigan, USA. Department of Psychology and Neuroscience Program, Michigan State University, East Lansing, Michigan, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16981011 Abstract: Kennedy disease, a degenerative disorder characterized by androgen-dependent neuromuscular weakness, is caused by a CAG/glutamine tract expansion in the androgen receptor (Ar) gene. We developed a mouse model of Kennedy disease, using gene targeting to convert mouse androgen receptor (AR) to human sequence while introducing 113 glutamines. AR113Q mice developed hormone and glutamine length-dependent neuromuscular weakness characterized by the early occurrence of myopathic and neurogenic skeletal muscle pathology and by the late development of neuronal intranuclear inclusions in spinal neurons. AR113Q males unexpectedly died at 2-4 months. We show that this androgen-dependent death reflects decreased expression of skeletal muscle chloride channel 1 (CLCN1) and the skeletal muscle sodium channel alpha-subunit, resulting in myotonic discharges in skeletal muscle of the lower urinary tract. AR113Q limb muscles show similar myopathic features and express decreased levels of mRNAs encoding neurotrophin-4 and glial cell line-derived neurotrophic factor. These data define an important myopathic contribution to the Kennedy disease phenotype and suggest a role for muscle in non-cell autonomous toxicity of lower motor neurons. Year: 2006 Title: Anesthesia and perioperative medical management of children with spinal muscular atrophy Authors: Graham, R. J. Athiraman, U. Laubach, A. E. Sethna, N. F. Auth Address: Associate in Division of Critical Care Medicine, Children's Hospital Boston, Boston, MA, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19558636 Abstract: Objective: To describe the perioperative medical care, anesthetic considerations, and the risk of postanesthetic respiratory failure in patients with pediatric spinal muscular atrophy (SMA). Methods: There is a retrospective chart review carried out at an urban, tertiary care pediatric hospital. All patients with ICD-9 codes corresponding to SMA diagnoses were identified, and records screened for anesthetic and perioperative medical management. Medical records were reviewed for demographic, clinical, and outcome data. Main results: Twenty-five children with SMA (10 type I, 8 type II, 7 type III) accounted for 56 general and regional anesthetic cases. Twenty-one (38%) cases had preexisting ventilator dependence. Total intravenous anesthesia with nitrous oxide was provided in 14 (25%) cases, balanced anesthesia with inhalational agents and epidural or systemic opioids were used in 41 (84%) cases, and one infant received a spinal anesthesia. Intraoperative and postoperative complications occurred in 2 (4%) and 2 (4%) cases respectively. One case required an unplanned postanesthetic intensive care unit admission, and there were two late deaths with redirection of care. Conclusions: Perioperative care can be provided for children with SMA safely and effectively with total intravenous or inhaled anesthetics along with the judicious use of opioids to improve patient comfort without increased morbidity. Further prospective studies of standardized anesthetic and perioperative respiratory medical management in this population are required to minimize the decompensation of tenuous preoperative respiratory function. Year: 2009 Title: Animal models of spinal muscular atrophy Authors: Monani, U. R. Coovert, D. D. Burghes, A. H. Auth Address: Department of Neurology, Means Hall, 1654 Upham Drive, The Ohio State University, Columbus, OH 43210, USA. Pages: 2451-7 Volume: 9 Number: 16 Keywords: Animals Cyclic AMP Response Element-Binding Protein *Disease Models, Animal Forecasting Gene Duplication Humans Mice *Muscular Atrophy, Spinal/genetics Nerve Tissue Proteins/genetics/metabolism RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11005801 Abstract: Proximal spinal muscular atrophy (SMA) is the second most common autosomal recessive inherited disorder in humans. It is the most common genetic cause of infant mortality. As yet, there is no cure for this neuromuscular disorder which affects the lower motor neurons and proximal muscles of the limbs and trunk. In the last decade, significant advances have been made in understanding this disease, from linkage analysis to isolating the defective gene and identifying its protein product. This review summarizes the most recent advance in SMA research: the development of animal models of the disease, in particular mouse models of SMA. The SMA mice that we describe here present with symptoms similar to those seen in SMA patients. They promise to further the understanding of the molecular basis of this disease and demonstrate the feasibility of using the intact SMN2 gene, found in all SMA patients, as a means of treating this disorder. Year: 2000 Month: 10 Title: Another step forward for SELEXive splicing Authors: Buratti, E. Baralle, F. E. Auth Address: International Centre for Genetic Engineering and Biotechnology, Padriciano 99, Trieste 34012, Italy. Pages: 5-9 Volume: 11 Number: 1 Keywords: *Alternative Splicing Animals Base Sequence Biomedical Research/methods Computational Biology Cyclic AMP Response Element-Binding Protein/*genetics Exons Humans Internet Molecular Sequence Data Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/*genetics RNA Precursors/*genetics RNA, Messenger/genetics RNA-Binding Proteins/*genetics Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15649816 Abstract: Splicing alterations are being increasingly reported to cause human diseases. However, predicting beforehand whether a given mutation might lead to aberrant splicing is often hampered by insufficient knowledge of which cis-acting sequences affect exon recognition. To better define these sequences, experimental methods have been developed that integrate pre-mRNA splicing with sequence selection assays. Recently, a novel in vivo selection method based on a partially randomized full-length exon has enabled the identification of new splicing-controlling elements in SMN exon 7. Skipping of this exon results in an aberrant protein and is involved in proximal spinal muscular atrophy (SMA). Hopefully, this approach will provide novel targets for nascent RNA molecular medicine approaches for the recovery of abnormal pre-mRNA splicing events. Year: 2005 Month: 1 Title: Antisense masking of an hnRNP A1/A2 intronic splicing silencer corrects SMN2 splicing in transgenic mice Authors: Hua, Y. Vickers, T. A. Okunola, H. L. Bennett, C. F. Krainer, A. R. Auth Address: Cold Spring Harbor Laboratory, PO Box 100, Cold Spring Harbor, NY 11724, USA. Pages: 834-48 Volume: 82 Number: 4 Keywords: Animals Base Sequence Cell Line Cyclic AMP Response Element-Binding Protein/*genetics Exons Gene Therapy Heterogeneous-Nuclear Ribonucleoprotein Group A-B/*metabolism Humans Introns Mice Mice, Transgenic Molecular Sequence Data Muscular Atrophy, Spinal/therapy Nerve Tissue Proteins/*genetics Oligonucleotides, Antisense/genetics/*pharmacology/therapeutic use RNA Splicing/*drug effects RNA-Binding Proteins/*genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18371932 Abstract: Survival of motor neuron 2, centromeric (SMN2) is a gene that modifies the severity of spinal muscular atrophy (SMA), a motor-neuron disease that is the leading genetic cause of infant mortality. Increasing inclusion of SMN2 exon 7, which is predominantly skipped, holds promise to treat or possibly cure SMA; one practical strategy is the disruption of splicing silencers that impair exon 7 recognition. By using an antisense oligonucleotide (ASO)-tiling method, we systematically screened the proximal intronic regions flanking exon 7 and identified two intronic splicing silencers (ISSs): one in intron 6 and a recently described one in intron 7. We analyzed the intron 7 ISS by mutagenesis, coupled with splicing assays, RNA-affinity chromatography, and protein overexpression, and found two tandem hnRNP A1/A2 motifs within the ISS that are responsible for its inhibitory character. Mutations in these two motifs, or ASOs that block them, promote very efficient exon 7 inclusion. We screened 31 ASOs in this region and selected two optimal ones to test in human SMN2 transgenic mice. Both ASOs strongly increased hSMN2 exon 7 inclusion in the liver and kidney of the transgenic animals. Our results show that the high-resolution ASO-tiling approach can identify cis-elements that modulate splicing positively or negatively. Most importantly, our results highlight the therapeutic potential of some of these ASOs in the context of SMA. Year: 2008 Month: 4 Title: Antisense oligonucleotide therapy for neurodegenerative disease Authors: Smith, R. A. Miller, T. M. Yamanaka, K. Monia, B. P. Condon, T. P. Hung, G. Lobsiger, C. S. Ward, C. M. McAlonis-Downes, M. Wei, H. Wancewicz, E. V. Bennett, C. F. Cleveland, D. W. Auth Address: Ludwig Institute for Cancer Research, University of California, San Diego, La Jolla, California, USA. Pages: 2290-6 Volume: 116 Number: 8 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16878173 Abstract: Neurotoxicity from accumulation of misfolded/mutant proteins is thought to drive pathogenesis in neurodegenerative diseases. Since decreasing levels of proteins responsible for such accumulations is likely to ameliorate disease, a therapeutic strategy has been developed to downregulate almost any gene in the CNS. Modified antisense oligonucleotides, continuously infused intraventricularly, have been demonstrated to distribute widely throughout the CNS of rodents and primates, including the regions affected in the major neurodegenerative diseases. Using this route of administration, we found that antisense oligonucleotides to superoxide dismutase 1 (SOD1), one of the most abundant brain proteins, reduced both SOD1 protein and mRNA levels throughout the brain and spinal cord. Treatment initiated near onset significantly slowed disease progression in a model of amyotrophic lateral sclerosis (ALS) caused by a mutation in SOD1. This suggests that direct delivery of antisense oligonucleotides could be an effective, dosage-regulatable means of treating neurodegenerative diseases, including ALS, where appropriate target proteins are known. Year: 2006 Month: 8 Title: Antisense oligonucleotide-induced exon skipping restores dystrophin expression in vitro in a canine model of DMD Authors: McClorey, G. Moulton, H. M. Iversen, P. L. Fletcher, S. Wilton, S. D. Auth Address: 1Experimental Molecular Medicine Group, Australian Neuromuscular Research Insitute, Centre for Neuromuscular and Neurological Disorders, University of Western Australia, QEII Medical Centre, Perth, Western Australia, Australia. Pages: 1373-81 Volume: 13 Number: 19 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16724091 Abstract: Manipulation of pre-mRNA splicing by antisense oligonucleotides (AOs) offers considerable potential for a number of genetic disorders. One of these is Duchenne muscular dystrophy (DMD), where mutations in the dystrophin gene typically result in premature termination of translation that causes a loss of functional protein. AOs can induce exon skipping such that the mutation is by-passed and the reading frame restored, producing an internally deleted protein similar to that found in the milder Becker muscular dystrophy. To date, this approach has been applied to the mdx mouse model in vitro and in vivo and in human myoblast cultures. Here, we report the application of AO-directed exon skipping to induce dystrophin expression in vitro in a canine model of DMD, golden retriever muscular dystrophy (GRMD). The efficacy of 2'-O-methyl phosphorothioate (2OMe), phosphorodiamidate morpholino oligomers (PMOs) and peptide-linked PMOs (PMO-Pep) to induce dystrophin expression was assessed. The 2OMe chemistry was only effective for short-term induction of corrected transcript and could not induce detectable dystrophin protein. The PMO chemistry generally induced limited exon skipping at only high concentrations; however, a low level of dystrophin protein was produced in treated cells. Use of the PMO-Pep, applied here for the first time to a DMD model, was able to induce high and sustained levels of exon skipping and induced the highest level of dystrophin expression with no apparent adverse effects upon the cells. The induction of dystrophin in the GRMD model offers the potential for further testing of AO delivery regimens in a larger animal model of DMD, in preparation for application in human clinical trials.Gene Therapy (2006) 13, 1373-1381. doi:10.1038/sj.gt.3302800; published online 25 May 2006. Year: 2006 Month: 10 Title: Antisense technologies. Improvement through novel chemical modifications Authors: Kurreck, J. Auth Address: Institut fur Chemie-Biochemie, Freie Universitat Berlin, Germany. jkurreck@chemie.fu-berlin.de Pages: 1628-44 Volume: 270 Number: 8 Keywords: Animals Base Sequence Clinical Trials Humans Oligodeoxyribonucleotides, Antisense/chemical synthesis/*chemistry/pharmacokinetics/therapeutic use Oligoribonucleotides, Antisense/chemical synthesis/chemistry/pharmacokinetics RNA, Catalytic/chemistry RNA, Small Interfering/genetics Research Support, Non-U.S. Gov't Tissue Distribution Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12694176 Abstract: Antisense agents are valuable tools to inhibit the expression of a target gene in a sequence-specific manner, and may be used for functional genomics, target validation and therapeutic purposes. Three types of anti-mRNA strategies can be distinguished. Firstly, the use of single stranded antisense-oligonucleotides; secondly, the triggering of RNA cleavage through catalytically active oligonucleotides referred to as ribozymes; and thirdly, RNA interference induced by small interfering RNA molecules. Despite the seemingly simple idea to reduce translation by oligonucleotides complementary to an mRNA, several problems have to be overcome for successful application. Accessible sites of the target RNA for oligonucleotide binding have to be identified, antisense agents have to be protected against nucleolytic attack, and their cellular uptake and correct intracellular localization have to be achieved. Major disadvantages of commonly used phosphorothioate DNA oligonucleotides are their low affinity towards target RNA molecules and their toxic side-effects. Some of these problems have been solved in 'second generation' nucleotides with alkyl modifications at the 2' position of the ribose. In recent years valuable progress has been achieved through the development of novel chemically modified nucleotides with improved properties such as enhanced serum stability, higher target affinity and low toxicity. In addition, RNA-cleaving ribozymes and deoxyribozymes, and the use of 21-mer double-stranded RNA molecules for RNA interference applications in mammalian cells offer highly efficient strategies to suppress the expression of a specific gene. Year: 2003 Month: 4 Title: Apoptosis-related proteins in skeletal muscle fibers of spinal muscular atrophy Authors: Tews, D. S. Goebel, H. H. Auth Address: Division of Neuropathology, Mainz University Medical Center, Germany. Pages: 150-6 Volume: 56 Number: 2 Keywords: Adult Aged Apoptosis/*physiology Caspase 1 Cysteine Endopeptidases/metabolism Female Humans Infant Male Middle Aged Muscle Fibers/metabolism Muscle Proteins/metabolism/*physiology Muscle, Skeletal/*metabolism/pathology Muscular Atrophy, Spinal/*metabolism/*pathology Peripheral Nervous System Diseases/metabolism/pathology Proto-Oncogene Proteins/metabolism Proto-Oncogene Proteins c-bcl-2/metabolism Reference Values bcl-2-Associated X Protein bcl-X Protein Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9034368 Abstract: There is evidence that apoptosis in spinal muscular atrophies (SMA) is not restricted to motor neurons but also affects muscle fibers. Studying the expression of several apoptosis-associated proteins we found constant expression of bax in muscle fibers, which promoted cell death. The expression of bax correlated with defective innervation of muscle fibers was also indicated by upregulation of N-CAM. While in early-onset SMA atrophic as well as normo- and hypertrophic muscle fibers displayed expression of bax, muscle fibers in late-onset SMA and peripheral neuropathies showed bax-expression only in atrophic fibers. Other investigated apoptosis-associated factors comprised interleukin-1 beta converting enzyme (ICE), mediating cell death by cleavage of actin filaments, as well as bcl-2 and bcl-x, both inhibiting apoptosis by acting as antioxidants. They were only expressed in atrophic muscle fibers, predominantly in late-onset SMA and peripheral neuropathies. We consider the lack of expression of these apoptosis-related proteins in early infantile SMA to be associated with muscle fiber immaturity due to defective innervation and suggest that immature muscle fibers are not able to produce sufficient levels of some proteins. A sufficient amount of expression of apoptosis-protecting factors such as bcl-2 is needed to neutralize high bax-levels, and a lack of this expression will secondarily promote muscle fiber death in defective innervation. Year: 1997 Month: 2 Title: Apoptosis: a mechanism contributing to remodeling of skeletal muscle in response to hindlimb unweighting Authors: Allen, D. L. Linderman, J. K. Roy, R. R. Bigbee, A. J. Grindeland, R. E. Mukku, V. Edgerton, V. R. Auth Address: Department of Physiological Science, University of California, Los Angeles 90095-1527, USA. Pages: C579-87 Volume: 273 Number: 2 Pt 1 Keywords: Animals *Apoptosis Female Hindlimb/*physiology Insulin-Like Growth Factor I/physiology Muscle, Skeletal/cytology/*physiology Physical Conditioning, Animal Rats Rats, Sprague-Dawley Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Time Factors *Weight-Bearing Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9277355 Abstract: The role of apoptosis in the elimination of myonuclei during hindlimb unloading-induced atrophy and the inhibition of apoptosis in the prevention of muscle atrophy were examined. The number of nuclei demonstrating double-stranded DNA fragmentation seen by terminal deoxynucleotidyl transferase (TDT) histochemical staining, an indicator of apoptosis, was significantly increased after 14 days of suspension. Double staining with TDT and antilaminin immunohistochemistry revealed that some TDT-positive nuclei were within the fiber lamina and were most likely myonuclei. The number of fibers containing morphologically abnormal nuclei was also significantly greater in suspended compared with control rats. Combined treatment with growth hormone and insulin-like growth factor I (GH/ IGF-I) and resistance exercise attenuated the increase in TDT-positive nuclei (approximately 26%, P > 0.05) and significantly decreased the number of fibers with morphologically abnormal nuclei. The data suggest that 1) "programmed nuclear death" contributes to the elimination of myonuclei and/or satellite cells from atrophying fibers, and 2) GH/IGF-I administration plus muscle loading ameliorates the apoptosis associated with hindlimb unloading. Year: 1997 Month: 8 Title: APP binds DR6 to trigger axon pruning and neuron death via distinct caspases Authors: Nikolaev, A. McLaughlin, T. O'Leary, D. D. Tessier-Lavigne, M. Auth Address: Division of Research, Genentech, Inc., 1 DNA Way, South San Francisco, California 94080, USA. Pages: 981-9 Volume: 457 Number: 7232 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19225519 Abstract: Naturally occurring axonal pruning and neuronal cell death help to sculpt neuronal connections during development, but their mechanistic basis remains poorly understood. Here we report that beta-amyloid precursor protein (APP) and death receptor 6 (DR6, also known as TNFRSF21) activate a widespread caspase-dependent self-destruction program. DR6 is broadly expressed by developing neurons, and is required for normal cell body death and axonal pruning both in vivo and after trophic-factor deprivation in vitro. Unlike neuronal cell body apoptosis, which requires caspase 3, we show that axonal degeneration requires caspase 6, which is activated in a punctate pattern that parallels the pattern of axonal fragmentation. DR6 is activated locally by an inactive surface ligand(s) that is released in an active form after trophic-factor deprivation, and we identify APP as a DR6 ligand. Trophic-factor deprivation triggers the shedding of surface APP in a beta-secretase (BACE)-dependent manner. Loss- and gain-of-function studies support a model in which a cleaved amino-terminal fragment of APP (N-APP) binds DR6 and triggers degeneration. Genetic support is provided by a common neuromuscular junction phenotype in mutant mice. Our results indicate that APP and DR6 are components of a neuronal self-destruction pathway, and suggest that an extracellular fragment of APP, acting via DR6 and caspase 6, contributes to Alzheimer's disease. Year: 2009 Title: Application of biomedical informatics to chronic pediatric diseases: A systematic review Authors: Moeinedin, F. Moineddin, R. Jadad, A. R. Hamid, J. S. To, T. Beyene, J. Auth Address: Pages: 22 Volume: 9 Number: 1 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19416540 Abstract: ABSTRACT: BACKGROUND: Chronic diseases affect millions of children worldwide leading to substantial disease burden to the children and their families as well as escalating health care costs. The increasing trend in the prevalence of complex pediatric chronic diseases requires innovative and optimal delivery of care. Health informatics (e-health) applications play an important role in improving health outcomes while being cost-effective. However, their utility in pediatric chronic diseases has not been studied in a comprehensive and systematic way. The objective of this study was to conduct a systematic review of the effects of biomedical informatics applications in pediatric chronic diseases. METHODS: A comprehensive literature search was conducted using MEDLINE, the Cochrane Library and EMBASE databases from inception of each database to September 2008. We included studies of any methodological type and any language that applied biomedical informatics to chronic conditions in children and adolescents 18 years of age or younger. Two independent reviewers carried out study selection and data extraction. Quality assessment was performed using a study design evaluation instrument to appraise the strength of the studies and their methodological adequacy. Because of heterogeneity in the conditions and outcomes we studied, a formal meta-analysis has not been performed. RESULTS: Based on our search strategy, 655 titles and abstracts were reviewed. From this set we identified 27 relevant articles that met our inclusion criteria. The results from these studies indicated that biomedical informatics applications have favourable clinical and patient outcomes including, but not limited to, reduced number of emergency room visits, improved knowledge on disease management, and enhanced satisfaction. Seventy percent of reviewed papers were published after year 2000, 89% of users were patients and 11% were either providers or caregivers. The majority (96%) of the selected studies reported improved outcomes. CONCLUSIONS: Published studies suggested positive impacts of informatics predominantly in pediatric asthma. As electronic tools become more widely adopted, there will be opportunities to improve patient care in a wide range of chronic illnesses through informatics solutions. Year: 2009 Title: Applying proteomics to the diagnosis and treatment of ALS and related diseases Authors: Bowser, R. Lacomis, D. Auth Address: Department of Pathology, University of Pittsburgh School of Medicine, BST S-420, 200 Lothrop Street, Pittsburgh, Pennsylvania 15261, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19670321 Abstract: Protein-based biomarkers for amyotrophic lateral sclerosis (ALS) and other motor neuron diseases (MNDs) have many potential clinical utilities, including diagnostic, prognostic, and drug development indications. During the past decade a number of potential protein biomarkers have been proposed for MNDs. Further verification studies, followed by large validation and qualification studies, are required to advance these initial discoveries toward clinical use. Study of additional patient populations, including disease mimics, is required during the validation phase of biomarker development. Important regulatory issues are discussed that will affect the timing and strategy for biomarker assay development in ALS and other MNDs. The continued development of protein biomarkers for MNDs requires extensive collaboration between academic clinicians and scientists in conjunction with the biotechnology and pharmaceutical industries. Muscle Nerve, 2009. Year: 2009 Title: Arimoclomol at dosages up to 300 mg/day is well tolerated and safe in amyotrophic lateral sclerosis Authors: Cudkowicz, M. E. Shefner, J. M. Simpson, E. Grasso, D. Yu, H. Zhang, H. Shui, A. Schoenfeld, D. Brown, R. H. Wieland, S. Barber, J. R. Auth Address: Neurology Clinical Trials Unit, Massachusetts General Hospital, 13th Street, Charlestown, MA 02129, USA. mcudkowicz@partners.org Pages: 837-44 Volume: 38 Number: 1 Keywords: Aged Amyotrophic Lateral Sclerosis/*drug therapy/metabolism Blood-Brain Barrier Dose-Response Relationship, Drug Double-Blind Method Female Humans Hydroxylamines/*administration & dosage/*adverse effects/pharmacokinetics Male Middle Aged Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18551622 Abstract: Arimoclomol is an investigational drug for amyotrophic lateral sclerosis (ALS) that amplifies heat shock protein gene expression during cell stress. The objectives of the present study were to assess the safety, tolerability, and pharmacokinetics of arimoclomol in ALS. Eighty-four participants with ALS received arimoclomol at one of three oral doses (25, 50, or 100 mg three times daily) or placebo. The primary outcome measure was safety and tolerability. A subset of 44 participants provided serum and cerebrospinal fluid (CSF) samples for pharmacokinetic analysis. Participants who completed 12 weeks of treatment could enroll in a 6-month open-label study. Arimoclomol at doses up to 300 mg/day was well tolerated and safe. Arimoclomol resulted in dose-linear pharmacologic exposures and the half-life did not change with continued treatment. Arimoclomol CSF levels increased with dose. Arimoclomol was shown to be safe, and it crosses the blood-brain barrier. Serum pharmacokinetic profiles support dosing of three times per day. An efficacy study in ALS is planned. Year: 2008 Month: 7 Title: Assessment of caregiver burden in families of persons with multiple sclerosis Authors: Buhse, M. Auth Address: School of Nursing, State University of New York at Stony Brook, Stony Brook, NY, USA. marijean.buhse@stonybrook.edu Pages: 25-31 Volume: 40 Number: 1 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18330407 Abstract: Multiple sclerosis (MS) is a chronic, unpredictable, progressive, disabling disease. It is generally diagnosed in young adult females between the ages of 20 and 40 years. Symptoms of MS may include profound fatigue, depression, gait disorder, spasticity, blurred vision, and bladder and bowel problems. It is an unpredictable disease and has the potential to create a stressful family life. Because MS is frequently diagnosed in early adulthood, it may affect developmental experiences such as raising a family and building and sustaining a career. Satisfaction with relationships can also be altered. MS has a significant social, psychological, and physical impact on the affected individual as well as his or her family. Partners of people with MS often become caregivers, adding to the demands and challenges of family life. As the individual's disease progresses, the capacity for self-care may decrease, and the individual may require daily assistance from family members. However, the daily assistance that family members provide to a disabled spouse, parent, partner, or child can take a physical and economic toll on the caregiver, causing caregiver burden. Caregiver burden is a multidimensional response to physical, psychological, emotional, social, and financial stressors associated with the caregiving experience. Caregivers who experience burden are more likely to have a higher risk of depression and a lower quality of life. Early recognition of caregiver burden is important in determining appropriate interventions. Year: 2008 Month: 2 Title: Assessment of disease progression in motor neuron disease Authors: Winhammar, J. M. Rowe, D. B. Henderson, R. D. Kiernan, M. C. Auth Address: Department of Neurology and Multidisciplinary Motor Neurone Disease Clinic, Royal North Shore Hospital, NSW, Australia. Pages: 229-38 Volume: 4 Number: 4 Keywords: Algorithms Biological Markers Brain/pathology Disease Progression Humans *Motor Neuron Disease/diagnosis/physiopathology Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15778102 Abstract: Motor neuron disease (MND) is characterised by progressive deterioration of the corticospinal tract, brainstem, and anterior horn cells of the spinal cord. There is no pathognomonic test for the diagnosis of MND, and physicians rely on clinical criteria-upper and lower motor neuron signs-for diagnosis. The presentations, clinical phenotypes, and outcomes of MND are diverse and have not been combined into a marker of disease progression. No single algorithm combines the findings of functional assessments and rating scales, such as those that assess quality of life, with biological markers of disease activity and findings from imaging and neurophysiological assessments. Here, we critically appraise developments in each of these areas and discuss the potential of such measures to be included in the future assessment of disease progression in patients with MND. Year: 2005 Month: 4 Title: Assessment of sleep-disordered breathing in pediatric neuromuscular diseases Authors: Katz, S. L. Auth Address: Children's Hospital of Eastern Ontario, Division of Respirology, University of Ottawa, Ottawa, Ontario, Canada K1H 8L1. skatz@cheo.on.ca Pages: S222-5 Volume: 123 Suppl 4 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19420148 Abstract: This is a summary of the presentation on the assessment of sleep-disordered breathing in pediatric neuromuscular diseases, presented as part of the program on pulmonary management of pediatric patients with neuromuscular disorders at the 30th annual Carrell-Krusen Neuromuscular Symposium on February 20, 2008. Year: 2009 Month: 5 Title: Astrocytes expressing ALS-linked mutated SOD1 release factors selectively toxic to motor neurons Authors: Nagai, M. Re, D. B. Nagata, T. Chalazonitis, A. Jessell, T. M. Wichterle, H. Przedborski, S. Auth Address: [1] Department of Neurology, Columbia University, 710 West 168th Street, New York, New York 10032, USA. [2] Center for Motor Neuron Biology and Disease, Columbia University, 701 West 168th Street, New York, New York 10032, USA. [3] These authors contributed equally to this work. Pages: 615-622 Volume: 10 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17435755 Abstract: Mutations in superoxide dismutase-1 (SOD1) cause a form of the fatal paralytic disorder amyotrophic lateral sclerosis (ALS), presumably by a combination of cell-autonomous and non-cell-autonomous processes. Here, we show that expression of mutated human SOD1 in primary mouse spinal motor neurons does not provoke motor neuron degeneration. Conversely, rodent astrocytes expressing mutated SOD1 kill spinal primary and embryonic mouse stem cell-derived motor neurons. This is triggered by soluble toxic factor(s) through a Bax-dependent mechanism. However, mutant astrocytes do not cause the death of spinal GABAergic or dorsal root ganglion neurons or of embryonic stem cell-derived interneurons. In contrast to astrocytes, fibroblasts, microglia, cortical neurons and myocytes expressing mutated SOD1 do not cause overt neurotoxicity. These findings indicate that astrocytes may play a role in the specific degeneration of spinal motor neurons in ALS. Identification of the astrocyte-derived soluble factor(s) may have far-reaching implications for ALS from both a pathogenic and therapeutic standpoint. Year: 2007 Month: 5 Title: Asymmetrical late onset motor neuropathy associated with a novel mutation in the small heat shock protein HSPB1 (HSP27) Authors: James, P. A. Rankin, J. Talbot, K. Auth Address: MRC Functional Genetics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK. Pages: 461-3 Volume: 