Lysosome-associated membrane glycoprotein (LAMP)--preliminary study on a hidden antigen target for vaccination against schistosomiasis

Our previously reported gene atlasing of schistosome tissues revealed transcripts that were highly enriched in the digestive tract of Schistosoma mansoni. From these, we selected two candidates, Sm-LAMP and Sm-NPC2 for testing as vaccine targets. The two molecules were selected on the basis of relatively high expression in the gastrodermis, their potentially important biological function, divergence from homologous molecules of the host and possible apical membrane expression in the gastrodermis. Bacterially expressed recombinant peptides corresponding to regions excluding trans-membrane domains of the selected vaccine targets were used in blinded vaccine trials in CBA mice using alum-CpG as adjuvant. Vaccine trials using the recombinant insoluble Sm-LAMP protein showed 16-25% significant reduction in total worm burden. Faecal egg count reduction was 52% and 60% in two trials, respectively, with similar results for the solubly expressed protein. Liver egg burden was reduced significantly (20% and 38%) with an insoluble recombinant Sm-LAMP in two trials, but not with the soluble recombinant form. Parasite fecundity was not affected by either Sm-LAMP protein preparations in the trials. It is concluded that Sm-LAMP may provide limited protection towards S. mansoni infections but could be used in combination with other vaccine candidates, to provide more comprehensive protection.

New Observations of Balmer Continuum Flux in Solar Flares:Instrument Description and First Results

Increase in the Balmer continuum radiation during solar flares was predicted by various authors, but has never been firmly confirmed observationally using ground-based slit spectrographs. Here we describe a new post-focal instrument, the image selector, with which the Balmer continuum flux can be measured from the whole flare area, in analogy to successful detections of flaring dMe stars. The system was developed and put into operation at the horizontal solar telescope HSFA2 of the Ondřejov Observatory. We measure the total flux by a fast spectrometer from a limited but well-defined region on the solar disk. Using a system of diaphragms, the disturbing contribution of a bright solar disk can be eliminated as much as possible. Light curves of the measured flux in the spectral range 350 – 440 nm are processed, together with the Hα images of the flaring area delimited by the appropriate diaphragm. The spectral flux data are flat-fielded, calibrated, and processed to be compared with model predictions. Our analysis of the data proves that the described device is sufficiently sensitive to detect variations in the Balmer continuum during solar flares. Assuming that the Balmer-continuum kernels have at least a similar size as those visible in Hα, we find the flux increase in the Balmer continuum to reach 230 – 550 % of the quiet continuum during the observed X-class flare. We also found temporal changes in the Balmer continuum flux starting well before the onset of the flare in Hα.

LOTUS: A low cost, ultraviolet spectrograph

We describe the design, construction and commissioning of LOTUS; a simple, low-cost long-slit spectrograph for the Liverpool Telescope. The design is optimized for near-UV and visible wavelengths and uses all transmitting optics. It exploits the instrument focal plane field curvature to partially correct axial chromatic aberration. A stepped slit provides narrow (2.5x95 arcsec) and wide (5x25 arcsec) options that are optimized for spectral resolution and flux calibration respectively. On sky testing shows a wavelength range of 3200-6300 Angstroms with a peak system throughput (including detector quantum efficiency) of 15 per cent and wavelength dependant spectral resolution of R=225-430. By repeated observations of the symbiotic emission line star AG Peg we demonstrate the wavelength stability of the system is less than 2 Angstroms rms and is limited by the positioning of the object in the slit. The spectrograph is now in routine operation monitoring the activity of comet 67P/Churyumov-Gerasimenko during its current post-perihelion apparition.

A Loop-mediated Isothermal Amplification Method for Rapid Direct Detection and Differentiation of Non-pathogenic and Verocytotoxigenic Escherichia coli in Beef and Bovine Faeces

The aim of this study was to develop a multiplex loop-mediated isothermal amplification (LAMP) method capable of detecting Escherichia coli generally and verocytotoxigenic E. coli (VTEC) specifically in beef and bovine faeces. The LAMP assay developed was highly specific (100%) and able to distinguish between E. coli and VTEC based on the amplification of the phoA, and stx1 and/or stx2 genes, respectively. In the absence of an enrichment step, the limit of detection 50% (LOD50) of the LAMP assay was determined to be 2.83, 3.17 and 2.83-3.17 log CFU/g for E. coli with phoA, stx1 and stx2 genes, respectively, when artificially inoculated minced beef and bovine faeces were tested. The LAMP calibration curves generated with pure cultures, and spiked beef and faeces, suggested that the assay had good quantification capability. Validation of the assay, performed using retail beef and bovine faeces samples, demonstrated good correlation between counts obtained by the LAMP assay and by a conventional culture method, but suggested the possibility of false negative LAMP results for 12.5-14.7% of samples tested. The multiplex LAMP assay developed potentially represents a rapid alternative to culture for monitoring E.coli levels in beef or faeces and it would provide additional information on the presence of VTEC. However, some further optimisation is needed to improve detection sensitivity.