The Enterococcus faecalis Exoproteome: Identification and Temporal Regulation by Fsr

Analysis of the culture supernatant exoproteins produced by two PFGE clusters of high-level gentamicin and ciprofloxacin-resistant clinical isolates of Enterococcus faecalis from the UK and Ireland revealed two distinct protein profiles. This grouping distinguished OG1RF and GelE metalloprotease-expressing isolates from JH2-2 and other GelE-negative isolates. The integrity of the fsrABDC operon was found to determine the exoproteome composition, since an fsrB mutant of strain OG1RF appeared very similar to that of strain JH2-2, and complementation of the latter with the fsrABDC operon produced an OG1RF-like exoproteome. The proteins present in the supernatant fraction of OG1RF were separated using 2D gels and identified by mass spectrometry and comprised many mass and pI variants of the GelE and SprE proteases. In addition cell wall synthesis and cell division proteins were identified. An OG1RF fsrB mutant had a distinct exoprotein fraction with an absence of the Fsr-regulated proteases and was characterised by general stress and glycolytic proteins. The exoproteome of the OG1RF fsrB mutant resembles that of a divIVA mutant of E. faecalis, suggestive of a stress phenotype

Wildlife and pollution: 2001-02 Annual Report

The Wildlife and Pollution contract covers a long-term monitoring programme, the Predatory Bird Monitoring Scheme (PBMS), that examines the levels of certain pollutants in selected wildlife species in Britain. The programme was started in the early 1960s, when there were serious concerns over the effects of organochlorine insecticides and organomercury fungicides on various species of birds and mammals. This early work demonstrated the effects of the organochlorines and eventually contributed to the ban on their use in the UK and abroad. The programme has subsequently assessed the success of these bans by measuring whether there has been a decline in the concentrations of organochlorine pesticides in the livers and eggs of predatory and freshwater fish-eating birds. Investigations have also been made into the levels of industrial polychlorinated biphenyls (PCBs), following their identification as pollutants in 1966. Mercury levels, derived from both agricultural and industrial sources, have also been tracked, although mercury concentrations were not measured in birds collected in 2001. In recent years, investigations have been made into the effects of the newest generation of rodenticides on barn owls Tyto alba. Northern gannet Morus bassanus eggs are also collected approximately biennially from two colonies and, when available, from other sites; eggs were last collected in 2000. This programme is now the longest-running of its kind anywhere in the world and the findings stimulate considerable interest internationally, as well as in Britain. Annual reports give an interim summary of results and every three years these annual results are gathered together into a more substantial report in which they are integrated with previous findings. The latest report of this type covers the period up to and including 2000 (Shore et al. 2005). Results are published periodically in the scientific literature. This current report presents the results of analyses carried out on material collected in 2001. It also includes a review of long-term trends in second-generation anticoagulant rodenticide residues in barn owls that occurred during the monitoring period up to and including the year 2001. The Wildlife and Pollution contract was the subject of scientific assessment within JNCC's rolling programme of peer review in autumn 1993 and was further assessed in 1996. As a result of the last two assessments, some monitoring was curtailed. Most notably, common kestrels Falco tinnunculus are no longer monitored for organochlorines. However, from 2001 onwards, kestrels will be monitored for second-generation anticoagulant rodenticides following the results from an individual study, carried out as part of the PBMS activities, which demonstrated that this species may be particularly vulnerable to exposure to these compounds (Shore et al. 2001). Carcasses and eggs of predatory bird species (such as peregrine falcon Falco peregrinus, common buzzard Buteo buteo, long-eared owl Asio otus, little owl Athene noctua, common kingfisher Alcedo atthis, great crested grebe Podiceps cristatus, and great bittern Botaurus stellaris) which do not form the core part of the PBMS, but are sent to the Centre for Ecology & Hydrology (CEH) by volunteers, are not analysed chemically. However, post-mortem examinations are carried out the carcasses, relevant information is recorded and the cause of death is determined (and reported back to the volunteer who submitted the carcass). Samples of the egg contents and body organs for these species, and samples for the species that do form part of the core monitoring, are all archived at -20°C as part of CEH's unique long-term tissue bank, and are often used in specific targeted research studies in subsequent years. Each section within the Wildlife and Pollution contract is summarised below. Each is dependent on the provision of material from amateur naturalists and other interested parties, and it is not always possible to obtain desired material for analysis, especially from remote areas

