Depletion of four nitrofuran antibiotics and their tissue-bound metabolites in porcine tissues and determination using LC-MS/MS and HPLC-UV

Depletion of the nitrofuran antibiotics furazolidone, furaltadone, nitrofurantoin and nitrofurazone and their tissue-bound metabolites AOZ, AMOZ, AHD and SEM from pig muscle, liver and kidney tissues is described. Groups of pigs were given feed medicated with one of the nitrofuran drugs at a therapeutic concentration (400 mg kg(-1)) for ten days. Animals were slaughtered at intervals and tissue samples collected for analysis for six weeks following withdrawal of medicated feed. These samples were analysed both for parent nitrofurans (using LC-MS/MS and HPLC-UV), and for tissue-bound metabolites (using LC-MS/MS). The parent drugs were detectable only sporadically and only in pigs subjected to no withdrawal period whatsoever. This confirms the instability of the four major nitrofuran antibiotics in edible tissues. In contrast, the metabolites accumulated to high concentrations in tissues (ppm levels) and had depletion half lives of between 5.5 and 15.5 days. The metabolites of all four drugs were still readily detectable in tissues six weeks after cessation of treatment. This emphasizes the benefits of monitoring for the stable metabolites of the nitrofurans.

Monoclonal antibody-based ELISA for the quantification of nitrofuran metabolite 3-amino-2-oxazolidinone in tissues using a simplified sample preparation

A sensitive and specific monoclonal ELISA for the determination of tissue bound furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) is described. The procedure enables the detection of AOZ in matrix supernatant after homogenisation, protease treatment, acid hydrolysis and derivatisation of AOZ released from the tissue by o-nitrobenzaldehyde. The formed p-nitrophenyl 3-amino-2-oxazolidinone (NPAOZ) is determined by ELISA calibrated with matrix-matched standards in the concentration range of 0.05-5.0 mu g l(-1). The assay was validated according to criteria set down by Commission Decision 2002/657/EC for the performance and validation of analytical methods for chemical residues. Detection capability, set on the basis of acceptance of no false negative results, was 0.4 mu g kg(-1) for shrimp, poultry, beef and pork muscle. This sensitivity approaches the established confirmatory LC-MS/MS able to quantify tissue-bound AOZ at levels as low as 0.3 mu g kg(-1). An excellent correlation of results obtained by ELISA and LC/MS-MS within the concentration range 0-32.1 mu g kg(-1) was found in the naturally contaminated shrimp samples (r = 0.999, n = 8). A similar con-elation was found for the incurred poultry samples within the concentration range of 0-10.5 mu g kg(-1) (r = 0.99, n = 8). (c) 2005 Elsevier B.V All rights reserved.

Risk assessment of potentially toxic elements in agricultural soils and maize tissues from selected districts in Tanzania

A field survey was conducted to investigate the contamination of potentially toxic elements (PTEs) arsenic (As), lead (Pb), chromium (Cr), and nickel (Ni) in Tanzanian agricultural soils and to evaluate their uptake and translocation in maize as proxy to the safety of maize used for human and animal consumption. Soils and maize tissues were sampled from 40 farms in Tanzania and analyzed using inductively coupled plasma-mass spectrometry in the United Kingdom. The results showed high levels of PTEs in both soils and maize tissues above the recommended limits. Nickel levels of up to 34.4 and 56.9mgkg(-1) respectively were found in some maize shoots and grains from several districts. Also, high Pb levels >0.2mgkg(-1) were found in some grains. The grains and shoots with high levels of Ni and Pb are unfit for human and animal consumption. Concentrations of individual elements in maize tissues and soils did not correlate and showed differences in uptake and translocation. However, Ni showed a more efficient transfer from soils to shoots than As, Pb and Cr. Transfer of Cr and Ni from shoots to grains was higher than other elements, implying that whatever amount is assimilated in maize shoots is efficiently mobilized and transferred to grains. Thus, the study recommended to the public to stop consuming and feeding their animals maize with high levels of PTEs for their safety.

Potential hazard to human health from exposure to fragments of lead bullets and shot in the tissues of game animals

Lead is highly toxic to animals. Humans eating game killed using lead ammunition generally avoid swallowing shot or bullets and dietary lead exposure from this source has been considered low. Recent evidence illustrates that lead bullets fragment on impact, leaving small lead particles widely distributed in game tissues. Our paper asks whether lead gunshot pellets also fragment upon impact, and whether lead derived from spent gunshot and bullets in the tissues of game animals could pose a threat to human health.

