Modeling long-term compost application effects on nitrate leaching

Application of compost to agricultural soils may be beneficial for crop production by increasing soil fertility and supplying plant nutrients, however, any raise of the soil organic matter content may increase the potential for unintended groundwater contamination by nitrate leaching. In this paper, the effect of long-term compost applications on nitrate leaching, soil organic matter content, and crop production is analyzed using results of simulation scenarios for agricultural sites with loamy and sandy soils. Simulations were carried out using the Danish Nitrogen Simulation System (DAISY) which describes the nitrogen balance in a one-dimensional soil-plant-atmosphere system and considers compost type and application rate as well as management and cropping practice. Estimations of hydraulic and solute transport parameters are based on pedotransfer functions. Data from a 4.5-year period of field experiments with compost applications in northern Germany as well as from laboratory experiments with compost amended soil are used for model calibration. Simulation results suggest that: (i) with respect to nitrogen turnover the differences between compost types (i.e., non-matured and matured) are small compared to site-specific properties (i.e., soil temperature and water balance) and management practices (i.e., crop rotation) when considering a compost application period of 50 years; (ii) with respect to nitrate leaching the effect of different compost application scenarios is highly sensitive at the sand and relatively small at the loam site; and (iii) relatively high crop yields and acceptably low nitrate concentrations in the drainage water are obtained at the sand site when applying a combination of 10 t compost/ha/yr and soil- N_min adjusted mineral nitrogen additions of about 20 kg/ha/yr to a winter-grain dominated crop rotation. Further optimization may be possible by applying reduced rates of the (economically interesting) non-matured compost. This revised version was published online in June 2006 with corrections to the Cover Date. This revised version was published online in June 2006 with corrections to the Cover Date. This revised version was published online in June 2006 with corrections to the Cover Date.     

The Genomic Impacts of Drift and Selection for Hybrid Performance in Maize

Although maize is naturally an outcrossing organism, modern breeding utilizes highly inbred lines in controlled crosses to produce hybrids. The U.S. Department of Agriculture’s reciprocal recurrent selection experiment between the Iowa Stiff Stalk Synthetic (BSSS) and the Iowa Corn Borer Synthetic No. 1 (BSCB1) populations represents one of the longest running experiments to understand the response to selection for hybrid performance. To investigate the genomic impact of this selection program, we genotyped the progenitor lines and >600 individuals across multiple cycles of selection using a genome-wide panel of ∼40,000 SNPs. We confirmed previous results showing a steady temporal decrease in genetic diversity within populations and a corresponding increase in differentiation between populations. Thanks to detailed historical information on experimental design, we were able to perform extensive simulations using founder haplotypes to replicate the experiment in the absence of selection. These simulations demonstrate that while most of the observed reduction in genetic diversity can be attributed to genetic drift, heterozygosity in each population has fallen more than expected. We then took advantage of our high-density genotype data to identify extensive regions of haplotype fixation and trace haplotype ancestry to single founder inbred lines. The vast majority of regions showing such evidence of selection differ between the two populations, providing evidence for the dominance model of heterosis. We discuss how this pattern is likely to occur during selection for hybrid performance and how it poses challenges for dissecting the impacts of modern breeding and selection on the maize genome.     

Ultrasonographic images of liver neoplasms--comparison with histopathological findings

Ultrasound pictures taken in 100 patients with histologically confirmed primary of metastatic neoplasms of the liver were analysed retrospectively. The most frequent were hyperechogenic foci (43%). Hypoechogenic foci were found in 33% of the patients whereas mixed changes in the liver scans (hyper- and hypoechogenic) in 22%. Thirty seven per cent of the hyperechogenic foci were attributable to metastatic process. Hypoechogenic disorders were most frequently due to the primary cancer of the liver (20%). Mixed lesions were mainly produced by the cancer of large intestine and liver (41% and 27%, respectively). Metastases into the liver produced all three types of ultrasound pictures. The obtained results confirm the opinion that ultrasound picture of metastatic liver tumours is not suitable for the diagnosis of the type and origin of the neoplasm.     

A highly sensitive FRET-based approach reveals secretion of the actin-binding protein toxofilin during Toxoplasma gondii infection

We have utilized a highly sensitive approach based on fluorescence resonance energy transfer (FRET) and β-lactamase (BLA), which we adapted for the detection of Toxoplasma gondii secreted proteins. This assay revealed that the actin-binding protein toxofilin appears to be secreted into host cells during invasion. To determine the function of toxofilin during infection, we engineered a type I (RH strain) parasite with a targeted deletion of the toxofilin gene and compared the phenotypes of control and toxofilin knockout (Δ txf ) parasites in several in vitro assays, including invasion, growth, gliding motility, and egress of the Δ txf parasites, as well as F-actin staining, phagocytosis and migration of cells infected with Δ txf parasites or wild-type controls. Despite its apparent secretion into host cells and its ability to bind to and modulate host actin, we observed that toxofilin does not appear to play a role in these processes, under the conditions we examined, and we report these findings here.     