Wildlife and pollution: 2000-01 Annual Report

The Wildlife and Pollution contract covers a long-term monitoring programme that examines the levels of certain pollutants in selected wildlife species in Britain. The programme was started in the early 1960s, when there were serious concerns over the effects of organochlorine insecticides and organomercury fungicides on various species of birds and mammals. This early work demonstrated the effects of the organochlorines, and eventually contributed to the ban on their use in the UK and abroad. The programme has measured levels of these compounds in predatory and fish-eating birds since then. Investigations have also been made into the levels of industrial polychlorinated biphenyls (PCBs), following their identification as pollutants in 1966. Mercury levels, derived from both agricultural and industrial sources, have also been tracked. In recent years, investigations have been made into the effects of the latest generation of rodenticides on barn owls Tyto alba. Northern gannet Morus bassanus eggs are also collected approximately biennially from two colonies and, when available, from other sites; eggs were collected from one site in 2000. This programme is now the longest-running of its kind anywhere in the world and the findings stimulate considerable interest internationally, as well as in Britain. Annual reports give an interim summary of results. This current report presents the results of analyses carried out on material collected in 2000. Every three years these annual results are gathered together into a more substantial report in which they are integrated with previous findings. The last report of this type covered the period up to and including 1997 (Newton et al. 1998) and is updated here. The present report summarises the long-term trends in all contaminants (except rodenticides, analysed in Shore et al. 2005) that occurred during the monitoring period up to and including the year 2000. Results are published periodically in the scientific literature, and recent key papers are listed in the references to the present report. The Wildlife and Pollution contract was the subject of scientific assessment within JNCC's rolling programme of peer review in autumn 1993 and was further assessed in 1996. As a result of the last two assessments, some monitoring was curtailed. Specifically, common kestrels Falco tinnunculus were no longer monitored for organochlorines, although from 2001 this species will be monitored annually for second-generation anticoagulant rodenticides. The Centre for Ecology & Hydrology (CEH) still collects specimens for studying other contaminants as part of its core research programme. Similarly, other species (peregrine falcon Falco peregrinus, common buzzard Buteo buteo, long-eared owl Asio otus, little owl Athene noctua, common kingfisher Alcedo atthis, great crested grebe Podiceps cristatus, and great bittern Botaurus stellaris) that were received in small numbers in occasional years were also not analysed routinely, although some were analysed in specific one-off studies and tissues from all birds received at CEH in 2000 are archived in deep-freeze for future potential analyses. Each section within the Wildlife and Pollution contract is summarised below. Each is dependent on the provision of material from amateur naturalists and other interested parties, and it is not always possible to obtain desired material for analysis, especially from remote areas

WESTT (Workload, Error, Situational Awareness, Time and Teamwork): An analytical prototyping system for command and control

Modern developments in the use of information technology within command and control allow unprecedented scope for flexibility in the way teams deal with tasks. These developments, together with the increased recognition of the importance of knowledge management within teams present difficulties for the analyst in terms of evaluating the impacts of changes to task composition or team membership. In this paper an approach to this problem is presented that represents team behaviour in terms of three linked networks (representing task, social network structure and knowledge) within the integrative WESTT software tool. In addition, by automating analyses of workload and error based on the same data that generate the networks, WESTT allows the user to engage in the process of rapid and iterative “analytical prototyping”. For purposes of illustration an example of the use of this technique with regard to a simple tactical vignette is presented

Combined chelation therapy in thalassemia major for the treatment of severe myocardial siderosis with left ventricular dysfunction

