RESUMO
Surface coating of plasmonic nanoparticles is of huge importance to suppress fluorescence quenching in plasmon-enhanced fluorescence sensing. Herein, a one-pot method for synthesizing polymer-coated silver nanoparticles was developed using a functional polymer conjugated with disulfide-containing anchoring groups. The disulfides played a crucial role in covalently bonding polymers to the surface of the silver nanoparticles. The covalent bond enabled the polymer layer to form a long-term stable coating on the silver nanoparticles. The polymer layer coated was adequately thin to efficiently achieve plasmonic enhancement of fluorescence and also thick enough to effectively suppress quenching of fluorescence, achieving a huge net enhancement of fluorescence. The polymer-coated plasmonic nanoparticles are a promising platform for demonstrating highly sensitive biosensing for medical diagnostics.
RESUMO
The sulfated polysaccharide dermatan sulfate (DS) forms proteoglycans with a number of distinct core proteins. Iduronic acid-containing domains in DS have a key role in mediating the functions of DS proteoglycans. Two tissue-specific DS epimerases, encoded by DSE and DSEL, and a GalNAc-4-O-sulfotransferase encoded by CHST14 are necessary for the formation of these domains. CHST14 mutations were previously identified for patients with the musculocontractural type of Ehlers-Danlos syndrome (MCEDS). We now identified a homozygous DSE missense mutation (c.803C>T, p.S268L) by the positional candidate approach in a male child with MCEDS, who was born to consanguineous parents. Heterologous expression of mutant full-length and soluble recombinant DSE proteins showed a loss of activity towards partially desulfated DS. Patient-derived fibroblasts also showed a significant reduction in epimerase activity. The amount of DS disaccharides was markedly decreased in the conditioned medium and the cell fraction from cultured fibroblasts of the patient when compared with a healthy control subject, whereas no apparent difference was observed in the chondroitin sulfate (CS) chains from the conditioned media. However, the total amount of CS disaccharides in the cell fraction from the patient was increased â¼1.5-fold, indicating an increased synthesis or a reduced conversion of CS chains in the cell fraction. Stable transfection of patient fibroblasts with a DSE expression vector increased the amount of secreted DS disaccharides. DSE deficiency represents a specific defect of DS biosynthesis. We demonstrate locus heterogeneity in MCEDS and provide evidence for the importance of DS in human development and extracellular matrix maintenance.
