The purification and properties of phosphonoacetate hydrolase, a novel carbon-phosphorus bond-cleavage enzyme from Pseudomonas fluorescens 23F

A novel, inducible, carbon-phosphorus bond-cleavage enzyme, phosphonoacetate hydrolase, was purified from cells of Pseudomonas fluorescens 23F grown phosphonoacetate. The native enzyme had a molecular mass of approximately 80 kDa and, upon SDS/PAGE, yielded a homogenous protein band with an apparent molecular mass of about 38 kDa. Activity of purified phosphonoacetate hydrolase was Zn2+ dependent and showed pH and temperature optima of approximately 7.8 and 37 degrees C, respectively. The purified enzyme had an apparent K-m of 1.25 mM for its sole substrate phosphonoacetate, and was inhibited by the structural analogues 3-phosphonopropionate and phosphonoformate. The NH2-terminal sequence of the first 19 amino acids displayed no significant similarity to other databank sequences.

The use and limitation of realistic evaluation as a tool for evidence-based practice: a critical realist perspective

In this paper, we assess realistic evaluation’s articulation with evidence-based practice (EBP) from the perspective of critical realism. We argue that the adoption by realistic evaluation of a realist causal ontology means that it is better placed to explain complex healthcare interventions than the traditional method used by EBP, the randomized controlled trial (RCT). However, we do not conclude from this that the use of RCTs is without merit, arguing that it is possible to use both methods in combination under the rubric of realist theory. More negatively, we contend that the rejection of critical theory and utopianism by realistic evaluation in favour of the pragmatism of piecemeal social engineering means that it is vulnerable to accusations that it promotes technocratic interpretations of human problems. We conclude that, insofar as realistic evaluation adheres to the ontology of critical realism, it provides a sound contribution to EBP, but insofar as it rejects the critical turn of Bhaskar’s realism, it replicates the technocratic tendencies inherent in EBP.

Low flow water quality in rivers; septic tank systems and high-resolution phosphorus signals

Rural point sources of phosphorus (P), including septic tank systems, provide a small part of the overall phosphorus budget to surface waters in agricultural catchments but can have a disproportionate impact on the low flow P concentration of receiving rivers. This has particular importance as the discharges are approximately constant into receiving waters and these have restricted dilution capacity during ecologically sensitive summer periods. In this study, a number of identified high impact septic systems were replaced with modern sequential batch reactors in three rural catchments during a monitoring period of 4 years. Sub-hourly P monitoring was conducted using bankside-analysers. Results show that strategic replacement of defective septic tank systems with modern systems and polishing filters decreased the low flow P concentration of one catchment stream by 0.032 mg TP L- 1 (0.018 mg TRP L- 1) over the 4 years. However two of the catchment mitigation efforts were offset by continued new-builds that increased the density of septic systems from 3.4 km- 2 to 4.6 km- 2 and 13.8 km- 2 to 17.2 km- 2 and subsequently increased low flow P concentrations. Future considerations for septic system mitigation should include catchment carrying capacity as well as technology changes.

The mgtC gene of Burkholderia cenocepacia is required for growth under magnesium limitation conditions and intracellular survival in macrophages

Burkholderia cenocepacia, a bacterium commonly found in the environment, is an important opportunistic pathogen in patients with cystic fibrosis (CF). Very little is known about the mechanisms by which B. cenocepacia causes disease, but chronic infection of the airways in CF patients may be associated, at least in part, with the ability of this bacterium to survive within epithelial cells and macrophages. Survival in macrophages occurs in a membrane-bound compartment that is distinct from the lysosome, suggesting that B. cenocepacia prevents phagolysosomal fusion. In a previous study, we employed signature-tagged mutagenesis and an agar bead model of chronic pulmonary infection in rats to identify B. cenocepacia genes that are required for bacterial survival in vivo. One of the most significantly attenuated mutants had an insertion in the mgtC gene. Here, we show that mgtC is also needed for growth of B. cenocepacia in magnesium-depleted medium and for bacterial survival within murine macrophages. Using fluorescence microscopy, we demonstrated that B. cenocepacia mgtC mutants, unlike the parental isolate, colocalize with the fluorescent acidotropic probe LysoTracker Red. At 4 h postinfection, mgtC mutants expressing monomeric red fluorescent protein cannot retain this protein within the bacterial cytoplasm. Together, these results demonstrate that, unlike the parental strain, an mgtC mutant does not induce a delay in phagolysosomal fusion and the bacterium-containing vacuoles are rapidly targeted to the lysosome, where bacteria are destroyed.