Analysis of nine NSAIDs in ungulate tissues available to critically endangered vultures in India

In 2006, India, Pakistan, and Nepal banned the manufacture of veterinary formulations of the nonsteroidal anti-inflammatory drug (NSAID) diclofenac. This action was taken to halt the unprecedented decline of three Gyps vulture species that were being poisoned by diclofenac residues commonly present in carcasses of domestic livestock upon which they scavenged. To assess the affect of this ban and evaluate residue prevelances of other NSAIDs, we present a method to detect diclofenac and eight more NSAIDs by liquid chromatography-mass spectrometry and apply this to 1488 liver samples from carcasses of livestock taken across seven Indian states. Diclofenac was present in 11.1% of samples taken between April and December 2006, and meloxicam (4%), ibuprofen (0.6%), and ketoprofen (0.5%) were also detected. Although meloxicam is safe for a range of avian scavengers, including Gypsvultures, data regarding the safety of other NSAIDs is currently limited. If wild Gyps on the Indian subcontinent are to survive, diclofenac bans must be completely effective, and NSAIDs that replace it within the veterinary drug market must be of low toxicity toward Gyps and other scavenging birds.

The distribution of arsenic in the body tissues of wood mice and bank voles

Arsenic is accumulated by free-living small mammals, but there is little information on the resultant concentrations in different tissues other than liver and kidney. Such information is important because the severity of toxicological effects may be related to the amount of arsenic accumulated in specific organs, and the availability of arsenic to predators is, in part, dependent on which tissues accumulate arsenic. The objective of this study was to quantify the arsenic concentrations and the percentage of the total body burden (%TBB) accumulated in different body tissues of free-living small mammals and to determine how these factors varied with severity of habitat contamination. Arsenic concentrations were measured in various tissues of wood mice (Apodemus sylvaticus) and bank voles (Clethrionomys glareolus) from a range of arsenic-contaminated sites in southwest Britain. Arsenic concentrations in the gastrointestinal (GI) tract (including contents), liver, kidneys, spleen, lung, femur, and fur of both species varied significantly between sites and were higher in mice and voles from heavily contaminated areas. Heart and brain arsenic concentrations did not vary with degree of environmental contamination. The GI tract and excised carcass contained roughly equal amounts of arsenic and, in sum, comprised 75-85% of the TBB on uncontaminated sites and 90-99% on contaminated sites. Although the excised carcass contains about half of the TBB, its importance in food-chain transfer of arsenic to predators may depend on the bioavailability of arsenic sequestered in fur. In contrast, the GI tract and its contents, provided that it is consumed, will always be a major transfer pathway for arsenic to predators, regardless of the severity of habitat contamination.

Optimisation of time/temperature treatment, for heat treated soft wheat flour

Chlorination of wheat flour in the EU countries has been replaced in recent years, to some extent, by heat treated flour which is used to produce high ratio cakes. Heat treated flour allows high ratio recipes to be developed which generate products with longer shelf life, finer texture, moist crumb and sweeter taste. The mechanism by which heat treatment improves the flour is not fully understood, but it is known that during the heat treatment process, protein denaturation and partial gelatinisation of the starch granules occurs, as well as an increase in batter viscosity. Therefore, it is important to optimize the flour heat treatment process, in order to enhance baking quality. Laboratory preparation of heat treated base wheat flour (culinary, soft, low protein) was carried out in a fluidised bed drier using a range of temperatures and times. The gluten was extracted from the final product and its quality was tested, to obtain objective and comparative information on the extent of protein denaturation. The results indicated that heat treatment of flour decreases gluten extensibility and partial gelatinisation of the starch granules occurred. After heat treatment the gluten appeared to retain moisture. The optimum time/temperature for the heat treatment of base flour was 120-130°C for 30 min with moisture content of ˜12.5%.© 2012 Elsevier Ltd. All rights reserved.

The (Soft) Power of Commitment:The EU and Conflict Resolution in Northern Ireland

This paper examines the potential of international actors to contribute to conflict resolution by analysing the evolving role of the European Union (EU) in embedding Northern Ireland's peace process. Scrutiny of the EU's approach to conflict resolution in Northern Ireland offers useful insights into the scope and potential of soft power for facilitating behavioural change from governmental to grass-roots levels. This paper traces the development of the EU's approach to conflict resolution in Northern Ireland from one concentrated on encouraging state-level agreement, to nurturing peace through multilevel funding, through now to consolidating the peace by facilitating regional-level empowerment. The core argument is that, in sum, the most critical element of the EU's contribution to peace in Northern Ireland has been, quite simply, that of enduring commitment.