79 Number: 4 Keywords: Aged *DNA Mutational Analysis Diagnosis, Differential Electrodiagnosis Electromyography Functional Laterality/*genetics Gait Disorders, Neurologic/diagnosis/*genetics Heat-Shock Proteins/*genetics Humans Male Motor Neuron Disease/diagnosis/*genetics Muscle Weakness/diagnosis/*genetics Neoplasm Proteins/*genetics Phenotype Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18344398 Abstract: Distal hereditary motor neuropathy, also known as distal spinal muscular atrophy, is characterised by slowly progressive weakness and wasting of the hands and feet and has a heterogeneous genetic basis. One form of distal hereditary motor neuropathy is associated with mutations in the gene for the small heat shock protein HSPB1 (hsp27). Families have been described in which slowly progressive, symmetrical, lower limb predominant motor weakness is usually evident by middle age. Here we report a novel mutation, G84R, in an elderly patient presenting with strikingly asymmetrical weakness. Expression of this and other known mutations in cell culture demonstrated enhanced aggregation of mutant HSPB1 protein compared with wild-type. Year: 2008 Month: 4 Title: Attrition and translation Authors: Duyk, G. Auth Address: Exelixis Inc., 170 Harbor Way, South San Francisco, CA 94083, USA. duyk@exelixis.com Pages: 603-5 Volume: 302 Number: 5645 Keywords: Access to Information *Biological Sciences/education *Biomedical Research Biotechnology Chemistry, Pharmaceutical/economics Clinical Trials Costs and Cost Analysis Drug Industry/economics Guidelines Humans Molecular Biology/education *Pharmaceutical Preparations Pharmacology/education Physiology/education Private Sector Public Sector Research Support Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14576424 Abstract: The recently published NIH Roadmap proposes that public-sector science should place increased emphasis on the development of new therapeutics and diagnostics based on the fruits of fundamental research. Such "translational research" activities, traditionally the province of the private sector, have long been compromised by high rates of attrition (failure during the course of preclinical or clinical development of therapeutics). Attrition has led to growing financial costs, as well as opportunity costs. The new focus offers a way to reverse these trends, especially if the scientific community can improve on its ability to reconcile molecular genetic research with integrative organ- and organism-based research. Year: 2003 Title: Atypical presentations of spinal muscular atrophy type III (Kugelberg-Welander disease) Authors: Kang, P. B. Krishnamoorthy, K. S. Jones, R. M. Shapiro, F. D. Darras, B. T. Auth Address: Department of Neurology, Children's Hospital Boston and Harvard Medical School, Fegan 11, 300 Longwood Avenue, Boston, MA 02115, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16797181 Abstract: Spinal muscular atrophy type III (SMA III, Kugelberg-Welander disease) typically presents with symmetric proximal weakness, areflexia, and hypotonia. We present four children with spinal muscular atrophy type III who had atypical phenotypes. Three patients clearly had asymmetric weakness at presentation and two had upper motor neuron signs in the lower extremities (one patient had both features). Two of the patients had prolonged evaluations before the diagnosis was made. All patients had Gowers signs and two had pes planus. In patients with proximal muscle weakness the presence of asymmetrical weakness, upper motor neuron signs, or both, may be compatible with spinal muscular atrophy type III. The diagnosis of spinal muscular atrophy should be considered when other possibilities have been excluded. Year: 2006 Title: Autophagy and access: understanding the role of androgen receptor subcellular localization in SBMA Authors: Montie, H. L. Merry, D. E. Auth Address: Thomas Jefferson University, Department of Biochemistry and Molecular Biology, Philadelphia, PA, USA. Pages: 1194-7 Volume: 5 Number: 8 Keywords: Animals *Autophagy/drug effects Bulbo-Spinal Atrophy, X-Linked/*metabolism/*pathology Cell Nucleus/drug effects/metabolism Mice Motor Neurons/drug effects/metabolism/pathology PC12 Cells Peptides/metabolism Protein Kinase Inhibitors/pharmacology Protein Transport/drug effects Rats Receptors, Androgen/*metabolism Subcellular Fractions/metabolism Trehalose/pharmacology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19770590 Abstract: Ridding neurons of toxic misfolded proteins is a critical feature of many neurodegenerative diseases. We have recently reported that lack of access of nuclear polyglutamine-expanded androgen receptor (AR) to the autophagic degradation pathway is a critical point in pathogenesis. When mutant AR is contained within the cytoplasm, it can be degraded by autophagy, resulting in amelioration of its toxic effects, as has been observed in other polyglutamine expansion diseases involving cytoplasmic mutant proteins. However, we have also found that pharmacological induction of autophagy protects SBMA motor neurons from the toxic effects of even nuclear localized mutant AR, albeit without affecting mutant nuclear AR levels. Thus, we have further investigated the mechanism by which autophagy elicits therapeutic benefit in cell culture. We found that endogenous autophagy only slightly alters nuclear mutant AR aggregation compared to substantial effects on cytoplasmic AR aggregation. Interestingly, pharmacological activation of mTOR-dependent autophagy did not significantly alter nuclear AR aggregation, whereas we observed that it protects SBMA motor neurons. Our findings indicate that therapeutic intervention to induce autophagy represents a potential potent benefit for SBMA, and that it likely does so by protecting SBMA motor neurons independent of a direct effect on mutant AR. Year: 2009 Month: 11 Title: Autophagy and amyotrophic lateral sclerosis: The multiple roles of lithium Authors: Fornai, F. Longone, P. Ferrucci, M. Lenzi, P. Isidoro, C. Ruggieri, S. Paparelli, A. Auth Address: Department of Human Morphology and Applied Biology, Faculty of Medicine, University of Pisa, Italy. f.fornai@med.unipi.it Pages: 527-30 Volume: 4 Number: 4 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18367867 Abstract: In a pilot clinical study that we recently published we found that lithium administration slows the progression of Amyotrophic Lateral Sclerosis (ALS) in human patients. This clinical study was published in addition with basic (in vitro) and pre-clinical (in vivo) data demonstrating a defect of autophagy as a final common pathway in the genesis of ALS. In fact, lithium was used as an autophagy inducer. In detailing the protective effects of lithium we found for the first time that this drug stimulates the biogenesis of mitochondria in the central nervous system and, uniquely in the spinal cord, it induces neuronogenesis and neuronal differentiation. In particular, the effects induced by lithium can be summarized as follows: (i) the removal of altered mitochondria and protein aggregates; (ii) the biogenesis of well-structured mitochondria; (iii) the suppression of glial proliferation; (iv) the differentiation of newly formed neurons in the spinal cord towards a specific phenotype. In this addendum we focus on defective autophagy as a "leit motif" in ALS and the old and novel features of lithium which bridge autophagy activation to concomitant effects that may be useful for the treatment of a variety of neurodegenerative disorders. In particular, the biogenesis of mitochondria and the increase of calbindin D 28K-positive neurons, which are likely to support powerful neuroprotection towards autophagy failure, mitochondriopathy and neuronal loss in the spinal cord. Year: 2008 Title: Autophagy for the avoidance of neurodegeneration Authors: Madeo, F. Eisenberg, T. Kroemer, G. Auth Address: Institute of Molecular Biosciences, University of Graz, 8010 Graz, Austria. frank.madeo@uni-graz.at Pages: 2253-9 Volume: 23 Number: 19 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19797764 Abstract: Cellular defense mechanisms, including the unfolded protein response (UPR) and autophagy, attempt to resolve toxic protein aggregates, which are common denominators of neurodegenerative diseases. In this issue of Genes & Development, Hetz and colleagues (pp. 2294-2306) surprisingly show that inhibition of the UPR by knockout of XBP-1 causes a massive increase in autophagy, enhances clearance of superoxide dismutase 1 (SOD1) aggregates, and delays the development of amyotrophic lateral sclerosis. These findings suggest the existence of a homeostatic-if not hormetic-balance between distinct cellular defense mechanisms. Year: 2009 Title: Autophagy in health and disease: 3. Autophagy Involvement in Muscle Atrophy Authors: Sandri, M. Auth Address: 1University of Padova. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20089936 Abstract: Loss of muscle mass aggravates a variety of diseases and understanding the molecular mechanisms which control muscle wasting is critical for developing new therapeutic approaches. Weakness is caused by loss of muscle proteins and recent studies have underlined a major role for the autophagy-lysosome system in regulating muscle mass. Some key components of the autophagy machinery are transcriptionally upregulated during muscle wasting and their induction precedes muscle loss. However, it is unclear whether autophagy is detrimental, causing atrophy, or beneficial, promoting survival during catabolic conditions. This review discusses recent findings on signaling pathways regulating autophagy. Title: Axon regeneration requires a conserved MAP kinase pathway Authors: Hammarlund, M. Nix, P. Hauth, L. Jorgensen, E. M. Bastiani, M. Auth Address: Department of Biology, University of Utah, 257 South 1400 East, Salt Lake City, UT 84112-0840, USA. Pages: 802-6 Volume: 323 Number: 5915 Keywords: Aging Animals Axons/*physiology/ultrastructure Axotomy Caenorhabditis elegans/genetics/*physiology Caenorhabditis elegans Proteins/genetics/*metabolism Growth Cones/physiology MAP Kinase Kinase 4/genetics/metabolism MAP Kinase Kinase Kinases/genetics/*metabolism *MAP Kinase Signaling System Mitogen-Activated Protein Kinases/genetics/metabolism Models, Biological Motor Neurons/*physiology Mutation Nerve Regeneration/physiology RNA Interference gamma-Aminobutyric Acid/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19164707 Abstract: Regeneration of injured neurons can restore function, but most neurons regenerate poorly or not at all. The failure to regenerate in some cases is due to a lack of activation of cell-intrinsic regeneration pathways. These pathways might be targeted for the development of therapies that can restore neuron function after injury or disease. Here, we show that the DLK-1 mitogen-activated protein (MAP) kinase pathway is essential for regeneration in Caenorhabditis elegans motor neurons. Loss of this pathway eliminates regeneration, whereas activating it improves regeneration. Further, these proteins also regulate the later step of growth cone migration. We conclude that after axon injury, activation of this MAP kinase cascade is required to switch the mature neuron from an aplastic state to a state capable of growth. Year: 2009 Title: Axonal defects in mouse models of motoneuron disease Authors: Jablonka, S. Wiese, S. Sendtner, M. Auth Address: Institute of Clinical Neurobiology, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Pages: 272-86 Volume: 58 Number: 2 Keywords: Aged Alternative Splicing/physiology Amyotrophic Lateral Sclerosis/genetics Animals Axonal Transport Axons/metabolism/*pathology Carrier Proteins/metabolism Caspases/genetics/metabolism Cell Survival Cells, Cultured Ciliary Neurotrophic Factor/deficiency/genetics Comparative Study Cyclic AMP Response Element-Binding Protein *Disease Models, Animal Embryo Fungal Proteins/genetics/metabolism Genes, rRNA/physiology HSP70 Heat-Shock Proteins/metabolism Heterogeneous-Nuclear Ribonucleoproteins/metabolism Humans Immunohistochemistry Mice Mice, Transgenic Middle Aged Motor Neuron Disease/genetics/metabolism/*pathology/physiopathology Mutation Nerve Growth Factor/metabolism Nerve Tissue Proteins/metabolism Precipitin Tests *Protein-Serine-Threonine Kinases Proto-Oncogene Proteins/metabolism Proto-Oncogene Proteins A-raf Proto-Oncogene Proteins c-akt Proto-Oncogene Proteins c-raf/metabolism RNA, Messenger/biosynthesis RNA-Binding Proteins Research Support, Non-U.S. Gov't Reverse Transcriptase Polymerase Chain Reaction/methods Sciatic Nerve/metabolism Spine/metabolism/pathology Superoxide Dismutase/genetics Transcription Factors/genetics/metabolism Tubulin/metabolism tau Proteins/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14704958 Abstract: Human motoneuron disease is characterized by loss of motor endplates, axonal degeneration, and cell death of motoneurons. The identification of the underlying gene defects for familial ALS, spinal muscular atrophy (SMA), and spinal muscular atrophy with respiratory distress (SMARD) has pointed to distinct pathophysiological mechanisms that are responsible for the various forms of the disease. Accumulating evidence from mouse models suggests that enhanced vulnerability and sensitivity to proapoptotic stimuli is only responsible for some but not all forms of motoneuron disease. Mechanisms that modulate microtubule assembly and the axonal transport machinery are defective in several spontaneous and ENU (ethylnitrososurea) mutagenized mouse models but also in patients with mutations in the p150 subunit of dynactin. Recent evidence suggests that axonal growth defects contribute significantly to the pathophysiology of spinal muscular atrophy. Reduced levels of the survival motoneuron protein that are responsible for SMA lead to disturbed RNA processing in motoneurons. This could also affect axonal transport of mRNAs for beta-actin and other proteins that play an essential role in axon growth and synaptic function. The local translation of specific proteins might be affected, because developing motoneurons contain ribosome-like structures in distal axons and growth cones. Altogether, the evidence from these mouse models and the new genetic data from patients suggest that axon growth and maintenance involves a variety of mechanisms, including microtubule assembly and axonal transport of proteins and ribonucleoproteins (RNPs). Thus, defects in axon maintenance could play a leading role in the development of several forms of human motoneuron disease. Year: 2004 Title: Axonal growth of embryonic stem cell-derived motoneurons in vitro and in motoneuron-injured adult rats Authors: Harper, J. M. Krishnan, C. Darman, J. S. Deshpande, D. M. Peck, S. Shats, I. Backovic, S. Rothstein, J. D. Kerr, D. A. Auth Address: Department of Neurology, Johns Hopkins University School of Medicine, Pathology 627C, 600 North Wolfe Street, Baltimore, MD 21287, USA. Pages: 7123-8 Volume: 101 Number: 18 Keywords: Animals Axons/*metabolism Genes, Reporter Motor Neurons/*metabolism Myelin Sheath/metabolism Rats Rats, Inbred Lew Research Support, Non-U.S. Gov't Stem Cells/metabolism Wound Healing/*physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15118094 Abstract: We generated spinal motoneurons from embryonic stem (ES) cells to determine the developmental potential of these cells in vitro and their capacity to replace motoneurons in the adult mammalian spinal cord. ES cell-derived motoneurons extended long axons, formed neuromuscular junctions, and induced muscle contraction when cocultured with myoblasts. We transplanted motoneuron-committed ES cells into the spinal cords of adult rats with motoneuron injury and found that approximately 3,000 ES cell-derived motoneurons (25% of input) survived for >1 month in the spinal cord of each animal. ES cell-derived axonal growth was inhibited by myelin, and this inhibition was overcome by administration of dibutyryl cAMP (dbcAMP) or a Rho kinase inhibitor in vitro and in vivo. In transplanted rats infused with dbcAMP, approximately 80 ES cell-derived motor axons were observed within the ventral roots of each animal, whereas none were observed in transplanted rats not treated with dbcAMP. Because these cells replicate many of the developmental and mature features of true motoneurons, they are an important biological tool to understand formation of motor units in vitro and a potential therapeutic tool to reconstitute neural circuits in vivo. Year: 2004 Title: Axonal-SMN (a-SMN), a protein isoform of the survival motor neuron gene, is specifically involved in axonogenesis Authors: Setola, V. Terao, M. Locatelli, D. Bassanini, S. Garattini, E. Battaglia, G. Auth Address: *Molecular Neuroanatomy Laboratory, Department of Experimental Neurophysiology and Epileptology, Istituto Neurologico "C. Besta," via Celoria 11, 20133 Milano, Italy. Pages: 1959-64 Volume: 104 Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17261814 Abstract: Spinal muscular atrophy (SMA) is an autosomal recessive disease of childhood due to loss of the telomeric survival motor neuron gene, SMN1. The general functions of the main SMN1 protein product, full-length SMN (FL-SMN), do not explain the selective motoneuronal loss of SMA. We identified axonal-SMN (a-SMN), an alternatively spliced SMN form, preferentially encoded by the SMN1 gene in humans. The a-SMN transcript and protein are down-regulated during early development in different tissues. In the spinal cord, the a-SMN protein is selectively expressed in motor neurons and mainly localized in axons. Forced expression of a-SMN stimulates motor neuron axonogenesis in a time-dependent fashion and induces axonal-like growth in non-neuronal cells. Exons 2b and 3 are essential for the axonogenic effects. This discovery indicates an unexpected complexity of the SMN gene system and may help in understanding the pathogenesis of SMA. Year: 2007 Title: Back to the future: the 'old-fashioned' way to new medications for neurodegeneration Authors: Lansbury, P. T., Jr. Auth Address: Harvard Center for Neurodegeneration and Repair and the Department of Neurology, Harvard Medical School, Center for Neurologic Diseases, Brigham and Women's Hospital, 65 Landsdowne St., Cambridge, Massachusetts 02139, USA. plansbury@rics.bwh.harvard.edu Pages: S51-7 Volume: 10 Suppl Number: Keywords: Alzheimer Disease/therapy Animals Case-Control Studies Disease Models, Animal Humans Models, Neurological Neural Networks (Computer) Neurodegenerative Diseases/diagnosis/*therapy Parkinson Disease/therapy Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15298008 Abstract: Despite the increasing prevalence of Alzheimer's disease, Parkinson's disease and less common neurodegenerative diseases-and despite the large amount of primary research that has been carried out into the causes and pathogenic features of these conditions-progress toward effective treatments has been remarkably slow. Why is this, and what can be done to accelerate it? There are a number of obstacles to effective drug discovery for neurodegeneration, but by considering these problems it is possible to identify lessons for the future. Year: 2004 Month: 7 Title: Baclofen toxicity in an 8-year-old with an intrathecal baclofen pump Authors: Yeh, R. N. Nypaver, M. M. Deegan, T. J. Ayyangar, R. Auth Address: Department of Emergency Medicine, University of Michigan, Ann Arbor, Michigan, USA. Pages: 163-7 Volume: 26 Number: 2 Keywords: Baclofen/*administration & dosage/*poisoning Cerebral Palsy/*drug therapy Child Equipment Design Equipment Failure *Equipment Failure Analysis Humans Infusion Pumps, Implantable/adverse effects Injections, Spinal/*instrumentation Male Muscle Relaxants, Central/*administration & dosage/*poisoning Muscle Spasticity/drug therapy Overdose/etiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14980337 Abstract: Baclofen delivered by intrathecal pumps (ITB) is increasingly being utilized in the pediatric population, however, resources and education to support problems with these devices are limited. Typical management strategies for systemic baclofen overdose include removal of baclofen from the device reservoir or removal of cerebrospinal fluid from the adjacent device catheter. Appropriate care of these patients requires awareness of the clinical patterns of toxicity and mechanics of the ITB pump delivery system. This report describes the clinical presentation, unfamiliar dilemmas, and the management of a pediatric patient with intrathecal baclofen toxicity, noting problems that may arise in the care of these patients. Year: 2004 Month: 2 Title: Balance control in patients with distal versus proximal muscle weakness Authors: Horlings, C. G. Kung, U. M. van Engelen, B. G. Voermans, N. C. Hengstman, G. J. van der Kooi, A. J. Bloem, B. R. Allum, J. H. Auth Address: Radboud University Nijmegen Medical Centre, Donders Institute for Brain, Cognition and Behaviour, Department of Neurology, The Netherlands; Department of ORL, University Hospital, Basel, Switzerland. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19796669 Abstract: Muscle weakness is consistently associated with falls in the elderly people, typically when present along with other risk factors. However, it remains unknown whether and how muscle weakness alone affects balance. This hampers development of more effective fall prevention strategies. Clinical observations suggest that the amount and distribution of muscle weakness influences balance control. We therefore investigated balance corrections in patients with either predominantly proximal (limb girdle muscular dystrophy (LGMD); n=8) or distal (distal spinal muscular atrophy; n=5) leg weakness, and 27 matched healthy controls. Balance was perturbed using surface tilt rotations that were delivered randomly in eight directions. Balance measures were full body kinematics and surface electromyographic activity (EMG) of leg, arm, and trunk muscles. Both patient groups were more unstable than controls, as reflected by greater excursions of the centre of mass (COM), especially in the pitch (anterior-posterior (AP)) plane. COM displacements were greater in distal weakness patients. Patients with distal weakness had excessive and unstable trunk, knee and ankle movements, and this was present following both forward and backward directed balance perturbations, possibly reflecting the greater use of distal leg muscles in these directions. In contrast, the less weak proximal weakness patients demonstrated unstable trunk and ankle movements only for backward directed balance perturbations. Both patient groups used arm movements to compensate for their instability. We conclude that primarily distal but also proximal muscle weakness leads to significant postural instability. This observation, together with the retained ability of patients to use compensatory arm movements, provides targets that may be amenable to improvement with therapeutic intervention. Year: 2009 Title: Bayesian analysis and risk assessment in genetic counseling and testing Authors: Ogino, S. Wilson, R. B. Auth Address: Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA. sogino@partners.org Pages: 1-9 Volume: 6 Number: 1 Keywords: *Bayes Theorem Cystic Fibrosis/diagnosis/genetics Female Genetic Counseling/*statistics & numerical data Genetic Predisposition to Disease/genetics Genetic Screening/*statistics & numerical data Heterozygote Detection Humans Male Models, Genetic Muscular Atrophy, Spinal/diagnosis/genetics Pedigree Risk Factors Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14736820 Abstract: Risk assessment is an essential component of genetic counseling and testing, and Bayesian analysis plays a central role in genetic risk assessment. Bayesian analysis allows calculation of the probability of a particular hypothesis, either disease or carrier status, based on family information and/or genetic test results. Genetic risk should be assessed as accurately as possible for family decision making. Additional information, from the pedigree and/or from genetic testing, can often dramatically improve the accuracy of genetic risk assessment. We illustrate herein the application of Bayes' theorem and describe important basic principles in genetic risk assessment. Year: 2004 Month: 2 Title: Behavioral and functional analysis of mouse phenotype: SHIRPA, a proposed protocol for comprehensive phenotype assessment Authors: Rogers, D.C. Fisher, E.M. Brown, S.D. Peters, J. Hunter, A.J. Martin, J.E. Auth Address: Pages: 711-713 Volume: 8 Number: Keywords: No keywords found Related urls: Abstract: Month: 1 Title: Behavioral changes after acetylcholinesterase inhibition with physostigmine in mice Authors: Mach, M. Grubbs, R. D. Price, W. A. Paton, S. J. Lucot, J. B. Auth Address: Wright State University, School of Medicine, Dayton, OH 45435, USA. mojmir.mach@sauba.sk Pages: 533-40 Volume: 79 Number: 3 Keywords: Animals Cholinesterase Inhibitors/*pharmacology Dose-Response Relationship, Drug Male Maze Learning/*drug effects/physiology Mice Mice, Inbred C57BL Motor Activity/*drug effects/physiology Physostigmine/*pharmacology Startle Reaction/*drug effects/physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15582025 Abstract: The effect of the central and peripheral acetylcholinesterase (AChE) inhibitor, physostigmine (PHY), was examined on spatial memory using a water maze, motor activity as well as acoustic startle response (ASR) and prepulse inhibition (PPI) in C57BL/6J mice. PHY was administered intraperitoneally (IP) at doses of 0.0, 0.01, 0.03, 0.1 and 0.3 mg/kg and the mice were tested 30 min after injection. Administration of PHY reduced motor activity in the open field in a dose-dependent fashion, with notable decreases in activity observed at 0.1 and 0.3 mg/kg. The results also showed that animals receiving 0.1 mg/kg spent more total time in the peripheral zone than in the central zone. The water maze data showed impairment of acquisition and performance of the task, accompanied by a reduced swimming time and enhanced thigmotaxis at a dose of 0.1 mg/kg. We also found that the ASR was significantly decreased after 0.03 and 0.1 mg/kg with no change in PPI. These results indicate that central plus peripheral cholinesterase inhibition (ChEI) decreased ASR, which is contrary to our previous experiments with the peripheral ChEI pyridostigmine bromide (PB), suggesting different involvement of cholinergic systems in modulating ASR in mice. Year: 2004 Month: 11 Title: Best practice guidelines for molecular analysis in spinal muscular atrophy Authors: Scheffer, H. Cobben, J. M. Matthijs, G. Wirth, B. Auth Address: Department of Medical Genetics, University of Groningen, The Netherlands. h.scheffer@medgen.azg.nl Pages: 484-91 Volume: 9 Number: 7 Keywords: Cyclic AMP Response Element-Binding Protein Humans Muscular Atrophy, Spinal/diagnosis/*genetics Mutation Nerve Tissue Proteins/genetics *Practice Guidelines RNA-Binding Proteins Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11464239 Abstract: With a prevalence of approximately 1/10 000, and a carrier frequency of 1/40-1/60 the proximal spinal muscular atrophies (SMAs) are among the most frequent autosomal recessive hereditary disorders. Patients can be classified clinically into four groups: acute, intermediate, mild, and adult (SMA types I, II, III, and IV, respectively). The complexity and instability of the genomic region at chromosome 5q13 harbouring the disease-causing survival motor neuron 1 (SMN1) gene hamper molecular diagnosis in SMA. In addition, affected individuals with SMA-like phenotypes not caused by SMN1, and asymptomatic individuals with two mutant alleles exist. The SMN gene is present in at least one telomeric (SMN1) and one centromeric copy (SMN2) per chromosome in normal (non-carrier) individuals, although chromosomes containing more copies of SMN1 and/or SMN2 exist. Moreover, the two SMN genes (SMN1 and SMN2) are highly homologous and contain only five base-pair differences within their 3' ends. Also, a relatively high de novo frequency is present in SMA. Guidelines for molecular analysis in diagnostic applications, carrier detection, and prenatal analysis using direct and indirect approaches are described. Overviews of materials used in the molecular diagnosis as well as Internet resources are included. Year: 2001 Month: 7 Title: Beta-adrenoceptor stimulation and neutrophil accumulation in mouse airways Authors: Miyamoto, M. Tomaki, M. Lotvall, J. Linden, A. Auth Address: Lung Pharmacology Group, Dept of Respiratory Medicine and Allergology, Institute of Internal Medicine, University of Goteborg, Gothenburg, Sweden. Pages: 231-7 Volume: 24 Number: 2 Keywords: Administration, Intranasal Airway Resistance/drug effects Albuterol/*pharmacology Animals Bronchoalveolar Lavage Fluid/chemistry/cytology Chemotaxis, Leukocyte/drug effects Cytokines/*metabolism Drug Interactions Endotoxins Male Mice Mice, Inbred C57BL Models, Animal Monokines/*metabolism Neutrophils/drug effects/*physiology Probability Receptors, Adrenergic, beta/*drug effects Sensitivity and Specificity Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15332390 Abstract: This study characterised the effect of beta-adrenoceptor stimulation on endotoxin-induced accumulation of neutrophilic granulocytes in mouse airways, where the cytokines interleukin (IL)-6 and macrophage inflammatory protein (MIP)-2 are involed as mediators. The beta2-adrenoceptor agonist salbutamol (0.025-250 fMol) was administered intranasally in mice 24 h prior to administration of endotoxin (10 microg) intranasally. Bronchoalveolar lavage (BAL) fluid and venous blood, respectively, was harvested (6 or 24 h) after administration of endotoxin. Salbutamol substantially decreased the number of neutrophils in BAL fluid from endotoxin-exposed (6 and 24 h) mice and this effect was dose dependent (24 h). Pretreatment with the beta-adrenoceptor antagonist propranolol attenuated the inhibitory effect of salbutamol on BAL neutrophils (6 and 24 h), an attenuation that was not due to any unspecific effect of propranolol. Salbutamol also substantially decreased IL-6, but not MIP-2 in BAL fluid (6 h). In contrast to BAL fluid, salbutamol caused a moderate increase in blood neutrophils (24 h). In conclusion, as indicated in mouse airways in vivo, beta-adrenoceptor stimulation prior to endotoxin exposure inhibits the induced accumulation of neutrophils at a time point much later than that anticipated from its bronchodilatory effect. Even though the detailed molecular mechanisms behind this sustained "anti-inflammatory" effect remain unknown, it seems likely that this effect is in part due to a decrease in the local concentration of interleukin-6. Year: 2004 Month: 8 Title: Bifunctional antisense oligonucleotides provide a trans-acting splicing enhancer that stimulates SMN2 gene expression in patient fibroblasts Authors: Skordis, L. A. Dunckley, M. G. Yue, B. Eperon, I. C. Muntoni, F. Auth Address: Dubowitz Neuromuscular Centre, Imperial College London, Hammersmith Hospital, London W12 0NN, United Kingdom. Pages: 4114-9 Volume: 100 Number: 7 Keywords: *Alternative Splicing Base Sequence Cyclic AMP Response Element-Binding Protein DNA Primers *Enhancer Elements (Genetics) Exons Fibroblasts/drug effects/physiology Gene Expression Regulation/drug effects/*physiology Humans Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/*genetics Oligodeoxyribonucleotides, Antisense/*pharmacology RNA, Messenger/genetics RNA-Binding Proteins Research Support, Non-U.S. Gov't Reverse Transcriptase Polymerase Chain Reaction Sequence Deletion *Trans-Activation (Genetics) Transcription, Genetic Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12642665 Abstract: The multiplicity of proteins compared with genes in mammals owes much to alternative splicing. Splicing signals are so subtle and complex that small perturbations may allow the production of new mRNA variants. However, the flexibility of splicing can also be a liability, and several genetic diseases result from single-base changes that cause exons to be skipped during splicing. Conventional oligonucleotide strategies can block reactions but cannot restore splicing. We describe here a method by which the use of a defective exon was restored. Spinal muscular atrophy (SMA) results from mutations of the Survival Motor Neuron (SMN) gene. Mutations of SMN1 cause SMA, whereas SMN2 acts as a modifying gene. The two genes undergo alternative splicing with SMN1, producing an abundance of full-length mRNA transcripts, whereas SMN2 predominantly produces exon 7-deleted transcripts. This discrepancy is because of a single nucleotide