Fasciola hepatica:development of monoclonal antibodies against somatic antigens and their characterization by ultrastructural localization of antibody binding

A series of monoclonal antibodies was prepared against tegumental and internal antigens of Fasciola hepatica by immunizing mice with whole adult-fluke homogenates prior to harvesting the splenic lymphocytes for fusion. Preliminary screening by the Indirect Fluorescent Antibody technique indicated the occurrence of discrete groups of monoclonals differing from one another in tissue-specificity but within which IFA labelling patterns were fairly consistent. Representative hybridomas for 5 of these groups were stabilized and used to produce ascites fluid in mice. By application of an immunogold labelling technique it was possible to map the distribution of antigens for which each monoclonal antibody had affinity throughout the tissues of 4-week and 12-week flukes. Several monoclonals specifically labelled antigenic determinants on the important tegumental antigen T1. However the distribution of gold colloid labelling suggested that epitopes other than that normally exposed to the infected host were recognized; and several monoclonals specifically attached to T1 antigen in the tegument of juvenile worms only. The glycocalyx of the gut and excretory system of flukes shared T1 antigenicity with the tegument. Monoclonal antibodies were produced against an internal immunogen associated with ribosomes and heterochromatin in active protein-producing cells, and against interstitial material of adult flukes. Monoclonals against antigens in parenchymal cell cytoplasm and in mature vitelline cells were recognized but the corresponding hybridomas were not stabilized.

Lucky Imaging of transiting planet host stars with LuckyCam

We obtained high-resolution, high-contrast optical imaging in the Sloan Digital Sky Survey i′ band with the LuckyCam camera mounted on the 2.56 m Nordic Optical Telescope, to search for faint stellar companions to 16 stars harbouring transiting exoplanets. The Lucky imaging technique uses very short exposures to obtain near diffraction-limited images yielding sub-arcsecond sensitivity, allowing us to search for faint stellar companions within the seeing disc of the primary planet host. Here, we report the detection of two candidate stellar companions to the planet host TrES-1 at separations <6.5 arcsec and we confirm stellar companions to CoRoT-2, CoRoT-3, TrES-2, TrES-4 and HAT-P-7 already known in the literature. We do not confirm the candidate companions to HAT-P-8 found via Lucky imaging by Bergfors et al., however, most probably because HAT-P-8 was observed in poor seeing conditions. Our detection sensitivity limits allow us to place constraints on the spectral types and masses of the putative bound companions to the planet host stars in our sample. If bound, the stellar companions identified in this work would provide stringent observational constraints to models of planet formation and evolution. In addition, these companions could affect the derived physical properties of the exoplanets in these systems.

The host galaxy of the super-luminous SN 2010gx and limits on explosive nickel-56 production

Super-luminous supernovae have a tendency to occur in faint host galaxies which are likely to have low mass and low metallicity. While these extremely luminous explosions have been observed from z=0.1 to 1.55, the closest explosions allow more detailed investigations of their host galaxies. We present a detailed analysis of the host galaxy of SN 2010gx (z=0.23), one of the best studied super-luminous type Ic supernovae. The host is a dwarf galaxy (M_g=-17.42+/-0.17) with a high specific star formation rate. It has a remarkably low metallicity of 12+log(O/H)=7.5+/-0.1 dex as determined from the detection of the [OIII] 4363 Angs line. This is the first reliable metallicity determination of a super-luminous stripped-envelope supernova host. We collected deep multi-epoch imaging with Gemini + GMOS between 240-560 days after explosion to search for any sign of radioactive nickel-56, which might provide further insights on the explosion mechanism and the progenitor's nature. We reach griz magnitudes of m_AB~26, but do not detect SN 2010gx at these epochs. The limit implies that any nickel-56 production was similar to or below that of SN 1998bw (a luminous type Ic SN that produced around 0.4 M_sun of nickel-56). The low volumetric rates of these supernovae (~10^-4 of the core-collapse population) could be qualitatively matched if the explosion mechanism requires a combination of low-metallicity (below 0.2 Z_sun), high progenitor mass (>60 M_sun) and high rotation rate (fastest 10% of rotators).

The poxvirus protein A52R targets Toll-like receptor signaling complexes to suppress host defense

Toll-like receptors (TLRs) are crucial in the innate immune response to pathogens, in that they recognize and respond to pathogen associated molecular patterns, which leads to activation of intracellular signaling pathways and altered gene expression. Vaccinia virus (VV), the poxvirus used to vaccinate against smallpox, encodes proteins that antagonize important components of host antiviral defense. Here we show that the VV protein A52R blocks the activation of the transcription factor nuclear factor kappa B (NF-kappa B) by multiple TLRs, including TLR3, a recently identified receptor for viral RNA. A52R associates with both interleukin 1 receptor-associated kinase 2 (IRAK2) and tumor necrosis factor receptor-associated factor 6 (TRAF6), two key proteins important in TLR signal transduction. Further, A52R could disrupt signaling complexes containing these proteins. A virus deletion mutant lacking the A52R gene was attenuated compared with wild-type and revertant controls in a murine intranasal model of infection. This study reveals a novel mechanism used by VV to suppress the host immunity. We demonstrate viral disabling of TLRs, providing further evidence for an important role for this family of receptors in the antiviral response.

Production of a broad specificity antibody for the development and validation of an optical SPR screening method for free and intracellular microcystins and nodularin in cyanobacteria cultures

A highly sensitive broad specificity monoclonal antibody was produced and characterised for microcystin detection through the development of a rapid surface plasmon resonance (SPR) optical biosensor based immunoassay. The antibody displayed the following cross-reactivity: MC-LR 100%; MC-RR 108%; MC-YR 68%; MC-LA 69%; MC-LW 71%; MC-LF 68%; and Nodularin 94%. Microcystin-LR was covalently attached to a CM5 chip and with the monoclonal antibody was employed in a competitive 4min injection assay to detect total microcystins in water samples below the WHO recommended limit (1µg/L). A 'total microcystin' level was determined by measuring free and intracellular concentrations in cyanobacterial culture samples as this toxin is an endotoxin. Glass bead beating was used to lyse the cells as a rapid extraction procedure. This method was validated according to European Commission Decision 96/23/EC criteria. The method was proven to measure intracellular microcystin levels, the main source of the toxin, which often goes undetected by other analytical procedures and is advantageous in that it can be used for the monitoring of blooms to provide an early warning of toxicity. It was shown to be repeatable and reproducible, with recoveries from spiked samples ranging from 74 to 123%, and had % CVs below 10% for intra-assay analysis and 15% for inter-assay analysis. The detection capability of the assay was calculated as 0.5ng/mL for extracellular toxins and 0.05ng/mL for intracellular microcystins. A comparison of the SPR method with LC-MS/MS was achieved by testing six Microcystis aeruginosa cultures and this study yielded a correlation R(2) value of 0.9989.