Sialyl Residues Modulate LPS-Mediated Signaling through the Toll-Like Receptor 4 Complex

We previously reported that neuraminidase (NA) pretreatment of human PBMCs markedly increased their cytokine response to lipopolysaccharide (LPS). To study the mechanisms by which this occurs, we transfected HEK293T cells with plasmids encoding TLR4, CD14, and MD2 (three components of the LPS receptor complex), as well as a NFκB luciferase reporting system. Both TLR4 and MD2 encoded by the plasmids are α-2,6 sialylated. HEK293T cells transfected with TLR4/MD2/CD14 responded robustly to the addition of LPS; however, omission of the MD2 plasmid abrogated this response. Addition of culture supernatants from MD2 (sMD2)-transfected HEK293T cells, but not recombinant, non-glycosylated MD2 reconstituted this response. NA treatment of sMD2 enhanced the LPS response as did NA treatment of the TLR4/CD14-transfected cell supplemented with untreated sMD2, but optimal LPS-initiated responses were observed with NA-treated TLR4/CD14-transfected cells supplemented with NA-treated sMD2. We hypothesized that removal of negatively charged sialyl residues from glycans on the TLR4 complex would hasten the dimerization of TLR4 monomers required for signaling. Co-transfection of HEK293T cells with separate plasmids encoding either YFP- or FLAG-tagged TLR4, followed by treatment with NA and stimulation with LPS, led to an earlier and more robust time-dependent dimerization of TLR4 monomers on co-immunoprecipitation, compared to untreated cells. These findings were confirmed by fluorescence resonance energy transfer (FRET) analysis. Overexpression of human Neu1 increased LPS-initiated TLR4-mediated NFκB activation and a NA inhibitor suppressed its activation. We conclude that (1) sialyl residues on TLR4 modulate LPS responsiveness, perhaps by facilitating clustering of the homodimers, and that (2) sialic acid, and perhaps other glycosyl species, regulate MD2 activity required for LPS-mediated signaling. We speculate that endogenous sialidase activity mobilized during cell activation may play a role in this regulation

Nonlinear optics and saturation behavior of quantum dot samples under continuous wave driving

The nonlinear optical response of self-assembled quantum dots is relevant to the application of quantum dot based devices in nonlinear optics, all-optical switching, slow light and self-organization. Theoretical investigations are based on numerical simulations of a spatially and spectrally resolved rate equation model, which takes into account the strong coupling of the quantum dots to the carrier reservoir created by the wetting layer states. The complex dielectric susceptibility of the ground state is obtained. The saturation is shown to follow a behavior in between the one for a dominantly homogeneously and inhomogeneously broadened medium. Approaches to extract the nonlinear refractive index change by fringe shifts in a cavity or self-lensing are discussed. Experimental work on saturation characteristic of InGa/GaAs quantum dots close to the telecommunication O-band (1.24-1.28 mm) and of InAlAs/GaAlAs quantum dots at 780 nm is described and the first demonstration of the cw saturation of absorption in room temperature quantum dot samples is discussed in detail

Structural basis of TIR-domain-assembly formation in MAL- and MyD88-dependent TLR4 signaling

Toll-like receptor (TLR) signaling is a key innate immunity response to pathogens. Recruitment of signaling adapters such as MAL (TIRAP) and MyD88 to the TLRs requires Toll/interleukin-1 receptor (TIR)-domain interactions, which remain structurally elusive. Here we show that MAL TIR domains spontaneously and reversibly form filaments in vitro. They also form cofilaments with TLR4 TIR domains and induce formation of MyD88 assemblies. A 7-Å-resolution cryo-EM structure reveals a stable MAL protofilament consisting of two parallel strands of TIR-domain subunits in a BB-loop-mediated head-to-tail arrangement. Interface residues that are important for the interaction are conserved among different TIR domains. Although large filaments of TLR4, MAL or MyD88 are unlikely to form during cellular signaling, structure-guided mutagenesis, combined with in vivo interaction assays, demonstrated that the MAL interactions defined within the filament represent a template for a conserved mode of TIR-domain interaction involved in both TLR and interleukin-1 receptor signaling

Rapid assessment of influenza vaccine effectiveness: analysis of an internet-based cohort.

The effectiveness of influenza vaccination programmes is seldom known during an epidemic. We developed an internet-based system to record influenza-like symptoms and response to infection in a participating cohort. Using self-reports of influenza-like symptoms and of influenza vaccine history and uptake, we estimated vaccine effectiveness (VE) without the need for individuals to seek healthcare. We found that vaccination with the 2010 seasonal influenza vaccine was significantly protective against influenza-like illness (ILI) during the 2010-2011 influenza season (VE 52%, 95% CI 27-68). VE for individuals who received both the 2010 seasonal and 2009 pandemic influenza vaccines was 59% (95% CI 27-77), slightly higher than VE for those vaccinated in 2010 alone (VE 46%, 95% CI 9-68). Vaccinated individuals with ILI reported taking less time off work than unvaccinated individuals with ILI (3.4 days vs. 5.3 days, P<0.001)

Preliminary Results from NASA/GSFC Ka-Band High Rate Demonstration for Near-Earth Communications

In early 2000, the National Aeronautics and Space Administration (NASA) commenced the Ka-Band Transition Project (KaTP) as another step towards satisfying wideband communication requirements of the space research and earth exploration-satellite services. The KaTP team upgraded the ground segment portion of NASA's Space Network (SN) in order to enable high data rate space science and earth science services communications. The SN ground segment is located at the White Sands Complex (WSC) in New Mexico. NASA conducted the SN ground segment upgrades in conjunction with space segment upgrades implemented via the Tracking and Data Relay Satellite (TDRS)-HIJ project. The three new geostationary data relay satellites developed under the TDRS-HIJ project support the use of the inter-satellite service (ISS) allocation in the 25.25-27.5 GHz band (the 26 GHz band) to receive high speed data from low earth-orbiting customer spacecraft. The TDRS H spacecraft (designated TDRS-8) is currently operational at a 171 degrees west longitude. TDRS I and J spacecraft on-orbit testing has been completed. These spacecraft support 650 MHz-wide Ka-band telemetry links that are referred to as return links. The 650 MHz-wide Ka-band telemetry links have the capability to support data rates up to at least 1.2 Gbps. Therefore, the TDRS-HIJ spacecraft will significantly enhance the existing data rate elements of the NASA Space Network that operate at S-band and Ku-band

Immunoblots with rhodopsin antisera suggest that a purified mu opioid binding protein has structural characteristics of a G-protein-coupled receptor.

A mu opioid binding protein (OBP), previously purified to homogeneity from bovine striatal membranes, was examined by immunoblotting with six antisera against bovine rhodopsin. An antibody against the carboxyl-terminal tail of rhodopsin and one against membrane-associated rhodopsin gave strong signals at the appropriate molecular mass (65 kDa). An antibody directed against the first cytoplasmic loop of rhodopsin was weakly reactive. Three other antibodies did not recognize OBP. This pattern of crossreactivity was identical to that previously seen with beta-adrenergic receptors. The existence of domains in the OBP, which are antigenically similar to those in two other guanine nucleotide regulatory protein-coupled receptors, supports the hypothesis that mu opioid receptors have the structure characteristic of this receptor family