difference in SMN2 exon 7, which disrupts an exonic splicing enhancer containing an SF2ASF binding site. We have designed oligoribonucleotides that are complementary to exon 7 and contain exonic splicing enhancer motifs to provide trans-acting enhancers. These tailed oligoribonucleotides increased SMN2 exon 7 splicing in vitro and rescued the incorporation of SMN2 exon 7 in SMA patient fibroblasts. This treatment also resulted in the partial restoration of gems, intranuclear structures containing SMN protein that are severely reduced in patients with SMA. The use of tailed antisense oligonucleotides to recruit positively acting factors to stimulate a splicing reaction may have therapeutic applications for genetic disorders, such as SMA, in which splicing patterns are altered. Year: 2003 Title: Bifunctional compounds eliciting anti-inflammatory and anti-cholinesterase activity as potential treatment of nerve and blister chemical agents poisoning Authors: Amitai, G. Adani, R. Fishbein, E. Meshulam, H. Laish, I. Dachir, S. Auth Address: Division of Medicinal Chemistry, Israel Institute for Biological Research, PO Box 19, Ness Ziona 74100, Israel. amitai@iibr.gov.il Pages: 81-7 Volume: 26 Number: 1 Keywords: Animals Anti-Inflammatory Agents, Non-Steroidal/*therapeutic use Blister/chemically induced/drug therapy Brain/drug effects/pathology Chemical Warfare Agents/poisoning Cholinesterase Inhibitors/*therapeutic use Diclofenac/*therapeutic use Edema/chemically induced/drug therapy Ibuprofen/*therapeutic use Mice Mice, Inbred Strains Mustard Gas/*poisoning Pyridostigmine Bromide/therapeutic use Skin/drug effects/pathology Soman/*poisoning Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16167317 Abstract: Certain organophosphorus (OP) nerve agents (e.g. soman) induce neuroinflammatory processes during acute poisoning. An increased level of typical inflammation markers was also observed in poisoning by alkylating agents such as sulfur mustard (HD). The therapeutic potential of new bifunctional compounds was investigated, eliciting activity of non-steroidal anti-inflammatory drug (NSAID) and anti-cholinesterase (anti-ChE) activity, as an antidotal treatment for both soman and HD poisoning in mice. Three bifunctional compounds were used that include the ChE inhibitor pyridostigmine (PYR) coupled to either ibuprofen (IBU) or diclofenac (DICLO) through an eight (octyl) or ten (decyl) hydrocarbon chain spacer: IBU-PO, IBU-PD and DICLO-PD. These compounds are 15-25 fold less toxic than PYR in mice and exert peripheral and central anti-inflammatory and anti-ChE activity in vivo. IBU-PO (4 mg kg(-1), i.p.), IBU-PD (4 mg kg(-1), i.p.) and PYR (0.13 mg kg(-1), i.p.) reduced to control levels the brain edema in soman-poisoned mice (1.1 LD50, s.c.). Pre-treatment with IBU-PO, IBU-PD and DICLO-PD 4-5 h before soman challenge (2.2-2.3 LD50, s.c.) combined with antidotal treatment (atropine, 11 mg kg(-1), 2-PAM-Cl, 25 mg kg(-1), i.m.) afforded a longer 24 h survival rate (SR) than with PYR pre-treatment. DICLO-PD exhibited the largest protection efficacy (SR = 70% vs 17% with PYR). These results indicate a longer duration of action of bifunctional compounds compared with PYR. DICLO-PD (5% in propyleneglycol) reduced significantly the HD-induced edema in mouse ear-skin (51% increase in biopsy weight compared with 100% without treatment). Quantitative evaluation of ear-skin sections showed that only following DICLO-PD treatment was there a marked decrease in edema. DICLO-PD also elicited a significant decrease in HD-induced vesication as displayed by the reduced sub-epidermal blister level. The data indicate possible use of NSAID-ChEI bifunctional compounds for the medical treatment of both nerve and alkylating chemical agents. Year: 2006 Title: Binding proteins for mRNA localization and local translation, and their dysfunction in genetic neurological disease Authors: Bassell, G. J. Kelic, S. Auth Address: Department of Neuroscience, Rose Kennedy Center for Mental Retardation, Albert Einstein College of Medicine, 1410 Pelham Parkway, Bronx, New York 10461, New York, USA. bassell@aecom.yu.edu Pages: 574-81 Volume: 14 Number: 5 Keywords: Active Transport, Cell Nucleus/genetics Animals Central Nervous System/*metabolism/physiopathology Fragile X Syndrome/genetics/metabolism Heredodegenerative Disorders, Nervous System/*genetics/metabolism/physiopathology Humans Muscular Atrophy, Spinal/genetics/metabolism Protein Biosynthesis/genetics RNA, Messenger/*metabolism RNA-Binding Proteins/genetics/*metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15464890 Abstract: Neurons utilize mRNA transport and local translation as a means to influence development and plasticity. The molecular mechanisms for this mRNA sorting involve the recognition of cis-acting sequences by distinct mRNA binding proteins that have a dual role, acting in both mRNA transport and translational regulation. Other proteins play a part in the assembly of messenger ribonucleoprotein complexes into transport granules. mRNA binding proteins are crucial targets of phosphorylation signals that regulate local translation. Fragile X syndrome and spinal muscular atrophy have emerged as two genetic neurological diseases that could result, in part, from impaired assembly, localization, and translational regulation of these messenger ribonucleoproteins. Year: 2004 Month: 10 Title: Biochemical and genetic evidence for a role of IGHMBP2 in the translational machinery Authors: de Planell-Saguer, M. Schroeder, D. G. Rodicio, M. C. Cox, G. A. Mourelatos, Z. Auth Address: Department of Pathology and Laboratory Medicine, Division of Neuropathology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6100, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19299493 Abstract: The human motor neuron degenerative disease Spinal Muscular Atrophy with Respiratory Distress type 1 (SMARD1) is caused by loss of function mutations of immunoglobulin mu-binding protein 2 (IGHMBP2), a protein of unknown function that contains DNA/RNA helicase and nucleic acid binding domains. Reduced IGHMBP2 protein levels in neuromuscular degeneration (nmd) mice, the mouse model of SMARD1, lead to motor neuron degeneration. We report the biochemical characterization of IGHMBP2 and the isolation of a modifier locus that rescues the phenotype and motor neuron degeneration of nmd mice. We find that a 166Kb BAC transgene derived from CAST/EiJ mice and containing tRNA genes and Activator of Basal Transcription 1 (Abt1), a protein coding gene that is required for ribosome biogenesis, contains the genetic modifier responsible for motor neuron rescue. Our biochemical investigations show that IGHMBP2 associates physically with tRNAs and in particular with tRNA(Tyr), which are present in the modifier and with the ABT1 protein. We find that Transcription factor IIIC-220kD (TFIIIC220), an essential factor required for tRNA transcription and the helicases Reptin and Pontin, which function in transcription and in ribosome biogenesis, are also part of IGHMBP2-containing complexes. Our findings strongly suggest that IGHMBP2 is a component of the translational machinery and that these components can be manipulated genetically to suppress motor neuron degeneration. Year: 2009 Title: Biological and clinical manifestations of Huntington's disease in the longitudinal TRACK-HD study: cross-sectional analysis of baseline data Authors: Tabrizi, S. J. Langbehn, D. R. Leavitt, B. R. Roos, R. A. Durr, A. Craufurd, D. Kennard, C. Hicks, S. L. Fox, N. C. Scahill, R. I. Borowsky, B. Tobin, A. J. Rosas, H. D. Johnson, H. Reilmann, R. Landwehrmeyer, B. Stout, J. C. Auth Address: UCL Institute of Neurology, University College London, Queen Square, London, UK. Pages: 791-801 Volume: 8 Number: 9 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19646924 Abstract: BACKGROUND: Huntington's disease (HD) is an autosomal dominant, fully penetrant, neurodegenerative disease that most commonly affects adults in mid-life. Our aim was to identify sensitive and reliable biomarkers in premanifest carriers of mutated HTT and in individuals with early HD that could provide essential methodology for the assessment of therapeutic interventions. METHODS: This multicentre study uses an extensive battery of novel assessments, including multi-site 3T MRI, clinical, cognitive, quantitative motor, oculomotor, and neuropsychiatric measures. Blinded analyses were done on the baseline cross-sectional data from 366 individuals: 123 controls, 120 premanifest (pre-HD) individuals, and 123 patients with early HD. FINDINGS: The first participant was enrolled in January, 2008, and all assessments were completed by August, 2008. Cross-sectional analyses identified significant changes in whole-brain volume, regional grey and white matter differences, impairment in a range of voluntary neurophysiological motor, and oculomotor tasks, and cognitive and neuropsychiatric dysfunction in premanifest HD gene carriers with normal motor scores through to early clinical stage 2 disease. INTERPRETATION: We show the feasibility of rapid data acquisition and the use of multi-site 3T MRI and neurophysiological motor measures in a large multicentre study. Our results provide evidence for quantifiable biological and clinical alterations in HTT expansion carriers compared with age-matched controls. Many parameters differ from age-matched controls in a graded fashion and show changes of increasing magnitude across our cohort, who range from about 16 years from predicted disease diagnosis to early HD. These findings might help to define novel quantifiable endpoints and methods for rapid and reliable data acquisition, which could aid the design of therapeutic trials. FUNDING: CHDI/High Q Foundation. Year: 2009 Month: 9 Title: Biomarker qualification via public-private partnerships Authors: Eck, S. L. Paul, S. M. Auth Address: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana, USA. Pages: 21-3 Volume: 87 Number: 1 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20019697 Abstract: Biomarkers linked to patient outcomes (safety and efficacy) have an increasingly important role in drug development. Consequently, validation and qualification of such biomarkers are essential, often requiring large data sets from well-controlled randomized clinical trials. In the December 2009 issue of Clinical Pharmacology & Therapeutics, investigators utilizing data from four pharmaceutical companies and working under the auspices of the Biomarkers Consortium described the utility of adiponectin as an early predictor of glycemic control in diabetic patients taking peroxisome proliferator-activated receptor (PPAR) agonists. This work illustrates the advantages of large public-private partnerships for biomarker qualification. Month: 1 Title: Biomarkers for predicting clinical responses to HDAC inhibitors Authors: Stimson, L. La Thangue, N. B. Auth Address: Laboratory of Cancer Biology, Dept of Clinical Pharmacology, Old Road Campus Research Building, Old Road Campus, Oxford OX3 7DQ, United Kingdom. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19362413 Abstract: Post-translational modifications of histone and non-histone proteins by acetylation are known to play a key role in tumourigenesis. Pharmacological manipulation of acetylation has been possible with the identification of small molecule inhibitors of histone deacetylases (HDAC), the enzymes responsible for deacetylating lysine residues. An explosion of drug discovery efforts in recent years has led to the development of an extensive group of HDAC inhibitors, many of which have been shown pre-clinically to have potent anti-tumour activity. Clinical trials using these agents are now underway, with Vorinostat (suberoylanilide hydroxamic acid) having been approved by the FDA for treating cutaneous T-cell lymphoma (CTCL) in patients with progressive, persistent or recurrent disease. This review discusses how biomarkers are being identified and used to expand our knowledge of the mechanisms by which HDAC inhibitors exhibit their anti-cancer effects. In the longer term, biomarkers will provide a means towards achieving patient stratification in tumour types that will respond favourably to HDAC inhibitors. Year: 2009 Title: Blood-based proteomics for personalized medicine: examples from neurodegenerative disease Authors: Goldknopf, I. L. Auth Address: Pages: 1-8 Volume: 5 Number: 1 Keywords: Alzheimer Disease/blood/classification/diagnosis/genetics Biological Markers Blood Proteins/*analysis/genetics Diagnosis, Differential *Electrophoresis, Gel, Two-Dimensional *Genetic Privacy Neurodegenerative Diseases/*blood/diagnosis/genetics Parkinson Disease/blood/diagnosis/genetics Protein Isoforms/blood Proteomics/*methods Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18282116 Abstract: Year: 2008 Month: 2 Title: Bone loss in survival motor neuron (Smn(-/-) SMN2) genetic mouse model of spinal muscular atrophy Authors: Shanmugarajan, S. Tsuruga, E. Swoboda, K. J. Maria, B. L. Ries, W. L. Reddy, S. V. Auth Address: Charles P. Darby Children's Research Institute, Charleston, SC, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19434631 Abstract: Spinal muscular atrophy (SMA) is characterized by degenerating lower motor neurons and an increased incidence of congenital bone fractures. Survival motor neuron (SMN) levels are significantly reduced due to deletions/mutations in the telomeric SMN1 gene in these patients. We utilized the Smn(-/-) SMN2 mouse model of SMA to determine the functional role for SMN in bone remodelling. microCT analysis of lumber vertebrae, tibia and femur bones from SMA mice revealed an osteoporotic bone phenotype. Histological analysis demonstrated a thin porous cortex of cortical bone and thin trabeculae at the proximal end of the growth plate in the vertebrae of SMA mice compared to wild-type mice. Histochemical staining of the vertebrae showed the presence of abundant activated osteoclasts on the sparse trabeculae and on the endosteal surface of the thin cortex in SMA mice. Histomorphometric analysis of vertebrae from SMA mice showed an increased number of osteoclasts. Serum TRAcP5b and urinary NTx levels were elevated, consistent with increased bone resorption in these mice. SMA mice showed a significant decrease in the levels of osteoblast differentiation markers, osteocalcin, osteopontin and osterix mRNA expression; however, there were no change in the levels of alkaline phosphatase expression compared to WT mice. SMA mouse bone marrow cultures revealed an increased rate of osteoclast formation (54%) and bone resorption capacity (46%) compared to WT mice. Pre-osteoclast cells from SMA mice showed constitutive up-regulation of RANK receptor signalling molecules critical for osteoclast differentiation. Our results implicate SMN function in bone remodelling and skeletal pathogenesis in SMA. Understanding basic mechanisms of SMN action in bone remodelling may uncover new therapeutic targets for preventing bone loss/fracture risk in SMA. Copyright (c) 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. Year: 2009 Title: Bone marrow transplantation attenuates the myopathic phenotype of a muscular mouse model of spinal muscular atrophy Authors: Salah-Mohellibi, N. Millet, G. Andre-Schmutz, I. Desforges, B. Olaso, R. Roblot, N. Courageot, S. Bensimon, G. Cavazzana-Calvo, M. Melki, J. Auth Address: Molecular Neurogenetics Laboratory, INSERM U798, 2 rue Gaston Cremieux CP5724, 91057 Evry, France. j.melki@genopole.inserm.fr. Pages: 2723-32 Volume: 24 Number: 12 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16888281 Abstract: Bone marrow (BM) transplantation was performed on a muscular mouse model of spinal muscular atrophy that had been created by mutating the survival of motor neuron gene (Smn) in myofibers only. This model is characterized by a severe myopathy and progressive loss of muscle fibers leading to paralysis. Transplantation of wild-type BM cells following irradiation at a low dose (6 Gy) improved motor capacity (+85%). This correlated with a normalization of myofiber number associated with a higher number of regenerating myofibers (1.6-fold increase) and an activation of CD34 and Pax7 satellite cells. However, BM cells had a very limited capacity to replace or fuse to mutant myofibers (2%). These data suggest that BM transplantation was able to attenuate the myopathic phenotype through an improvement of skeletal muscle regeneration of recipient mutant mice, a process likely mediated by a biological activity of BM-derived cells. This hypothesis was further supported by the capacity of muscle protein extracts from transplanted mutant mice to promote myoblast proliferation in vitro (1.6-fold increase). In addition, a tremendous upregulation of hepatocyte growth factor (HGF), which activates quiescent satellite cells, was found in skeletal muscle of transplanted mutants compared with nontransplanted mutants. Eventually, thanks to the Cre-loxP system, we show that BM-derived muscle cells were strong candidates harboring this biological activity. Taken together, our data suggest that a biological activity is likely involved in muscle regeneration improvement mediated by BM transplantation. HGF may represent an attractive paracrine mechanism to support this activity. Year: 2006 Month: 12 Title: Bulbar and spinal muscular atrophy (Kennedy's disease): a review Authors: Finsterer, J. Auth Address: Krankenanstalt Rudolfstiftung, Vienna, Austria. fifigs1@yahoo.de Pages: 556-61 Volume: 16 Number: 5 Keywords: Animals *Bulbo-Spinal Atrophy, X-Linked Humans Receptors, Androgen/genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19405197 Abstract: Bulbar and spinal muscular atrophy (BSMA) is an adult-onset, X-linked recessive trinucleotide, polyglutamine disorder, caused by expansion of a polymorphic CAG tandem-repeat in exon 1 of the androgen-receptor (AR) gene on chromosome Xq11-12. Pathogenetically, mutated AR accumulates in nuclei and cytoplasm of motor neurons, resulting in their degeneration and loss. Phenotypically, patients present with amyotrophic, proximal or distal weakness and wasting of the facial, bulbar and limb muscles, occasionally sensory disturbances, and endocrinologic disturbances, such as androgen resistance, gynecomastia, elevated testosterone or progesterone, and reduced fertility. There may be mild hyper-CK-emia, abnormal motor and sensory nerve conduction studies, and neuropathic and myopathic alterations on muscle biopsy. The golden standard for diagnosing BSMA is genetic analysis, demonstrating a CAG-repeat number >40. No causal therapy is available, but symptomatic therapy should be provided for tremor, endocrinologic abnormalities, sensory disturbances, or muscle cramps. The course is slowly progressive, the ability to walk lost only late in life, only few patients require ventilatory support, and life expectancy only slightly reduced. Year: 2009 Month: 5 Title: Butyrate and acetyl-carnitine inhibit the cytogenetic expression of the fragile X in vitro Authors: Pomponi, M. G. Neri, G. Auth Address: Istituto di Genetica Medica, Facolta di Medicina A. Gemelli, Universita Cattolica, Rome, Italy. Pages: 447-50 Volume: 51 Number: 4 Keywords: Acetylation Acetylcarnitine/*pharmacology Adolescent Adult Aged Butyric Acid Butyric Acids/*pharmacology Cells, Cultured/drug effects Child Child, Preschool Chromosome Fragile Sites Chromosome Fragility Fragile X Syndrome/*genetics Gene Expression/*drug effects Histones/metabolism Humans Lymphocytes/drug effects Male Middle Aged Mutagenesis/drug effects Mutation Repetitive Sequences, Nucleic Acid/*drug effects Transcription, Genetic/drug effects X Chromosome/drug effects Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7943015 Abstract: Cytogenetic expression of the fragile site at Xq27.3 is only found in those patients who have a full mutation of the FMR1 gene, i.e., a large amplification of the CGG repeat. However, an expansion of this repeat, although necessary, does not seem to be sufficient to cause expression of fra(X)(q27.3). Other factors are clearly needed, e.g., thymidylate stress. Little or no attention has been paid to the possible role of histones in the expression of the fragile sites, in spite of their structural and regulatory role in the chromatin complex. Histones can be modified by treating intact cells in vitro with butyrate, a substance that causes histone acetylation. The purpose of the present work is to test the effect of butyrate and of the acetylating compound acetyl-L-carnitine on the expression of fra(X)(q27.3) by treating peripheral lymphocytes of fragile X syndrome patients with these substances in vitro. We show that this treatment causes a significant inhibition of fra(X)(q27.3) expression. Year: 1994 Title: Caenorhabditis elegans in the study of SMN-interacting proteins: a role for SMI-1, an orthologue of human Gemin2 and the identification of novel components of the SMN complex Authors: Burt, E. C. Towers, P. R. Sattelle, D. B. Auth Address: Centre for Haematology, Bart's and The London, Queen Mary's School of Medicine, Institute of Cell and Molecular Science, 4 Newark Street, London, E1 2AT, UK. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16964508 Abstract: Spinal muscular atrophy is a common neuromuscular disorder caused by mutations in the survival motor neuron (SMN) gene. In mammals, SMN is tightly associated with Gemin2. To gain further insight into the functions of SMN and Gemin2, we have cloned and sequenced smi-1 (Survival of Motor neuron-Interacting protein 1), a C. elegans homologue of the human Gemin2 gene. We show that the SMI-1 expression pattern and RNA interference phenotype show considerable overlap with that previously reported for SMN-1. Finally, we demonstrate that the SMN-1 and SMI-1 proteins directly interact. Having demonstrated the utility of the C. elegans genetic model for investigating genes encoding SMN-interacting proteins, we have undertaken a yeast two-hybrid screen of a C. elegans cDNA library to identify novel proteins that interact with SMN-1. We show the direct interaction of SMN-1 with nine novel proteins, several of which may be involved in RNA metabolism. Year: 2006 Title: Cajal-body formation correlates with differential coilin phosphorylation in primary and transformed cell lines Authors: Hearst, S. M. Gilder, A. S. Negi, S. S. Davis, M. D. George, E. M. Whittom, A. A. Toyota, C. G. Husedzinovic, A. Gruss, O. J. Hebert, M. D. Auth Address: Department of Biochemistry, The University of Mississippi Medical Center, Jackson, MS 39216, USA. Pages: 1872-81 Volume: 122 Number: Pt 11 Keywords: Amino Acid Sequence Animals *Cell Line, Tumor/cytology/metabolism *Cells, Cultured/cytology/metabolism Coiled Bodies/*metabolism Humans Molecular Sequence Data Nuclear Proteins/chemistry/genetics/*metabolism Phosphoprotein Phosphatases/metabolism Phosphorylation Recombinant Fusion Proteins/genetics/metabolism Signal Transduction/physiology Survival of Motor Neuron 1 Protein/genetics/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19435804 Abstract: Cajal bodies (CBs) are nuclear structures that are thought to have diverse functions, including small nuclear ribonucleoprotein (snRNP) biogenesis. The phosphorylation status of coilin, the CB marker protein, might impact CB formation. We hypothesize that primary cells, which lack CBs, contain different phosphoisoforms of coilin compared with that found in transformed cells, which have CBs. Localization, self-association and fluorescence recovery after photobleaching (FRAP) studies on coilin phosphomutants all suggest this modification impacts the function of coilin and may thus contribute towards CB formation. Two-dimensional gel electrophoresis demonstrates that coilin is hyperphosphorylated in primary cells compared with transformed cells. mRNA levels of the nuclear phosphatase PPM1G are significantly reduced in primary cells and expression of PPM1G in primary cells induces CBs. Additionally, PPM1G can dephosphorylate coilin in vitro. Surprisingly, however, expression of green fluorescent protein alone is sufficient to form CBs in primary cells. Taken together, our data support a model whereby coilin is the target of an uncharacterized signal transduction cascade that responds to the increased transcription and snRNP demands found in transformed cells. Year: 2009 Title: Calorie restriction promotes mammalian cell survival by inducing the SIRT1 deacetylase Authors: Cohen, H. Y. Miller, C. Bitterman, K. J. Wall, N. R. Hekking, B. Kessler, B. Howitz, K. T. Gorospe, M. de Cabo, R. Sinclair, D. A. Auth Address: Department of Pathology, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA. Pages: 390-2 Volume: 305 Number: 5682 Keywords: Acetylation Adipose Tissue/metabolism Alleles Animals Antigens, Nuclear/metabolism *Apoptosis *Caloric Restriction Cell Line *Cell Survival DNA-Binding Proteins/metabolism Histone Deacetylases/genetics/*metabolism Humans Insulin/metabolism/pharmacology Insulin-Like Growth Factor I/metabolism/pharmacology Kidney/metabolism Liver/metabolism Male Mitochondria/metabolism Mutation Proto-Oncogene Proteins/metabolism *Proto-Oncogene Proteins c-bcl-2 RNA, Small Interfering Rats Rats, Inbred F344 Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sirtuins/genetics/*metabolism bcl-2-Associated X Protein Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15205477 Abstract: A major cause of aging is thought to result from the cumulative effects of cell loss over time. In yeast, caloric restriction (CR) delays aging by activating the Sir2 deacetylase. Here we show that expression of mammalian Sir2 (SIRT1) is induced in CR rats as well as in human cells that are treated with serum from these animals. Insulin and insulin-like growth factor 1 (IGF-1) attenuated this response. SIRT1 deacetylates the DNA repair factor Ku70, causing it to sequester the proapoptotic factor Bax away from mitochondria, thereby inhibiting stress-induced apoptotic cell death. Thus, CR could extend life-span by inducing SIRT1 expression and promoting the long-term survival of irreplaceable cells. Year: 2004 Title: Can regenerating axons recapitulate developmental guidance during recovery from spinal cord injury? Authors: Harel, N. Y. Strittmatter, S. M. Auth Address: Program in Cellular Neuroscience, Neurodegeneration and Repair, Department of Neurology, Yale University School of Medicine, New Haven, Connecticut 06520, USA. Pages: 603-16 Volume: 7 Number: 8 Keywords: Animals Axons/*physiology Feedback, Biochemical/physiology Humans Nerve Regeneration/*physiology Neuronal Plasticity/physiology Recovery of Function/*physiology Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Spinal Cord Injuries/metabolism/*physiopathology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16858389 Abstract: The precise wiring of the adult mammalian CNS originates during a period of stunning growth, guidance and plasticity that occurs during and shortly after development. When injured in adults, this intricate system fails to regenerate. Even when the obstacles to regeneration are cleared, growing adult CNS fibres usually remain misdirected and fail to reform functional connections. Here, we attempt to fill an important niche related to the topics of nervous system development and regeneration. We specifically contrast the difficulties faced by growing fibres within the adult context to the precise circuit-forming capabilities of developing fibres. In addition to focusing on methods to stimulate growth in the adult, we also expand on approaches to recapitulate development itself. Year: 2006 Month: 8 Title: Cardiac manifestations of neuromuscular disorders in children Authors: Hsu, D. T. Auth Address: Children's Hospital at Montefiore and the Albert Einstein College of Medicine, Bronx, New York 10467, USA. dhsu@montefiore.org Pages: 35-8 Volume: 11 Number: 1 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20113990 Abstract: Cardiac abnormalities occur in association with many of the neuromuscular disorders that present in childhood. Genetic defects involving the cytoskeleton, nuclear membrane, and mitochondrial function have all been described in patients with skeletal myopathy and cardiac involvement. The most common classes of neuromuscular disorders with cardiac manifestations are the muscular dystrophies- Duchenne, Becker, limb-girdle and Emery Dreifuss. Friedreich Ataxia and myotonic dystrophy also have important cardiac involvement. The type and extent of cardiac manifestations are specific to the type of neuromuscular disorder. The most common cardiac findings include dilated or hypertrophic cardiomyopathy, atrioventricular conduction defects, atrial fibrillation and ventricular arrhythmias. Screening for cardiac involvement should be performed in all children with neuromuscular disorders that have the potential for cardiac involvement. This review discusses the cardiac findings associated with specific neuromuscular disorders and outlines the indications for evaluation and treatment. Month: 3 Title: Carnitine and acyltransferase in experimental neurogenic atrophies: changes with treatment Authors: Bresolin, N. Freddo, L. Tegazzin, V. Bet, L. Armani, M. Angelini, C. Auth Address: Pages: 170-5 Volume: 231 Number: 4 Keywords: Acyltransferases/*metabolism Adult Animals Carnitine/*metabolism/pharmacology Denervation Humans Infant Infant, Newborn Male Muscular Atrophy/enzymology/etiology/*metabolism/pathology Nerve Regeneration Rats Rats, Inbred Strains Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6512569 Abstract: Carnitine level and carnitine palmityl transferase (CPT) activity were investigated in muscles of patients with infantile and juvenile spinal muscular atrophy and polyneuropathies. A significant decrease of both carnitine and CPT was found in the infantile spinal muscular atrophy, but not in the other neurogenic muscle atrophies. These findings were compared with the experimental effect of denervation and reinnervation upon the lipid metabolism in soleus and extensor digitorum longus (EDL) of adult and newborn rats. Twenty-one days after denervation free and total carnitine decreased significantly in both EDL (P less than 0.001) and soleus (P less than 0.05) of adult animals. CPT activity was significantly decreased in the soleus 50 days after denervation (P less than 0.005). Long-term reinnervation restored the level of carnitine fraction and CPT activity. L-carnitine treatment for 21 days restored the level of free carnitine to normal in the soleus of denervated adult animals. Denervation in newborn rats influenced carnitine concentration in soleus and EDL to a lesser extent; the treatment with L-carnitine raised short-chain acylcarnitines in denervated muscles, while reinnervation restored carnitine level within 50 days. Year: 1984 Month: 1 Title: Carrier frequency of SMA by quantitative analysis of the SMN1 deletion in the Iranian population Authors: Hasanzad, M. Azad, M. Kahrizi, K. Saffar, B. S. Nafisi, S. Keyhanidoust, Z. Azimian, M. Refah, A. A. Also, E. Urtizberea, J. A. Tizzano, E. F. Najmabadi, H. Auth Address: Genetics Research Center, University of Social Welfare & Rehabilitation Sciences, Tehran, Iran. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19538222 Abstract: Background and purpose: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder. Carrier frequency studies of SMA have been reported for various populations. Although no large-scale population-based studies of SMA have been performed in Iran, previous estimates have indicated that the incidence of autosomal recessive disorder partly because of the high prevalence of consanguineous marriage is much higher in the Iranian population than in other populations. Methods: In this study, we used a reliable and highly sensitive quantitative real-time PCR assay with SYBR green I dye to detect the copy number of the SMN1 gene to determine the carrier frequency of SMA in 200 healthy unrelated, non-consanguineous couples from different part of Iran. Results: To validate the method in our samples, we determined the relative quantification (RQ) of patients with homozygous deletion (0.00) and hemyzygous carriers (0.29-0.55). The RQ in 10 of 200 normal individuals were within the carrier range of 0.31-0.57, estimating a carrier frequency of 5% in the Iranian population. Conclusions: Our data show that the SMA carrier frequency in Iran is higher than in the European population and that further programs of population carrier detection and prenatal testing should be implemented. Year: 2009 Title: Carrier incidence for spinal muscular atrophy in southern Chinese Authors: Chan, V. Yip, B. Yam, I. Au, P. Lin, C. K. Wong, V. Chan, T. K. Auth Address: University Department of Medicine, Queen Mary Hospital, Pokfulam Road, Hong Kong. vnychana@hkucc.hku.hk Pages: 1089-93 Volume: 251 Number: 9 Keywords: Asian Continental Ancestry Group/*genetics/statistics & numerical data China/epidemiology Cyclic AMP Response Element-Binding Protein *Heterozygote Detection Humans Incidence Muscular Atrophy, Spinal/*epidemiology/*genetics Nerve Tissue Proteins/genetics RNA-Binding Proteins Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15372251 Abstract: A real time quantitative PCR (QPCR) method using TaqMan technology was used to assess the copy number of the two survival motor neuron genes (SMN1 and SMN2) on chromosome 5q13. This allows the accurate determination of carriers for spinal muscular atrophy (SMA), with one copy of SMN1. Analysis of 569 normal southern Chinese individuals revealed a carrier incidence of 1.6%, similar to that found in the western society. Study of 42 obligatory carriers showed a (2 + 0) genotype in two (4.8 %). In 27 SMA patients with homozygous deletion of the SMN1 gene, the number of SMN2 gene correlated with disease phenotype, with 68% of type II and III patients carrying three or more SMN2 genes, whilst the incidence of three or more SMN2 genes in the normal population was 1.57%. Year: 2004 Month: 9 Title: Carrier screening for spinal muscular atrophy Authors: Prior, T. W. Auth Address: From the Department of Pathology, The Ohio State University, Columbus, Ohio. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18941424 Abstract: Year: 2008 Title: Case report: birth after preimplantation genetic diagnosis of a subtle mutation in SMN1 gene Authors: Moutou, C. Machev, N. Gardes, N. Viville, S. Auth Address: Service de Biologie de la Reproduction-SIHCUS-CMCO, CHU de Strasbourg, 19, rue Louis Pasteur, BP120, 67303 Schiltigheim cedex, France. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16941715 Abstract: Spinal muscular atrophy (SMA) preimplantation genetic diagnosis (PGD) has been available since 1998. Protocols are based on the detection of the homozygous deletion of exon 7, which are present in 90-98% of SMA patients. A couple where the woman was a heterozygous carrier of the usual SMN1 Del7 mutation and the man was a heterozygous carrier of pMet263Arg substitution in exon 6 of SMN1 gene was referred for PGD. The usual PGD test being unsuitable for this couple, we developed a novel duplex polymerase chain reaction (PCR)-based PGD test for the detection of the mutation pMet263Arg by allele specific amplification, combined with the amplification of D5S641 extragenic polymorphic marker. PCR conditions were established using single control lymphoblasts and lymphocytes from the pMet263Arg substitution carrier. Amplification was obtained in 100% of the 86 single cells tested, amplification refractory mutation system (ARMS) PCR was specific in 100% of single cells tested and a complete genotype (mutation plus D5S641) was achieved in 88% of them. A PGD cycle was performed successfully and a pregnancy was obtained. An unaffected girl was born and postnatal diagnosis confirmed PGD results. This is the first PGD described for SMA because of another mutation than the major homozygous exon 7 deletion of SMN1. In the future, a similar strategy could be adopted for other subtle mutations of this gene. Copyright (c) 2006 John Wiley & Sons, Ltd. Year: 2006 Title: cDNA isolation, expression, and chromosomal localization of the mouse survival motor neuron gene (Smn) Authors: Viollet, L. Bertrandy, S. Bueno Brunialti, A. L. Lefebvre, S. Burlet, P. Clermont, O. Cruaud, C. Guenet, J. L. Munnich, A. Melki, J. Auth Address: Unite de Recherches sur les Handicaps Genetiques de l'Enfant, INSERM, Unite 393, IFREM, Institut Necker, Hopital des Enfants Malades, Paris, France. Pages: 185-8 Volume: 40 Number: 1 Keywords: Amino Acid Sequence Animals Blotting, Northern Blotting, Southern *Chromosome Mapping Chromosomes, Human, Pair 5 Cyclic AMP Response Element-Binding Protein DNA, Complementary Gene Expression Humans Mice Molecular Sequence Data Nerve Tissue Proteins/*genetics Poly A RNA-Binding Proteins Research Support, Non-U.S. Gov't Tissue Distribution Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9070939 Abstract: Spinal muscular atrophy (SMA) is a frequent autosomal recessive disease in human characterized by degeneration of motor neurons of the spinal cord. The genomic region containing the defective gene (5q13) is particularly unstable and prone to large-scale deletions whose characterization led to the identification of the survival motor neuron (SMN) gene, the SMA determining gene encoding a hitherto unknown protein. As an initial step toward the generation of a murine model for SMA, we identified and characterized a full-length murine Smn cDNA. The coding sequence of the mouse Smn gene was found to be 82% identical, at the amino acid level, with the human SMN coding sequence. The Smn locus was mapped to the segment of mouse chromosome 13 exhibiting conservation of synteny with human chromosome 5q11-q23, which contains the SMN gene. However, no evidence for a duplication of the Smn gene was found in the mouse, suggesting that the duplication reported in human is a recent evolutionary event. Year: 1997 Title: Celastrol blocks neuronal cell death and extends life in transgenic mouse model of amyotrophic lateral sclerosis Authors: Kiaei, M. Kipiani, K. Petri, S. Chen, J. Calingasan, N. Y. Beal, M. F. Auth Address: Department of Neurology and Neuroscience, Weill Medical College of Cornell University, New York-Presbyterian Hospital, New York, NY 10021, USA. mak2026@med.cornell.edu Pages: 246-54 Volume: 2 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16909005 Abstract: There is substantial evidence that both inflammation and oxidative damage contribute to the pathogenesis of motor neuron degeneration in the G93A SOD1 transgenic mouse model of amyotrophic lateral sclerosis (ALS). Celastrol is a natural product from Southern China, which exerts potent anti-inflammatory and antioxidative effects. It also acts potently to increase expression of heat shock proteins including HSP70. We administered it in the diet to G93A SOD1 mice starting at 30 days of age. Celastrol treatment significantly improved weight loss, motor performance and delayed the onset of ALS. Survival of celastrol-treated G93A mice increased by 9.4% and 13% for 2 mg/kg/day and 8 mg/kg/day doses, respectively. Cell counts of lumbar spinal cord neurons confirmed a protective effect, i.e. 30% increase in neuronal number in the lumbar spinal cords of celastrol-treated animals. Celastrol treatment reduced TNF-alpha, iNOS, CD40, and GFAP immunoreactivity in the lumbar spinal cord sections of celastrol-treated G93A mice compared to untreated G93A mice. TNF-alpha immunoreactivity co-localized with SMI-32 (neuronal marker) and GFAP (astrocyte marker). HSP70 immunoreactivity was increased in lumbar spinal cord neurons of celastrol-treated G93A mice. Celastrol has been widely used in treating inflammatory diseases in man, and is well tolerated; therefore, it may be a promising therapeutic candidate for the treatment of human ALS. Year: 2005 Month: 1 Title: Cell cycle-regulated trafficking of human telomerase to telomeres Authors: Tomlinson, R. L. Ziegler, T. D. Supakorndej, T. Terns, R. M. Terns, M. P. Auth Address: Departments of Biochemistry and Molecular Biology and Genetics, University of Georgia, Athens, GA 30602, USA. Pages: 955-65 Volume: 17 Number: 2 Keywords: Cell Cycle/*physiology Cell Nucleolus/enzymology/ultrastructure Coiled Bodies/chemistry/ultrastructure DNA-Binding Proteins/analysis/*metabolism Hela Cells Humans Models, Biological Protein Transport RNA, Untranslated/analysis/*metabolism Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't S Phase/physiology Telomerase/analysis/*metabolism Telomere/*metabolism/ultrastructure Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16339074 Abstract: Telomerase synthesizes telomeres at the ends of human chromosomes during S phase. The results presented here suggest that telomerase activity may be regulated by intranuclear trafficking of the key components of the enzyme in human cells. We examined the subcellular localization of endogenous human telomerase RNA (hTR) and telomerase reverse transcriptase (hTERT) in HeLa cervical carcinoma cells. Throughout most of the cell cycle, we found that the two essential components of telomerase accumulate at intranuclear sites separate from telomeres. However, during S phase, both hTR and hTERT are specifically recruited to subsets of telomeres. The localization of telomerase to telomeres is dynamic, peaking at mid-S phase. We also found complex associations of both hTR and hTERT with nucleoli and Cajal bodies during S phase, implicating both structures in the biogenesis and trafficking of telomerase. Our results mark the first observation of human telomerase at telomeres and provide a mechanism for the cell cycle-dependent regulation of telomere synthesis in human cells. Year: 2006 Month: 2 Title: Cell therapy and stem cells in animal models of motor neuron disorders Authors: Hedlund, E. Hefferan, M. P. Marsala, M. Isacson, O. Auth Address: Neuroregeneration Laboratory, Center for Neuroregeneration Research, McLean Hospital/Harvard Medical School, Belmont, MA 02478, USA. Pages: 1721-37 Volume: 26 Number: 7 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17897390 Abstract: Amyotrophic lateral sclerosis (ALS), spinal bulbar muscular atrophy (or Kennedy's disease), spinal muscular atrophy and spinal muscular atrophy with respiratory distress 1 are neurodegenerative disorders mainly affecting motor neurons and which currently lack effective therapies. Recent studies in animal models as well as primary and embryonic stem cell models of ALS, utilizing over-expression of mutated forms of Cu/Zn superoxide dismutase 1, have shown that motor neuron degeneration in these models is in part a non cell-autonomous event and that by providing genetically non-compromised supporting cells such as microglia or growth factor-excreting cells, onset can be delayed and survival increased. Using models of acute motor neuron injury it has been shown that embryonic stem cell-derived motor neurons implanted into the spinal cord can innervate muscle targets and improve functional recovery. Thus, a rationale exists for the development of cell therapies in motor neuron diseases aimed at either protecting and/or replacing lost motor neurons, interneurons as well as non-neuronal cells. This review evaluates approaches used in animal models of motor neuron disorders and their therapeutic relevance. Year: 2007 Month: 10 Title: Cell-lineage Regulated Myogenesis for Dystrophin Replacement: a Novel Therapeutic Approach for Treatment of Muscular Dystrophy Authors: Kimura, E. Han, J. J. Li, S. Fall, B. Ra, J. Haraguchi, M. Tapscott, S. J. Chamberlain, J. S. Auth Address: Department of Neurology, Senator Paul D Wellstone Muscular Dystrophy Cooperative Research Center University of Washington School of Medicine, Seattle, WA, 98195-7720 USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18511457 Abstract: Duchenne muscular dystrophy (DMD) is characterized in skeletal muscle by cycles of myofiber necrosis and regeneration leading to loss of muscle fibers and replacement with fibrotic connective and adipose tissue. The ongoing activation and recruitment of muscle satellite cells for myofiber regeneration results in loss of regenerative capacity in part due to proliferative senescence. We explored a method whereby new myoblasts could be generated in dystrophic muscles by transplantation of primary fibroblasts engineered to express a micro-dystrophin/eGFP (microDys/eGFP) fusion gene together with a tamoxifen-inducible form of the myogenic regulator MyoD [MyoD-ER(T)]. Fibroblasts isolated from mdx(4cv) mice, a mouse model for DMD, were efficiently transduced with lentiviral vectors expressing microDys/eGFP and MyoD-ER(T) and underwent myogenic conversion when exposed to tamoxifen. These cells could also be induced to differentiate into microDys/eGFP-expressing myocytes and myotubes. Transplantation of transduced mdx(4cv) fibroblasts into mdx(4cv) muscles enabled tamoxifen-dependent regeneration of myofibers that express micro-dystrophin. This lineage control method therefore allows replenishment of myogenic stem cells using autologous fibroblasts carrying an exogenous dystrophin gene. This strategy carries several potential advantages over conventional myoblast transplantation methods including: 1) the relative simplicity of culturing fibroblasts compared with myoblasts, 2) a readily available cell source and ease of expansion, and 3) the ability to induce MyoD gene expression in vivo via administration of a medication. Our study provides a proof of concept for a novel gene/stem cell therapy technique and opens another potential therapeutic approach for degenerative muscle disorders. Year: 2008 Title: Cell-penetrating DNA-binding protein as a safe and efficient naked DNA delivery carrier in vitro and in vivo Authors: Kim, E. S. Yang, S. W. Hong, D. K. Kim, W. T. Kim, H. G. Lee, S. K. Auth Address: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20043881 Abstract: Non-viral gene delivery is a safe and suitable alternative to viral vector-mediated delivery to overcome the immunogenicity and tumorigenesis associated with viral vectors. Using the novel, human-origin Hph-1 protein transduction domain that can facilitate the transduction of protein into cells, we developed a new strategy to deliver naked DNA in vitro and in vivo. The new DNA delivery system contains Hph-1-GAL4 DNA-binding domain (DBD) fusion protein and enhanced green fluorescent protein (EGFP) reporter plasmid that includes the five repeats of GAL4 upstream activating sequence (UAS). Hph-1-GAL4-DBD protein formed complex with plasmid DNA through the specific interaction between GAL4-DBD and UAS, and delivered into the cells via the Hph-1-PTD. The pEGFP DNA was successfully delivered by the Hph-1-GAL4 system, and the EGFP was effectively expressed in mammalian cells such as HeLa and Jurkat, as well as in Bright Yellow-2 (BY-2) plant cells. When 10mug of pEGFP DNA was intranasally administered to mice using Hph-1-GAL4 protein, a high level of EGFP expression was detected throughout the lung tissue for 7days. These results suggest that an Hph-1-PTD-mediated DNA delivery strategy may be an useful non-viral DNA delivery system for gene therapy and DNA vaccines. Year: 2009 Title: Cellular patterns of the atrophic response in murine soleus and gastrocnemius muscles submitted to simulated weightlessness Authors: Ferreira, R. Vitorino, R. Neuparth, M. J. Appell, H. J. Amado, F. Duarte, J. A. Auth Address: CIAFEL, Laboratory of Sport Biochemistry, Faculty of Sport Sciences, University of Porto, Rua Dr. Placido Costa, 91, 4200, Porto, Portugal. rmferreira@fcdef.up.pt Pages: 331-40 Volume: 101 Number: 3 Keywords: Animals Apoptosis Biological Markers/analysis/metabolism Caspase 3/metabolism Cell Proliferation Hindlimb Suspension/physiology Male Mice Mice, Inbred Strains Muscle, Skeletal/*pathology/physiology/ultrastructure Muscular Atrophy/metabolism/*pathology Myosin Light Chains/metabolism Necrosis/metabolism Protein Isoforms/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17624543 Abstract: The purpose of the present study was to investigate the mechanisms of cell death (apoptosis vs. necrosis) during muscle atrophy induced by 1 week of hindlimb suspension. Biochemical and morphological parameters were examined in murine soleus and gastrocnemius muscles. A total of 70 male Charles River CD1 mice were randomly assigned to seven groups (n = 10/group): Cont (loading control conditions) and 6HS, 12HS, 24HS, 48HS, 72HS and 1wkHS with respect to the period of hindlimb suspension (HS). Compared to the Cont, skeletal muscle atrophy was confirmed by a significant decrease of 44 and of 17% in fiber cross-sectional areas in the gastrocnemius and soleus, respectively. A significant increase in caspase-3 activity was noticed in 6HS (196%, P < 0.05) and in 12HS (201%, P < 0.05), as well as the amount of cytosolic mono- and oligonucleosomes at 12HS (142%, P < 0.05) and 24HS (203%, P < 0.05) in the gastrocnemius and soleus, respectively. The profile of necrotic markers showed a peak of myeloperoxidase activity at 24HS (170%, P < 0.05) and at 72HS (114%, P < 0.05) in the gastrocnemius and soleus, respectively. The analysis of N-acetylglucosaminidase activity evidenced more increment in the soleus at 72HS (60%, P < 0.05). The analysis of the basal values of these parameters suggested that apoptosis prevailed in the slow-twitch muscle analyzed, whereas lysosomic activity seemed to be more pronounced in the gastrocnemius. The morphological data supported the biochemical results pointing towards a shift from apoptosis to necrosis, which seems to corroborate the aponecrosis theory. Year: 2007 Month: 10 Title: Cellular therapies in motor neuron diseases Authors: Nayak, M. S. Kim, Y. S. Goldman, M. Keirstead, H. S. Kerr, D. A. Auth Address: Department of Neurology, Johns Hopkins University School of Medicine, 600 North Wolfe Street, Baltimore, MD 21287, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16872810 Abstract: Amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) are prototypical motor neuron diseases that result in progressive weakness as a result of motor neuron dysfunction and death. Though much work has been done in both diseases to identify the cellular mechanisms of motor neuron dysfunction, once motor neurons have died, one of potential therapies to restore function would be through the use of cellular transplantation. In this review, we discuss potential strategies whereby cellular therapies, including the use of stem cells, neural progenitors and cells engineered to secrete trophic factors, may be used in motor neuron diseases. We review pre-clinical data in rodents with each of these approaches and discuss advances and regulatory issues regarding the use of cellular therapies in human motor neuron diseases. Year: 2006 Title: Cep-1347 (KT7515), a semisynthetic inhibitor of the mixed lineage kinase family Authors: Maroney, A. C. Finn, J. P. Connors, T. J. Durkin, J. T. Angeles, T. Gessner, G. Xu, Z. Meyer, S. L. Savage, M. J. Greene, L. A. Scott, R. W. Vaught, J. L. Auth Address: Cephalon Inc., 145 Brandywine Pkwy., West Chester, Pennsylvania 19380, USA. amaroney@cephalon.com Pages: 25302-8 Volume: 276 Number: 27 Keywords: Animals CHO Cells Carbazoles/*pharmacology Cell Death Cricetinae Enzyme Activation Enzyme Inhibitors/*pharmacology Indoles/*pharmacology JNK Mitogen-Activated Protein Kinases MAP Kinase Kinase Kinases/*antagonists & inhibitors Mitogen-Activated Protein Kinase 8 Mitogen-Activated Protein Kinases/antagonists & inhibitors/metabolism Models, Chemical PC12 Cells *Protein Kinase Inhibitors Rats Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11325962 Abstract: CEP-1347 (KT7515) promotes neuronal survival at dosages that inhibit activation of the c-Jun amino-terminal kinases (JNKs) in primary embryonic cultures and differentiated PC12 cells after trophic withdrawal and in mice treated with 1-methyl-4-phenyl tetrahydropyridine. In an effort to identify molecular target(s) of CEP-1347 in the JNK cascade, JNK1 and known upstream regulators of JNK1 were co-expressed in Cos-7 cells to determine whether CEP-1347 could modulate JNK1 activation. CEP-1347 blocked JNK1 activation induced by members of the mixed lineage kinase (MLK) family (MLK3, MLK2, MLK1, dual leucine zipper kinase, and leucine zipper kinase). The response was selective because CEP-1347 did not inhibit JNK1 activation in cells induced by kinases independent of the MLK cascade. CEP-1347 inhibition of recombinant MLK members in vitro was competitive with ATP, resulting in IC(50) values ranging from 23 to 51 nm, comparable to inhibitory potencies observed in intact cells. In addition, overexpression of MLK3 led to death in Chinese hamster ovary cells, and CEP-1347 blocked this death at doses comparable to those that inhibited MLK3 kinase activity. These results identify MLKs as targets of CEP-1347 in the JNK signaling cascade and demonstrate that CEP-1347 can block MLK-induced cell death. Year: 2001 Title: Cerebrospinal Fluid from sporadic Amyotrophic Lateral Sclerosis patients induces degeneration of a cultured motor neuron cell line Authors: Vijayalakshmi, K. Alladi, P. A. Sathyaprabha, T. N. Subramaniam, J. R. Nalini, A. Raju, T. R. Auth Address: Department of Neurophysiology, National Institute of Mental Health and Neuro Sciences Hosur Road, Bangalore-29, India. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19368830 Abstract: We investigated the effect of cerebrospinal fluid (CSF) from sporadic Amyotrophic Lateral Sclerosis patients (SALS-CSF) on motor neuron-like cells to delineate the pathomechanism of SALS. Exposure of NSC-34 cells to SALS-CSF caused lower viability, reduction in differentiation and enhanced lactate dehydrogenase activity. Additionally, reduced choline acetyl transferase expression alongside intracellular aggregation of phosphorylated neurofilaments was prominently seen. The aggregates were immunopositive for ubiquitin. These findings are comparable to the pathological changes seen in the postmortem tissue of ALS patients. Unlimited supply of NSC-34 cells and their vulnerability to SALS-CSF render them to be a good bioassay system to identify new therapeutic agents conferring protection to motor neurons. Year: 2009 Title: Cervical spinal cord therapeutics delivery: preclinical safety validation of a stabilized microinjection platform Authors: Riley, J. Federici, T. Park, J. Suzuki, M. Franz, C. K. Tork, C. McHugh, J. Teng, Q. Svendsen, C. Boulis, N. M. Auth Address: Cleveland Clinic Foundation, The Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio, USA. Pages: 754-61; discussion 761-2 Volume: 65 Number: 4 Keywords: Animals Anterior Horn Cells/cytology/physiology/transplantation Cell Differentiation/physiology Cell Survival/physiology Cervical Vertebrae/anatomy & histology/surgery Female Graft Rejection/drug therapy/prevention & control Graft Survival/physiology Hematoma, Epidural, Spinal/etiology/pathology/physiopathology Humans Immunosuppressive Agents/therapeutic use Infusion Pumps Laminectomy Microinjections/adverse effects/*instrumentation/methods Neurogenesis/physiology Postoperative Complications/etiology/physiopathology/prevention & control Spinal Cord/*cytology/physiology/*surgery Spinal Cord Diseases/*surgery Stem Cell Transplantation/adverse effects/*instrumentation/methods Stem Cells/*cytology/physiology Stereotaxic Techniques Sus scrofa Syringes/adverse effects/*standards Transplantation, Heterologous/adverse effects/instrumentation/methods Treatment Outcome Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19834381 Abstract: OBJECTIVE: The current series represents a preclinical safety validation study for direct parenchymal microinjection of cellular grafts into the ventral horn of the porcine cervical spinal cord. METHODS: Twenty-four 30- to 40-kg female Yorkshire farm pigs immunosuppressed with cyclosporine underwent a cervical laminectomy and ventral horn human neural progenitor cell injection. Cell transplantation in groups 1 to 3 (n = 6 pigs each) was undertaken with the intent of assessing the safety of varied injection volumes: 10, 25, and 50 microL injected at 1, 2.5, and 5 microL/min, respectively. Groups 4 and 5 (n = 3 pigs each) received prolonged immunosuppressant pretreatment in an attempt to demonstrate graft viability. The latter was undertaken in an alternate species (mini-pig versus Yorkshire pig). RESULTS: Neurological morbidity was observed in 1 animal and was attributable to the presence of a resolving epidural hematoma noted at necropsy. Although instances of ventral horn targeting were achieved in all injection groups with a coordinate-based approach, opportunities exist for improvement in accuracy and precision. A relationship between injection volume and graft site cross-sectional area suggested limited reflux. Only animals from group 5 achieved graft survival at a survival end point (t = 1 week). CONCLUSION: This series demonstrated the functional safety of targeted ventral horn microinjection despite evidence for graft site immune rejection. Improvements in graft delivery may be augmented with an adapter to improve control of the cannula entry angle, intraoperative imaging, or larger graft volumes. Finally, demonstration of long-term graft viability in future preclinical toxicity studies may require tailored immunosuppressive therapies, an allograft construct, or tailored choice of host species. Year: 2009 Month: 10 Title: Challenges and opportunities in clinical trials for spinal muscular atrophy Authors: Hirtz, D. Iannaccone, S. Heemskerk, J. Gwinn-Hardy, K. Moxley, R., 3rd Rowland, L. P. Auth Address: National Institute of Neurological Disorders and Stroke, NIH, Bethesda, MD, USA. hirtzd@ninds.nih.gov Pages: 1352-7 Volume: 65 Number: 9 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16275820 Abstract: Spinal muscular atrophy (SMA) is the most common fatal neuromuscular disease of infancy. SMA type I is the most severe and mortality is usually due to respiratory failure. In type II the disability is of later onset and less severe, and prognosis has improved primarily due to supportive care. Type III is the mildest form with onset usually of weakness in adolescence or young adulthood. SMA is an autosomal recessive disorder with deletions or mutations of the gene at the 5 q11 locus. There is no specific prevention or treatment, but current progress toward potential therapies has been substantial and several candidates including histone deacetylase (HDAC) inhibitors are under consideration for further evaluation. The authors sought to address the challenges and opportunities for testing new therapies for SMA. Year: 2005 Title: Changes in cerebral neurotransmitters and metabolites induced by acute donepezil and memantine administrations: A microdialysis study Authors: Shearman, E. Rossi, S. Szasz, B. Juranyi, Z. Fallon, S. Pomara, N. Sershen, H. Lajtha, A. Auth Address: Nathan Kline Institute, Orangeburg, NY 10962, USA. Pages: 204-13 Volume: 69 Number: 2 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16533671 Abstract: Cholinesterase inhibitors including donepezil, rivastigmine, and galantamine and the N-methyl-d-aspartate (NMDA) antagonist, memantine are the medications currently approved for the treatment of Alzheimer's disease (AD). In addition to their beneficial effects on cognitive and functional domains typically disrupted in AD, these agents have also been shown to slow down the emergence of behavioral and psychotic symptoms associated with this disease. However, the underlying mechanisms for these therapeutic effects remain poorly understood and could involve effects of these medications on non-cholinergic or non-glutamatergic neurotransmitter systems respectively. These considerations prompted us to initiate a series of investigations to examine the acute and chronic effects of donepezil (Aricept (+/-)-2,3-dihydro-5,6-dimethoxy-2-[[1-(phenylmethyl)-4-piperidinyl]methyl] -1H-inden-1-1 hydrochloride and memantine (1-amino-3,5-dimethyladamantane hydrochloride C(12)H(21)N.HCl)). The present study focuses on the acute effects of donepezil and memantine on brain extracellular levels of acetylcholine, dopamine, serotonin, norepinephrine and their metabolites. We assayed changes in the ventral and dorsal hippocampus and the prefrontal and medial temporal cortex by microdialysis. Memantine resulted in significant increases in extracellular dopamine (DA), norepinephrine (NE), and their metabolites, in the cortical regions, and in a reduction of DA in the hippocampus. Donepezil produced an increase in extracellular DA in the cortex and in the dorsal hippocampus. Norepinephrine increased in the cortex; with donepezil it increased in the dorsal hippocampus and the medial temporal cortex, and decreased in the ventral hippocampus. Interestingly both compounds decreased extracellular serotonin (5HT) levels. The metabolites of the neurotransmitters were increased in most areas. We also found an increase in extracellular acetylcholine (ACh) by memantine in the nucleus accumbens and the ventral tegmental area. Our results suggest both region and drug specific neurotransmitter effects of these agents as well as some similarities. We conclude that drugs influencing cognitive mechanisms induce changes in a number of neurotransmitters with the changes being both region and drug specific. Release and metabolism are altered and extracellular neurotransmitter levels can be increased or decreased by the drugs. Other studies are in progress to determine the pharmacological effects associated with chronic treatment with these compounds, which may be more pertinent to the clinical situation in which patients take these medications for months or years. Year: 2006 Title: Chaperoning ribonucleoprotein biogenesis in health and disease Authors: Pellizzoni, L. Auth Address: Dulbecco Telethon Institute, Institute of Cell Biology (CNR), Via E. Ramarini 32, 00016 Monterotondo Scalo, Rome, Italy. livio.pellizzoni@ibc.cnr.it Pages: 340-5 Volume: 8 Number: 4 Keywords: Animals Cell Survival Cyclic AMP Response Element-Binding Protein/genetics/*metabolism Health Humans Molecular Chaperones/*metabolism Motor Neurons/metabolism/*physiology Muscular Atrophy, Spinal/*etiology/genetics Nerve Tissue Proteins/genetics/*metabolism Nervous System/cytology/*growth & development/metabolism RNA, Messenger/metabolism RNA-Binding Proteins/genetics/*metabolism Ribonucleoproteins, Small Nuclear/*metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17401408 Abstract: The survival motor neuron (SMN) protein is part of a macromolecular complex that functions in the biogenesis of small nuclear ribonucleoproteins (snRNPs)--the essential components of the pre-messenger RNA splicing machinery--as well as probably other RNPs. Reduced levels of SMN expression cause the inherited motor neuron disease spinal muscular atrophy (SMA). Knowledge of the composition, interactions and functions of the SMN complex has advanced greatly in recent years. The emerging picture is that the SMN complex acts as a macromolecular chaperone of RNPs to increase the efficiency and fidelity of RNA-protein interactions in vivo, and to provide an opportunity for these interactions to be regulated. In addition, it seems that RNA metabolism deficiencies underlie SMA. Here, a dual dysfunction hypothesis is presented in which two mechanistically and temporally distinct defects--that are dependent on the extent of SMN reduction in SMA--affect the homeostasis of specific messenger RNAs encoding proteins essential for motor neuron development and function. Year: 2007 Month: 4 Title: Characterization of the survival motor neuron (SMN) promoter provides evidence for complex combinatorial regulation in undifferentiated and differentiated P19 cells Authors: Rouget, R. Vigneault, F. Codio, C. Rochette, C. Paradis, I. Drouin, R. Simard, L. R. Auth Address: Centre de Recherche, Hopital Sainte-Justine and Universite de Montreal, Montreal, QC, Canada H3T 1C5. Pages: 433-43 Volume: 385 Number: Pt 2 Keywords: Animals Base Sequence/genetics Cell Line Cloning, Molecular/methods Conserved Sequence/genetics Cyclic AMP Response Element-Binding Protein/*genetics DNA Footprinting Electrophoretic Mobility Shift Assay/methods Embryo/cytology/innervation Enhancer Elements (Genetics)/genetics Genomics/methods Humans Hybrid Cells/chemistry/metabolism Mice Molecular Sequence Data Motor Neurons/chemistry/cytology/metabolism Mutagenesis, Site-Directed/genetics Nerve Tissue Proteins/*genetics Phylogeny Promoter Regions (Genetics)/*genetics RNA-Binding Proteins/*genetics Research Support, Non-U.S. Gov't Stem Cells/chemistry/metabolism Transcription, Genetic/genetics Transfection/methods Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15361068 Abstract: There exist two SMN (survival motor neuron) genes in humans, the result of a 500 kb duplication in chromosome 5q13. Deletions/mutations in the SMN1 gene are responsible for childhood spinal muscular atrophy, an autosomal recessive neurodegenerative disorder. While the SMN1 and SMN2 genes are not functionally equivalent, up-regulation of the SMN2 gene represents an important therapeutic target. Consequently, we exploited in silico, in vitro and in vivo approaches to characterize the core human and mouse promoters in undifferentiated and differentiated P19 