Methods for nanoparticle labeling of ricin and effect on toxicity

The unique optical properties associated with nanostructured materials that support the excitation of surface plasmons offer many new opportunities for the enhanced optical investigation of biological materials that pose a security threat. In particular, ricin is considered a significant bioterrorism risk due to its high toxicity combined with its ready availability as a byproduct in castor oil production. Therefore, the development of optical techniques capable of rapid on-site toxin detection with high molecular specificity and sensitivity continues to be of significant importance. Furthermore, understanding of the ricin cell entry and intracellular pathways remains poor due to a lack of suitable bioanalytical techniques. Initial work aimed at simultaneously tackling both these issues is described where different approaches for the nanoparticle labeling of ricin are investigated along with changes in ricin toxicity associated with the labeling process

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Web‐Teaching ‐ A Guide to Interactive Teaching for the World‐Wide Web by David W. Brooks, New York: Plenum, 1997. ISBN: 0–306–45552–8. Paperback, 214 pages. $30

Seabed corrugations beneath an Antarctic ice shelf revealed by autonomous underwater vehicle survey: Origin and implications for the history of Pine Island Glacier

Ice shelves are critical features in the debate about West Antarctic ice sheet change and sea level rise, both because they limit ice discharge and because they are sensitive to change in the surrounding ocean. The Pine Island Glacier ice shelf has been thinning rapidly since at least the early 1990s, which has caused its trunk to accelerate and retreat. Although the ice shelf front has remained stable for the past six decades, past periods of ice shelf collapse have been inferred from relict seabed "corrugations" (corrugated ridges), preserved 340 km from the glacier in Pine Island Trough. Here we present high-resolution bathymetry gathered by an autonomous underwater vehicle operating beneath an Antarctic ice shelf, which provides evidence of long-term change in Pine Island Glacier. Corrugations and ploughmarks on a sub-ice shelf ridge that was a former grounding line closely resemble those observed offshore, interpreted previously as the result of iceberg grounding. The same interpretation here would indicate a significantly reduced ice shelf extent within the last 11 kyr, implying Holocene glacier retreat beyond present limits, or a past tidewater glacier regime different from today. The alternative, that corrugations were not formed in open water, would question ice shelf collapse events interpreted from the geological record, revealing detail of another bed-shaping process occurring at glacier margins. We assess hypotheses for corrugation formation and suggest periodic grounding of ice shelf keels during glacier unpinning as a viable origin. This interpretation requires neither loss of the ice shelf nor glacier retreat and is consistent with a "stable" grounding-line configuration throughout the Holocene

The sedimentary legacy of a palaeo-ice stream on the shelf of the southern Bellingshausen Sea: Clues to West Antarctic glacial history during the Late Quaternary