Investigating the of Water treatment residuals on Phosphorus mobility derived from biosolid application to Willow coppice

Short rotation willow coppice (SRWC) treatment of biosolids is limited by the oversupply of biosolid derived phosphorus; this can lead to eventual losses of phosphorus to water. Water treatment residuals (WTR), a by-product of potable water treatment, have been identified as a viable soil amendment for mitigation of phosphorus loss. WTR exploit the capacity of internally held aluminium oxide-hydroxide complexes to immobilise labile phosphorus. However indiscriminate additions to plots can result in inadequate control or excessive immobilization of soluble P, leading to crop deficiencies. Four commercially grown common willow (Salix) genotypes (Terra Nova, Endeavour, Resolution and Tora) were grown in soil amended with WTR at five different application rates (0, 10, 25, 50 and 100 tonne ha-1 air-dry basis) in a glasshouse pot experiment. The effects of application rates on plant yields, tissue P concentrations, P uptake and soil labile P availability were measured. Results indicate labile P was reduced with increasing WTR application rates, without any negative agronomic impacts.

A PLATE ASSAY FOR THE DETECTION OF ORGANOPHOSPHONATE MINERALIZATION BY ENVIRONMENTAL BACTERIA, AND ITS MODIFICATION AS AN ACTIVITY STAIN FOR IDENTIFICATION OF THE CARBON-PHOSPHORUS BOND-CLEAVAGE ENZYME PHOSPHONOACETATE HYDROLASE

A replica plate screening technique, based on the acid molybdate assay for detection of phosphate has been developed to permit the detection of microorganisms capable of mineralizing organophosphonates. The method was further adapted as the basis of an activity stain for the detection of the carbon - phosphorus bond cleavage enzyme phosphonoacetate hydrolase in PAGE gels.

Neoadjuvant hormone therapy for radical prostate radiotherapy:bicalutamide monotherapy vs. luteinizing hormone-releasing hormone agonist monotherapy: a single-institution matched-pair analysis

The purpose of this study was to compare the prostate-specific antigen (PSA) response to either neoadjuvant bicalutamide (BC) monotherapy or neoadjuvant luteinizing hormone-releasing hormone agonist (LHRHa) monotherapy and the subsequent effect on biochemical failure-free survival (BFFS) in men receiving radical radiotherapy (RT) for localized prostate cancer.

Arsenic accumulation and phosphorus status in two rice (Oryza sativa L.) cultivars surveyed from fields in South China

The consumption of paddy rice (Oryza sativa L.) is a major inorganic arsenic exposure pathway in S.E. Asia. A multi-location survey was undertaken in Guangdong Province, South China to assess arsenic accumulation and speciation in 2 rice cultivars, one an Indica and the other a hybrid Indica. The results showed that arsenic concentrations in rice tissue increased in the order grain <husk <straw <root. Rice grain arsenic content of 2 rice cultivars was significant different and correlated with phosphorus concentration and molar ratio of P/As in shoot, being higher for the Indica cultivar than for the hybrid Indica, which suggests altering shoot phosphorus status as a promising route for breeding rice cultivars with reduced grain arsenic. Speciation of grain arsenic, performed using HPLC-ICP-MS, identified inorganic arsenic as the dominant arsenic species present in the rice grain.

