41 research outputs found
Dominant mutations of the Notch ligand Jagged1 cause peripheral neuropathy
Notch signaling is a highly conserved intercellular pathway with tightly regulated and pleiotropic roles in normal tissue development and homeostasis. Dysregulated Notch signaling has also been implicated in human disease, including multiple forms of cancer, and represents an emerging therapeutic target. Successful development of such therapeutics requires a detailed understanding of potential on-target toxicities. Here, we identify autosomal dominant mutations of the canonical Notch ligand Jagged1 (or JAG1) as a cause of peripheral nerve disease in 2 unrelated families with the hereditary axonal neuropathy Charcot-Marie-Tooth disease type 2 (CMT2). Affected individuals in both families exhibited severe vocal fold paresis, a rare feature of peripheral nerve disease that can be life-threatening. Our studies of mutant protein posttranslational modification and localization indicated that the mutations (p.Ser577Arg, p.Ser650Pro) impair protein glycosylation and reduce JAG1 cell surface expression. Mice harboring heterozygous CMT2-associated mutations exhibited mild peripheral neuropathy, and homozygous expression resulted in embryonic lethality by midgestation. Together, our findings highlight a critical role for JAG1 in maintaining peripheral nerve integrity, particularly in the recurrent laryngeal nerve, and provide a basis for the evaluation of peripheral neuropathy as part of the clinical development of Notch pathway–modulating therapeutics
Selective amplification of Brucella melitensis mRNA from a mixed host-pathogen total RNA
<p>Abstract</p> <p>Background</p> <p>Brucellosis is a worldwide anthropozoonotic disease caused by an in vivo intracellular pathogen belonging to genus <it>Brucella</it>. The characterization of brucelae transcriptome's during host-pathogen interaction has been limited due to the difficulty of obtaining an adequate quantity of good quality eukaryotic RNA-free pathogen RNA for downstream applications.</p> <p>Findings</p> <p>Here, we describe a combined protocol to prepare RNA from intracellular <it>B. melitensis </it>in a quantity and quality suitable for pathogen gene expression analysis. Initially, <it>B. melitensis </it>total RNA was enriched from a host:pathogen mixed RNA sample by reducing the eukaryotic RNA..Then, to increase the <it>Brucella </it>RNA concentration and simultaneously minimize the contaminated host RNA in the mixed sample, a specific primer set designed to anneal to all <it>B. melitensis </it>ORF allows the selective linear amplification of sense-strand prokaryotic transcripts in a previously enriched RNA sample.</p> <p>Conclusion</p> <p>The novelty of the method we present here allows analysis of the gene expression profile of <it>B. melitensis </it>when limited amounts of pathogen RNA are present, and is potentially applicable to both <it>in vivo </it>and <it>in vitro </it>models of infection, even at early infection time points.</p
Charcot-Marie-Tooth–Linked Mutant GARS Is Toxic to Peripheral Neurons Independent of Wild-Type GARS Levels
Charcot-Marie-Tooth disease type 2D (CMT2D) is a dominantly inherited peripheral neuropathy caused by missense mutations in the glycyl-tRNA synthetase gene (GARS). In addition to GARS, mutations in three other tRNA synthetase genes cause similar neuropathies, although the underlying mechanisms are not fully understood. To address this, we generated transgenic mice that ubiquitously over-express wild-type GARS and crossed them to two dominant mouse models of CMT2D to distinguish loss-of-function and gain-of-function mechanisms. Over-expression of wild-type GARS does not improve the neuropathy phenotype in heterozygous Gars mutant mice, as determined by histological, functional, and behavioral tests. Transgenic GARS is able to rescue a pathological point mutation as a homozygote or in complementation tests with a Gars null allele, demonstrating the functionality of the transgene and revealing a recessive loss-of-function component of the point mutation. Missense mutations as transgene-rescued homozygotes or compound heterozygotes have a more severe neuropathy than heterozygotes, indicating that increased dosage of the disease-causing alleles results in a more severe neurological phenotype, even in the presence of a wild-type transgene. We conclude that, although missense mutations of Gars may cause some loss of function, the dominant neuropathy phenotype observed in mice is caused by a dose-dependent gain of function that is not mitigated by over-expression of functional wild-type protein
Motor neuron intrinsic and extrinsic mechanisms contribute to the pathogenesis of FUS-associated amyotrophic lateral sclerosis
GARS axonopathy: not every neuron's cup of tRNA.