On the relativistic iron line and soft excess in the Seyfert 1 galaxy Markarian 335

We report a 133-ks XMM-Newton observation of the Seyfert 1 galaxy Markarian 335. The 0.4-12 keV spectrum contains an underlying power-law continuum, a soft excess below 2 keV, and a double-peaked iron emission feature in the 6-7 keV range. We investigate the possibility that the double-peaked emission might represent the characteristic signature of the accretion disc. Detailed investigations show that a moderately broad accretion disc line is most likely present, but that the peaks may be due to narrower components from more distant material. The peaks at 6.4 and 7 keV can be identified, respectively, with the molecular torus in active galactic nucleus unification schemes, and very highly ionized, optically thin gas filling the torus. The X-ray variability spectra on both long (~100 ks) and short (~1 ks) time-scales do not support the recent suggestion that the soft excess is an artefact of variable, moderately ionized absorption. © 2007 The Authors. Journal compilation © 2007 RAS.

Spatial autocorrelation in the response of soft-bottom marine benthos to gas extraction activities: The case of amphipods in the Ionian Sea

The spatial distributions of marine fauna and of pollution are both highly structured, and thus the resulting high levels of autocorrelation may invalidate conclusions based on classical statistical approaches. Here we analyse the close correlation observed between proxies for the disturbance associated with gas extraction activities and amphipod distribution patterns around four hydrocarbon platforms. We quantified the amount of variation independently accounted for by natural environmental variables, proxies for the disturbance caused by platforms, and spatial autocorrelation. This allowed us to demonstrate how each of these three factors significantly affects the community structure of amphipods. Sophisticated statistical techniques are required when taking into account spatial autocorrelation: nevertheless our data demonstrate that this approach not only enables the formulation of robust statistical inferences but also provides a much deeper understanding of the subtle interactions between human disturbance and natural factors affecting the structure of marine invertebrates communities. (C) 2012 Elsevier Ltd. All rights reserved.

RUNX3 acts as a tumor suppressor in breast cancer by targeting estrogen receptor α

Transcription factor RUNX3 is inactivated in a number of malignancies, including breast cancer, and is suggested to function as a tumor suppressor. How RUNX3 functions as a tumor suppressor in breast cancer remains undefined. Here, we show that about 20% of female Runx3(+/-) mice spontaneously developed ductal carcinoma at an average age of 14.5 months. Additionally, RUNX3 inhibits the estrogen-dependent proliferation and transformation potential of ERa-positive MCF-7 breast cancer cells in liquid culture and in soft agar and suppresses the tumorigenicity of MCF-7 cells in severe combined immunodeficiency mice. Furthermore, RUNX3 inhibits ERa-dependent transactivation by reducing the stability of ERa. Consistent with its ability to regulate the levels of ERa, expression of RUNX3 inversely correlates with the expression of ERa in breast cancer cell lines, human breast cancer tissues and Runx3(+/-) mouse mammary tumors. By destabilizing ERa, RUNX3 acts as a novel tumor suppressor in breast cancer.

Evaluation of a substructured soft Real-time Hybrid Test for performing Seismic analysis of complex structural systems

The hybrid test method is a relatively recently developed dynamic testing technique that uses numerical modelling combined with simultaneous physical testing. The concept of substructuring allows the critical or highly nonlinear part of the structure that is difficult to numerically model with accuracy to be physically tested whilst the remainder of the structure, that has a more predictable response, is numerically modelled. In this paper, a substructured soft-real time hybrid test is evaluated as an accurate means of performing seismic tests of complex structures. The structure analysed is a three-storey, two-by-one bay concentrically braced frame (CBF) steel structure subjected to seismic excitation. A ground storey braced frame substructure whose response is critical to the overall response of the structure is tested, whilst the remainder of the structure is numerically modelled. OpenSees is used for numerical modelling and OpenFresco is used for the communication between the test equipment and numerical model. A novel approach using OpenFresco to define the complex numerical substructure of an X-braced frame within a hybrid test is also presented. The results of the hybrid tests are compared to purely numerical models using OpenSees and a simulated test using a combination of OpenSees and OpenFresco. The comparative results indicate that the test method provides an accurate and cost effective procedure for performing
full scale seismic tests of complex structural systems.