Field and Laboratory Tests of Flow-Proportional Passive Samplers for Determining Average Phosphorus and Nitrogen Concentration in Rivers

Flow responsive passive samplers offer considerable potential in nutrient monitoring in catchments; bridging the gap between the intermittency of grab sampling and the high cost of automated monitoring systems. A commercially available passive sampler was evaluated in a number of river systems encapsulating a gradient in storm response, combinations of diffuse and point source pressures, and levels of phosphorus and nitrogen concentrations. Phosphorus and nitrogen are sequestered to a resin matrix in a permeable cartridge positioned in line with streamflow. A salt tracer dissolves in proportion to advective flow through the cartridge. Multiple deployments of different cartridge types were undertaken and the recovery of P and N compared with the flow-weighted mean concentration (FWMC) from high-resolution bank-side analysers at each site. Results from the passive samplers were variable and largely underestimated the FWMC derived from the bank-side analysers. Laboratory tests using ambient river samples indicated good replication of advective throughflow using pumped water, although this appeared not to be a good analogue of river conditions where flow divergence was possible. Laboratory tests also showed good nutrient retention but not elution and these issues appeared to combine to limit the utility in ambient river systems at the small catchment scale.

TEENAGERS AND ILLICIT DRUG-USE - EXPANDING THE USER VS NONUSER DICHOTOMY

The present study examines proximal and distal factors associated with the use and non-use of illegal substances within a sample of 860 teenagers in North Wales. Arguing that there is predictive utility in expanding the traditional 'users vs non-users' design dichotomy, four groups are identified-resistant and vulnerable non-users and experimental and repeated users. 'Person' variables (life satisfaction, deviance, hopelessness and drug-related attributions) appeared to primarily differentiate the vulnerable group from their resistant counterparts and identify this, as yet non-using group, with user samples. It is suggested that these variables might represent 'risk' factors for illicit substance use and that the group design employed suggests they precede, rather than follow as a consequence of, illicit drug use. Like their resistant counterparts however, the vulnerable group are differentiated from user samples on some lifestyle and context indices. It is argued that these represent 'protective' influences in an otherwise at-risk group of non-users. Variables associated with an escalation of illicit drug use are discussed in considering the differences between the experimental and repeated user groups. Apart from the more proximal factor of drug-related attributions, 'person' variables appeared less involved here. Repeated users did however, tend to use a greater number of drugs, have a greater proportion of friends who also used illegal substances and significantly fewer had a Welsh cultural identity.

Equal Security: Europe and the Strategic Arms Limitation Talks, 1968-1976

SALT threatened to institutionalize a bipolar world order. NWS and NNWS alike feared that the US and SU will prioritize global security principles such as systemic stability and conflict stability to Atlantic and European security. Endangered was Europe’s security and position in the future world order. Parity in strategic weapons invalidated the US nuclear umbrella. An ABM deployment and a non-transfer regime threatened Europe’s nuclear defence options. The danger of a Limited War or a denuclearization of Central Europe led to a European co-ordination on nuclear arms control to assure the preservation of the West and the future of Europe.

Influences of phosphorus starvation on OsACR2.1 expression and arsenic metabolism in rice seedlings

The effects of phosphorus (P) status on arsenate reductase gene (OsACR2.1) expression, arsenate reductase activity, hydrogen peroxide (H(2)O(2)) content, and arsenic (As) species in rice seedlings which were exposed to arsenate after -P or +P pretreatments were investigated in a series of hydroponic experiments. OsACR2.1 expression increased significantly with decreasing internal P concentrations; more than 2-fold and 10-fold increases were found after P starvation for 30 h and 14 days, respectively. OsACR2.1 expression exhibited a significant positive correlation with internal root H(2)O(2) accumulation, which increased upon P starvation or exposure to H(2)O(2) without P starvation. Characterization of internal and effluxed As species showed the predominant form of As was arsenate in P-starved rice root, which contrasted with the +P pretreated plants. Additionally, more As was effluxed from P-starved rice roots than from non-starved roots. In summary, an interesting relationship was observed between P-starvation induced H(2)O(2) and OsACR2.1 gene expression. However, the up-regulation of OsACR2.1 did not increase arsenate reduction in P-starved rice seedlings when exposed to arsenate.