A multiorganism pipeline for antiseizure drug discovery:Identification of chlorothymol as a novel γ-aminobutyric acidergic anticonvulsant

OBJECTIVE:Current medicines are ineffective in approximately one-third of people with epilepsy. Therefore, new antiseizure drugs are urgently needed to address this problem of pharmacoresistance. However, traditional rodent seizure and epilepsy models are poorly suited to high-throughput compound screening. Furthermore, testing in a single species increases the chance that therapeutic compounds act on molecular targets that may not be conserved in humans. To address these issues, we developed a pipeline approach using four different organisms. METHODS:We sequentially employed compound library screening in the zebrafish, Danio rerio, chemical genetics in the worm, Caenorhabditis elegans, electrophysiological analysis in mouse and human brain slices, and preclinical validation in mouse seizure models to identify novel antiseizure drugs and their molecular mechanism of action. RESULTS:Initially, a library of 1690 compounds was screened in an acute pentylenetetrazol seizure model using D rerio. From this screen, the compound chlorothymol was identified as an effective anticonvulsant not only in fish, but also in worms. A subsequent genetic screen in C elegans revealed the molecular target of chlorothymol to be LGC-37, a worm γ-aminobutyric acid type A (GABAA ) receptor subunit. This GABAergic effect was confirmed using in vitro brain slice preparations from both mice and humans, as chlorothymol was shown to enhance tonic and phasic inhibition and this action was reversed by the GABAA receptor antagonist, bicuculline. Finally, chlorothymol exhibited in vivo anticonvulsant efficacy in several mouse seizure assays, including the 6-Hz 44-mA model of pharmacoresistant seizures. SIGNIFICANCE:These findings establish a multiorganism approach that can identify compounds with evolutionarily conserved molecular targets and translational potential, and so may be useful in drug discovery for epilepsy and possibly other conditions

A phloem‐localized Arabidopsis metacaspase (AtMC3) improves drought tolerance

Increasing drought phenomena pose a serious threat to agricultural productivity. Although plants have multiple ways to respond to the complexity of drought stress, the underlying mechanisms of stress sensing and signaling remain unclear. The role of the vasculature, in particular the phloem, in facilitating inter-organ communication is critical and poorly understood.Combining genetic, proteomic and physiological approaches, we investigated the role of AtMC3, a phloem-specific member of the metacaspase family, in osmotic stress responses in Arabidopsis thaliana. Analyses of the proteome in plants with altered AtMC3 levels revealed differential abundance of proteins related to osmotic stress pointing into a role of the protein in water-stress-related responses.Overexpression of AtMC3 conferred drought tolerance by enhancing the differentiation of specific vascular tissues and maintaining higher levels of vascular-mediated transportation, while plants lacking the protein showed an impaired response to drought and inability to respond effectively to the hormone abscisic acid.Overall, our data highlight the importance of AtMC3 and vascular plasticity in fine-tuning early drought responses at the whole plant level without affecting growth or yield.ISSN:0028-646XISSN:1469-813

The immediate effect of musculoskeletal physiotherapy techniques and massage on pain and ease of breathing in adults with cystic fibrosis

AbstractThe optimal treatment approach to musculoskeletal pain in cystic fibrosis remains unclear. This study aimed to examine the effect of a combination of musculoskeletal physiotherapy techniques and massage therapy on musculoskeletal pain and ease of breathing. A single treatment session was associated with reduction in pain and improvement in ease of breathing in adults with cystic fibrosis

The Effects of Irradiation on Structure and Leaching of Pure and Doped Thin-Film Ceria SIMFUEL Models Prepared via Polymer-Templated Deposition

When studying hazardous materials such as spent nuclear fuel (SNF), minimisation of sample volumes is essential, together with the use of chemically-similar surrogates where possible. For example, the bulk behaviour of urania (UO2) can be mimicked by appropriately-engineered thin films of sufficient thickness and inactive materials such as ceria (CeO2) can be used to study effects within radioactive systems used to fuel nuclear fission. However, thin film properties are sensitive to the preparative method, many of which require the use of highly toxic precursors and specialised apparatus e.g. chemical vapour deposition. To address this, we present the development of a flexible, tuneable, scalable method for the preparation of thin-film CeO2 SIMFUEL models with a thickness of ≈ 5 μm. The effects of γ irradiation (up to 100 kGy) and dopants including trivalent lanthanides (Ln3+) and simulant ε-particles on the structure and long-term leaching of these systems under SNF storage conditions are explored, alongside the context of this within further work. It was found that the sensitivity of CeO2 films to reduction upon irradiation, particularly in the presence of simulant ε-particles results in increased leaching of Ce (as CeIII), while trivalent lanthanides (Nd3+ and Eu3+) have minimal effect on Ce leaching