Annexin A8 displays unique phospholipid and F-actin binding properties

Annexin A8 is a poorly characterized member of the annexin family of Ca2+-regulated membrane binding proteins. Initially only identified at the cDNA level it had been tentatively linked to acute promyelocytic leukaemia (APL) due to its high and regulated expression in APL-derived cells. Here we identify unique properties of the annexin A8 protein. We show that it binds Ca2+-dependently and with high specificity to phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P2) and is also capable of interacting with F-actin. In line with these characteristics annexin A8 is recruited to F-actin-associated PtdIns(4,5)P2-rich membrane domains formed in HeLa cells upon infection with non-invading enteropathogenic Escherichia coli. These properties suggest a role of annexin A8 in the organization of certain actin-associated membrane domains.     

Characterization of the Ca2+ -regulated ezrin-S100P interaction and its role in tumor cell migration

Ezrin is a multidomain protein providing regulated membrane-cytoskeleton contacts that play a role in cell differentiation, adhesion, and migration. Within the cytosol of resting cells ezrin resides in an autoinhibited conformation in which the N- and C-terminal ezrin/radixin/moesin (ERM) association domains (ERMADs) interact with one another. Activation of the ezrin membrane-cytoskeleton linker function requires an opening of this interdomain association that can result from phosphatidylinositol 4,5-bisphosphate binding to the N-ERMAD and threonine 567 phosphorylation in the C-ERMAD. We have shown that ezrin can also be activated by Ca(2+)-dependent binding of the EF-hand protein S100P. We now provide a quantitative analysis of this interaction and map the respective binding sites to the F2 lobe in the ezrin N-ERMAD and a stretch of hydrophobic residues in the C-terminal extension of S100P. Phospholipid binding assays reveal that S100P and phosphatidylinositol 4,5-bisphosphate compete to some extent for at least partially overlapping binding sites in N-ERMAD. Using interaction-competent as well as interaction-incompetent S100P derivatives and permanently active ezrin mutants, we also show that the protein interaction and a resulting activation of ezrin promote the transendothelial migration of tumor cells. Thus, a prometastatic role of ezrin and S100P that had been proposed based on their overexpression in highly metastatic cancers is probably due to a direct interaction between the two proteins and the S100P-mediated activation of ezrin.     

The effects of 2-phenylalanine 8-lysine vasopressin (octapressin) on blood vessels in the rat tail.

Isolated perfused segments of the ventral rat tail artery have been used in this study to investigate the effects of octapressin on vascular smooth muscle and its interaction with other vasoconstrictor agents. It was found that repeated octapressin administration was associated with tachyphylaxis and that the onset and rate of development of this phenomenon could be modified by cocaine. The constrictor action of catecholamines, electrical stimulation and serotonin on the rat tail artery was potentiated by octapressin. Mechanisms which may be involved in octapressin tachyphylaxis are discussed.     

Production of a Shigella sonnei Vaccine Based on Generalized Modules for Membrane Antigens (GMMA), 1790GAHB

Recently, we developed a high yield production process for outer membrane particles from genetically modified bacteria, called Generalized Modules of Membrane Antigens (GMMA), and the corresponding simple two step filtration purification, enabling economic manufacture of these particles for use as vaccines. Using a Shigella sonnei strain that was genetically modified to produce penta-acylated lipopolysaccharide (LPS) with reduced endotoxicity and to maintain the virulence plasmid encoding for the immunodominant O antigen component of the LPS, scale up of the process to GMP pilot scale was straightforward and gave high yields of GMMA with required purity and consistent results. GMMA were formulated with Alhydrogel and were highly immunogenic in mice and rabbits. In mice, a single immunization containing 29 ng protein and 1.75 ng of O antigen elicited substantial anti-LPS antibody levels. As GMMA contain LPS and lipoproteins, assessing potential reactogenicity was a key aspect of vaccine development. In an in vitro monocyte activation test, GMMA from the production strain showed a 600-fold lower stimulatory activity than GMMA with unmodified LPS. Two in vivo tests confirmed the low potential for reactogenicity. We established a modified rabbit pyrogenicity test based on the European Pharmacopoeia pyrogens method but using intramuscular administration of the full human dose (100 μg of protein). The vaccine elicited an average temperature rise of 0.5°C within four hours after administration, which was considered acceptable and showed that the test is able to detect a pyrogenic response. Furthermore, a repeat dose toxicology study in rabbits using intramuscular (100 μg/dose), intranasal (80 μg/dose), and intradermal (10 μg/dose) administration routes showed good tolerability of the vaccine by all routes and supported its suitability for use in humans. The S. sonnei GMMA vaccine is now in Phase 1 dose-escalation clinical trials.