A major trough ("Belgica Trough") eroded by a palaeo-ice stream crosses the continental shelf of the southern Bellingshausen Sea (West Antarctica) and is associated with a trough mouth fan ("Belgica TMF") on the adjacent continental slope. Previous marine geophysical and geological studies investigated the bathymetry and geomorphology of Belgica Trough and Belgica TMF, erosional and depositional processes associated with bedform formation, and the temporal and spatial changes in clay mineral provenance of subglacial and glaciomarine sediments. Here, we present multi-proxy data from sediment cores recovered from the shelf and uppermost slope in the southern Bellingshausen Sea and reconstruct the ice-sheet history since the last glacial maximum (LGM) in this poorly studied area of West Antarctica. We combined new data (physical properties, sedimentary structures, geochemical and grain-size data) with published data (shear strength, clay mineral assemblages) to refine a previous facies classification for the sediments. The multi-proxy approach allowed us to distinguish four main facies types and to assign them to the following depositional settings: 1) subglacial, 2) proximal grounding-line, 3) distal sub-ice shelf/sub-sea ice, and 4) seasonal open-marine. In the seasonal open-marine fades we found evidence for episodic current-induced winnowing of near-seabed sediments on the middle to outer shelf and at the uppermost slope during the late Holocene. In addition, we obtained data on excess Pb-210 activity at three core sites and 44 AMS C-14 dates from the acid-insoluble fraction of organic matter (AIO) and calcareous (micro-) fossils, respectively, at 12 sites. These chronological data enabled us to reconstruct, for the first time, the timing of the last advance and retreat of the West Antarctic Ice Sheet (WAIS) and the Antarctic Peninsula Ice Sheet (APIS) in the southern Bellingshausen Sea. We used the down-core variability in sediment provenance inferred from clay mineral changes to identify the most reliable AIO C-14 ages for ice-sheet retreat. The palaeo-ice stream advanced through Belgica Trough after similar to 36.0 corrected C-14 ka before present (B.P.). It retreated from the outer shelf at similar to 25.5 ka B.P, the middle shelf at similar to 19.8 ka B.P., the inner shelf in Eltanin Bay at similar to 12.3 ka B.P., and the inner shelf in Ronne Entrance at similar to 6.3 ka B.P. The retreat of the WAIS and APIS occurred slowly and stepwise, and may still be in progress. This dynamical ice-sheet behaviour has to be taken into account for the interpretation of recent and the prediction of future mass-balance changes in the study area. The glacial history of the southern Bellingshausen Sea is unique when compared to other regions in West Antarctica, but some open questions regarding its chronology need to be addressed by future work. (C) 2010 Elsevier Ltd. All rights reserved

LoCuSS:the connection between brightest cluster galaxy activity, gas cooling and dynamical disturbance of X-ray cluster cores

We study the distribution of projected offsets between the cluster X-ray centroid and the brightest cluster galaxy (BCG) for 65 X-ray-selected clusters from the Local Cluster Substructure Survey, with a median redshift of z= 0.23. We find a clear correlation between X-ray/BCG projected offset and the logarithmic slope of the cluster gas density profile at 0.04r500(α), implying that more dynamically disturbed clusters have weaker cool cores. Furthermore, there is a close correspondence between the activity of the BCG, in terms of detected Hα and radio emission, and the X-ray/BCG offset, with the line-emitting galaxies all residing in clusters with X-ray/BCG offsets of ≤15 kpc. Of the BCGs with α < −0.85 and an offset <0.02r500, 96 per cent (23/24) have optical emission and 88 per cent (21/24) are radio active, while none has optical emission outside these criteria. We also study the cluster gas fraction (fgas) within r500 and find a significant correlation with X-ray/BCG projected offset. The mean fgas of the ‘small offset’ clusters (<0.02r500) is 0.106 ± 0.005 (σ= 0.03) compared to 0.145 ± 0.009 (σ= 0.04) for those with an offset >0.02r500, indicating that the total mass may be systematically underestimated in clusters with larger X-ray/BCG offsets. Our results imply a link between cool core strength and cluster dynamical state consistent with the view that cluster mergers can significantly perturb cool cores, and set new constraints on models of the evolution of the intracluster medium

Assigning function to genome wide association study variants associated with complex gastrointestinal disease

PhDThe genome‐wide association study era has identified numerous loci associated with many common polygenic diseases. The next challenge is to identify the functional consequences of these variants and elicit how they impact on disease risk. Using a combination of protein based assays, large scale microarrays and high‐throughput generation sequencing platforms this thesis aims to identify the functional effects of disease loci, with particular focus on Crohn’s disease and coeliac disease, two common complex gastrointestinal diseases. Variants located within the Interleukin 23 receptor are associated with both susceptibility and protection from Crohn’s disease, a debilitating chronic inflammatory disease of the bowel. A study was undertaken to investigate the effect of these variants, at the mRNA as well as the protein level, on both cytokine and receptor levels. Coeliac disease is a dietary intolerance to the gluten component of wheat, barley and rye and has an estimated prevalence of approximately 1%. Genome‐wide association studies have identified eight genomic different loci as associated with coeliac disease but none have been functionally characterised. To investigate the effect that genotype has on gene transcript levels, a genetical genomics study was undertaken in patients with coeliac disease generating results with relevance to a range of autoimmune disorders. Before disease based effects can be identified, it is first important to fully characterise the normal human transcriptome and methylome. To this end CD4 + T cells were studied using novel high‐throughput sequencing techniques, with the aim of providing some insight into novel genomic properties that may illuminate current and future disease associated loci. Given the base pair resolution approach of high‐throughput sequencing, a novel method of assaying for SNP effects on gene expression was developed. This allele specific method, using whole transcriptome sequencing, is capable of identifying alterations in transcript expression on a genome‐wide scale

