Real-world evidence: Risdiplam in a patient with spinal muscular atrophy type I with a novel splicing mutation and one SMN2 copy.

Spinal muscular atrophy (SMA), which results from the deletion or/and mutation in the SMN1 gene, is an autosomal recessive neuromuscular disorder that leads to weakness and muscle atrophy. SMN2 is a paralogous gene of SMN1. SMN2 copy number affects the severity of SMA, but its role in patients treated with disease modifying therapies is unclear. The most appropriate individualized treatment for SMA has not yet been determined. Here, we reported a case of SMA type I with normal breathing and swallowing function. We genetically confirmed that this patient had a compound heterozygous variant: one deleted SMN1 allele and a novel splice mutation c.628-3T>G in the retained allele, with one SMN2 copy. Patient-derived sequencing of 4 SMN1 cDNA clones showed that this intronic single transversion mutation results in an alternative exon (e)5 3' splice site, which leads to an additional 2 nucleotides (AG) at the 5' end of e5, thereby explaining why the patient with only one copy of SMN2 had a mild clinical phenotype. Additionally, a minigene assay of wild type and mutant SMN1 in HEK293T cells also demonstrated that this transversion mutation induced e5 skipping. Considering treatment cost and goals of avoiding pain caused by injections and starting treatment as early as possible, risdiplam was prescribed for this patient. However, the patient showed remarkable clinical improvements after treatment with risdiplam for 7 months despite carrying only one copy of SMN2. This study is the first report on the treatment of risdiplam in a patient with one SMN2 copy in a real-world setting. These findings expand the mutation spectrum of SMA and provide accurate genetic counseling information, as well as clarify the molecular mechanism of careful genotype-phenotype correlation of the patient.

Recent advances in the application of ionic liquids in antimicrobial material for air disinfection and sterilization.

Airborne transmission is one of the most unpredictable routes of infection. Nowadays, airborne diseases increase ever than before because of the complex living air environment. Apart from the inorganic particles, active microorganisms including bacteria, viruses, and fungi are incorporated in the pathogens acting as threaten to public health, which can hardly be treated by the traditional air purification methods based on adsorption. Therefore, effective filtration material with antimicrobial activity is demanded to solve the problem. Ionic liquids (ILs) are a category of salts that remain liquid at room temperature. The stable physico-chemical properties and extremely low vapor pressure make them suitable for a wide range of applications. Thanks to the numerous combinations of cations and anions, as well as the ability of inheriting properties from the parent ions, Ils are believed to be a promising industrial material. In recent decades, several Ils, such as imidazolium, pyridinium, pyrrolidinium, phosphonium, and choline, have been found to have antimicrobial activity in their monomeric or polymeric forms. This work focuses on the antimicrobial activity and safety of the latest types of ionic liquids, discussing the synthesis or manufacturing methods of Ils for air purification and filtration. Furthermore, possible applications of Ils antimicrobial materials in medical instruments and indoor environments are mentioned to encourage the scientific community to further explore the potential applications of Ils.

Effect of Singing on Symptoms in Stable COPD: A Systematic Review and Meta-Analysis.

Background: Chronic obstructive pulmonary disease (COPD) is a chronic lung disease which feature is progressive airflow obstruction. Singing is a popular and convenient activity that requires people to manage their lung volumes and airflow actively. Despite the well-known benefits of singing to healthy people, the specific effect still remains unclear. Objective: To investigate the mental and psychological benefits of singing in patients with stable COPD. Search Methods: We used Preferred Reporting Items for Systematic Reviews and Meta-Analyses Guidelines (PRISMA) on randomized controlled trials (RCTs) including singing exercise as the main intervention in stable COPD. We searched 8 electronic databases, including Web of Science, PubMed, Embase, Cochrane Library, Clinical Trials.gov, and the Physical Therapy Evidence Database (PEDro), CNKI, and Wanfang Database from inception until May 2022. The searching languages was English or Chinese. Data extraction using standardized templates was performed by two independent reviewers. The quality of the studies was assessed using the PEDro scale. Data synthesis was performed with Revman 5.4. The pooled effect sizes are reported by MD and 95% CI. Results: Five RCTs involving 333 patients with stable COPD were included in this meta-analysis. Singing was regarded as the main intervention in the experimental group. Meta-analysis revealed that singing improves quality of life on Short Form 36 physical component summary (SF-36 PCS) (MD = 12.63, 95% CI: 5.52 to 19.73, P < 0.01) and respiratory muscle in maximal expiratory pressure (PEmax) (MD = 14.30, 95% CI: 0.87 to 27.73, P = 0.04) in patients with COPD. However, it has limited effects on Short Form 36 mental component summary (SF-36 MCS), lung function, exercise capability, and adverse mental state. Conclusion: Based on results of the meta-analysis, singing could be used to improve quality of life (SF-36 PCS) and respiratory muscles (PEmax) in patients with COPD.

Rhythmic Auditory Stimulation as an Adjuvant Therapy Improved Post-stroke Motor Functions of the Upper Extremity: A Randomized Controlled Pilot Study.

