Characterization and proteome profiling of extracellular vesicles in a murine model of Staphylococcus aureus endophthalmitis.

Endophthalmitis is a vision-threatening complication of intraocular surgery or penetrating injury of which Staphylococcus aureus is an important etiological agent. Extracellular vesicles (EVs) hold a tremendous possibility for developing diagnostic and therapeutic biomarkers due to their role in the pathogenesis of various infections. The aim of this study was to characterise the protein cargo of EVs, isolated from a murine (C57BL/6) model of S. aureus endophthalmitis by LC-MS/MS. Contralateral eye injected with sterile media served as control and both eyes were enucleated after 24 h, followed by extraction of EVs by ultracentrifugation. EVs were characterized by DLS and western blotting with tetraspanin markers, CD9 and CD81 and quantified by ExoCet quantification kit. Proteomic analysis identified 1964 proteins (FDR ≤ 0.01) in EVs from infected mice eyes, of which 40 proteins varied significantly in their amounts in comparison to EVs obtained from control eyeballs (P-value ≤ 0.05). The results of this study provide insight into the global EV proteome of S. aureus endophthalmitis with their functional correlation and differential protein amounts between infected and control set. Annexin A5, cathepsin D and C5a play a pivotal role in disease pathogenesis and could possibly play a role as a prognostic marker in endophthalmitis.

Proteomic profiling of exosomes in a mouse model of Candida albicans endophthalmitis.

Exosomes play pivotal roles in intercellular communication, and pathophysiological functions. In this study, we aimed to understand the role of exosomal proteome derived from C. albicans infected mice (C57BL/6) eyeball. Exosomes were characterized by Dynamic Light Scattering and Western blot, quantified and subjected to LC-MS/MS and cytokine quantification by ELISA. The average size of exosomes was 170-200 nm with number of exosomes amounted to 1.42 × 1010 in infected set compared to control (1.24 × 109). Western blot was positive for CD9, CD63 and CD81 confirming the presence of exosomes. IL-6, IL1ß, TNF-α, and IFN-γ levels were significantly elevated in infected eye at 72 h.p.i. Proteomic analysis identified 42 differentially expressed proteins, of these 37 were upregulated and 5 were downregulated. Gene Ontology (GO) revealed enrichment of cell adhesion, cytoskeleton organisation, and cellular response proteins such as aquaporin-5, gasdermin-A, CD5 antigen-like, Catenin, V-ATPase, and vesicle associated protein. Additionally, KEGG pathway analysis indicated the association of metabolic and carbon signalling pathways with exosomes from C. albicans infected eye. The protein cargo in exosomes released during endophthalmitis with C. albicans seems to play a unique role in the pathogenesis of the disease and further validations with larger cohort of patients is required to confirm them as biomarkers.

ZnO nanoparticles embedded silk fibroin-a piezoelectric composite for nanogenerator applications.

This paper demonstrates a flexible nanogenerator (NG) using Silk-Zinc Oxide (ZnO) composite by exploiting the inherent piezoelectric properties of silk and ZnO. A direct precipitation method was employed to synthesize Zinc Oxide nanoparticles (NPs). Silk-ZnO composite film was then prepared by spin-coating the homogenous silk-ZnO solution. The composition and morphology of silk-ZnO composite were analyzed using various standard characterization procedures. The biocompatibility study of the composite film was also performed through cell viability testing. The utility of as prepared composites was demonstrated through the fabrication of piezoelectric nanogenerator. This hybrid nanogenerator was capable to generate a maximum open circuit voltage of 25 V (peak to peak value) in the bending state for a specific ZnO concentration. The output response of the nanogenerator exhibited a good correlation with the bending angle of the device. A peak outputpower density of 6.67 mW cm-3was achieved from the nanogenerator. The fabricated prototype is efficient to light-up commercial red LEDs and to harvest energy from human body movement. The piezoelectric coefficient (d33) of silk-ZnO composite film was also experimentally figured out.