cells. Phylogenetic comparison revealed four highly conserved regions that contained a number of cis-elements, only some of which were shown to activate/repress SMN promoter activity. Interestingly, the effect of two Sp1 cis-elements varied depending on the state of P19 cells and was only observed in combination with a neighbouring Ets cis-element. Electrophoretic mobility-shift assay and in vivo DNA footprinting provided evidence for DNA-protein interactions involving Sp, NF-IL6 and Ets cis-elements, whereas transient transfection experiments revealed complex interactions involving these recognition sites. SMN promoter activity was strongly regulated by an NF-IL6 response element and this regulation was potentiated by a downstream Ets element. In vivo results suggested that the NF-IL6 response must function either via a protein-tethered transactivation mechanism or a transcription factor binding an upstream element. Our results provide strong evidence for complex combinatorial regulation and suggest that the composition or state of the basal transcription complex binding to the SMN promoter is different between undifferentiated and differentiated P19 cells. Year: 2005 Title: Chemical genetics and orphan genetic diseases Authors: Lunn, M. R. Stockwell, B. R. Auth Address: Department of Biological Sciences, Sherman Fairchild Center for the Life Sciences, Columbia University, New York, New York 10027, USA. Pages: 1063-73 Volume: 12 Number: 10 Keywords: Drug Design Drug Industry/*trends Humans Models, Molecular Muscular Atrophy, Spinal/drug therapy/genetics/pathology Rare Diseases/*drug therapy/*genetics/pathology Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16242649 Abstract: Many orphan diseases have been identified that individually affect small numbers of patients but cumulatively affect approximately 6%-10% of the European and United States populations. Human genetics has become increasingly effective at identifying genetic defects underlying such orphan genetic diseases, but little progress has been made toward understanding the causal molecular pathologies and creating targeted therapies. Chemical genetics, positioned at the interface of chemistry and genetics, can be used for elucidation of molecular mechanisms underlying diseases and for drug discovery. This review discusses recent advances in chemical genetics and how small-molecule tools can be used to study and ultimately treat orphan genetic diseases. We focus here on a case study involving spinal muscular atrophy, a pediatric neurodegenerative disease caused by homozygous deletion of the SMN1 (survival of motor neuron 1) gene. Year: 2005 Month: 10 Title: Chemopreventive effect of peroxisome proliferator-activated receptor gamma on gastric carcinogenesis in mice Authors: Lu, J. Imamura, K. Nomura, S. Mafune, K. Nakajima, A. Kadowaki, T. Kubota, N. Terauchi, Y. Ishii, G. Ochiai, A. Esumi, H. Kaminishi, M. Auth Address: Department of Gastrointestinal Surgery, Faculty of Medicine, University of Tokyo, Japan. jlu@east.ncc.go.jp Pages: 4769-74 Volume: 65 Number: 11 Keywords: Animals Anticarcinogenic Agents/*pharmacology Carcinogens Chromans/*pharmacology Female Gastric Mucosa/metabolism Ligands Male Methylnitrosourea Mice Mice, Knockout PPAR gamma/biosynthesis/deficiency/genetics/*physiology Stomach Neoplasms/chemically induced/metabolism/pathology/*prevention & control Thiazolidinediones/*pharmacology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15930296 Abstract: Peroxisome proliferator-activated receptor gamma (PPARgamma) is known to be expressed in several cancers, and the treatment of these cancer cells with PPARgamma ligands often induces cell differentiation and apoptosis. Recently, the chemopreventive potential of PPARgamma ligands on colon carcinogenesis was reported, although the effect of PPARgamma on colon carcinogenesis and the mechanism of the effect remain controversial. In this study, we attempted to elucidate the role of PPARgamma in gastric carcinogenesis and explored the possible use of PPARgamma ligand as a chemopreventive agent for gastric cancer. N-methyl-N-nitrosourea (MNU, 240 ppm) was given in drinking water for 10 weeks to induce gastric cancer in PPARgamma wild-type (+/+) and heterozygous-deficient (+/-) mice, followed by treatment with PPARgamma ligand [troglitazone, 0.15% (w/w) in powder food] or the vehicle alone for 42 weeks. At the end of the experiment, PPARgamma (+/-) mice were more susceptible to MNU-induced gastric cancer than wild-type (+/+) mice (89.5%/55.5%), and troglitazone significantly reduced the incidence of gastric cancer in PPARgamma (+/+) mice (treatment 55.5%/vehicle 9%) but not in PPARgamma (+/-) mice. The present study showed that (a) PPARgamma suppresses gastric carcinogenesis, (b) the PPARgamma ligand troglitazone is a potential chemopreventive agent for gastric carcinogenesis, and (c) troglitazone's chemopreventive effect is dependent on PPARgamma. Year: 2005 Title: Childhood spinal muscular atrophy induces alterations in contractile and regulatory protein isoform expressions Authors: Stevens, L. Bastide, B. Maurage, C. A. Dupont, E. Montel, V. Cieniewski-Bernard, C. Cuisset, J. M. Vallee, L. Mounier, Y. Auth Address: Laboratoire de Plasticite Neuromusculaire, Universite des Sciences et Technologies de Lille, Villeneuve d'Ascq cedex, France. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18363640 Abstract: Aims: Although modifications of the survival motor neurone gene are responsible for most spinal muscular atrophy (SMA) cases, the molecular pathophysiology and the muscular target proteins involved are still unknown. The aim of this study was to compare the expression of contractile and regulatory protein isoforms in quadriceps muscles from SMA children with age-matched control quadriceps. Methods: The isoform patterns of myosin heavy chains (MHC), troponin subunits (T, C and I) and tropomyosin were determined by immunoblotting, reverse transcription-polymerase chain reaction and mass spectrometry analyses. Depending on the disease severity, their expression levels were followed in specific variants of SMA populations (types I, II and III), with comparison with age-matched control muscles. Results: The isoform transitions in SMA muscles were different from the fast-to-faster transitions occurring in normal muscles from children aged 1 month to 5 years old. Moreover, the expression of the neonatal MHC isoform was not repressed in SMA muscles. Conclusions: The presence of the neonatal MHC isoform in SMA muscles indicates an alteration of the phenotype in these diseased muscles. It is strongly suggested that MHC and troponin T proteins may be good markers for the SMA pathology. Year: 2008 Title: Choline acetyltransferase expression does not identify early pathogenic events in fetal SMA spinal cord Authors: Soler-Botija, C. Cusco, I. Lopez, E. Clua, A. Gich, I. Baiget, M. Ferrer, I. Tizzano, E. F. Auth Address: Servei de Genetica and Institut de Recerca, Hospital de Sant Pau, Av. Padre Claret 167, 08025 Barcelona, Spain. Pages: 253-8 Volume: 15 Number: 3 Keywords: Age Factors Blotting, Western/methods Cell Count/methods Choline O-Acetyltransferase/*metabolism Comparative Study Fetus Gestational Age Humans Immunohistochemistry/methods Infant, Newborn Motor Neurons/metabolism Muscular Atrophy, Spinal/*enzymology/metabolism Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15725587 Abstract: We investigated the expression of choline acetyltransferase, a specific marker for cholinergic neurons, in control and spinal muscular atrophy fetuses and newborns. By immunoblot we observed at 12 and 15 weeks a similar pattern of choline acetyltransferase expression in spinal muscular atrophy with respect to controls, although at 22 weeks this expression was reduced, probably due to a smaller number of motor neurons in the spinal muscular atrophy spinal cord. By immunohistochemistry, the counting of positive and negative motor neurons for choline acetyltransferase immunostaining in control and spinal muscular atrophy fetuses showed a similar proportion at all stages analyzed. The choline acetyltransferase-negative motor neurons were of similar appearance in both groups. After birth, chromatolytic motor neurons were detected in spinal muscular atrophy, all of which were choline acetyltransferase-negative. Our results in spinal muscular atrophy fetuses indicate that choline acetyltransferase immunostaining does not identify early events in neuronal pathogenesis and suggest that the spinal muscular atrophy surviving motor neurons may not be dysfunctional during this period. Furthermore, spinal muscular atrophy choline acetyltransferase-negative motor neurons showed detectable pathological changes only after birth, indicating that choline acetyltransferase is a late marker for motor neuron degeneration and not a primary contributing factor in this process. Year: 2005 Month: 3 Title: Chromatin: the interface between extrinsic cues and the epigenetic regulation of muscle regeneration Authors: Guasconi, V. Puri, P. L. Auth Address: Istituto Dulbecco Telethon at Istituto Di Ricovero e Cura a Carattere Scientifico, Santa Lucia Fondazione and European Brain Research Institute, 64 Via del Fosso di Fiorano, 00143 Rome, Italy. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19394225 Abstract: Muscle regeneration provides a paradigm by which to study how extrinsic signals coordinate gene expression in somatic stem cells (satellite cells) by directing the genome distribution of chromatin-modifying complexes. Understanding the signal-dependent control of the epigenetic events underlying the transition of muscle stem cells through sequential regeneration stages holds the promise to reveal new targets for selective interventions toward repairing diseased muscles. This review describes the latest findings on how regeneration cues are integrated at the chromatin level to build the transcription network that regulates progression of endogenous muscle progenitors throughout the myogenic program. In particular, we describe how specific epigenetic signatures can confer responsiveness to extrinsic cues on discrete regions of the muscle stem cell genome. Year: 2009 Title: Chromogranin-mediated secretion of mutant superoxide dismutase proteins linked to amyotrophic lateral sclerosis Authors: Urushitani, M. Sik, A. Sakurai, T. Nukina, N. Takahashi, R. Julien, J. P. Auth Address: Department of Anatomy and Physiology, Laval University, Centre de Recherche du Centre Hospitalier de l' Universite Laval, 2705 boulevard Laurier, Sainte-Foy, Quebec G1V 4G2, Canada. Pages: 108-18 Volume: 9 Number: 1 Keywords: 3T3 Cells Amyotrophic Lateral Sclerosis/*enzymology/*genetics Animals Astrocytes/metabolism Blotting, Western COS Cells Cells, Cultured Cercopithecus aethiops Chromogranins/*physiology Fluorescent Antibody Technique Gliosis/pathology Humans Immunoprecipitation Interneurons/metabolism/physiology Mice Mice, Transgenic Microscopy, Immunoelectron Microsomes/enzymology Molecular Chaperones/genetics Motor Neurons/metabolism/physiology Mutation/*physiology Plasmids/genetics Proteasome Endopeptidase Complex/antagonists & inhibitors Protein Folding Research Support, Non-U.S. Gov't Spinal Cord/cytology/metabolism Subcellular Fractions/physiology Superoxide Dismutase/*genetics/*metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16369483 Abstract: Here we report that chromogranins, components of neurosecretory vesicles, interact with mutant forms of superoxide dismutase (SOD1) that are linked to amyotrophic lateral sclerosis (ALS), but not with wild-type SOD1. This interaction was confirmed by yeast two-hybrid screen and by co-immunoprecipitation assays using either lysates from Neuro2a cells coexpressing chromogranins and SOD1 mutants or lysates from spinal cord of ALS mice. Confocal and immunoelectron microscopy revealed a partial colocalization of mutant SOD1 with chromogranins in spinal cord of ALS mice. Mutant SOD1 was also found in immuno-isolated trans-Golgi network and in microsome preparations, suggesting that it can be secreted. Indeed we report evidence that chromogranins may act as chaperone-like proteins to promote secretion of SOD1 mutants. From these results, and our finding that extracellular mutant SOD1 can trigger microgliosis and neuronal death, we propose a new ALS pathogenic model based on the toxicity of secreted SOD1 mutants. Year: 2006 Month: 1 Title: Chronic activation in presymptomatic amyotrophic lateral sclerosis (ALS) mice of a feedback loop involving Fas, Daxx, and FasL Authors: Raoul, C. Buhler, E. Sadeghi, C. Jacquier, A. Aebischer, P. Pettmann, B. Henderson, C. E. Haase, G. Auth Address: Ecole Polytechnique Federale de Lausanne (EPFL), Integrative Biosciences Institute, SV IBI LEN, AAB 1 32, CH-1015 Lausanne, Switzerland; Institut de Neurobiologie de la Mediterranee (INMED), Institut National de la Sante et de la Recherche Medicale (INSERM), Equipe Avenir, F-13273 Marseille Cedex 09, France; Universite de la Mediterranee, F-13288 Marseille, France. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16581901 Abstract: The reasons for the cellular specificity and slow progression of motoneuron diseases such as ALS are still poorly understood. We previously described a motoneuron-specific cell death pathway downstream of the Fas death receptor, in which synthesis of nitric oxide (NO) is an obligate step. Motoneurons from ALS model mice expressing mutant SOD1 showed increased susceptibility to exogenous NO as compared with controls. Here, we report a signaling mechanism whereby NO leads to death of mutant, but not control, motoneurons. Unexpectedly, exogenous NO triggers expression of Fas ligand (FasL) in cultured motoneurons. In mutant SOD1(G93A) and SOD1(G85R), but not in control motoneurons, this up-regulation results in activation of Fas, leading through Daxx to phosphorylation of p38 and further NO synthesis. This Fas/NO feedback amplification loop is required for motoneuron death in vitro. In vivo, mutant SOD1(G93A) and SOD1(G85R) mice show increased numbers of positive motoneurons and Daxx nuclear bodies weeks before disease onset. Moreover, FasL up-regulation is reduced in the presence of transgenic dominant-negative Daxx. We propose that chronic low-level activation of the Fas/NO feedback loop may underlie the motoneuron loss that characterizes familial ALS and may help to explain its slowly progressive nature. Year: 2006 Title: Ciliary neurotrophic factor fused to a protein transduction domain retains full neuroprotective activity in the absence of cytokine-like side effects Authors: Rezende, A. C. Peroni, D. Vieira, A. S. Rogerio, F. Talaisys, R. L. Costa, F. T. Langone, F. Skaper, S. D. Negro, A. Auth Address: Department of Physiology and Biophysics, State University of Campinas, Campinas, Brazil. Pages: 1680-90 Volume: 109 Number: 6 Keywords: Adipose Tissue, Brown/drug effects/metabolism Animals Animals, Newborn Axotomy/methods Body Weight/drug effects Cell Count/methods Cells, Cultured Chick Embryo Ciliary Neurotrophic Factor/metabolism/*therapeutic use Cricetinae Cricetulus Dose-Response Relationship, Drug Ganglia, Spinal/cytology Green Fluorescent Proteins/genetics Humans Motor Neurons/drug effects/physiology Neuroprotective Agents/*therapeutic use Rats Rats, Wistar Recombinant Fusion Proteins/metabolism/therapeutic use STAT3 Transcription Factor/metabolism Sciatic Neuropathy/*drug therapy/etiology/physiopathology Sensory Receptor Cells/drug effects/physiology Transduction, Genetic/methods tat Gene Products, Human Immunodeficiency Virus/genetics/therapeutic use Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19457136 Abstract: Ciliary neurotrophic factor (CNTF) is a multifunctional cytokine that can regulate the survival and differentiation of many types of developing and adult neurons. CNTF prevents the degeneration of motor neurons after axotomy and in mouse mutant progressive motor neuronopathy, which has encouraged trials of CNTF for human motor neuron disease. Given systemically, however, CNTF causes severe side effects, including cachexia and a marked immune response, which has limited its clinical application. The present work describes a novel approach for administering recombinant human CNTF (rhCNTF) while conserving neurotrophic activity and avoiding deleterious side effects. rhCNTF was fused to a protein transduction domain derived from the human immunodeficiency virus-1 TAT (transactivator) protein. The resulting fusion protein (TAT-CNTF) crosses the plasma membrane within minutes and displays a nuclear localization. TAT-CNTF was equipotent to rhCNTF in supporting the survival of cultured chicken embryo dorsal root ganglion neurons. Local or subcutaneous administration of TAT-CNTF, like rhCNTF rescued motor neurons from death in neonatal rats subjected to sciatic nerve transection. In contrast to subcutaneous rhCNTF, which caused a 20-30% decrease in body weight in neonatal rats between postnatal days 2 and 7 together with a considerable fat mobilization in brown adipose tissue, TAT-CNTF lacked such side effects. Together, these results indicate that rhCNTF fused with the protein transduction domain/TAT retains neurotrophic activity in the absence of CNTFs cytokine-like side effects and may be a promising candidate for the treatment of motor neuron and other neurodegenerative diseases. Year: 2009 Month: 6 Title: Ciliary neurotrophic factor prevents degeneration of motor neurons in mouse mutant progressive motor neuronopathy Authors: Sendtner, M. Schmalbruch, H. Stockli, K. A. Carroll, P. Kreutzberg, G. W. Thoenen, H. Auth Address: Department of Neurochemistry, Max-Planck-Institute for Psychiatry, Martinsried, Germany. Pages: 502-4 Volume: 358 Number: 6386 Keywords: Animals Cell Count Cell Line Ciliary Neurotrophic Factor Gene Therapy Mice Mice, Neurologic Mutants Motor Neuron Disease/*drug therapy/pathology Motor Neurons/pathology Nerve Tissue Proteins/genetics/*therapeutic use Phrenic Nerve/pathology Pons/pathology Research Support, Non-U.S. Gov't Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1641039 Abstract: Ciliary neurotrophic factor (CNTF) supports the survival of embryonic motor neurons in vitro and in vivo, and prevents lesion-mediated degeneration of rat motor neurons during early post-natal stages. Here we report that CNTF greatly reduces all the functional and morphological changes in pmn/pmn mice, an autosomal recessive mutant leading to progressive caudo-cranial motor neuron degeneration. The first manifestations of progressive motor neuronopathy in homozygous pmn/pmn mice become apparent in the hind limbs at the end of the third post-natal week, and all the mice die up to 6 or 7 weeks after birth from respiratory paralysis. Treatment with CNTF prolongs survival and greatly improves motor function of these mice. Moreover, morphological manifestations, such as loss of motor axons in the phrenic nerve and degeneration of facial motor neurons, were greatly reduced by CNTF, although the treatment did not start until the first symptoms of the disease had already become apparent and substantial degenerative changes were already present. The protective and restorative effects of CNTF in this mouse mutant give new perspectives for the treatment of human degenerative motor neuron diseases with CNTF. Year: 1992 Title: Ciliary neurotrophic factor-induced sprouting preserves motor function in a mouse model of mild spinal muscular atrophy Authors: Simon, C. M. Jablonka, S. Ruiz, R. Tabares, L. Sendtner, M. Auth Address: Institute for Clinical Neurobiology, Josef-Schneider-Str. 11, 97080 Wuerzburg, Germany. Pages: 973-86 Volume: 19 Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20022887 Abstract: Proximal spinal muscular atrophy (SMA) is caused by homozygous loss or mutation of the SMN1 gene on human chromosome 5. Depending on the levels of SMN protein produced from a second SMN gene (SMN2), different forms of the disease are distinguished. In patients with milder forms of the disease, type III or type IV SMA that normally reach adulthood, enlargement of motor units is regularly observed. However, the underlying mechanisms are not understood. Smn(+/-) mice, a mouse model of type III/IV SMA, reveal progressive loss of motor neurons and denervation of motor endplates starting at 4 weeks of age. Loss of spinal motor neurons between 1 month and 12 months reaches 40%, whereas muscle strength is not reduced. In these animals, amplitude of single motor unit action potentials in the gastrocnemic muscle is increased more than 2-fold. Confocal analysis reveals pronounced sprouting of innervating motor axons. As ciliary neurotrophic factor (CNTF) is highly expressed in Schwann cells, we investigated its role for a compensatory sprouting response and maintenance of muscle strength in this mouse model. Genetic ablation of CNTF results in reduced sprouting and decline of muscle strength in Smn(+/-) mice. These findings indicate that CNTF is necessary for a sprouting response and thus enhances the size of motor units in skeletal muscles of Smn(+/-) mice. This compensatory mechanism could guide the way to new therapies for this motor neuron disease. Title: Class IIA HDACs in the regulation of neurodegeneration Authors: Majdzadeh, N. Morrison, B. E. D'Mello, S. R. Auth Address: University of Texas at Dallas, Department of Molecular and Cell Biology, Richardson, Texas 75080, USA. Pages: 1072-82 Volume: 13 Number: Keywords: Animals Apoptosis Catalysis Cell Cycle Cell Differentiation Enzyme Inhibitors/pharmacology Histone Deacetylases/*antagonists & inhibitors/*metabolism Humans Models, Biological Nerve Degeneration Neurodegenerative Diseases/*metabolism Neurons/metabolism Phosphorylation Protein Structure, Tertiary Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17981613 Abstract: Neurodegenerative diseases affect millions of patients annually and are a significant burden on the health care systems around the world. While there are symptomatic remedies for patients suffering from various neurodegenerative diseases, there are no cures as of today. Cell death by apoptosis is a common hallmark of neurodegeneration. Therefore, deciphering the molecular pathways regulating this process is of significant value to scientists' endeavor to understand neurodegenerative disorders. Efforts along these lines have uncovered a number of molecular pathways that regulate neuronal apoptosis. Recently, a family of proteins known as histone deacetylases (HDACs) has been linked to regulation of cell survival as well as death. The focus of this review is to summarize our current understanding of the role of HDACs and in particular a subgroup of proteins in this family classified as class IIa HDACs in the regulation of neuronal cell death. It is apparent based on the information presented in this review that although very similar in their primary sequence, members of this family of proteins often have distinct roles in orchestrating apoptotic cell death in the brain. Year: 2008 Month: 1 Title: Classical infantile spinal muscular atrophy with SMN deficiency causes sensory neuronopathy Authors: Rudnik-Schoneborn, S. Goebel, H. H. Schlote, W. Molaian, S. Omran, H. Ketelsen, U. Korinthenberg, R. Wenzel, D. Lauffer, H. Kreiss-Nachtsheim, M. Wirth, B. Zerres, K. Auth Address: Institute for Human Genetics, University of Technology Aachen, Germany. srudnik-schoeneborn@ukaachen.de Pages: 983-7 Volume: 60 Number: 6 Keywords: Axons/ultrastructure Cyclic AMP Response Element-Binding Protein Exons/genetics Female Genes, Recessive Genetic Heterogeneity Humans Infant Infant, Newborn Male Nerve Fibers, Myelinated/pathology Nerve Tissue Proteins/*deficiency/genetics/physiology Neuronal Apoptosis-Inhibitory Protein Neurons, Afferent/*pathology Phenotype RNA-Binding Proteins Research Support, Non-U.S. Gov't Sensation Disorders/*etiology Spinal Muscular Atrophies of Childhood/classification/*complications/genetics Sural Nerve/pathology Wallerian Degeneration Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12654964 Abstract: OBJECTIVE: Classic infantile spinal muscular atrophy (SMA) is believed to be a purely motor disorder, affecting neurons of the spinal anterior horn and nuclei of the lower cranial nerves. Other organ malformations or peripheral nerve involvement have been regarded as exclusion criteria for infantile SMA. Whether SMN protein deficiency can also lead to loss of sensory neurons has not been systematically addressed. METHODS: The authors evaluated the sural nerve biopsies of 19 patients with infantile SMA of varying severity. The diagnosis of SMA was confirmed by the presence of a homozygous deletion of the SMN1 gene in all patients. RESULTS: In seven unrelated infants with SMA type I, axonal degeneration of the sural nerve was noted. Five patients showed abnormal sensory conduction, thus prompting sural nerve biopsy. Sural nerves showed different degrees of axonal loss: fiber density ranged from 3.482 to 22.076/mm2 and was markedly reduced in four patients. There was no evidence of primary demyelination: the ratio of total myelinated fiber thickness to axon diameter (g-ratio) was normal in the patients examined. In seven patients with SMA II and five patients with SMA III, no sural nerve alterations were seen, and conduction velocity was normal. In addition to SMN1 gene deletions, homozygous NAIP gene deletions were detected in six out of seven infants with peripheral neuropathy, whereas there was no evidence of a large deletion including the multicopy markers C212 and Ag1-CA in two out of three families tested. CONCLUSIONS: In this series of patients with SMA I through III who underwent sural nerve biopsy, there was significant sensory nerve pathology in severely affected patients with SMA type I, whereas there were no sensory nerve alterations clinically or morphologically in patients with milder SMA type II or III. Year: 2003 Title: Classification of spinal muscular atrophies Authors: Pearn, J. Auth Address: Pages: 919-22 Volume: 1 Number: 8174 Keywords: Adolescent Adult Age Factors Child Child, Preschool Diagnosis, Differential England Genes, Dominant Genes, Recessive Humans Infant London Middle Aged Motor Neurons Muscular Atrophy/*classification/diagnosis/genetics Nerve Degeneration Neuromuscular Diseases/diagnosis Spinal Diseases/*classification/diagnosis/genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6103267 Abstract: Clinical heterogeneity within the spinal muscular atrophies (SMA) has long been a source of confusion for questions of prognosis and genetic counselling. Comprehensive clinical and genetic analyses of 240 consecutive index cases from two English centres (The English SMA Study) have enabled some nosological questions to be resolved. The different SMA syndromes can be discriminated by (a) age at the first clinical signs of the disease, (b) pattern of muscle involvement, (c) age at death of other patients within an affected kindred, and (d) genetic evidence. Seven different SMA syndromes can be defined clinically and genetically; thirteen mutant genes are incriminated. Prevalence and incidence figures are presented. SMA type I (Werdnig-Hoffman disease) and chronic childhood SMA together comprise 74% of all SMA cases. The classification of the spinal muscular atrophies presented also provides the differential diagnosis for newly presenting cases. Year: 1980 Title: Clenbuterol retards loss of motor function in motor neuron degeneration mice Authors: Zeman, R. J. Peng, H. Etlinger, J. D. Auth Address: Department of Cell Biology and Anatomy, New York Medical College, Valhalla, NY 10595, USA. Zeman@nymc.edu Pages: 460-7 Volume: 187 Number: 2 Keywords: Adrenergic beta-Agonists/*therapeutic use Animals Clenbuterol/*therapeutic use Diagnostic Techniques, Neurological Disease Models, Animal Disease Progression Female Mice Mice, Inbred C57BL Mice, Neurologic Mutants Motor Activity/drug effects Motor Neuron Disease/*drug therapy/etiology/physiopathology Motor Neurons/drug effects/pathology Neuronal Ceroid-Lipofuscinosis/complications/pathology/*physiopathology Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Treatment Outcome Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15144872 Abstract: Motor neuron degeneration (mnd) mice exhibit lysosomal accumulation of lipofuscin-like material that is associated with progressive loss of motor function and strength. Motor dysfunction scores at 8.5-9 months of age were highly correlated with the occurrence of abnormal spinal motor neurons with eccentric nuclei, although the total numbers of motor neurons were not significantly reduced. Nuclear eccentricity is a characteristic of the axon reaction that results from injury and subsequent compensatory axonal sprouting indicating axonal/synaptic dysfunction in mnd motor neurons. Treatment with clenbuterol, a beta(2)-adrenoceptor agonist that can enhance regeneration of motor neuron axons, opposed the development of motor deficits in parallel with a reduced proportion of motor neurons with eccentric nuclei consistent with improved synaptic function. Clenbuterol also opposed decreases in grip strength and muscle mass suggesting beta(2)-agonist treatment as a potential therapeutic modality for lipofuscinoses. Year: 2004 Month: 6 Title: Clinical and genetic study of chronic (types II and III) childhood onset spinal muscular atrophy Authors: Souchon, F. Simard, L. R. Lebrun, S. Rochette, C. Lambert, J. Vanasse, M. Auth Address: Department de pediatrie, Hopital Sainte-Justine, Universite de Montreal, Quebec, Canada. Pages: 419-24 Volume: 6 Number: 6 Keywords: Adolescent Age of Onset Child Child, Preschool Chromosome Deletion Chronic Disease Demography Follow-Up Studies Homozygote Humans Infant Motor Neurons/physiology Muscle Weakness/*physiopathology Nerve Tissue Proteins/genetics Neuronal Apoptosis-Inhibitory Protein Research Support, Non-U.S. Gov't Retrospective Studies Spinal Muscular Atrophies of Childhood/genetics/mortality/*physiopathology Telomere Vital Capacity/*physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9027849 Abstract: We have conducted a retrospective study of 63 patients affected by chronic forms of spinal muscular atrophy (SMA) to better document the natural history of this disease. Thirty-nine patients had type II and 24 type III SMA. These patients had manual muscle testing (MMT) and forced vital capacity (FVC) studies done every six to 12 months over follow up period ranging from six to 140 months. A decline in FVC was seen in both types of SMA but there was no significant change in MMT in either group. Genetic studies were also done in a subset of 17 families (23 patients) included in this study. Homozygous deletions in the telomeric survival motor neuron (SMN) and the neuronal apoptosis inhibitory protein (NAIP) genes were observed in 100% and 11.8% of the patients tested respectively. Year: 1996 Month: 12 Title: Clinical and pathological studies of an inheritary neuropathy in mice (dystonia musculorum) Authors: Duchen, L.W. Strich, S.J. Falconer, D.S. Auth Address: Pages: Volume: 87 Number: 367-378 Keywords: No keywords found Related urls: Abstract: Year: 1964 Month: 1 Title: Clinical commentary: obstetric and respiratory management of pregnancy with severe spinal muscular atrophy Authors: Flunt, D. Andreadis, N. Menadue, C. Welsh, A. W. Auth Address: Department of Respiratory and Sleep Medicine, The Royal Prince Alfred Hospital, Missenden Road, Camperdown, NSW 2050, Australia. Pages: 942301 Volume: 2009 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19960049 Abstract: We present a combined obstetric and respiratory perspective on two pregnancies for a woman with severe Type 2 Spinal Muscular Atrophy (SMA). Our patient had the lowest prepregnancy weight (20 kg) and vital capacity of 0.34 L (VC 11% predicted) yet to be reported in the sparse literature on pregnancy with SMA. She delivered two live healthy infants via planned caesarean section without pregnancy or neonatal complication. We describe the respiratory and obstetric management techniques used for a pregnancy with this degree of respiratory compromise. Year: 2009 Month: 1 Title: Clinical features of spinal and bulbar muscular atrophy Authors: Rhodes, L. E. Freeman, B. K. Auh, S. Kokkinis, A. D. La Pean, A. Chen, C. Lehky, T. J. Shrader, J. A. Levy, E. W. Harris-Love, M. Di Prospero, N. A. Fischbeck, K. H. Auth Address: Neurogenetics Branch, NINDS, NIH, Bethesda, MD, USA. Pages: 3242-51 Volume: 132 Number: Pt 12 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19846582 Abstract: Spinal and bulbar muscular atrophy is an X-linked motor neuron disease caused by a CAG repeat expansion in the androgen receptor gene. To characterize the natural history and define outcome measures for clinical trials, we assessed the clinical history, laboratory findings and muscle strength and function in 57 patients with genetically confirmed disease. We also administered self-assessment questionnaires for activities of daily living, quality of life and erectile function. We found an average delay of over 5 years from onset of weakness to diagnosis. Muscle strength and function correlated directly with serum testosterone levels and inversely with CAG repeat length, age and duration of weakness. Motor unit number estimation was decreased by about half compared to healthy controls. Sensory nerve action potentials were reduced in nearly all subjects. Quantitative muscle assessment and timed 2 min walk may be useful as meaningful indicators of disease status. The direct correlation of testosterone levels with muscle strength indicates that androgens may have a positive effect on muscle function in spinal and bulbar muscular atrophy patients, in addition to the toxic effects described in animal models. Year: 2009 Month: 12 Title: Clinical grade mesenchymal stem cells transdifferentiated under xenofree conditions alleviates motor deficiencies in a rat model of Parkinson's disease Authors: Shetty, P. Ravindran, G. Sarang, S. Thakur, A. Rao, H. S. Viswanathan, C. Auth Address: Regenerative Medicine Reliance Life Sciences Pvt Ltd Dhirubhai Ambani Life Sciences Centre R-282, TTC Industrial Area of MIDC, Thane Belapur Road, Rabale, Navi Mumbai 400701, India. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19465139 Abstract: Bone marrow derived mesenchymal stem cells (BMMSCs) is a valid, definitive candidate for repair of damaged tissues in degenerative disorders in general and neurological diseases in particular. We have standardized the processing conditions for proliferation of BMMSCs using xenofree medium and checked their in vitro and in vivo neurogenic potential. METHOD: The proliferative potential of BMMSCs was analyzed using xenofree media and functionality checked by transplantation into Parkinson's disease (PD) animal models. In vitro neuronal differentiation was investigated by neuronal induction media supplemented with growth factors. Differentiated cells were characterized at cellular and molecular levels. In vitro functionality estimated by dopamine secretion. RESULTS: A pure population of BMMSCs showing an 8-10 fold expansion was obtained using xenofree media. On differentiation to neuronal lineage, they exhibited neuronal morphology. Detectable levels of dopamine (1.93ng/ml) were secreted into the culture media of differentiated cells. There was a significant behavioural improvement in PD models 3 months post transplantation. CONCLUSION: Our study demonstrates that BMMSCs can be transdifferentiated efficiently into functional dopaminergic neurons both in vitro and in vivo. This holds immense clinical potential as a replacement therapy for PD and other neurodegenerative diseases. Year: 2009 Title: Clinical Outcome Measures in Spinal Muscular Atrophy Authors: Montes, J. Gordon, A. M. Pandya, S. De Vivo, D. C. Kaufmann, P. Auth Address: Department of Neurology, Columbia University Medical Center, New York. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19509409 Abstract: Spinal muscular atrophy is one of the most devastating neurological diseases of childhood. Affected infants and children suffer from often severe muscle weakness caused by degeneration of lower motor neurons in the spinal cord and brainstem. Identification of the causative genetic mutation in most cases has resulted in development of potential treatment strategies. To test these new drugs, clinically feasible outcomes are needed. Several different assessments, validated in spinal muscular atrophy or similar disorders, are being used by national and international research groups; however, their sensitivity to detect change is unknown. Acceptance of a few standardized, easily administered, and functionally meaningful outcomes, applicable to the phenotypic spectrum of spinal muscular atrophy, is needed. Consensus is imperative to facilitate collaboration and explore the ability of these measures to identify the therapeutic effect of disease-modifying agents. Following is an evidence-based review of available clinical outcome measures in spinal muscular atrophy. Year: 2009 Title: Clinical outcomes after long-term treatment with alglucosidase alfa in infants and children with advanced Pompe disease Authors: Nicolino, M. Byrne, B. Wraith, J. E. Leslie, N. Mandel, H. Freyer, D. R. Arnold, G. L. Pivnick, E. K. Ottinger, C. J. Robinson, P. H. Loo, J. C. Smitka, M. Jardine, P. Tato, L. Chabrol, B. McCandless, S. Kimura, S. Mehta, L. Bali, D. Skrinar, A. Morgan, C. Rangachari, L. Corzo, D. Kishnani, P. S. Auth Address: Division of Pediatric Endocrinology, Diabetology and Metabolism, Hopital Debrousse, University Lyon, Lyon, France. Pages: 210-9 Volume: 11 Number: 3 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19287243 Abstract: PURPOSE: A clinical trial was conducted to evaluate the safety and efficacy of alglucosidase alfa in infants and children with advanced Pompe disease. METHODS: Open-label, multicenter study of IV alglucosidase alfa treatment in 21 infants 3-43 months old (median 13 months) with minimal acid alpha-glucosidase activity and abnormal left ventricular mass index by echocardiography. Patients received IV alglucosidase alfa every 2 weeks for up to 168 weeks (median 120 weeks). Survival results were compared with an untreated reference cohort. RESULTS: At study end, 71% (15/21) of patients were alive and 44% (7/16) of invasive-ventilator free patients remained so. Compared with the untreated reference cohort, alglucosidase alfa reduced the risk of death by 79% (P < 0.001) and the risk of invasive ventilation by 58% (P = 0.02). Left ventricular mass index improved or remained normal in all patients evaluated beyond 12 weeks; 62% (13/21) achieved new motor milestones. Five patients were walking independently at the end of the study and 86% (18/21) gained functional independence skills. Overall, 52% (11/21) of patients experienced infusion-associated reactions; 95% (19/20) developed IgG antibodies to recombinant human lysosomal acid alpha-glucosidase; no patients withdrew from the study because of safety concerns. CONCLUSIONS: In this population of infants with advanced disease, biweekly infusions with alglucosidase alfa prolonged survival and invasive ventilation-free survival. Treatment also improved indices of cardiomyopathy, motor skills, and functional independence. Year: 2009 Month: 3 Title: Clinical perspectives of high-resolution mass spectrometry-based proteomics in neuroscience-Exemplified in amyotrophic lateral sclerosis biomarker discovery research Authors: Ekegren, T. Hanrieder, J. Bergquist, J. Auth Address: Analytical Chemistry, Department of Physical and Analytical Chemistry, Uppsala University, Uppsala, Sweden. Pages: 559-71 Volume: 43 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18416436 Abstract: Biomarker discovery is a central application in today's proteomic research. There is an urgent need for valid biomarkers to improve diagnostic tools and treatment in many disorders, such as the rapidly progressing neurodegenerative disorder amyotrophic lateral sclerosis (ALS) that has a fatal outcome in about 3 years and yet no curative treatment. Screening for clinically relevant biomarkers puts high demands on high-throughput, rapid and precise proteomic techniques. There is a large variety in the methods of choice involving mainly gel-based approaches as well as chromatographic techniques for multi-dimensional protein and peptide separations followed by mass spectrometry (MS) analysis. This special feature article will discuss some important aspects of MS-based clinical proteomics and biomarker discovery in the field of neurodegenerative diseases and ALS research respectively, with the aim to provide a prospective view on current and future research aspects in the field. Furthermore, examples for application of high-resolution MS-based proteomic strategies for ALS biomarker discovery will be demonstrated with two studies previously reported by our group. These studies include among others, utilization of capillary liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS) for advanced protein pattern classification in cerebrospinal fluid (CSF) samples of ALS patients as well as highly sensitive protein identification in minimal amounts of postmortem spinal cord tissue and laser micro-dissected motor neurons using FT-ICR-MS in conjunction with nanoflow LC coupled to matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (LC-MALDI-TOF-TOF-MS). Copyright (c) 2008 John Wiley & Sons, Ltd. Year: 2008 Month: 5 Title: Clinical proteomics in neurodegenerative disorders Authors: Zetterberg, H. Ruetschi, U. Portelius, E. Brinkmalm, G. Andreasson, U. Blennow, K. Brinkmalm, A. Auth Address: Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy at Goteborg University, Sweden. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18279484 Abstract: Neurodegenerative disorders are characterized by neuronal impairment that eventually leads to neuronal death. In spite of the brain's known capacity for regeneration, lost neurons are difficult to replace. Therefore, drugs aimed at inhibiting neurodegenerative processes are likely to be most effective if the treatment is initiated as early as possible. However, clinical manifestations in early disease stages are often numerous, subtle and difficult to diagnose. This is where biomarkers that specifically reflect onset of pathology, directly or indirectly, may have a profound impact on diagnosis making in the future. A triplet of biomarkers for Alzheimer's disease (AD), total and hyperphosphorylated tau and the 42 amino acid isoform of beta-amyloid, has already been established for early detection of AD before the onset of dementia. However, more biomarkers are needed both for AD and for other neurodegenerative disorders, such as Parkinson's disease, frontotemporal dementia and amyotrophic lateral sclerosis. This review provides an update on recent advances in clinical neuroproteomics, a biomarker discovery field that has expanded immensely during the last decade, and gives an overview of the most commonly used techniques and the major clinically relevant findings these techniques have lead to. Year: 2008 Title: Clinical research. Advocating, the clinical way Authors: Couzin, J. Auth Address: Pages: 940-2 Volume: 308 Number: 5724 Keywords: Animals Blindness/prevention & control *Clinical Trials/economics/methods Cystic Fibrosis/therapy *Foundations/economics/organization & administration Gene Therapy Humans Investments Patient Advocacy Rare Diseases/*therapy *Research Support Retinal Diseases/therapy Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15890856 Abstract: Year: 2005 Title: Clinical studies of histone deacetylase inhibitors Authors: Prince, H. M. Bishton, M. J. Harrison, S. J. Auth Address: Division of Haematology and Medical Oncology, Peter MacCallum Cancer Centre, East Melbourne, Melbourne, Australia and University of Melbourne, Parkville, Victoria, Australia. miles.prince@petermac.org Pages: 3958-69 Volume: 15 Number: 12 Keywords: Antibiotics, Antineoplastic/adverse effects/pharmacology/*therapeutic use Benzamides/administration & dosage/adverse effects/therapeutic use Clinical Trials as Topic Depsipeptides/administration & dosage/adverse effects/therapeutic use Drug Therapy, Combination Enzyme Inhibitors/adverse effects/pharmacology/*therapeutic use Hematologic Neoplasms/*drug therapy/pathology Histone Deacetylases/*antagonists & inhibitors Humans Hydroxamic Acids/administration & dosage/adverse effects/therapeutic use Pyridines/administration & dosage/adverse effects/therapeutic use Pyrimidines/administration & dosage/adverse effects/therapeutic use Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19509172 Abstract: Over the last 5 years, a plethora of histone deacetylase inhibitors (HDACi) have been evaluated in clinical trials. These drugs have in common the ability to hyperacetylate both histone and nonhistone targets, resulting in a variety of effects on cancer cells, their microenvironment, and immune responses. To date, responses with single agent HDACi have been predominantly observed in advanced hematologic malignancies including T-cell lymphoma, Hodgkin lymphoma, and myeloid malignancies. Therefore, in this review we focus upon hematologic malignancies. Generally HDACi are well tolerated with the most common acute toxicities being fatigue, gastrointestinal, and transient cytopenias. Of note, few patients have been treated for prolonged periods of time and little is known about long-term toxicities. The use of the biomarker of histone hyperacetylation has been useful as a guide to target specificity, but generally does not predict for response and the search for more clinically relevant biomarkers must continue. Year: 2009 Title: Clinical trials in spinal muscular atrophy Authors: Darras, B. T. Kang, P. B. Auth Address: Department of Neurology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts, USA. Pages: 675-679 Volume: 19 Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18025935 Abstract: PURPOSE OF REVIEW: Spinal muscular atrophy is a neuromuscular disorder manifesting as weakness and hypotonia across a broad spectrum of severity. Mutations in the telomeric copy of the survival motor neuron gene (SMN1) cause the autosomal recessive form. Disease severity is modified by the number of centromeric copies of the gene (SMN2) and the quantity of survival motor neuron protein. This has given rise to a number of treatment strategies. RECENT FINDINGS: Histone deacetylase inhibitors appear to increase the expression of SMN2, with an increase in survival motor neuron protein in various cell types. Clinical trials have been performed with three histone deacetylase inhibitors which are already licensed in the USA. Phenylbutyrate showed promise in a mouse model and an open-label pilot study, but was not effective in a phase 2 trial. Valproate may enhance transcription and reverse SMN2 splicing pattern, and has induced promising motor-function improvement in patients. Hydroxyurea may enhance splice function and increase the number of nuclear 'gems', small nuclear organelles in which survival motor neuron protein concentrates. SUMMARY: Discoveries regarding the genetics and pathogenesis of spinal muscular atrophy have identified potential targets for pharmacotherapy, raising hope that better treatments will eventually be developed. Year: 2007 Month: 12 Title: Clinical variability in distal spinal muscular atrophy type 1 (DSMA1): determination of steady-state IGHMBP2 protein levels in five patients with infantile and juvenile disease Authors: Guenther, U. P. Handoko, L. Varon, R. Stephani, U. Tsao, C. Y. Mendell, J. R. Lutzkendorf, S. Hubner, C. von Au, K. Jablonka, S. Dittmar, G. Heinemann, U. Schuetz, A. Schuelke, M. Auth Address: Department of Neuropediatrics, Charite University Medical School of Berlin, Augustenburger Platz 1, 13353, Berlin, Germany. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18802676 Abstract: Distal spinal muscular atrophy type 1 (DSMA1) is caused by mutations in the immunoglobulin mu-binding protein 2 (IGHMBP2) gene. Patients with DSMA1 present between 6 weeks and 6 months of age with progressive muscle weakness and respiratory failure due to diaphragmatic palsy. Contrary to this "classic" infantile disease, we have previously described a DSMA1 patient with juvenile disease onset. In this paper, we present (1) a second juvenile case and (2) the first study of DSMA1 on protein level in patients with infantile (n = 3) as well as juvenile (n = 2) disease onset observing elevated residual steady-state IGHMBP2 protein levels in the patients with late onset DSMA1 as compared to those with classic DSMA1. Mutation screening in IGHMBP2 revealed two patients compound heterozygous for a novel missense mutation (c.1478C-->T; p.T493I) and another previously described mutation. In lymphoblastoid cells of both patients, steady-state IGHMBP2 protein levels were reduced. In comparison to wild-type IGHMBP2, the p.T493I variant protein had an increased tendency to aggregate and spontaneously degrade in vitro. We verified a change in the physicochemical properties of the p.T493I variant which may explain the pathogenicity of this mutation. Our data further suggest that the age of onset of DSMA1 is variable, and we discuss the effect of residual IGHMBP2 protein levels on the clinical course and the severity of the disease. Year: 2008 Title: Clonal analysis of Drosophila embryonic neuroblasts: neural cell types, axon projections and muscle targets Authors: Schmid, A. Chiba, A. Doe, C. Q. Auth Address: Institute of Neuroscience and Institute of Molecular Biology, Eugene OR 97403, USA. Pages: 4653-89 Volume: 126 Number: 21 Keywords: Animals Axons Cell Lineage Cell Size Clone Cells Comparative Study Drosophila/*embryology/genetics Embryo, Nonmammalian/innervation Embryonic Induction Insects/embryology Interneurons/physiology Motor Neurons/physiology Muscles/*cytology/*embryology/innervation Neuroglia/cytology/physiology Neurons/*cytology/*physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10518486 Abstract: An experimental analysis of neurogenesis requires a detailed understanding of wild-type neural development. Recent DiI cell lineage studies have begun to elucidate the family of neurons and glia produced by each Drosophila embryonic neural precursor (neuroblast). Here we use DiI labeling to extend and clarify previous studies, but our analysis differs from previous studies in four major features: we analyze and compare lineages of every known embryonic neuroblast; we use an in vivo landmark (engrailed-GFP) to increase the accuracy of neuroblast identification; we use confocal fluorescence and Nomarski microscopy to collect three-dimensional data in living embryos simultaneously for each DiI-labeled clone, the engrailed-GFP landmark, and the entire CNS and muscle target field (Nomarski images); and finally, we analyze clones very late in embryonic development, which reveals novel cell types and axon/dendrite complexity. We identify the parental neuroblasts for all the cell types of the embryonic CNS: motoneurons, intersegmental interneurons, local interneurons, glia and neurosecretory cells (whose origins had never been determined). We identify muscle contacts for every thoracic and abdominal motoneuron at stage 17. We define the parental neuroblasts for neurons or glia expressing well-known molecular markers or neurotransmitters. We correlate Drosophila cell lineage data with information derived from other insects. In addition, we make the following novel conclusions: (1) neuroblasts at similar dorsoventral positions, but not anteroposterior positions, often generate similar cell lineages, and (2) neuroblasts at similar dorsoventral positions often produce the same motoneuron subtype: ventral neuroblasts typically generate motoneurons with dorsal muscle targets, while dorsal neuroblasts produce motoneurons with ventral muscle targets. Lineage data and movies can be found at http://www.biologists. com/Development/movies/dev8623.html http://www.neuro.uoregon. edu/doelab/lineages/ Year: 1999 Month: 11 Title: Cloning, characterization, and copy number of the murine survival motor neuron gene: homolog of the spinal muscular atrophy-determining gene Authors: DiDonato, C. J. Chen, X. N. Noya, D. Korenberg, J. R. Nadeau, J. H. Simard, L. R. Auth Address: Service de Genetique Medicale, Hopital Ste-Justine, Quebec, Canada. Pages: 339-52 Volume: 7 Number: 4 Keywords: Amino Acid Sequence Animals Base Sequence Blotting, Northern Chromosome Mapping/methods Chromosomes, Artificial, Yeast Chromosomes, Bacterial Chromosomes, Human, Pair 5 Cloning, Molecular Cyclic AMP Response Element-Binding Protein Female Gene Amplification Humans In Situ Hybridization, Fluorescence Mice Mice, Inbred C57BL Mice, Inbred Strains Molecular Sequence Data Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins/*genetics *Polymorphism, Single-Stranded Conformational RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Sequence Analysis, DNA *Sequence Homology, Amino Acid Tissue Distribution Transcription, Genetic Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9110173 Abstract: Because of a 500-kb inverted duplication, there are two copies of the survival motor neuron (SMN) gene in humans, cenSMN and telSMN. Both genes produce identical ubiquitously expressed transcripts; however, only mutations in telSMN are responsible for spinal muscular atrophy (SMA), the second most common autosomal recessive childhood disease. We have cloned the murine homolog Smn and mapped the gene to Chromosome 13 within the conserved syntenic region of human chromosome 5q13. We show that the Smn transcript (1.4 kb) is expressed as early as embryonic day 7. In contrast to humans, we found no evidence of alternative splicing. The predicted amino acid sequence between mouse and human SMN is 82% identical, and a putative nuclear localization signal is conserved. FISH data indicate that the duplication of the SMA region observed in humans is not present in the mouse. We also found no evidence of multiple Smn genes using Southern blot hybridization and single-strand conformation analysis. Using these methods, we detected at least four copies of Naip exon 5 clustering distal to Smn. Finally, three biallelic markers were identified within the Smn coding region; two are silent polymorphisms, whereas the third changes a cysteine residue to a tyrosine residue in exon 7. Overall, our results indicate that Smn is single copy within the mouse genome, which should facilitate gene disruption experiments to create an animal model of SMA. Year: 1997 Month: 4 Title: CNS immune privilege: hiding in plain sight Authors: Carson, M. J. Doose, J. M. Melchior, B. Schmid, C. D. Ploix, C. C. Auth Address: Division of Biomedical Sciences, University of California, Riverside, Riverside, CA, USA. Pages: 48-65 Volume: 213 Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16972896 Abstract: Central nervous system (CNS) immune privilege is an experimentally defined phenomenon. Tissues that are rapidly rejected by the immune system when grafted in sites, such as the skin, show prolonged survival when grafted into the CNS. Initially, CNS immune privilege was construed as CNS isolation from the immune system by the blood-brain barrier (BBB), the lack of draining lymphatics, and the apparent immunoincompetence of microglia, the resident CNS macrophage. CNS autoimmunity and neurodegeneration were presumed automatic consequences of immune cell encounter with CNS antigens. Recent data have dramatically altered this viewpoint by revealing that the CNS is neither isolated nor passive in its interactions with the immune system. Peripheral immune cells can cross the intact BBB, CNS neurons and glia actively regulate macrophage and lymphocyte responses, and microglia are immunocompetent but differ from other macrophage/dendritic cells in their ability to direct neuroprotective lymphocyte responses. This newer view of CNS immune privilege is opening the door for therapies designed to harness autoreactive lymphocyte responses and also implies (i) that CNS autoimmune diseases (i.e. multiple sclerosis) may result as much from neuronal and/or glial dysfunction as from immune system dysfunctions and (ii) that the severe neuronal and glial dysfunction associated with neurodegenerative disorders (i.e. Alzheimer's disease) likely alters CNS-specific regulation of lymphocyte responses affecting the utility of immune-based therapies (i.e. vaccines). Year: 2006 Month: 10 Title: CNS-targeted gene therapy improves survival and motor function in a mouse model of spinal muscular atrophy Authors: Passini, M. A. Bu, J. Roskelley, E. M. Richards, A. M. Sardi, S. P. O'Riordan, C. R. Klinger, K. W. Shihabuddin, L. S. Cheng, S. H. Auth Address: Genzyme Corporation, 49 New York Avenue, Room 2410, Framingham, MA 01701, USA. marco.passini@genzyme.com Pages: 1253-64 Volume: 120 Number: 4 Keywords: Animals Disease Models, Animal *Gene Therapy Humans Mice Motor Neurons/*physiology Muscle Strength Muscle, Skeletal/pathology Muscular Atrophy, Spinal/mortality/physiopathology/*therapy Neurites/metabolism Neuromuscular Junction/pathology Survival of Motor Neuron 1 Protein/*genetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20234094 Abstract: Spinal muscular atrophy (SMA) is a neuromuscular disease caused by a deficiency of survival motor neuron (SMN) due to mutations in the SMN1 gene. In this study, an adeno-associated virus (AAV) vector expressing human SMN (AAV8-hSMN) was injected at birth into the CNS of mice modeling SMA. Western blot analysis showed that these injections resulted in widespread expression of SMN throughout the spinal cord, and this translated into robust improvement in skeletal muscle physiology, including increased myofiber size and improved neuromuscular junction architecture. Treated mice also displayed substantial improvements on behavioral tests of muscle strength, coordination, and locomotion, indicating that the neuromuscular junction was functional. Treatment with AAV8-hSMN increased the median life span of mice with SMA-like disease to 50 days compared with 15 days for untreated controls. Moreover, injecting mice with SMA-like disease with a human SMN-expressing self-complementary AAV vector - a vector that leads to earlier onset of gene expression compared with standard AAV vectors - led to improved efficacy of gene therapy, including a substantial extension in median survival to 157 days. These data indicate that CNS-directed, AAV-mediated SMN augmentation is highly efficacious in addressing both neuronal and muscular pathologies in a severe mouse model of SMA. Month: 4 Title: Co-regulation of survival of motor neuron (SMN) protein and its interactor SIP1 during development and in spinal muscular atrophy Authors: Jablonka, S. Bandilla, M. Wiese, S. Buhler, D. Wirth, B. Sendtner, M. Fischer, U. Auth Address: Klinische Forschergruppe Neuroregeneration, Department of Neurology, University of Wurzburg, Josef-Schneider-Strasse 11, D-97080 Wurzburg, Germany. Pages: 497-505 Volume: 10 Number: 5 Keywords: Amino Acid Sequence Animals Base Sequence Blotting, Western Cell Line Cloning, Molecular Cyclic AMP Response Element-Binding Protein Humans Mice Molecular Sequence Data Motor Neurons/metabolism Muscular Atrophy, Spinal/*genetics Nerve Tissue Proteins RNA-Binding Proteins Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11181573 Abstract: Spinal muscular atrophy (SMA) is a neuromuscular disease characterized by the degeneration of motor neurons in the spinal cord. The disease is caused by mutations of the survival of motor neuron 1 gene (SMN1), resulting in a reduced production of functional SMN protein. A major question unanswered thus far is why reduced amounts of ubiquitously expressed SMN protein specifically cause the degeneration of motor neurons without affecting other somatic cell types. In a first attempt to address this issue we have investigated the Smn interacting protein 1 (Sip1), with an emphasis on its developmental expression and subcellular distribution in spinal motor neurons in relation to Smn. By confocal immunofluorescence studies we provide evidence that a significant amount of Smn does not co-localize with Sip1 in neurites of motor neurons, indicating that Smn may exert motor neuron-specific functions that are not dependent on Sip1. Sip1 is highly expressed in the spinal cord during early development and expression decreases in parallel with Smn during postnatal development. Strikingly, reduced production of Smn as observed in cell lines derived from SMA patients or in a mouse model for SMA coincides with a simultaneous reduction of Sip1. The finding that expression of Sip1 and Smn is tightly co-regulated, together with the unique localization of Smn in neurites, may help in understanding the motor neuron-specific defects observed in SMA patients. Year: 2001 Title: Coilin phosphorylation mediates interaction with SMN and SmB' Authors: Toyota, C. G. Davis, M. D. Cosman, A. M. Hebert, M. D. Auth Address: Department of Biochemistry, University of Mississippi Medical Center, Jackson, MS, 39216, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19997741 Abstract: Cajal bodies (CBs) are subnuclear domains that participate in spliceosomal small nuclear ribonucleoprotein (snRNP) biogenesis and play a part in the assembly of the spliceosomal complex. The CB marker protein, coilin, interacts with survival of motor neuron (SMN) and Sm proteins. Several coilin phosphoresidues have been identified by mass spectrometric analysis. Phosphorylation of coilin affects its self-interaction and localization in the nucleus. We hypothesize that coilin phosphorylation also impacts its binding to SMN and Sm proteins. In vitro binding studies with a C-terminal fragment of coilin and corresponding phosphomimics show that SMN binds preferentially to dephosphorylated analogs and that SmB' binds preferentially to phosphomimetic constructs. Bacterially expressed full-length coilin binds more SMN and SmB' than does the C-terminal fragment. Co-immunoprecipitation and phosphatase experiments show that SMN also binds dephosphorylated coilin in vivo. These data show that phosphorylation of coilin influences interaction with its target proteins and, thus, may be significant in managing the flow of snRNPs through the CB. Year: 2009 Title: Coinheritance of mutated SMN1 and MECP2 genes in a child with phenotypic features of spinal muscular atrophy (SMA) type II and Rett syndrome Authors: Voutoufianakis, S. Psoni, S. Vorgia, P. Tsekoura, F. Kekou, K. Traeger-Synodinos, J. Kitsiou, S. Kanavakis, E. Fryssira, H. Auth Address: Pediatric Department of Venizelion General Hospital Iraklion, Crete, Greece. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17276711 Abstract: Spinal muscular atrophy (SMA) is a neuromuscular autosomal recessive disease characterized by progressive muscle weakness and atrophy combined with motor neuron degeneration caused by mutations in the SMN 1 gene locus (5q11.2-13.2). Rett syndrome (RS) is an X-linked dominant neurodevelopmental disorder caused by mutations in MECP2 (Xq28) and characterized by normal development until 6-12 months of age, followed by regression with loss of acquired skills, gradual onset of microcephaly, stereotypic hand movements and psychomotor delay. We report a 6-year-old girl who, at 2 years of age, presented with hypotonia, psychomotor delay, amyotrophy and areflexia of the lower extremities. Molecular DNA analysis (PCR-RFLP's) for SMA type II revealed that both exons 7 and 8 of SMN 1 gene were deleted. Over the past 4 years, onset of stereotypic hand-washing movements, epileptic seizures, microcephaly, hyperventilation/breath-holding attacks and severe psychomotor delay raised the suspicion of the coexistence of RS. DNA analysis (DGGE and sequencing) identified the hotspot missense mutation R306C (c.916C>T) in exon 4 of the MECP2 gene. The coinheritance of SMA and RS, two rare monogenic syndromes in the same patient, has not been previously reported. Thorough clinical evaluation in combination with DNA analysis, allowed accurate diagnosis, providing valuable information for the genetic counseling of the family. Year: 2007 Title: Combination brain and systemic injections of AAV provide maximal functional and survival benefits in the Niemann-Pick mouse Authors: Passini, M. A. Bu, J. Fidler, J. A. Ziegler, R. J. Foley, J. W. Dodge, J. C. Yang, W. W. Clarke, J. Taksir, T. V. Griffiths, D. A. Zhao, M. A. O'Riordan, C. R. Schuchman, E. H. Shihabuddin, L. S. Cheng, S. H. Auth Address: Genzyme Corporation, Framingham, MA 01701, USA. marco.passini@genzyme.com Pages: 9505-10 Volume: 104 Number: 22 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17517638 Abstract: Niemann-Pick disease (NPD) is caused by the loss of acid sphingomyelinase (ASM) activity, which results in widespread accumulation of undegraded lipids in cells of the viscera and CNS. In this study, we tested the effect of combination brain and systemic injections of recombinant adeno-associated viral vectors encoding human ASM (hASM) in a mouse model of NPD. Animals treated by combination therapy exhibited high levels of hASM in the viscera and brain, which resulted in near-complete correction of storage throughout the body. This global reversal of pathology translated to normal weight gain and superior recovery of motor and cognitive functions compared to animals treated by either brain or systemic injection alone. Furthermore, animals in the combination group did not generate antibodies to hASM, demonstrating the first application of systemic-mediated tolerization to improve the efficacy of brain injections. All of the animals treated by combination therapy survived in good health to an investigator-selected 54 weeks, whereas the median lifespans of the systemic-alone, brain-alone, or untreated ASM knockout groups were 47, 48, and 34 weeks, respectively. These data demonstrate that combination therapy is a promising therapeutic modality for treating NPD and suggest a potential strategy for treating disease indications that cause both visceral and CNS pathologies. Year: 2007 Title: Combination of SMN2 copy number and NAIP deletion predicts disease severity in spinal muscular atrophy Authors: Watihayati, M. S. Fatemeh, H. Marini, M. Atif, A. B. Zahiruddin, W. M. Sasongko, T. H. Tang, T. H. Zabidi-Hussin, Z. Nishio, H. Zilfalil, B. A. Auth Address: Human Genome Center, School of Medical Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kota Bharu, Kelantan, Malaysia. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18842367 Abstract: Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder caused by mutations in the SMN1 gene. The SMN2 gene is highly homologous to SMN1 and has been reported to be correlated with severity of the disease. The clinical presentation of SMA varies from severe to mild, with three clinical subtypes (type I, type II, and type III) that are assigned according to age of onset and severity of the disease. Here, we aim to investigate the potential association between the number of copies of SMN2 and the deletion in the NAIP gene with the clinical severity of SMA in patients of Malaysian origin. Forty-two SMA patients (14 of type I, 20 type II, and 8 type III) carrying deletions of the SMN1 gene were enrolled in this study. SMN2 copy number was determined by fluorescence-based quantitative polymerase chain reaction assay. Twenty-nine percent of type I patients carried one copy of SMN2, while the remaining 71% carried two copies. Among the type II and type III SMA patients, 29% of cases carried two copies of the gene, while 71% carried three or four copies of SMN2. Deletion analysis of NAIP showed that 50% of type I SMA patients had a homozygous deletion of exon 5 of this gene and that only 10% of type II SMA cases carried a homozygous deletion, while all type III patients carried intact copies of the NAIP gene. We conclude that there exists a close relationship between SMN2 copy number and SMA disease severity, suggesting that the determination of SMN2 copy number may be a good predictor of SMA disease type. Furthermore, NAIP gene deletion was found to be associated with SMA severity. In conclusion, combining the analysis of deletion of NAIP with the assessment of SMN2 copy number increases the value of this tool in predicting the severity of SMA. Year: 2008 Title: Combined lithium and valproate treatment delays disease onset, reduces neurological deficits and prolongs survival in an amyotrophic lateral sclerosis mouse model Authors: Feng, H. L. Leng, Y. Ma, C. H. Zhang, J. Ren, M. Chuang, D. M. Auth Address: Neurology Department, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, PR China. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18640245 Abstract: Lithium and valproic acid (VPA) are two primary drugs used to treat bipolar disorder, and have been shown to have neuroprotective properties in vivo and in vitro. A recent study demonstrated that combined treatment with lithium and VPA elicits synergistic neuroprotective effects against glutamate excitotoxicity in cultured brain neurons, and the synergy involves potentiated inhibition of glycogen synthase kinase-3 (GSK-3) activity through enhanced GSK-3 serine phosphorylation [Leng Y, Liang MH, Ren M, Marinova Z, Leeds P, Chuang DM (2008) Synergistic neuroprotective effects of lithium and valproic acid or other histone deacetylase inhibitors in neurons: roles of glycogen synthase kinase-3 inhibition. J Neurosci 28:2576-2588]. We therefore investigated the effects of lithium and VPA cotreatment on the disease symptom onset, survival time and neurological deficits in cooper zinc superoxide dismutase (SOD1) G93A mutant mice, a commonly used mouse model of amyotrophic lateral sclerosis (ALS). The G93A ALS mice received twice daily i.p. injections with LiCl (60 mg/kg), VPA (300 mg/kg) or lithium plus VPA, starting from the 30(th) day after birth and continuing until death. We found that combined treatment with lithium and VPA produced a greater and more consistent effect in delaying the onset of disease symptoms, prolonging the lifespan and decreasing the neurological deficit scores, compared with the results of monotreatment with lithium or VPA. Moreover, lithium in conjunction with VPA was more effective than lithium or VPA alone in enhancing the immunostaining of phospho-GSK-3beta(Ser9) in brain and lumbar spinal cord sections. To our knowledge, this is the first demonstration of enhanced neuroprotection by a combinatorial approach using mood stabilizers in a mouse ALS model. Our results suggest that clinical trials using lithium and VPA in combination for ALS patients are a rational strategy. Year: 2008 Title: Combining growth factor and stem cell therapy for amyotrophic lateral sclerosis Authors: Suzuki, M. Svendsen, C. N. Auth Address: The Waisman Center and Departments of Anatomy and Neurology, University of Wisconsin-Madison, Madison, WI 53707-2280, USA. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18329734 Abstract: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease where motor neurons within the brain and spinal cord are lost, leading to paralysis and death. Certain growth factors should, in principle, be able to protect dying motor neurons. However, targeted delivery to the spinal cord or brain has been a constant problem. There is also accumulating evidence that glial cells might play a crucial role in maintaining motor neuron function and survival in ALS. Stem cells isolated and expanded in culture can be modified to release growth factors and generate glial cells following transplantation into the spinal cord or brain. As such, they might be able to both detoxify the local environment around dying motor neurons and deliver trophic factors. Here we examine the feasibility of translating these findings into new treatments for ALS patients. Year: 2008 Title: Comparative Analysis of Adeno-Associated Viral Vector Serotypes 1, 2, 5, 7, And 8 in Mouse Brain Authors: Taymans, J. M. Vandenberghe, L. H. Haute, C. V. Thiry, I. Deroose, C. M. Mortelmans, L. Wilson, J. M. Debyser, Z. Baekelandt, V. Auth Address: Laboratory for Neurobiology and Gene Therapy, Division of Molecular Medicine, Department of Molecular and Cellular Medicine, Katholieke Universiteit Leuven, 3000 Leuven, Flanders, Belgium. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17343566 Abstract: Recombinant adeno-associated virus serotype 2 (rAAV2) vectors have been shown to deliver genes effectively to neurons in the brain, retina, and spinal cord. The characterization of new AAV serotypes revealed different patterns of transduction in a diverse array of tissues (Gao, G., Vandenberghe, L.H., and Wilson, J.M. [2005]. Curr. Gene Ther. 5, 285-297). Here, we extensively compare the neural tropism of human-derived rAAVs (types 2/1, 2, and 2/5) with nonhuman primate-derived rAAVs (types 2/7 and 2/8) in adult mouse brain. Mice were injected with rAAV type 2/1, 2, 2/5, 2/7, or 2/8 via the caudate-putamen and substantia nigra. Intrahippocampal injections were also performed for rAAV2/7 and rAAV2/8. In all regions injected, the vectors transduced neurons almost exclusively. Retrograde transduction of all rAAV pseudotypes was also observed in particular CNS areas. At high titers, all rAAV pseudotypes transduced comparable brain volumes in all targeted regions except for rAAV2, which transduced much smaller brain volumes. A dose-range comparison of intrastriatally injected rAAV types 2/5, 2/7, and 2/8 highlighted that the transduction efficiency, as determined by transduced volume and biophotonic imaging of green fluorescent protein expression intensity, was significantly higher for rAAV2/5 and rAAV2/7 compared with rAAV2/8 at low titers, whereas all three serotypes performed equally well at higher doses. These results demonstrate the use and efficiency of both human- and nonhuman primate-derived rAAV vectors for disease modeling and their potential for gene therapy. Year: 2007 Title: Comparative Analysis of Antisense Oligonucleotide Sequences for Targeted Skipping of Exon 51 During Dystrophin Pre-mRNA Splicing in Human Muscle Authors: Arechavala-Gomeza, V. Graham, I. R. Popplewell, L. J. Adams, A. M. Aartsma-Rus, A. Kinali, M. Morgan, J. E. van Deutekom, J. C. Wilton, S. D. Dickson, G. Muntoni, F. Auth Address: Department of Paediatrics, Imperial College, London W12ONN, United Kingdom. Pages: 798-810 Volume: 18 Number: 9 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17767400 Abstract: Duchenne muscular dystrophy (DMD) is caused by mutations in the dystrophin gene that result in the absence of functional protein. In the majority of cases these are out-of-frame deletions that disrupt the reading frame. Several attempts have been made to restore the dystrophin mRNA reading frame by modulation of pre-mRNA splicing with antisense oligonucleotides (AOs), demonstrating success in cultured cells, muscle explants, and animal models. We are preparing for a phase I/IIa clinical trial aimed at assessing the safety and effect of locally administered AOs designed to inhibit inclusion of exon 51 into the mature mRNA by the splicing machinery, a process known as exon skipping. Here, we describe a series of systematic experiments to validate the sequence and chemistry of the exon 51 AO reagent selected to go forward into the clinical trial planned in the United Kingdom. Eight specific AO sequences targeting exon 51 were tested in two different chemical forms and in three different preclinical models: cultured human muscle cells and explants (wild type and DMD), and local in vivo administration in transgenic mice harboring the entire human DMD locus. Data have been validated independently in the different model systems used, and the studies describe a rational collaborative path for the preclinical selection of AOs for evaluation in future clinical trials. Year: 2007 Month: 9 Title: Comparative study of mesenchymal stem cells from C57BL/10 and mdx mice Authors: Li, Y. Zhang, C. Xiong, F. Yu, M. J. Peng, F. L. Shang, Y. C. Zhao, C. P. Xu, Y. F. Zhou, C. Liu, Z. S. Wu, J. L. Auth Address: Pages: 24 Volume: 9 Number: 1 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18489762 Abstract: ABSTRACT: BACKGROUND: Human mesenchymal stem cells (MSCs) have been studied and applied extensively because of their ability to self-renew and differentiate into various cell types. Since most human diseases models are murine, mouse MSCs should have been studied in detail. However, knowledge concerning mouse MSCs remains limited. The mdx mouse--a Duchenne muscular dystrophy model--was produced by introducing a point mutation in the dystrophin gene. To understand the role of dystrophin in MSCs, we compared MSCs from mdx and C57BL/10 mice, focusing particularly on the aspects of light and electron microscopic morphology, immunophenotyping, and differentiation potential. RESULTS: Our study showed that at passage 10, mdx-derived MSCs exhibited increased heterochromatin, larger vacuoles, and more lysosomes under electron microscopy compared to C57BL/10-derived MSCs. At passage 13, C57BL/10-derived MSCs formed a few myotubes, while mdx-derived MSCs did not. By passage 21, mdx-derived MSCs but not C57BL/10-derived MSCs had gradually lost their proliferative ability. In addition, a significant difference in the expression of CD34, not Sca-1 and CD11b, was observed between the MSCs from the 2 mice. CONCLUSIONS: Our current study reveals that the MSCs from the 2 mice, namely, C57BL/10 and mdx, exhibit differences in proliferative and myogenic abilities. The results suggest that the changes in mouse MSC behavior may be influenced by the point mutation in the dystrophin gene. Year: 2008 Title: Comparison of the growth hormone, IGF-1 and insulin in cerebrospinal fluid and serum between patients with motor neuron disease and healthy controls Authors: Bilic, E. Bilic, E. Rudan, I. Kusec, V. Zurak, N. Delimar, D. Zagar, M. Auth Address: Department of Neurology, University Hospital Centre, Zagreb, Croatia. Pages: 1340-5 Volume: 13 Number: 12 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17116217 Abstract: Neurotrophic effects of the growth hormone (GH), insulin-like growth factor-1 (IGF-1) and insulin on the central nervous system have become more apparent in the past decade. In this study, we measured serum and cerebrospinal fluid (CSF) concentrations of GH, IGF-1 and insulin in 35 patients with motor neuron disease (MND) [24 patients with definite amyotrophic lateral sclerosis (ALS) and 11 patients with progressive bulbar palsy] and in 40 healthy controls. Levels of serum concentrations of GH and IGF-1 did not significantly differ between the MND patient group and the healthy controls, while the level of insulin was significantly decreased (P = 0.0033) in the MND patient group. However, levels of all three examined parameters in CSF were significantly lower in the MND group than in the healthy controls with the statistical significance for IGF-1 and insulin of P < 0.001. This finding has not been reported previously, and further investigations into its association with ALS should establish whether it can be used as an early marker of the disease, or whether it merely represents a consequence of ALS development. Year: 2006 Month: 12 Title: Comparison of valproate concentrations in human plasma, CSF and brain tissue after administration of different formulations of valproate or valpromide Authors: Wieser, H. G. Auth Address: Department of Neurology, University Hospital Zurich, Switzerland. Pages: 154-9 Volume: 9 Number: 2 Keywords: Adolescent Adult Brain/*metabolism Comparative Study Electroencephalography/drug effects Female Humans Male Seizures/drug therapy Valproic Acid/administration & dosage/*analogs & derivatives/*pharmacokinetics Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1794353 Abstract: The concentration of valproate was measured in plasma, CSF and brain tissue of patients who underwent resective surgical treatment because of severe temporal lobe epilepsy after pretreatment with either a sustained release formulation of valproate (Depakine Chrono; 5 patients), the conventional formulation of valproate (Depakine; 6 patients) or valpromide (Depamide; 2 patients). With a mean serum value for all 13 patients of 32.3 micrograms/g valproate, the mean brain/serum ratio was 15.1% (SD 6.1%). The valproate concentration of the hippocampus was significantly higher than that of the amygdala and patients who had the sustained release formulation had significantly higher valproate concentration in the CSF and in the hippocampal formation than those patients who had the conventional valproate. Since a few patients had tumors, whereas others had varying degrees of gliosis, it cannot be ruled out that these differences are the result of different histopathological conditions with related differences in blood-brain barrier functions. Year: 1991 Month: 7 Title: Compensatory mechanisms during walking in response to muscle weakness in spinal muscular atrophy, type III Authors: Matjacic, Z. Olensek, A. Krajnik, J. Eymard, B. Zupan, A. Praznikar, A. Auth Address: Institute for Rehabilitation, Republic of Slovenia, Linhartova 51, SI-1000 Ljubljana, Slovenia. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17980600 Abstract: Our knowledge on altered neurological control of walking due to weakness of various muscle groups of the lower extremities is limited. The aim of this study was to assess kinematic, kinetic and electromyographic (EMG) walking patterns in a functionally homogeneous group of seven subjects with spinal muscular atrophy, type III (SMA group) and compare them with normal data obtained from nine healthy subjects (CONTROL group) in order to identify characteristic compensatory changes. Muscle strength at the ankle and knee joints was assessed using isokinetic dynamometry to determine variability in muscle strength: this was found to be similar in the two groups. Kinematic, kinetic and EMG patterns were assessed during walking in the SMA and CONTROL groups. The results showed changes in the activity of ankle plantarflexors and associated control of the center of pressure during loading response and midstance, which facilitated minimization of the external flexion moment acting on the knee and hip in the SMA group. Additionally, we identified distinct and consistent changes in the control of hip rotators that act to rapidly extend the hip early in stance phase and in the control of contralateral hip abductors that act delay weight shift onto the leg entering the stance phase. From these results we can conclude that the most important muscle groups compensating for reduced strength in knee and hip muscles are the ankle plantarflexors, hip rotators and hip abductors. This finding would have direct application in rehabilitation treatment programs. Year: 2007 Title: Competitive inhibition of histone deacetylase activity by trichostatin A and butyrate Authors: Sekhavat, A. Sun, J. M. Davie, J. R. Auth Address: MB Institute of Cell Biology, University of Manitoba, 675 McDermot Avenue, Winnipeg, MB, R3E 0V9 Canada. Pages: 751-758 Volume: 85 Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18059533 Abstract: Histone deacetylases (HDACs) play a pivotal role in gene expression through their involvement in chromatin remodeling. The abnormal targeting or retention of HDACs to DNA regulatory regions is observed in many cancers, and hence HDAC inhibitors are being tested as promising anti-tumor agents. The results of previous kinetic studies, characterizing trichostatin A (TSA), as well as butyrate, as HDAC noncompetitive inhibitors, conflict with crystallographic and homology modeling data suggesting that TSA should act as a competitive inhibitor. Our results demonstrate that each of the HDAC inhibitors TSA and butyrate inhibits HDAC activity in a competitive fashion. Co-immunoprecipitation studies show that the inhibition of HDAC1 and HDAC2 activity by TSA does not disturb the extensive level of their association in the human breast cancer cell line MCF-7. Moreover, the inhibition of HDAC activity by TSA does not interfere with the interaction of HDAC1 and HDAC2 with Sin3A, a core component of the Sin3 complex. Thus, repressor complexes such as Sin3, appear to be stable in the presence of TSA. The association of HDAC2 with transcription factor Sp1 is also not affected by TSA. Year: 2007 Month: 12 Title: Competitive or noncompetitive, that's the question: research toward histone deacetylase inhibitors Authors: Su, H. Altucci, L. You, Q. Auth Address: Department of Medicinal Chemistry, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, Jiangsu, 210009 People's Republic of China. youqidong@gmail.com. Pages: 1007-12 Volume: 7 Number: 5 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18483291 Abstract: Histone deacetylase (HDAC) catalyze deacetylation of acetylated lysine residues on histones and a growing number of nonhistone proteins including many transcription factors, playing an important role in the upstream control of gene transcription, cell cycle progression, and apoptosis. It has been wildly recognized that HDACs are promising targets for cancer therapy. At least 10 HDAC inhibitors are currently in clinical evaluation. However, none of them is practically isoform selective. More and more evidence suggests that acetylation modification occurring in approximately 85% of eukaryotic proteins should be a general mechanism for altering protein structures or protein-protein interactions. Unselectively inhibiting the deacetylation activity of HDACs and the consequent modulation of the acetylation status of so many substrates might have multiple mechanisms of action in vivo, resulting in both therapeutic responses and unanticipated side effects. Lack of selectivity for the existing HDAC inhibitors is somewhat logical for the highly conserved residues in the catalytic site and the malleable structure in the rim of the active site of HDAC enzymes. For further advancements in the development of HDAC inhibitors, clues for selectivity will have to be considered. [Mol Cancer Ther 2008;7(5):1007-12]. Year: 2008 Month: 5 Title: Complement C3c and related protein biomarkers in amyotrophic lateral sclerosis and Parkinson's disease Authors: Goldknopf, I. L. Sheta, E. A. Bryson, J. Folsom, B. Wilson, C. Duty, J. Yen, A. A. Appel, S. H. Auth Address: Power3 Medical Products, Inc., The Woodlands, TX, USA. Pages: 1034-9 Volume: 342 Number: 4 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16516157 Abstract: We have used quantitative 2D gel electrophoresis to analyze serum proteins from 422 patients with neurodegenerative diseases and normal individuals in an unbiased approach to identify biomarkers. Differences in abnormal serum levels were found between amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), and related disorders for 34 protein biomarker spots, nine of which were related to the complement system. Of these nine, four spots originated from the Complement C3b-alpha-chain (C3c(1), C3c(2a), C3c(2b), and C3dg). The C3c spots (C3c(1), C3c(2a), and C3c(2b)) had the same amino acid sequence and glycosylation, though only C3c(1) was phosphorylated. In addition, Complement Factors H, Bb, and Pre-Serum amyloid protein displayed different serum concentrations in ALS, PD, and normal sera, whereas Complement C4b gamma-chain and Complement Factor I did not. The differential expression of the complement proteins provides potentially useful biomarkers as well as evidence for the involvement of inflammatory processes in the pathogenesis of ALS and PD. Year: 2006 Title: Complete dissociation of motor neuron death from motor dysfunction by Bax deletion in a mouse model of ALS Authors: Gould, T. W. Buss, R. R. Vinsant, S. Prevette, D. Sun, W. Knudson, C. M. Milligan, C. E. Oppenheim, R. W. Auth Address: Department of Neurobiology and Anatomy and Program in Neuroscience, Wake Forest University, Winston-Salem, North Carolina 27157-1010, USA. Pages: 8774-86 Volume: 26 Number: 34 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16928866 Abstract: The death of cranial and spinal motoneurons (MNs) is believed to be an essential component of the pathogenesis of amyotrophic lateral sclerosis (ALS). We tested this hypothesis by crossing Bax-deficient mice with mice expressing mutant superoxide dismutase 1 (SOD1), a transgenic model of familial ALS. Although Bax deletion failed to prevent neuromuscular denervation and mitochondrial vacuolization, MNs were completely rescued from mutant SOD1-mediated death. However, Bax deficiency extended lifespan and delayed the onset of motor dysfunction of SOD1 mutants, suggesting that Bax acts via a mechanism distinct from cell death activation. Consistent with this idea, Bax elimination delayed the onset of neuromuscular denervation, which began long before the activation of cell death proteins in SOD1 mutants. Additionally, we show that denervation preceded accumulation of mutant SOD1 within MNs and astrogliosis in the spinal cord, which are also both delayed in Bax-deficient SOD1 mutants. Interestingly, MNs exhibited mitochondrial abnormalities at the innervated neuromuscular junction at the onset of neuromuscular denervation. Additionally, both MN presynaptic terminals and terminal Schwann cells expressed high levels of mutant SOD1 before MNs withdrew their axons. Together, these data support the idea that clinical symptoms in the SOD1 G93A model of ALS result specifically from damage to the distal motor axon and not from activation of the death pathway, and cast doubt on the utility of anti-apoptotic therapies to combat ALS. Furthermore, they suggest a novel, cell death-independent role for Bax in facilitating mutant SOD1-mediated motor denervation. Year: 2006 Title: Complete nucleotide sequence, genomic organization, and promoter analysis of the murine survival motor neuron gene (Smn) Authors: DiDonato, C. J. Brun, T. Simard, L. R. Auth Address: Centre de Recherche, Hopital Ste-Justine, 3175 Cote Ste-Catherine, Montreal, Quebec, Canada H3T 1C5. Pages: 638-41 Volume: 10 Number: 6 Keywords: Amino Acid Sequence Animals Cyclic AMP Response Element-Binding Protein Databases, Factual Exons Humans Mice Mice, Inbred Strains Molecular Sequence Data Motor Neurons/metabolism Nerve Tissue Proteins/*genetics/metabolism Promoter Regions (Genetics) Protein Biosynthesis RNA-Binding Proteins Recombinant Proteins/genetics/metabolism Recombination, Genetic Repetitive Sequences, Nucleic Acid Research Support, Non-U.S. Gov't Sequence Analysis Sequence Homology, Amino Acid Software Transcription, Genetic Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10341102 Abstract: Year: 1999 Month: 6 Title: Complications of intrathecal baclofen pumps in children Authors: Gooch, J. L. Oberg, W. A. Grams, B. Ward, L. A. Walker, M. L. Auth Address: Primary Children's Medical Center, Salt Lake City, Utah 84113, USA. pcjgooch@ihc.com Pages: 1-6 Volume: 39 Number: 1 Keywords: Adolescent Adult Age Factors Baclofen/*administration & dosage/*adverse effects/therapeutic use Child Equipment Design Equipment Failure Humans Infusion Pumps, Implantable/*adverse effects Infusions, Parenteral/adverse effects Muscle Relaxants, Central/*administration & dosage/*adverse effects/therapeutic use Muscle Spasticity/*drug therapy Retrospective Studies Severity of Illness Index Time Factors Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12784068 Abstract: Intrathecal baclofen is increasingly being used to manage severe spasticity in children. Although substantial tone reduction with this treatment has been documented, complications also occur. In this study, we describe the device- and major non-device-related complications in a group of 100 consecutive children and young adults who received 117 intrathecal baclofen pumps for the management of severe spasticity. Twenty-four patients (24%) experienced a total of 48 complications. The most common complication was disconnection of the catheter at its connection to the pump, occurring in 9% of pumps implanted. This complication occurred more frequently in pumps with catheter access ports (16%) than in those without ports (2%). Catheter dislodgement from the intrathecal space was the next most common complication, occurring in 8% of pumps implanted (13% of pumps with ports, 4% of pumps without ports). To decrease the occurrence of the most common complications of intrathecal pumps, we now typically implant pumps without catheter access ports, and we use 2-piece catheters. Although the lack of an access port may be a disadvantage for troubleshooting, most complications can be detected in pumps without a port. Patient and family education is critical in preventing serious consequences of baclofen withdrawal resulting from catheter-related complications. Year: 2003 Month: 7 Title: Concerns about the design of clinical trials for spinal muscular atrophy Authors: Crawford, T. O. Auth Address: Department of Neurology and Pediatrics, Johns Hopkins University, Jefferson 123, 600 N Wolfe Street, Baltimore, MD 21287, USA. tcrawfo@welchlink.welch.jhu.edu Pages: 456-60 Volume: 14 Number: 8-9 Keywords: Animals Clinical Trials/*methods Humans Muscular Atrophy, Spinal/*therapy Research Design/*standards Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15336685 Abstract: The distinctive clinical course of SMA, characterized by slowing of the rate of degeneration with the passage of time, presents a special challenge to therapeutic clinical trial planning. Much of the actual functional decline may represent either an inevitable consequence of growth or the result of various secondary complications of weakness, making the study of agents intended to improve the course by increasing the level of SMN protein that much more difficult. Studies intended to demonstrate a slowing of the rate of degeneration, modeled upon clinical trials for ALS, are problematic. In contrast, short-term trials designed to demonstrate improved strength have substantial design advantages, but depend upon the demonstration of salutary effects of increased SMN that are plausible but at present only theoretical. This form of study thus has some potential for type II error, falsely rejecting a useful drug. Despite this limitation, logistic and statistical concerns suggest that the best strategy for evaluating any promising new therapy will be to use first a short-term study. Year: 2004 Month: 9 Title: Concurrent administration of Neu2000 and lithium produces marked improvement of motor neuron survival, motor function, and mortality in a mouse model of ALS Authors: Shin, J. H. Cho, S. I. Lim, H. R. Lee, J. K. Lee, Y. A. Noh, J. S. Joo, I. S. Kim, K. W. Gwag, B. J. Auth Address: Ajou. