Attomolar Detection of Botulinum Toxin Type A in Complex Biological Matrices

BACKGROUND: A highly sensitive, rapid and cost efficient method that can detect active botulinum neurotoxin (BoNT) in complex biological samples such as foods or serum is desired in order to 1) counter the potential bioterrorist threat 2) enhance food safety 3) enable future pharmacokinetic studies in medical applications that utilize BoNTs. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a botulinum neurotoxin serotype A assay with a large immuno-sorbent surface area (BoNT/A ALISSA) that captures a low number of toxin molecules and measures their intrinsic metalloprotease activity with a fluorogenic substrate. In direct comparison with the "gold standard" mouse bioassay, the ALISSA is four to five orders of magnitudes more sensitive and considerably faster. Our method reaches attomolar sensitivities in serum, milk, carrot juice, and in the diluent fluid used in the mouse assay. ALISSA has high specificity for the targeted type A toxin when tested against alternative proteases including other BoNT serotypes and trypsin, and it detects the holotoxin as well as the multi-protein complex form of BoNT/A. The assay was optimized for temperature, substrate concentration, size and volume proportions of the immuno-sorbent matrix, enrichment and reaction times. Finally, a kinetic model is presented that is consistent with the observed improvement in sensitivity. CONCLUSIONS/SIGNIFICANCE: The sensitivity, specificity, speed and simplicity of the BoNT ALISSA should make this method attractive for diagnostic, biodefense and pharmacological applications

International Ocean Discovery Program Expedition 360 preliminary report: Southwest Indian Ridge Lower Crust and Moho the nature of the lower crust and Moho at slower spreading ridges (SloMo Leg 1)

International Ocean Discovery Program (IODP) Expedition 360 was the first leg of Phase I of the SloMo (shorthand for "The nature of the lower crust and Moho at slower spreading ridges") Project, a multiphase drilling program that proposes to drill through the outermost of the global seismic velocity discontinuities, the Mohorovičić seismic discontinuity (Moho). The Moho corresponds to a compressional wave velocity increase, typically at ∼7 km beneath the oceans, and has generally been regarded as the boundary between crust and mantle. An alternative model, that the Moho is a hydration front in the mantle, has recently gained credence upon the discovery of abundant partially serpentinized peridotite on the seafloor and on the walls of fracture zones, such as at Atlantis Bank, an 11-13 My old elevated oceanic core complex massif adjacent to the Atlantis II Transform on the Southwest Indian Ridge. Hole U1473A was drilled on the summit of Atlantis Bank during IODP Expedition 360, 1-2 km away from two previous Ocean Drilling Program (ODP) holes: Hole 735B (drilled during ODP Leg 118 in 1987 and ODP Leg 176 in 1997) and Hole 1105A (drilled during ODP Leg 179 in 1998). A mantle peridotite/gabbro contact has been traced by dredging and diving along the transform wall for 40 km. The contact is located at ∼4200 m depth at the drill sites but shoals considerably 20 km to the south, where it was observed in outcrop at 2563 m depth. Moho reflections have, however, been found at ∼5-6 km beneath Atlantis Bank an

The Molecular Identification of Organic Compounds in the Atmosphere: State of the Art and Challenges

SSCI-VIDE+ATARI:CARE+BNO:BDAInternational audienc

The genome of the domesticated apple (Malus x domestica Borkh.)

We report a high-quality draft genome sequence of the domesticated apple (Malus x domestica). We show that a relatively recent (> 50 million years ago) genome-wide duplication (GWD) has resulted in the transition from nine ancestral chromosomes to 17 chromosomes in the Pyreae. Traces of older GWDs partly support the monophyly of the ancestral paleohexaploidy of eudicots. Phylogenetic reconstruction of Pyreae and the genus Malus, relative to major Rosaceae taxa, identified the progenitor of the cultivated apple as M. sieversii. Expansion of gene families reported to be involved in fruit development may explain formation of the pome, a Pyreae-specific false fruit that develops by proliferation of the basal part of the sepals, the receptacle. In apple, a subclade of MADS-box genes, normally involved in flower and fruit development, is expanded to include 15 members, as are other gene families involved in Rosaceae-specific metabolism, such as transport and assimilation of sorbitol