Project description:The cellular vesicle is a fluid-filled structure separated from the surrounding environment by a biological membrane. Here, we isolated nanovesicles (NVs) from the juice of clementines using a discontinuous density gradient ultracentrifugation method. To gain information about the protein content of vesicles, mass spectrometry-based organelle proteomics and bioinformatics were applied to the exosome-like vesicle fraction isolated in the 1 mol/L sucrose/D2O cushion. Analysis of 1018 identified proteins revealed a highly complex mixture of different intra, extracellular and artificially-formed vesicle populations. In particular, clathrin-coated vesicles were significantly expressed in this sample. Membrane transporters are significantly represented in clementines nanovesicles. We have found 162 proteins associated with the transport Gene Ontology term (GO: 0006810) which includes; 71 transmembrane transport related, 53 vesicle mediated and 50 intracellular transporters. Platellin-3 like carrier protein containing a Sec14 domain is known to have a role in plant-virus interaction and that is one of the most abundant proteins in our dataset. The presence of transmembrane transporters like ATPases, aquaporins, ATP Binding Cassette (ABC) transporters and tetraspanins, regulators of protein trafficking suggests that nanovesicles of clementines can actively interact with their environment in a controlled way.
Project description:A novel antiinfective approach is to exploit stresses already imposed on invading organisms by the in vivo environment. Fe metabolism is a key vulnerability of infecting bacteria because organisms require Fe for growth, and it is critical in the pathogenesis of infections. Furthermore, humans have evolved potent Fe-withholding mechanisms that can block acute infection, prevent biofilm formation leading to chronic infection, and starve bacteria that succeed in infecting the host. Here we investigate a "Trojan horse" strategy that uses the transition metal gallium to disrupt bacterial Fe metabolism and exploit the Fe stress of in vivo environments. Due to its chemical similarity to Fe, Ga can substitute for Fe in many biologic systems and inhibit Fe-dependent processes. We found that Ga inhibits Pseudomonas aeruginosa growth and biofilm formation and kills planktonic and biofilm bacteria in vitro. Ga works in part by decreasing bacterial Fe uptake and by interfering with Fe signaling by the transcriptional regulator pvdS. We also show that Ga is effective in 2 murine lung infection models. These data, along with the fact that Ga is FDA approved (for i.v. administration) and there is the dearth of new antibiotics in development, make Ga a potentially promising new therapeutic for P. aeruginosa infections.
Project description:In this study, we report a green and economical hydrothermal synthesis of fluorescent-nitrogen-doped carbon quantum dots (NCQDs) using citrus lemon as a carbon source. The prepared NCQDs possess high water solubility, high ionic stability, resistance to photobleaching, and bright blue color under ultraviolet radiation with a high quantum yield (∼31%). High-resolution transmission electron microscopy (HRTEM) results show that the prepared NCQDs have a narrow size distribution (1-6 nm) with an average particle size of 3 nm. The mercury ion (Hg2+) sensing efficiency of the NCQDs was studied, and the result indicated that the material has high sensitivity, high precision, and good selectivity for Hg2+. The limit of detection (LOD) is 5.3 nM and the limit of quantification (LOQ) is 18.3 nM at a 99% confidence level. The cytotoxicity was evaluated using MCF7 cells, and the cell viabilities were determined to be greater than 88% upon the addition of NCQDs over a wide concentration range from 0 to 2 mg/mL. Based on the low cytotoxicity, good biocompatibility, and other revealed interesting merits, we also applied the prepared NCQDs as an effective fluorescent probe for multicolor live cell imaging.