RESUMO
Weak interfacial strength restricts the mechanical properties of carbon fiber-reinforced composites. Here, inspired by natural hook-groove microstructure system (HGMS) of black kite (Milvus migrans), we detail the steps to construct a biomimetic HGMS based on dopamine-functionalized carbon fibers (CFs) and zinc oxide nanorods (ZnO NRs) using a two-step modification approach. We describe the fabrication of biomimetic carbon fiber composites using vacuum-assisted contact molding (VACM) and subsequent characterization using standard comprehensive mechanical tests techniques. For complete details on the use and execution of this protocol, please refer to Wang et al. (2022).
Assuntos
Biomimética , Óxido de Zinco , Fibra de Carbono , Propriedades de SuperfícieRESUMO
Chloride-proton exchange by the lysosomal anion transporter ClC-7/Ostm1 is of pivotal importance for the physiology of lysosomes and bone resorption. Mice lacking either ClC-7 or Ostm1 develop a lysosomal storage disease and mutations in either protein have been found to underlie osteopetrosis in mice and humans. Some human disease-causing CLCN7 mutations accelerate the usually slow voltage-dependent gating of ClC-7/Ostm1. However, it has remained unclear whether the fastened kinetics is indeed causative for the disease. Here we identified and characterized a new deleterious ClC-7 mutation in Belgian Blue cattle with a severe symptomatology including perinatal lethality and in most cases gingival hamartomas. By autozygosity mapping and genome-wide sequencing we found a handful of candidate variants, including a cluster of three private SNPs causing the substitution of a conserved tyrosine in the CBS2 domain of ClC-7 by glutamine. The case for ClC-7 was strengthened by subsequent examination of affected calves that revealed severe osteopetrosis. The Y750Q mutation largely preserved the lysosomal localization and assembly of ClC-7/Ostm1, but drastically accelerated its activation by membrane depolarization. These data provide first evidence that accelerated ClC-7/Ostm1 gating per se is deleterious, highlighting a physiological importance of the slow voltage-activation of ClC-7/Ostm1 in lysosomal function and bone resorption.