Charcot-Marie-Tooth disease type 2D, a hereditary axonal neuropathy, is caused by mutations in glycyl-tRNA synthetase (GARS). The mutations are distributed throughout the protein in multiple functional domains. In biochemical and cell culture experiments, some mutant forms of GARS have been indistinguishable from wild-type protein, suggesting that these in vitro tests might not adequately assess the aberrant activity responsible for axonal degeneration. Recently, mouse and fly models have offered new insights into the disease mechanism. There are still gaps in our understanding of how mutations in a ubiquitously expressed component of the translation machinery result in axonal neuropathy. Here, we review recent reports, weigh the evidence for and against possible mechanisms and suggest areas of focus for future work
Dominant, toxic gain-of-function mutations in gars lead to non-cell autonomous neuropathology
Charcot-Marie-Tooth (CMT) neuropathies are collectively the most common hereditary neurological condition and a major health burden for society. Dominant mutations in the gene GARS, encoding the ubiquitous enzyme, glycyl-tRNA synthetase (GlyRS), cause peripheral nerve degeneration and lead to CMT disease type 2D. This genetic disorder exemplifies a recurring motif in neurodegeneration, whereby mutations in essential, widely expressed genes have selective deleterious consequences for the nervous system. Here, using novel Drosophila models, we show a potential solution to this phenomenon. Ubiquitous expression of mutant GlyRS leads to motor deficits, progressive neuromuscular junction (NMJ) denervation and pre-synaptic build-up of mutant GlyRS. Intriguingly, neuronal toxicity is, at least in part, non-cell autonomous, as expression of mutant GlyRS in mesoderm or muscle alone results in similar pathology. This mutant GlyRS toxic gain-of-function, which is WHEP domain-dependent, coincides with abnormal NMJ assembly, leading to synaptic degeneration, and, ultimately, reduced viability. Our findings suggest that mutant GlyRS gains access to ectopic sub-compartments of the motor neuron, providing a possible explanation for the selective neuropathology caused by mutations in a widely expressed gene
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Fast ion profiles during neutral beam and lower hybrid heating
Profiles of the d(d,p)t fusion reaction are measured in the PLT Tokamak using an array of collimated 3 MeV proton detectors. During deuterium neutral beam injection, the emission profile indicates that the beam deposition is as narrow as predicted by a bounce-averaged Fokker-Planck code. The fast ion tail formed by lower hybrid waves (at densities above the critical density for current drive) also peaks strongly near the magnetic axis
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Fast ion profiles during neutral beam and lower hybrid heating
Profiles of the d(d,p)t fusion reaction are measured in the PLT Tokamak using an array of collimated 3 MeV proton detectors. During deuterium neutral beam injection, the emission profile indicates that the beam deposition is as narrow as predicted by a bounce-averaged Fokker-Planck code. The fast ion tail formed by lower hybrid waves (at densities above the critical density for current drive) also peaks strongly near the magnetic axis
Fast ion profiles during neutral beam and lower hybrid heating
Profiles of the d(d,p)t fusion reaction are measured in the PLT Tokamak using an array of collimated 3 MeV proton detectors. During deuterium neutral beam injection, the emission profile indicates that the beam deposition is as narrow as predicted by a bounce-averaged Fokker-Planck code. The fast ion tail formed by lower hybrid waves (at densities above the critical density for current drive) also peaks strongly near the magnetic axis
Successful treatment of a large choroidal abscess in an immunocompetent child
Background We report the case of a systemically well 4-year-old Aboriginal boy who developed a choroidal abscess after being poked in the left eye with a blunt object. Case report This boy presented with redness and reduced vision in the left eye after a blunt object was poked into his eye by his sibling. He was noted to have a choroidal mass which finally manifested as a choroidal abscess. Results His initial visual acuity was 6/60, and dilated fundus examination demonstrated a localised solid-appearing choroidal elevation involving the posterior pole, including the macula. An ultrasound of the eye revealed a choroidal haematoma with an atypical appearance, whose height was 8 mm with a base of 12 mm × 10 mm. The lesion failed to resolve, and eventually resulted in orbital cellulitis that did not respond to intravenous and topical antibiotic treatment. He then went on to achieve complete visual recovery after successful management by transcleral incision, drainage and systemic antibiotic therapy. Conclusion Choroidal abscess has been described in patients who are debilitated, immunocompromised or suffer with systemic disease such as cystic fibrosis or endocarditis. This case represents a unique report of staphylococcal choroidal abscess in a healthy child that completely resolved after transcleral drainage and systemic antibiotics