Mitigation options for sediment and phosphorus loss from winter-sown arable crops

Sediment and P inputs to freshwaters from agriculture are a major problem in the United Kingdom (UK). This study investigated mitigation options for diffuse pollution losses from arable land. Field trials were undertaken at the hillslope scale over three winters at three UK sites with silt (Oxyaquic Hapludalf), sand (Udic Haplustept), and clay (Typic Haplaquept) soils. None of the mitigation treatments was effective in every year trialled, but each showed overall average reductions in losses. Over five site years, breaking up the compaction in tramlines (tractor wheel tracks) using a tine reduced losses of sediment and P to losses similar to those observed from areas without tramlines, with an average reduction in P loss of 1.06 kg TP ha(-1) Compared to traditional plowing, TP losses under minimum tillage were reduced by 0.30 kg TT ha(-1) over five site years, TP losses under contour cultivation were reduced by 0.30 kg TP ha(-1) over two site years, and TP losses using in-field barriers were reduced by 0.24 kg TP ha(-1) over two site years. In one site year, reductions in losses due to crop residue incorporation were nor significant. Each of the mitigation options trialled. is associated with a small cost at the farm-scale of up to 5 pound ha(-1), or with cost savings. The results indicate that each of the treatments his the potential to be a cost-effective mitigation option, but that tramline management is the most promising treatment, because tramlines dominate sediment and P transfer in surface runoff from arable hillslopes.

The effects of minimal tillage, contour cultivation, in-field vegetative barriers and crop residues on phosphorus and sediment yields

Runoff, sediment, total phosphorus and total dissolved phosphorus losses in overland flow were measured for two years on unbounded plots cropped with wheat and oats. Half of the field was cultivated with minimum tillage (shallow tillage with a tine cultivator) and half was conventionally ploughed. Within each cultivation treatment there were different treatment areas (TAs). In the first year of the experiment, one TA was cultivated up and down the slope, one TA was cultivated on the contour, with a beetle bank acting as a vegetative barrier partway up the slope, and one had a mixed direction cultivation treatment, with cultivation and drilling conducted up and down the slope and all subsequent operations conducted on the contour. In the second year, this mixed treatment was replaced with contour cultivation. Results showed no significant reduction in runoff, sediment losses or total phosphorus losses from minimum tillage when compared to the conventional plough treatment, but there were increased losses of total dissolved phosphorus with minimum tillage. The mixed direction cultivation treatment increased surface runoff and losses of sediment and phosphorus. Increasing surface roughness with contour cultivation reduced surface runoff compared to up and down slope cultivation in both the plough and minimum tillage treatment areas, but this trend was not significant. Sediment and phosphorus losses in the contour cultivation treatment followed a very similar pattern to runoff. Combining contour cultivation with a vegetative barrier in the form of a beetle bank to reduce slope length resulted in a non-significant reduction in surface runoff, sediment and total phosphorus when compared to up and down slope cultivation, but there was a clear trend towards reduced losses. However, the addition of a beetle bank did not provide a significant reduction in runoff, sediment losses or total phosphorus losses when compared to contour cultivation, suggesting only a marginal additional benefit. The economic implications for farmers of the different treatment options are investigated in order to assess their suitability for implementation at a field scale.

Lack of myostatin results in excessive muscle growth but impaired force generation

The lack of myostatin promotes growth of skeletal muscle, and blockade of its activity has been proposed as a treatment for various muscle-wasting disorders. Here, we have examined two independent mouse lines that harbor mutations in the myostatin gene, constitutive null (Mstn(-/-)) and compact (Berlin High Line, BEH(c/c)). We report that, despite a larger muscle mass relative to age-matched wild types, there was no increase in maximum tetanic force generation, but that when expressed as a function of muscle size (specific force), muscles of myostatin-deficient mice were weaker than wild-type muscles. In addition, Mstn(-/-) muscle contracted and relaxed faster during a single twitch and had a marked increase in the number of type IIb fibers relative to wild-type controls. This change was also accompanied by a significant increase in type IIB fibers containing tubular aggregates. Moreover, the ratio of mitochondrial DNA to nuclear DNA and mitochondria number were decreased in myostatin-deficient muscle, suggesting a mitochondrial depletion. Overall, our results suggest that lack of myostatin compromises force production in association with loss of oxidative characteristics of skeletal muscle.

