Wnt3a-mediated formation of phosphatidylinositol 4,5-bisphosphate regulates LRP6 phosphorylation

The canonical Wnt-beta-catenin signaling pathway is initiated by inducing phosphorylation of one of the Wnt receptors, low-density lipoprotein receptor-related protein 6 (LRP6), at threonine residue 1479 (Thr1479) and serine residue 1490 (Ser1490). By screening a human kinase small interfering RNA library, we identified phosphatidylinositol 4-kinase type II alpha and phosphatidylinositol-4-phosphate 5-kinase type I (PIP5KI) as required for Wnt3a-induced LRP6 phosphorylation at Ser1490 in mammalian cells and confirmed that these kinases are important for Wnt signaling in Xenopus embryos. Wnt3a stimulates the formation of phosphatidylinositol 4,5-bisphosphates [PtdIns (4,5)P2] through frizzled and dishevelled, the latter of which directly interacted with and activated PIP5KI. In turn, PtdIns (4,5)P2 regulated phosphorylation of LRP6 at Thr1479 and Ser1490. Therefore, our study reveals a signaling mechanism for Wnt to regulate LRP6 phosphorylation.     

Relatedness of cytotoxins from geographically diverse isolates of Moraxella bovis

To determine whether amino acid sequence variation exists in the Moraxella bovis (M. bovis) cytotoxin (MbxA) from geographically diverse M. bovis isolated in the United States, mbxA was amplified and sequenced. The MbxA deduced amino acid sequence from M. bovis originally isolated in California, Washington, North Carolina, and Georgia, as well as reference strains of M. bovis isolated at the National Animal Disease Laboratory, Ames, IA, USA, all encoded a nearly identical 927 amino acid protein. MbxA from two of the four California isolates (SFS 9a and SFS 100a) differed from all other isolates at two sites at which the polar amino acids glutamine (position 666) and asparagine (position 823) were replaced by ionized amino acids glutamic acid and aspartic acid, respectively. Rabbit antiserum to the expressed carboxy terminus (amino acids 590-927) of MbxA from M. bovis (Tifton I) neutralized the hemolytic activity of SFS 9a and SFS 100a. The M. bovis cytotoxin appears to be conserved amongst geographically diverse isolates of M. bovis from the USA. Antiserum against the carboxy terminus of MbxA common to the majority of isolates neutralized the hemolytic activity of two strains with a divergent MbxA deduced amino acid sequence. Vaccines against IBK that incorporate MbxA as antigen may offer protection against geographically diverse strains of M. bovis.     

Examination of mink (Mustela vison) fed a sulphuric acid preserved fish during lactation and growth period. I. A clinical-chemical examination

Addition of sulphuric acid preserved fish in great amounts as fur animal feed has a serious influence on the metabolism of the animals. The present experimental work shows that several clinical-chemical parameters are changed during feeding of acid preserved feed resulting in changes of the fluid-, electrolyte- and acid-base balance as well as in the immunological status. The changes are most probably due to changes also found in the organ functions. The present work together with earlier investigations by the authors has formed the scientific background for the use of acid preserved fish silage in mink during the last 10 years. This has made it possible to out-distance the competitors further, both with respect to fur quality and feeding costs.     

Influence of different disease control pesticide strategies on multiple pesticide residue levels in apple

Summary

Seven pesticide application strategies were investigated to control apple scab (Venturia inaequalis) and powdery mildew (Podosphaera leucotricha) and, at the same time, fulfil the new quality standards implemented by some German retailers. These demand that pesticide residues should be below 80% of European Maximum Residue Levels (EU MRL) and that the number of residues present at levels above 0.01 mg kg^?1 should be limited to a maximum of four. The strategies fulfilled the requirement to use combinations of different active substances in order to prevent the emergence of resistance to pesticides. The trials were conducted at two sites in Switzerland, in 2007, and all strategies and applications were in accordance with actual practice. Four replicates of apple samples from each strategy were then analysed for pesticide residues. The incidence of infection with apple scab and powdery mildew were monitored during the season in order to evaluate the efficacy of the different strategies. The efficacies of the different strategies against apple scab and powdery mildew were between 84% and 100% successful. In general, the level of pesticide residues found correlated with application rate and time, and no measured residue level exceeded the EU–MRL. The numbers of residues present at > 0.01 mg kg^?1 were between two and five.     