OBJECTIVES: To explore whether rhythmic auditory stimulation (RAS) could improve motor functions of post-stroke hemiparetic upper extremity. DESIGN: A prospective, randomized controlled, assessor-blinded pilot study. METHODS: Thirty stroke patients were randomly distributed into the RAS group (n = 15) and the control group (n = 15). Both groups received regular therapies. The RAS group received additional 30 min of RAS training, while the control group received additional 30 min of regular therapies for 5 days per week for 4 weeks. The Fugl-Meyer Assessment-Upper Extremity (FMA-UE), Wolf Motor Function Test (WMFT), and Barthel Index (BI) were used. The co-activation interval and co-contraction index were calculated from surface electromyography (sEMG) recordings on the affected biceps and triceps during elbow flexion and extension. Assessments were performed before and after the treatments. RESULTS: Significant improvements in motor functions were observed within both groups (p < 0.05 in the FMA-UE, WMFT, and BI, respectively), as well as between groups after the treatments (higher scores in the RAS group, all p < 0.05 except for p = 0.052 in the FMA-UE; group × time interaction, all p < 0.05). Statistical significance was found in the co-activation interval between groups after the treatments (lower in the RAS group; p = 0.022 during elbow extension; p = 0.001 during elbow flexion; group × time interaction, p < 0.05 only during elbow extension). No statistical significance was found in the co-contraction index between groups; an inversed pattern of changes was observed between groups supported by relatively higher increments in the triceps recruitments to the biceps. CONCLUSION: Using RAS in task-oriented exercises was effective in moderating co-contraction, facilitating task-oriented movements of the hemiparetic upper extremity, and improving ADLs among those who had emerging isolated joint movements. The effects were evident on sEMG possibly by adjusting the balance of recruitments between the agonist and the antagonist. CLINICAL TRIAL REGISTRATION: The study was registered at the Chinese Clinical Trial Registry (No. 1900026665).

Alexander disease mutant glial fibrillary acidic protein compromises glutamate transport in astrocytes.

Alexander disease (AxD) is a leukodystrophy caused by heterozygous mutations in the gene for glial fibrillary acidic protein, an intermediate filament protein expressed by astrocytes. The mutation causes prominent protein aggregates inside astrocytes; there is also loss of myelin and oligodendrocytes and neuronal degeneration. We show that immunohistochemical staining for glutamate transporter 1, the major brain glutamate transporter expressed primarily in astrocytes suggests decreased levels in the hippocampi of infantile AxD patients. A knock-in mouse model of AxD also shows significant reduction of glutamate transporter 1 in the hippocampus. To explore this phenomenon at the cellular level, wild-type and R239C mutant glial fibrillary acidic proteins (the most common mutation) were overexpressed in astrocytes in culture. Western blotting and whole-cell patch clamp recordings demonstrated that the R239C astrocytes exhibited markedly reduced glutamate transporter 1 protein levels; this resulted in attenuated or abolished glutamate-induced inward transporter current. Neurons cocultured with the R239C astrocytes exhibited increased death after glutamate challenge. These results indicate that aberrant astrocytes have decreased glutamate uptake, which may play an important role in the pathogenesis of neuronal and oligodendrocyte injury and death in AxD.

Plectin regulates the organization of glial fibrillary acidic protein in Alexander disease.

Alexander disease (AxD) is a rare but fatal neurological disorder caused by mutations in the astrocyte-specific intermediate filament protein glial fibrillary acidic protein (GFAP). Histologically, AxD is characterized by cytoplasmic inclusion bodies called Rosenthal fibers (RFs), which contain GFAP, small heat shock proteins, and other undefined components. Here, we describe the expression of the cytoskeletal linker protein plectin in the AxD brain. RFs displayed positive immunostaining for plectin and GFAP, both of which were increased in the AxD brain. Co-localization, co-immunoprecipitation, and in vitro overlay analyses demonstrated direct interaction of plectin and GFAP. GFAP with the most common AxD mutation, R239C (RC GFAP), mainly formed abnormal aggregates in human primary astrocytes and murine plectin-deficient fibroblasts. Transient transfection of full-length plectin cDNA converted these aggregates to thin filaments, which exhibited diffuse cytoplasmic distribution. Compared to wild-type GFAP expression, RC GFAP expression lowered plectin levels in astrocytoma-derived stable transfectants and plectin-positive fibroblasts. A much higher proportion of total GFAP was found in the Triton X-insoluble fraction of plectin-deficient fibroblasts than in wild-type fibroblasts. Taken together, our results suggest that insufficient amounts of plectin, due to RC GFAP expression, promote GFAP aggregation and RF formation in AxD.

Alexander-disease mutation of GFAP causes filament disorganization and decreased solubility of GFAP.

Alexander disease is a fatal neurological illness characterized by white-matter degeneration and the formation of astrocytic cytoplasmic inclusions called Rosenthal fibers, which contain the intermediate filament glial fibrillary acidic protein (GFAP), the small heat-shock proteins HSP27 and alphaB-crystallin, and ubiquitin. Many Alexander-disease patients are heterozygous for one of a set of point mutations in the GFAP gene, all of which result in amino acid substitutions. The biological effects of the most common alteration, R239C, were tested by expressing the mutated protein in cultured cells by transient transfection. In primary rat astrocytes and Cos-7 cells, the mutant GFAP was incorporated into filament networks along with the endogenous GFAP and vimentin, respectively. In SW13Vim(-) cells, which have no endogenous cytoplasmic intermediate filaments, wild-type human GFAP frequently formed filamentous bundles, whereas the R239C GFAP formed 'diffuse' and irregular patterns. Filamentous bundles of R239C GFAP were sometimes formed in SW13Vim(-) cells when wild-type GFAP was co-transfected. Although the presence of a suitable coassembly partner (vimentin or GFAP) reduced the potential negative effects of the R239C mutation on GFAP network formation, the mutation affected the stability of GFAP in cells in a dominant fashion. Extraction of transfected SW13Vim(-) cells with Triton-X-100-containing buffers showed that the mutant GFAP was more resistant to solubilization at elevated KCl concentrations. Both wild-type and R239C GFAP assembled into 10 nm filaments with similar morphology in vitro. Thus, although the R239C mutation does not appear to affect filament formation per se, the mutation alters the normal solubility and organization of GFAP networks.