Care of Adult Patients in Acute Care Facilities with a Tracheostomy

Adult patients with a tracheostomy tube are a vulnerable patient group because of changes to their airway. Additionally, their care may be highly complex involving a number of clinicians across different healthcare specialties; highlighting the need for good communication, coordination, team functioning and documentation. In 2009, the Clinical Excellence Commission released a report that found that patients with a tracheostomy were experiencing significant adverse events as a result of deficits in their care. Specifically, these deficits were related to the competencies of clinical staff and lack of appropriate action especially after hours. The guideline has been developed using a multi-stage process to ensure that: • the recommendations reflect best available evidence and current accepted standards of clinical practice • the scope of practice of all healthcare professionals (HCP) involved in the care of the patient has been considered • broad consultation with all HCP • recommendations are flexible and applicable across all acute care settings

The Effects of Irradiation on Structure and Leaching of Pure and Doped Thin-Film Ceria SIMFUEL Models Prepared via Polymer-Templated Deposition

When studying hazardous materials such as spent nuclear fuel (SNF), minimisation of sample volumes is essential, together with the use of chemically-similar surrogates where possible. For example, the bulk behaviour of urania (UO2) can be mimicked by appropriately-engineered thin films of sufficient thickness and inactive materials such as ceria (CeO2) can be used to study effects within radioactive systems used to fuel nuclear fission. However, thin film properties are sensitive to the preparative method, many of which require the use of highly toxic precursors and specialised apparatus e.g. chemical vapour deposition. To address this, we present the development of a flexible, tuneable, scalable method for the preparation of thin-film CeO2 SIMFUEL models with a thickness of ≈ 5 μm. The effects of γ irradiation (up to 100 kGy) and dopants including trivalent lanthanides (Ln3+) and simulant ε-particles on the structure and long-term leaching of these systems under SNF storage conditions are explored, alongside the context of this within further work. It was found that the sensitivity of CeO2 films to reduction upon irradiation, particularly in the presence of simulant ε-particles results in increased leaching of Ce (as CeIII), while trivalent lanthanides (Nd3+ and Eu3+) have minimal effect on Ce leaching

Robust Replication of Genotype-Phenotype Associations across Multiple Diseases in an Electronic Medical Record

Large-scale DNA databanks linked to electronic medical record (EMR) systems have been proposed as an approach for rapidly generating large, diverse cohorts for discovery and replication of genotype-phenotype associations. However, the extent to which such resources are capable of delivering on this promise is unknown. We studied whether an EMR-linked DNA biorepository can be used to detect known genotype-phenotype associations for five diseases. Twenty-one SNPs previously implicated as common variants predisposing to atrial fibrillation, Crohn disease, multiple sclerosis, rheumatoid arthritis, or type 2 diabetes were successfully genotyped in 9483 samples accrued over 4 mo into BioVU, the Vanderbilt University Medical Center DNA biobank. Previously reported odds ratios (ORPR) ranged from 1.14 to 2.36. For each phenotype, natural language processing techniques and billing-code queries were used to identify cases (n = 70–698) and controls (n = 808–3818) from deidentified health records. Each of the 21 tests of association yielded point estimates in the expected direction. Previous genotype-phenotype associations were replicated (p < 0.05) in 8/14 cases when the ORPR was > 1.25, and in 0/7 with lower ORPR. Statistically significant associations were detected in all analyses that were adequately powered. In each of the five diseases studied, at least one previously reported association was replicated. These data demonstrate that phenotypes representing clinical diagnoses can be extracted from EMR systems, and they support the use of DNA resources coupled to EMR systems as tools for rapid generation of large data sets required for replication of associations found in research cohorts and for discovery in genome science