Functional studies of signaling pathways in peri-implantation development of the mouse embryo by RNAi.

BACKGROUND: Studies of gene function in the mouse have relied mainly on gene targeting via homologous recombination. However, this approach is difficult to apply in specific windows of time, and to simultaneously knock-down multiple genes. Here we report an efficient method for dsRNA-mediated gene silencing in late cleavage-stage mouse embryos that permits examination of phenotypes at post-implantation stages. RESULTS: We show that introduction of Bmp4 dsRNA into intact blastocysts by electroporation recapitulates the genetic Bmp4 null phenotype at gastrulation. It also reveals a novel role for Bmp4 in the regulation the anterior visceral endoderm specific gene expression and its positioning. We also show that RNAi can be used to simultaneously target several genes. When applied to the three murine isoforms of Dishevelled, it leads to earlier defects than previously observed in double knock-outs. These include severe delays in post-implantation development and defects in the anterior midline and neural folds at headfold stages. CONCLUSION: Our results indicate that the BMP4 signalling pathway contributes to the development of the anterior visceral endoderm, and reveal an early functional redundancy between the products of the murine Dishevelled genes. The proposed approach constitutes a powerful tool to screen the functions of genes that govern the development of the mouse embryo.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Systemic inflammatory response syndrome after major abdominal surgery predicted by early upregulation of TLR4 and TLR5

OBJECTIVES To study innate immune pathways in patients undergoing hepatopancreaticobiliary surgery to understand mechanisms leading to enhanced inflammatory responses and identifying biomarkers of adverse clinical consequences. BACKGROUND Patients undergoing major abdominal surgery are at risk of life-threatening systemic inflammatory response syndrome (SIRS) and sepsis. Early identification of at-risk patients would allow tailored postoperative care and improve survival. METHODS Two separate cohorts of patients undergoing major hepatopancreaticobiliary surgery were studied (combined n = 69). Bloods were taken preoperatively, on day 1 and day 2 postoperatively. Peripheral blood mononuclear cells and serum were separated and immune phenotype and function assessed ex vivo. RESULTS Early innate immune dysfunction was evident in 12 patients who subsequently developed SIRS (postoperative day 6) compared with 27 who did not, when no clinical evidence of SIRS was apparent (preoperatively or days 1 and 2). Serum interleukin (IL)-6 concentration and monocyte Toll-like receptor (TLR)/NF-κB/IL-6 functional pathways were significantly upregulated and overactive in patients who developed SIRS (P < 0.0001). Interferon α-mediated STAT1 phosphorylation was higher preoperatively in patients who developed SIRS. Increased TLR4 and TLR5 gene expression in whole blood was demonstrated in a separate validation cohort of 30 patients undergoing similar surgery. Expression of TLR4/5 on monocytes, particularly intermediate CD14CD16 monocytes, on day 1 or 2 predicted SIRS with accuracy 0.89 to 1.0 (areas under receiver operator curves). CONCLUSIONS These data demonstrate the mechanism for IL-6 overproduction in patients who develop postoperative SIRS and identify markers that predict patients at risk of SIRS 5 days before the onset of clinical signs