Generation of candidate human influenza vaccine strains in cell culture: rehearsing the European response to an H7N1 pandemic threat

Background: Although H5N1 avian influenza viruses pose the most obvious imminent pandemic threat, there have been several recent zoonotic incidents involving transmission of H7 viruses to humans. Vaccines are the primary public health defense against pandemics, but reliance on embryonated chickens eggs to propagate vaccine and logistic problems posed by the use of new technology may slow our ability to respond rapidly in a pandemic situation. Objectives: We sought to generate an H7 candidate vaccine virus suitable for administration to humans whose generation and amplification avoided the use of eggs. Methods: We generated a suitable H7 vaccine virus by reverse genetics. This virus, known as RD3, comprises the internal genes of A/Puerto Rico/8/34 with surface antigens of the highly pathogenic avian strain A/Chicken/Italy/13474/99 (H7N1). The multi-basic amino acid site in the HA gene, associated with high pathogenicity in chickens, was removed. Results: The HA modification did not alter the antigenicity of the virus and the resultant single basic motif was stably retained following several passages in Vero and PER. C6 cells. RD3 was attenuated for growth in embryonated eggs, chickens, and ferrets. RD3 induced an antibody response in infected animals reactive against both the homologous virus and other H7 influenza viruses associated with recent infection by H7 viruses in humans. Conclusions: This is the first report of a candidate H7 vaccine virus for use in humans generated by reverse genetics and propagated entirely in mammalian tissue culture. The vaccine has potential use against a wide range of H7 strains.

Generation of anti-complement "prodrugs": cleavable reagents for specific delivery of complement regulators to disease sites

Expression of biologically active molecules as fusion proteins with antibody Fc can substantially extend the plasma half-life of the active agent but may also influence function. We have previously generated a number of fusion proteins comprising a complement regulator coupled to Fc and shown that the hybrid molecule has a long plasma half-life and retains biological activity. However, several of the fusion proteins generated had substantially reduced biological activity when compared with the native regulator or regulator released from the Fc following papain cleavage. We have taken advantage of this finding to engineer a prodrug with low complement regulatory activity that is cleaved at sites of inflammation to release active regulator. Two model prodrugs, comprising, respectively, the four short consensus repeats of human decay accelerating factor (CD55) linked to IgG4 Fc and the three NH2-terminal short consensus repeats of human decay accelerating factor linked to IgG2 Fc have been developed. In each, specific cleavage sites for matrix metalloproteinases and/or aggrecanases have been incorporated between the complement regulator and the Fc. These prodrugs have markedly decreased complement inhibitory activity when compared with the parent regulator in vitro. Exposure of the prodrugs to the relevant enzymes, either purified, or in supernatants of cytokine-stimulated chondrocytes or in synovial fluid, efficiently cleaved the prodrug, releasing active regulator. Such agents, having negligible systemic effects but active at sites of inflammation, represent a paradigm for the next generation of anti-C therapeutics.

Specificity grafting of human antibody frameworks selected from a phage display library: generation of a highly stable humanized anti-CD22 single-chain Fv fragment

A prerequisite for the enrichment of antibodies screened from phage display libraries is their stable expression on a phage during multiple selection rounds. Thus, if stringent panning procedures are employed, selection is simultaneously driven by antigen affinity, stability and solubility. To take advantage of robust pre-selected scaffolds of such molecules, we grafted single-chain Fv (scFv) antibodies, previously isolated from a human phage display library after multiple rounds of in vitro panning on tumor cells, with the specificity of the clinically established murine monoclonal anti-CD22 antibody RFB4. We show that a panel of grafted scFvs retained the specificity of the murine monoclonal antibody, bound to the target antigen with high affinity (6.4-9.6 nM), and exhibited exceptional biophysical stability with retention of 89-93% of the initial binding activity after 6 days of incubation in human serum at 37degreesC. Selection of stable human scaffolds with high sequence identity to both the human germline and the rodent frameworks required only a small number of murine residues to be retained within the human frameworks in order to maintain the structural integrity of the antigen binding site. We expect this approach may be applicable for the rapid generation of highly stable humanized antibodies with low immunogenic potential.

Stress generation in the tension wood of poplar is based on the lateral swelling power of the G-layer

The mechanism of active stress generation in tension wood is still not fully understood. To characterize the functional interdependency between the G-layer and the secondary cell wall, nanostructural characterization and mechanical tests were performed on native tension wood tissues of poplar (Populus nigra x Populus deltoids) and on tissues in which the G-layer was removed by an enzymatic treatment. In addition to the well-known axial orientation of the cellulose fibrils in the G-layer, it was shown that the microfibril angle of the S2-layer was very large (about 36 degrees). The removal of the G-layer resulted in an axial extension and a tangential contraction of the tissues. The tensile stress-strain curves of native tension wood slices showed a jagged appearance after yield that could not be seen in the enzyme-treated samples. The behaviour of the native tissue was modelled by assuming that cells deform elastically up to a critical strain at which the G-layer slips, causing a drop in stress. The results suggest that tensile stresses in poplar are generated in the living plant by a lateral swelling of the G-layer which forces the surrounding secondary cell wall to contract in the axial direction.