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17105868 Abstract: The Fas pathway and oxidative stress mediate neuronal death in stroke and may contribute to neurodegenerative disease. We tested the hypothesis that these two factors synergistically produce spinal motor neuron degeneration in amyotrophic lateral sclerosis (ALS). Levels of reactive oxygen species were increased in motor neurons from ALS mice compared to wild-type mice at age 10 weeks, before symptom onset. The proapoptotic proteins Fas, FADD, caspase 8, and caspase 3 were also elevated. Oral administration of Neu2000, a potent antioxidant, blocked the increase in reactive oxygen species but only slightly reduced activation of proapoptotic proteins. Administration of lithium carbonate (Li(+)), a mood stabilizer that prevents apoptosis, blocked the apoptosis machinery without preventing oxidative stress. Neu2000 or Li(+) alone significantly enhanced survival time and motor function and together had an additive effect. These findings provide evidence that jointly targeting oxidative stress and Fas-mediated apoptosis can prevent neuronal loss and motor dysfunction in ALS. Year: 2007 Title: Conditional activation of akt in adult skeletal muscle induces rapid hypertrophy Authors: Lai, K. M. Gonzalez, M. Poueymirou, W. T. Kline, W. O. Na, E. Zlotchenko, E. Stitt, T. N. Economides, A. N. Yancopoulos, G. D. Glass, D. J. Auth Address: Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill River Rd., Tarrytown, NY 10591-6707, USA. Pages: 9295-304 Volume: 24 Number: 21 Keywords: Adipose Tissue/metabolism Aging/physiology Animals Enzyme Activation Female Hypertrophy/*enzymology/genetics/metabolism/*pathology Male Mice Mice, Inbred C57BL Mice, Transgenic Microscopy, Confocal Microscopy, Fluorescence Muscle, Skeletal/drug effects/*enzymology/metabolism/*pathology Protein-Serine-Threonine Kinases/*genetics/*metabolism Proto-Oncogene Proteins/*genetics/*metabolism Proto-Oncogene Proteins c-akt Research Support, Non-U.S. Gov't Tamoxifen/pharmacology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15485899 Abstract: Skeletal muscle atrophy is a severe morbidity caused by a variety of conditions, including cachexia, cancer, AIDS, prolonged bedrest, and diabetes. One strategy in the treatment of atrophy is to induce the pathways normally leading to skeletal muscle hypertrophy. The pathways that are sufficient to induce hypertrophy in skeletal muscle have been the subject of some controversy. We describe here the use of a novel method to produce a transgenic mouse in which a constitutively active form of Akt can be inducibly expressed in adult skeletal muscle and thereby demonstrate that acute activation of Akt is sufficient to induce rapid and significant skeletal muscle hypertrophy in vivo, accompanied by activation of the downstream Akt/p70S6 kinase protein synthesis pathway. Upon induction of Akt in skeletal muscle, there was also a significant decrease in adipose tissue. These findings suggest that pharmacologic approaches directed toward activating Akt will be useful in inducing skeletal muscle hypertrophy and that an increase in lean muscle mass is sufficient to decrease fat storage. Year: 2004 Month: 11 Title: Congenital axonal neuropathy caused by deletions in the spinal muscular atrophy region Authors: Korinthenberg, R. Sauer, M. Ketelsen, U. P. Hanemann, C. O. Stoll, G. Graf, M. Baborie, A. Volk, B. Wirth, B. Rudnik-Schoneborn, S. Zerres, K. Auth Address: Department of Neuropediatrics and Muscular Diseases, Albert-Ludwigs-Universitat, Freiburg, Germany. Pages: 364-8 Volume: 42 Number: 3 Keywords: Axons/pathology Brain Stem/pathology Chromosomes, Human, Pair 5 Comparative Study Female Gene Deletion Hereditary Sensory and Autonomic Neuropathies/*complications/genetics/pathology Humans Infant, Newborn Male Microscopy, Electron Muscle, Skeletal/ultrastructure Neurons, Afferent/pathology Phenotype Spinal Muscular Atrophies of Childhood/*complications/genetics/pathology Sural Nerve/ultrastructure Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9307259 Abstract: Three newborn siblings presented with generalized weakness, asphyxia, facial diplegia, and external ophthalmoplegia. Electrophysiological testing showed inexcitability of motor and sensory nerves and myographic signs of denervation. Nerve biopsies and postmortem examination showed loss of myelinated fibers and axonal damage in sensory and mixed nerves. Many spinal motor neurons were chromatolytic although their number was normal. Molecular genetic investigations revealed a homozygous deletion of the survival motor neuron (SMN) gene and a loss of markers Ag1-CA and C212 in the paternal haplotype. These findings are consistent with the diagnosis of an unusually severe type of spinal muscular atrophy. Given the large extent of the deletion, it must be considered that the unusual severe phenotype with involvement of brainstem nuclei and afferent nerves might also be due to changes of yet unknown genes neighboring the SMN gene. Year: 1997 Month: 9 Title: Congenital bone fractures in spinal muscular atrophy: functional role for SMN protein in bone remodeling Authors: Shanmugarajan, S. Swoboda, K. J. Iannaccone, S. T. Ries, W. L. Maria, B. L. Reddy, S. V. Auth Address: Charles P. Darby Children's Research Institute, Medical University of South Carolina, Charleston, South Carolina 29425, USA. Pages: 967-73 Volume: 22 Number: 8 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17761651 Abstract: Spinal muscular atrophy is the second most common fatal childhood disorder. Core clinical features include muscle weakness caused by degenerating lower motor neurons and a high incidence of bone fractures and hypercalcemia. Fractures further compromise quality of life by progression of joint contractures or additional loss of motor function. Recent observations suggest that bone disease in spinal muscular atrophy may not be attributed entirely to lower motor neuron degeneration. The presence of the spinal muscular atrophy disease-determining survival motor neuron gene (SMN), SMN expression, and differential splicing in bone-resorbing osteoclasts was recently discovered. Its ubiquitous expression and the differential expression of splice variants suggest that SMN has specific roles in bone cell function. SMN protein also interacts with osteoclast stimulatory factor. Mouse models of human spinal muscular atrophy disease suggest a potential role of SMN protein in skeletal development. Dual energy x-ray absorptiometry analysis demonstrated a substantial decrease in total bone area and poorly developed caudal vertebra in the mouse model. These mice also had pelvic bone fractures. Studies delineating SMN signaling mechanisms and gene transcription in a cell-specific manner will provide important molecular insights into the pathogenesis of bone disease in children with spinal muscular atrophy. Moreover, understanding bone remodeling in spinal muscular atrophy may lead to novel therapeutic approaches to enhance skeletal health and quality of life. This article reviews the skeletal complications associated with spinal muscular atrophy and describes a functional role for SMN protein in osteoclast development and bone resorption activity. Year: 2007 Month: 8 Title: Congenital form of spinal muscular atrophy predominantly affecting the lower limbs: a clinical and muscle MRI study Authors: Mercuri, E. Messina, S. Kinali, M. Cini, C. Longman, C. Battini, R. Cioni, G. Muntoni, F. Auth Address: Dubowitz Neuromuscular Centre, Department of Paediatrics, Imperial College London, Hammersmith Hospital, Du Cane Road, London, W12 ONN, UK. e.mercuri@imperial.ac.uk Pages: 125-9 Volume: 14 Number: 2 Keywords: Adolescent Adult Child Chromosomes, Human, Pair 12/genetics Cyclic AMP Response Element-Binding Protein DNA Mutational Analysis Diagnosis, Differential Female Gait Disorders, Neurologic/genetics/pathology/physiopathology Genetic Screening Humans Leg/pathology/*physiopathology Magnetic Resonance Imaging Male Muscle, Skeletal/pathology/*physiopathology Muscular Atrophy, Spinal/congenital/*diagnosis/*physiopathology Mutation/genetics Nerve Tissue Proteins/deficiency/genetics RNA-Binding Proteins Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14733958 Abstract: We describe clinical and muscle magnetic resonance imaging (MRI) findings in 11 cases (three familial and eight sporadic) with the form of spinal muscular atrophy characterised by predominant involvement of the lower limbs with weakness of the proximal and distal muscles and marked atrophy of the distal leg and foot muscles. All patients presented at birth with talipes, which were in extension in seven of the 11. Arm muscle and function were preserved and lower limbs appeared to be disproportionately shorter compared to trunk and upper limbs. Functional abilities were markedly affected and only one of the 11 is able to walk independently for long distances, while six require support of crutches and two use callipers for walking. One child lost ambulation following a fall. The course of the disease is relatively stable and the progression of disability appeared to be related mostly to increased contractures rather than to loss of muscle strength. Respiratory and cardiac function were well preserved. A neurogenic disorder was suggested by electromyography and/or muscle biopsy in all patients, while motor nerve conduction was consistently normal. Muscle MRI of the thighs revealed diffuse atrophic appearance with relative hypertrophy of the adductor longus and of the semitendinosus. Genetic studies excluded the involvement of the survival motor neuron gene but none of these families was sufficiently informative to study linkage to the locus on chromosome 12q23-q24 previously found to be involved in patients with similar phenotype. In our experience this form of spinal muscular atrophy affecting predominantly the lower limbs is a relatively common form and should be considered in the differential diagnosis of infants with talipes and weakness in the lower limbs. The identical clinical and imaging features of the sporadic and familial cases suggest that these cases are likely to be affected by the same condition. Year: 2004 Month: 2 Title: Congenital heart defects in spinal muscular atrophy type I: A clinical report of two siblings and a review of the literature Authors: Menke, L. A. Poll-The, B. T. Clur, S. A. Bilardo, C. M. van der Wal, A. C. Lemmink, H. H. Cobben, J. M. Auth Address: Department of Pediatric Genetics, Academic Medical Center, Amsterdam, The Netherlands. Pages: 740-744 Volume: 146A Number: 6 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18266240 Abstract: A newborn girl presented with asphyxia, joint contractures and diminished spontaneous movements. Echocardiography showed hypoplastic left heart. Spinal muscular atrophy type I (SMA I) was diagnosed by detecting a homozygous deletion in the survival motor neuron 1 gene (SMN1). In the first trimester of a subsequent pregnancy, SMA I, hypoplastic left heart, and contractures were identified again. Congenital heart defects (CHD) have now been reported in 20 patients with SMA I, including three previously reported siblings and our two siblings, leading us to hypothesize that SMA I/CHD represents a unique phenotype of SMA I rather than a coincidental association. The homozygous SMN1 deletion may play a role in the development of CHD when it occurs in the presence of mutations or polymorphisms in other genes important for cardiac development. (c) 2008 Wiley-Liss, Inc. Year: 2008 Title: Congenital heart disease is a feature of severe infantile spinal muscular atrophy Authors: Rudnik-Schoneborn, S. Heller, R. Berg, C. Betzler, C. Grimm, T. Eggermann, T. Eggermann, K. Wirth, R. Wirth, B. Zerres, K. Auth Address: Institute of Human Genetics RWTH Aachen University, Germany. Pages: Volume: Number: Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18662980 Abstract: Objective Homozygous deletions/mutations of the SMN1 gene cause infantile spinal muscular atrophy (SMA). The presence of at least one SMN2 gene copy is required for normal embryogenesis. Lack of SMN protein results in degeneration of motor neurons, while extraneuronal manifestations have been regarded as a chance association with SMA. We report on heart defects in the subgroup of congenital SMA type I patients. Methods Data were recruited from 65 unselected SMA I patients whose diagnosis had been confirmed genetically within the first 6 months of age. SMN2 copy numbers were analyzed retrospectively and correlated with clinical findings including heart malformations. Results Four (6%) patients had one copy of SMN2, 56 (86%) had two and 5 (8%) had three SMN2 copies. Three out of 4 (75%) patients with a single SMN2 copy had congenital SMA with hemodynamically relevant atrial or ventricular septal defects. Discussion Previous case reports of SMA I patients with congenital heart defects did not clarify whether the cardiac malformations were coincidental. Given the respective incidences of congenitally lethal SMA with a single SMN2 copy and of cardiac septal defects in humans, a chance association of both conditions would occur in less than one out of 50 million individuals. Our findings suggest that the SMN protein is relevant for normal cardiogenesis. Year: 2008 Title: Consensus statement for standard of care in spinal muscular atrophy Authors: Wang, C. H. Finkel, R. S. Bertini, E. S. Schroth, M. Simonds, A. Wong, B. Aloysius, A. Morrison, L. Main, M. Crawford, T. O. Trela, A. Auth Address: Department of Neurology and Neurological Sciences, Stanford University Medical Center, Stanford, California 94305-5235, USA. wangch@stanford.edu Pages: 1027-49 Volume: 22 Number: 8 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17761659 Abstract: Spinal muscular atrophy is a neurodegenerative disease that requires multidisciplinary medical care. Recent progress in the understanding of molecular pathogenesis of spinal muscular atrophy and advances in medical technology have not been matched by similar developments in the care for spinal muscular atrophy patients. Variations in medical practice coupled with differences in family resources and values have resulted in variable clinical outcomes that are likely to compromise valid measure of treatment effects during clinical trials. The International Standard of Care Committee for Spinal Muscular Atrophy was formed in 2005, with a goal of establishing practice guidelines for clinical care of these patients. The 12 core committee members worked with more than 60 spinal muscular atrophy experts in the field through conference calls, e-mail communications, a Delphi survey, and 2 in-person meetings to achieve consensus on 5 care areas: diagnostic/new interventions, pulmonary, gastrointestinal/nutrition, orthopedics/rehabilitation, and palliative care. Consensus was achieved on several topics related to common medical problems in spinal muscular atrophy, diagnostic strategies, recommendations for assessment and monitoring, and therapeutic interventions in each care area. A consensus statement was drafted to address the 5 care areas according to 3 functional levels of the patients: nonsitter, sitter, and walker. The committee also identified several medical practices lacking consensus and warranting further investigation. It is the authors' intention that this document be used as a guideline, not as a practice standard for their care. A practice standard for spinal muscular atrophy is urgently needed to help with the multidisciplinary care of these patients. Year: 2007 Month: 8 Title: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets Authors: Lewis, B. P. Burge, C. B. Bartel, D. P. Auth Address: Pages: 15-20 Volume: 120 Number: 1 Keywords: 3' Untranslated Regions/genetics Adenosine/genetics/*metabolism Amino Acid Sequence Animals Chickens Dogs Gene Expression Regulation/genetics Gene Targeting/methods Humans Mice MicroRNAs/*genetics Molecular Sequence Data Nucleic Acid Hybridization/physiology Nucleotides/*metabolism RNA, Messenger/genetics Rats Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15652477 Abstract: We predict regulatory targets of vertebrate microRNAs (miRNAs) by identifying mRNAs with conserved complementarity to the seed (nucleotides 2-7) of the miRNA. An overrepresentation of conserved adenosines flanking the seed complementary sites in mRNAs indicates that primary sequence determinants can supplement base pairing to specify miRNA target recognition. In a four-genome analysis of 3' UTRs, approximately 13,000 regulatory relationships were detected above the estimate of false-positive predictions, thereby implicating as miRNA targets more than 5300 human genes, which represented 30% of our gene set. Targeting was also detected in open reading frames. In sum, well over one third of human genes appear to be conserved miRNA targets. Year: 2005 Title: Constitutive muscular abnormalities in culture in spinal muscular atrophy Authors: Braun, S. Croizat, B. Lagrange, M. C. Warter, J. M. Poindron, P. Auth Address: Departement d'Immunologie et d'Immunopharmacologie, Universite Louis Pasteur de Strasbourg, Illkirch, France. Pages: 694-5 Volume: 345 Number: 8951 Keywords: Culture Techniques Humans Muscle Fibers/pathology Muscles/innervation/pathology Muscular Atrophy, Spinal/*pathology Neuromuscular Diseases/pathology Neurons/pathology Research Support, Non-U.S. Gov't Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7741893 Abstract: To explore the cause of spinal muscular atrophy (SMA), we used an in-vitro model of nerve-muscle co-cultures in which motoneurons were normal and satellite cells were obtained from SMA patients. In co-cultures initiated with satellite cells from type I and type II SMA patients only, we observed degeneration of the innervated fibres after 1-3 weeks of nerve-muscle co-culture. This process involved vacuolisation, disorganisation, and death of the innervated muscle fibres. This observation points to a muscular implication in the severe forms of SMAs. Year: 1995 Title: Constitutively expressed catalytic proteasomal subunits are up-regulated during neuronal differentiation and required for axon initiation, elongation and maintenance Authors: Klimaschewski, L. Hausott, B. Ingorokva, S. Pfaller, K. Auth Address: Division of Neuroanatomy, Innsbruck Medical University, Innsbruck, Austria. lars.klimaschewski@i-med.ac.at Pages: 1708-17 Volume: 96 Number: 6 Keywords: Acetylcysteine/analogs & derivatives/pharmacology Animals Animals, Newborn Catalytic Domain/drug effects/*physiology Cell Differentiation/*physiology Cell Survival/drug effects/physiology Cysteine Proteinase Inhibitors/pharmacology Enzyme Inhibitors/pharmacology Ganglia, Autonomic/drug effects/growth & development/metabolism Ganglia, Sensory/drug effects/growth & development/metabolism Growth Cones/drug effects/*metabolism/ultrastructure Microscopy, Electron, Scanning Nerve Degeneration/chemically induced/metabolism/physiopathology Nerve Growth Factor/metabolism/pharmacology Nerve Net/drug effects/growth & development/metabolism Nervous System/drug effects/*growth & development/*metabolism PC12 Cells Proteasome Endopeptidase Complex/antagonists & inhibitors/genetics/*metabolism RNA, Messenger/drug effects/metabolism Rats Research Support, Non-U.S. Gov't Up-Regulation/drug effects/physiology Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16539686 Abstract: Inhibition of the proteasome by lactacystin, a specific blocker of the catalytic beta-subunits, results in transient neurite outgrowth by neuronal cell lines. Vice versa, as demonstrated in this study, treatment of pheochromocytoma (PC12) cells with nerve growth factor (NGF) or other differentiating agents reduces proteasomal activity. This is accompanied by an increase in mRNA and protein levels of the catalytically active subunits beta1, beta2 and beta5, but not of their inducible counterparts, indicating changes in subunit composition of the proteasome during neuronal differentiation. In contrast to neuronal cell lines, however, pre-treatment of primary neurons with proteasome inhibitors completely prevents axon formation, and lower concentrations of lactacystin (0.5-5 microm) significantly reduce axonal elongation and branching in vitro. Furthermore, established axonal networks degenerate rapidly and long-term survival of peripheral neurons is impaired in the presence of proteasome inhibitors. Axonal pathology is reminiscent of the morphological changes observed in neurodegenerative disorders and supports a crucial role of the constitutive catalytic subunits in axon initiation, maintenance and regeneration. Year: 2006 Month: 3 Title: Continuity of care for children with complex chronic health conditions: parents' perspectives Authors: Miller, A. R. Condin, C. J. McKellin, W. H. Shaw, N. Klassen, A. F. Sheps, S. Auth Address: Department of Pediatrics, University of British Columbia, British Columbia, Canada. amiller@cw.bc.ca Pages: 242 Volume: 9 Number: Keywords: Adult Anecdotes as Topic Canada Caregivers/psychology Child Child Health Services/standards Child, Preschool Chronic Disease/*therapy Community Networks/utilization Continuity of Patient Care/*standards Female *Health Knowledge, Attitudes, Practice Humans Interviews as Topic Male Middle Aged Needs Assessment Parents/*psychology *Professional-Family Relations Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20025770 Abstract: BACKGROUND: Continuity of care has been explored largely from academic and service provider perspectives, and in relation to adult patient/client groups. We interviewed parents of children with complex chronic health conditions to examine how their experiences and perceptions of continuity of care fit with these perspectives; and to identify the salient factors in the experience of, and factors contributing to, continuity in this population. METHODS: Parents of 47 elementary school-aged children with spina bifida, Down syndrome, attention-deficit/hyperactivity disorder, Duchenne muscular dystrophy or cystic fibrosis participated in semi-structured interviews. Parents described and mapped the pattern of their interactions with service providers over time in all domains relevant to their child's health, well-being, and development (medical, rehabilitational, educational, and social supportive services), with particular attention paid to their perceptions of connectedness or coherency in these interactions. Verbatim transcripts were analyzed thematically using a framework approach to impose structure regarding parents' perspectives on continuity of care. RESULTS: Existing academic concepts of relational, informational and management continuity were all discernable in parents' narratives. A thorough knowledge of the child on the part of service providers emerged as extremely important to parents; such knowledge was underpinned by continuity of personal relationships, principally, and also by written information. For this population, notions of continuity extend to the full range of service providers these children and families need to achieve optimal health status, and are not limited to physicians and nurses. Communication among providers was seen as integral to perceived continuity. Compartmentalization of services and information led to parents assuming a necessary, though at times, uncomfortable, coordinating role. Geographic factors, institutional structures and practices, provider attitudes, and, on occasion, parent preferences and judgments, were all found to create barriers to "seamless" management and provision of care continuity across providers, settings, and sectors. CONCLUSIONS: These findings add new perspectives to the understanding of continuity within chronically ill children's health care. They are relevant to contemporary initiatives to improve continuity of services to children with special health care needs, demonstrate the need for parental support of their important role in maintaining continuity, and suggest avenues for further research. Year: 2009 Month: 1 Title: Continuous critical care and long-term noninvasive ventilatory support for patients with neuromuscular disease Authors: Cheng, G. Bach, J. R. Auth Address: Pages: 246-7; author reply 247 Volume: 135 Number: 1 Keywords: Humans Lung Diseases, Obstructive/physiopathology/*therapy Reproducibility of Results Respiration, Artificial/*methods Treatment Outcome Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19136418 Abstract: Year: 2009 Month: 1 Title: Copy number analysis of survival motor neuron genes by multiplex ligation-dependent probe amplification Authors: Huang, C. H. Chang, Y. Y. Chen, C. H. Kuo, Y. S. Hwu, W. L. Gerdes, T. Ko, T. M. Auth Address: From the 1Genephile Bioscience Laboratory, Ko's Obstetrics and Gynecology, Taipei, Taiwan; 2Department of Pediatrics, National Taiwan University, Taipei, Taiwan; and 3Department of Clinical Genetics, Rigshospitalet, Copenhagen, Denmark. Pages: 241-248 Volume: 9 Number: 4 Keywords: No keywords found Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17438389 Abstract: PURPOSE:: To determine the copy number of survival motor genes using multiplex ligation-dependent probe amplification. METHODS:: Three hundred seventy-three subjects were recruited and divided into three groups. Group 1 included 310 subjects without a history of muscular atrophy, Group 2 consisted of 18 patients and 45 carriers of spinal muscular atrophy, and Group 3 included 20 subjects who were previously tested with denatured high-performance liquid chromatography. The copy number of survival motor neuron 1 and survival motor neuron 2 genes was determined with a commercially available multiplex ligation-dependent probe amplification kit. RESULTS:: Twenty-one genotypes of the survival motor neuron genes could be clearly defined in this series. The whole process of genotyping took <48 hours. In Group 1, 2:2 (survival motor neuron 1:survival motor neuron 2) was most common (52.90%), followed by 2:1 (30.32%); six (1.94%) subjects were found to be carriers of 1:2 or 1:3. In Group 2, all 18 patients had zero copies of the survival motor neuron 1 gene and variable copies of the survival motor neuron 2 gene. In Group 3, three subjects who had been told they were carriers of spinal muscular atrophy turned out to be noncarriers by multiplex ligation-dependent probe amplification. All 51 carriers from Groups 1 and 2 had one copy of the survival motor neuron 1 gene and one to four copies of the survival motor neuron 2 gene. CONCLUSION:: Multiplex ligation-dependent probe amplification is a simple and efficient method for copy number analysis of survival motor neuron genes. It can be used to detect the homozygous and heterozygous survival motor neuron deletion of spinal muscular atrophy. Year: 2007 Month: 4 Title: Correction of aberrant FGFR1 alternative RNA splicing through targeting of intronic regulatory elements Authors: Bruno, I. G. Jin, W. Cote, G. J. Auth Address: 1Department of Endocrine Neoplasia and Hormonal Disorders, The University of Texas M. D. Anderson Cancer Center, Houston, TX 77030, USA. Pages: 2409-20 Volume: 13 Number: 20 Keywords: Alternative Splicing/*genetics Caspases/analysis Cell Survival Exons/genetics Glioblastoma/genetics Humans Introns/*genetics Oligoribonucleotides, Antisense/*genetics RNA, Messenger/analysis/metabolism Receptor Protein-Tyrosine Kinases/*genetics Receptor, Fibroblast Growth Factor, Type 1 Receptors, Fibroblast Growth Factor/*genetics Regulatory Sequences, Ribonucleic Acid/*genetics Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Tumor Cells, Cultured Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15333583 Abstract: Alternative RNA splicing is now known to be pervasive throughout the genome and a target of human disease. We evaluated if targeting intronic splicing regulatory sequences with antisense oligonucleotides could be used to correct aberrant exon skipping. As a model, we targeted the intronic silencing sequence (ISS) elements flanking the alternatively spliced alpha-exon of the endogenous fibroblast growth factor receptor 1 (FGFR1) gene, which is aberrantly skipped in human glioblastoma. Antisense morpholino oligonucleotides targeting either upstream or downstream ISS elements increased alpha-exon inclusion from 10% up to 70% in vivo. The effect was dose dependent, sequence specific and reproducible in several human cell lines, but did not necessarily correlate with blocking of protein association in vitro. Simultaneous targeting of the ISS elements had no additive effect, suggesting that splicing regulation occurred through a shared mechanism. Broad applicability of this approach was demonstrated by similar targeting of the ISS elements of the human hnRNPA1 gene. The correction of FGFR1 gene splicing to >90% alpha-exon inclusion in glioblastoma cells had no discernable effect on cell growth in culture, but was associated with an increase in unstimulated caspase-3 and -7 activity. The ability to manipulate endogenously expressed mRNA variants allows exploration of their functional relevance under normal and diseased physiological states. Year: 2004 Title: Correction of disease-associated exon skipping by synthetic exon-specific activators Authors: Cartegni, L. Krainer, A. R. Auth Address: Cold Spring Harbor Laboratory, New York 11724, USA. Pages: 120-5 Volume: 10 Number: 2 Keywords: *Alternative Splicing Cyclic AMP Response Element-Binding Protein *Exons Genes, BRCA1 Genetic Diseases, Inborn/genetics/therapy Hela Cells Humans In Vitro Nerve Tissue Proteins/genetics Point Mutation RNA-Binding Proteins Research Support, U.S. Gov't, P.H.S. Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12524529 Abstract: Differential exon use is a hallmark of alternative splicing, a prevalent mechanism for generating protein isoform diversity. Many disease-associated mutations also affect pre-mRNA splicing, usually causing inappropriate exon skipping. SR proteins are essential splicing factors that recognize exonic splicing enhancers and drive exon inclusion. To emulate this function of SR proteins, we designed small chimeric effectors comprising a minimal synthetic RS domain covalently linked to an antisense moiety that targets an exon by Watson-Crick base pairing. Here we show that such synthetic effectors can mimic the functions of SR proteins and specifically restore wild type splicing when directed to defective BRCA1 or SMN2 pre-mRNA transcripts. This general approach can be used as a tool to investigate splicing mechanisms and modulate alternative splicing of specific genes, and as a therapeutic strategy to correct splicing defects responsible for numerous diseases. Year: 2003 Month: 2 Title: Correction of SMN2 Pre-mRNA splicing by antisense U7 small nuclear RNAs Authors: Madocsai, C. Lim, S. R. Geib, T. Lam, B. J. Hertel, K. J. Auth Address: Department of Microbiology and Molecular Genetics, College of Medicine, University of California at Irvine, Irvine, CA 92697-4025, USA. Pages: 1013-22 Volume: 12 Number: 6 Keywords: Blotting, Western Cyclic AMP Response Element-Binding Protein/*genetics DNA/chemistry/metabolism Exons Gene Therapy/*methods Genetic Techniques Genetic Vectors Hela Cells Homozygote Humans Models, Genetic Muscular Atrophy, Spinal/genetics/*therapy Mutation Nerve Tissue Proteins/*genetics Oligonucleotides, Antisense/chemistry Protein Isoforms *RNA Splicing RNA, Small Nuclear/*metabolism RNA-Binding Proteins/*genetics Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Transfection Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16226920 Abstract: Mutations in one of the duplicated survival of motor neuron (SMN) genes lead to the progressive loss of motor neurons and subsequent development of spinal muscular atrophy (SMA), a common, and usually fatal, hereditary disease. Homozygous absence of the telomeric copy (SMN1) correlates with development of SMA because differential splicing of the centromeric copy (SMN2) leads to exon 7 skipping and predominantly produces a biologically inactive protein isoform. To increase exon 7 inclusion of SMN2, we have designed a series of vectors that express modified U7 snRNAs containing antisense sequences complementary to the 3' splice site of SMN exon 8. Over 20 anti-SMN U7 snRNAs were tested for their ability to promote exon 7 inclusion in the SMN2 gene. Transient expression of anti-SMN U7 snRNAs in HeLa cells modulated SMN2 splicing to approximately 70% exon 7 inclusion in a sequence-specific and dose-dependent manner. Significantly, the administration of anti-SMN U7 snRNPs results in an increase in the concentration of SMN protein. These results suggest that modulation of SMN2 pre-mRNA splicing by modified U7 snRNAs provides a promising form of gene therapy for the treatment of SMA. Year: 2005 Month: 12 Title: Correlation between severity and SMN protein level in spinal muscular atrophy Authors: Lefebvre, S. Burlet, P. Liu, Q. Bertrandy, S. Clermont, O. Munnich, A. Dreyfuss, G. Melki, J. Auth Address: Unite de Recherches sur les Handicaps Genetiques de L'Enfant, INSERM, Unite 393, IFREM, Institut Necker, Hopital des Enfants Malades, Paris, France. Pages: 265-9 Volume: 16 Number: 3 Keywords: Blotting, Western Cell Line Cyclic AMP Response Element-Binding Protein Gene Expression Regulation Humans Liver/embryology/metabolism Muscular Atrophy, Spinal/embryology/*genetics/*metabolism Nerve Tissue Proteins/genetics/*metabolism Neuronal Apoptosis-Inhibitory Protein Point Mutation RNA-Binding Proteins Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sequence Deletion Spinal Cord/embryology/metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9207792 Abstract: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder characterized by degeneration of motor neurons of the spinal cord. Three different forms of childhood SMA have been recognized on the basis of age at onset and clinical course: Werdnig-Hoffmann disease (type-1), the intermediate form (type-II) and Kugelberg-Welander disease (type-III). A gene termed 'survival of motor neuron' (SMN) has been recognized as the disease-causing gene in SMA. SMN encodes a protein located within a novel nuclear structure and interacts with RNA-binding proteins. To elucidate the molecular mechanism underlying the pathogenesis of the disease, we examined the expression of the SMN gene in both controls and SMA patients by western blot and immunohistochemical analyses using antibodies raised against the SMN protein. The present study shows a marked deficiency of the SMN protein in SMA. Year: 1997 Month: 7 Title: Correlation of survival motor neuron expression in leukocytes and spinal cord in spinal muscular atrophy Authors: Tsai, L. K. Yang, C. C. Ting, C. H. Su, Y. N. Hwu, W. L. Li, H. Auth Address: Department of Neurology, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan. Pages: 303-5 Volume: 154 Number: 2 Keywords: Adult Age of Onset Animals Female Humans Leukocytes/*metabolism Male Mice Muscular Atrophy, Spinal/classification/*metabolism RNA, Messenger/metabolism Reverse Transcriptase Polymerase Chain Reaction SMN Complex Proteins/*metabolism Spinal Cord/*metabolism Related urls: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19150680 Abstract: Survival motor neuron (SMN) messenger RNA and protein levels in spinal muscular atrophy (SMA) model mice and in patients with SMA were measured. There was a high correlation between leukocyte and spinal cord SMN expression in SMA model mice and a moderate correlation between leukocyte SMN expression and age of disease onset in patients with SMA. Year: 2009 Month: 2 Title: Cortical excitability testing distinguishes Kennedy's disease from amyotrophic lateral sclerosis Authors: Vucic, S. Kiernan, M. C. Auth Address: Prince of Wales Medical Research Institute, Barker Street, Randwick, Sydney, NSW 2031, Australia. Pages: 1088-96 Volume: 119 Number: 5 Keywords: Adult Aged Amyotrophic Lateral Sclerosis/*physiopathology Cerebral Cortex/*physiopathology Diagnosis, Differen