<p>Abstract</p> <p>Background</p> <p>In thalassemia major (TM), severe cardiac siderosis can be treated by continuous parenteral deferoxamine, but poor compliance, complications and deaths occur. Combined chelation therapy with deferiprone and deferoxamine is effective for moderate myocardial siderosis, but has not been prospectively examined in severe myocardial siderosis.</p> <p>Methods</p> <p>T2* cardiovascular magnetic resonance (CMR) was performed in 167 TM patients receiving standard subcutaneous deferoxamine monotherapy, and 22 had severe myocardial siderosis (T2* < 8 ms) with impaired left ventricular (LV) function. Fifteen of these patients received combination therapy with subcutaneous deferoxamine and oral deferiprone with CMR follow-up.</p> <p>Results</p> <p>At baseline, deferoxamine was prescribed at 38 ± 10.2 mg/kg for 5.3 days/week, and deferiprone at 73.9 ± 4.0 mg/kg/day. All patients continued both deferiprone and deferoxamine for 12 months. There were no deaths or new cardiovascular complications. The myocardial T2* improved (5.7 ± 0.98 ms to 7.9 ± 2.47 ms; p = 0.010), with concomitant improvement in LV ejection fraction (51.2 ± 10.9% to 65.6 ± 6.7%; p < 0.001). Serum ferritin improved from 2057 (CV 7.6%) to 666 (CV 13.2%) μg/L (p < 0.001), and liver iron improved (liver T2*: 3.7 ± 2.9 ms to 10.8 ± 7.3 ms; p = 0.006).</p> <p>Conclusion</p> <p>In patients with severe myocardial siderosis and impaired LV function, combined chelation therapy with subcutaneous deferoxamine and oral deferiprone reduces myocardial iron and improves cardiac function. This treatment is considerably less onerous for the patient than conventional high dose continuous subcutaneous or intravenous deferoxamine monotherapy, and may be considered as an alternative. Very prolonged tailored treatment with iron chelation is necessary to clear myocardial iron, and alterations in chelation must be guided by repeated myocardial T2* scans.</p> <p>Trial registration</p> <p>This trial is registered as NCT00103753</p

Right ventricular volumes and function in thalassemia major patients in the absence of myocardial iron overload

<p>Abstract</p> <p>Aim</p> <p>We aimed to define reference ranges for right ventricular (RV) volumes, ejection fraction (EF) in thalassemia major patients (TM) without myocardial iron overload.</p> <p>Methods and results</p> <p>RV volumes, EF and mass were measured in 80 TM patients who had no myocardial iron overload (myocardial T2* > 20 ms by cardiovascular magnetic resonance). All patients were receiving deferoxamine chelation and none had evidence of pulmonary hypertension or other cardiovascular comorbidity. Forty age and sex matched healthy non-anemic volunteers acted as controls. The mean RV EF was higher in TM patients than controls (males 66.2 ± 4.1% vs 61.6 ± 6%, p = 0.0009; females 66.3 ± 5.1% vs 62.6 ± 6.4%, p = 0.017), which yielded a raised lower threshold of normality for RV EF in TM patients (males 58.0% vs 50.0% and females 56.4% vs 50.1%). RV end-diastolic volume index was higher in male TM patients (mean 98.1 ± 17.3 mL vs 88.4 ± 11.2 mL/m2, p = 0.027), with a higher upper limit (132 vs 110 mL/m2) but this difference was of borderline significance for females (mean 86.5 ± 13.6 mL vs 80.3 ± 12.8 mL/m2, p = 0.09, with upper limit of 113 vs 105 mL/m2). The cardiac index was raised in TM patients (males 4.8 ± 1.0 L/min vs 3.4 ± 0.7 L/min, p < 0.0001; females 4.5 ± 0.8 L/min vs 3.2 ± 0.8 L/min, p < 0.0001). No differences in RV mass index were identified.</p> <p>Conclusion</p> <p>The normal ranges for functional RV parameters in TM patients with no evidence of myocardial iron overload differ from healthy non-anemic controls. The new reference RV ranges are important for determining the functional effects of myocardial iron overload in TM patients.</p

Development and geometry of isotropic and directional shrinkage crack patterns

We have studied shrinkage crack patterns which form when a thin layer of an alumina/water slurry dries. Both isotropic and directional drying were studied. The dynamics of the pattern formation process and the geometric properties of the isotropic crack patterns are similar to what is expected from recent models, assuming weak disorder. There is some evidence for a gradual increase in disorder as the drying layer become thinner, but no sudden transition, in contrast to what has been seen in previous experiments. The morphology of the crack patterns is influenced by drying gradients and front propagation effects, with sharp gradients having a strong orienting and ordering effect.Comment: 8 pages, 11 figures, 8 in jpg format, 3 in postscript. See also http://mobydick.physics.utoronto.ca/mud.htm