Plasmonic Nanotrough Networks for Scalable Bacterial Raman Biosensing.

We demonstrate a novel approach for fabricating surface enhanced Raman scattering (SERS) substrates for single bacterial biosensing based on Ag cylindrical nanotrough networks (CNNs). This approach is developed with large scalability by leveraging a cellulose nanofiber template fabrication via facile electrospinning. Specifically, a concave nanotrough structure consisting of interconnected concave Ag nanoshells is demonstrated by depositing a thin layer of Ag atop a sacrificial electrospun nanofiber template and then completely removing the cellulose core in water. Our investigations of the scattering properties and SERS performances of single isolated Ag nanotroughs of different diameters reveal that nanotrough-based substrates provide tunable optical responses and enhanced SERS intensities. Further, Ag CNNs are fabricated in highly interconnected networks that yield reproducible SERS signals for molecular monolayers and whole bacterial cells, enabling rapid spectral discrimination between different bacterial strains. Finally, by performing principal component analysis on a large number of measured SERS spectra (40 spectra per bacterium), we demonstrate successful spectral discrimination between two types of Escherichia coli ( E. coli) bacteria, that is, E. coli K12 with its derivative E. coli DH 5α and E. coli BL21(DE3). The demonstrated cost-effective substrates feature several advantages over conventional SERS substrates including environmentally friendly and scalable fabrication compatible with versatile devices and provide an alternative approach to rapid SERS detection and screening of biochemicals.

Functional Interplay Between Murine Leukemia Virus Glycogag, Serinc5, and Surface Glycoprotein Governs Virus Entry, with Opposite Effects on Gammaretroviral and Ebolavirus Glycoproteins.

Gammaretroviruses, such as murine leukemia viruses (MLVs), encode, in addition to the canonical Gag, Pol, and Env proteins that will form progeny virus particles, a protein called "glycogag" (glycosylated Gag). MLV glycogag contains the entire Gag sequence plus an 88-residue N-terminal extension. It has recently been reported that glycogag, like the Nef protein of HIV-1, counteracts the antiviral effects of the cellular protein Serinc5. We have found, in agreement with prior work, that glycogag strongly enhances the infectivity of MLVs with some Env proteins but not those with others. In contrast, however, glycogag was detrimental to MLVs carrying Ebolavirus glycoprotein. Glycogag could be replaced, with respect to viral infectivity, by the unrelated S2 protein of equine infectious anemia virus. We devised an assay for viral entry in which virus particles deliver the Cre recombinase into cells, leading to the expression of a reporter. Data from this assay showed that both the positive and the negative effects of glycogag and S2 upon MLV infectivity are exerted at the level of virus entry. Moreover, transfection of the virus-producing cells with a Serinc5 expression plasmid reduced the infectivity and entry capability of MLV carrying xenotropic MLV Env, particularly in the absence of glycogag. Conversely, Serinc5 expression abrogated the negative effects of glycogag upon the infectivity and entry capability of MLV carrying Ebolavirus glycoprotein. As Serinc5 may influence cellular phospholipid metabolism, it seems possible that all of these effects on virus entry derive from changes in the lipid composition of viral membranes. IMPORTANCE: Many murine leukemia viruses (MLVs) encode a protein called "glycogag." The function of glycogag is not fully understood, but it can assist HIV-1 replication in the absence of the HIV-1 protein Nef under some circumstances. In turn, Nef counteracts the cellular protein Serinc5. Glycogag enhances the infectivity of MLVs with some but not all MLV Env proteins (which mediate viral entry into the host cell upon binding to cell surface receptors). We now report that glycogag acts by enhancing viral entry and that, like Nef, glycogag antagonizes Serinc5. Surprisingly, the effects of glycogag and Serinc5 upon the entry and infectivity of MLV particles carrying an Ebolavirus glycoprotein are the opposite of those observed with the MLV Env proteins. The unrelated S2 protein of equine infectious anemia virus (EIAV) is functionally analogous to glycogag in our experiments. Thus, three retroviruses (HIV-1, MLV, and EIAV) have independently evolved accessory proteins that counteract Serinc5.