Pharmacokinetics of oral omeprazole in llamas

Gastrogard, an oral formulation of omeprazole, was given to six llamas at a dose of 4 mg/kg once a day for 6 days. Plasma samples were collected at 0, 15, 30, 45, and 60 min and 2, 3, 4, 6, 8, 12, and 24 h on days 1 and 6. Plasma omeprazole concentrations were measured by high-pressure liquid chromatography with ultraviolet detection. Pharmacokinetic parameters calculated included the area under the curve (AUC(0-infinity)), peak plasma concentration (Cmax), time of peak plasma concentration (Tmax), and terminal half-life (t(1/2)). On day 6, plasma omeprazole concentrations reached a Cmax of 0.12 microg/mL at a Tmax of 45 min. The t(1/2) of omeprazole was 2.3 h and the AUC(0-infinity) was 0.38 h x microg/mL. Plasma concentrations remained above the minimum concentration for inhibition of gastric acid secretion projected from other studies on day 6 in all the llamas for approximately 6 h. However, the AUC(0-infinity) was below the concentrations associated with clinical efficacy. It was not possible to measure oral systemic bioavailability because there was no i.v. data collected from these animals. However, using data published on the i.v. pharmacokinetics of omeprazole in llamas, oral absorption was estimated to be only 2.95%. Due to low absorption the oral dose was increased to 8 and 12 mg/kg and studies were repeated. There were no significant differences in Cmax, Tmax, or AUC(0-infinity) for either of the increased doses. These results indicate that after 6 days of treatment with doses up to 12 mg/kg, oral omeprazole produced plasma drug concentrations which are not likely to be associated with clinical efficacy in camelids.     

Nematode diversity,food web condition,and chemical and physical properties in different soil habitats of an organic farm

The aim of this paper was to assess biodiversity among different habitats of an organic farm and the relationships between some soil properties, nematode taxonomic diversity, and soil food web condition. Eight habitats were studied in the farm: ponds, ditches, a riparian corridor, hedgerows, and four agricultural fields (mustard, oats, fallow, and legumes). The undisturbed riparian corridor had higher soil and concentrations, and potentially mineralizable N and higher abundances of bacterivore nematodes and longer food webs. Canonical correlation analysis showed associations between habitats and nematode trophic groups: predatory and bacterial-feeding nematodes in the riparian corridor and hedgerows, omnivore nematodes in the ponds and ditches, and fungal-feeding nematodes in the legume field. Soil chemical and physical properties mirrored the aboveground farm patterns and were more similar among habitats that were or had been cultivated, compared to the riparian corridor. Soil food web indices, based on functional analysis of nematode faunal composition, reflected the aboveground landscape heterogeneity. Discriminant analysis indicated that soil food web indices separated the two most disturbed habitats (ponds and tailwater ditches) from the two least disturbed habitats (the riparian corridor and hedgerows). The indices correlated with soil functioning as inferred by soil properties. Abundance of nematode taxa was not associated with aboveground landscape patterns. The complexity of the soil food web may have been influenced by (1) environmental factors that differed between years, (2) different time periods since disturbance in the various habitats, and (3) movement of nutrients and organisms by water flow between habitats in the farmscale.     

Estimation of detection probability in aerial surveys of antarctic pack-ice seals

We use line transect detection functions together with generalized linear and additive models to estimate detection probability when detection on the line (“g(0)”) may not be certain. The methods provide a flexible way of modeling detection probability for independent observer surveys, and for investigating the effects of explanatory variables. Analysis of data from an aerial survey of pack-ice seals produced g(0) estimates substantially below 1 for some observers (it varied from 0.80 to 0.98), demonstrated a fairly complex dependence of detection probability on covariates, and showed negative correlation between observers’ search width and their g(0). In addition to illustrating the utility of generalized additive models for capturing the effect of covariates on detection probability, the analysis suggests that detection functions may be sufficiently variable that use of g(0) correction factors obtained from other surveys would be inadvisable. We recommend that estimation of g(0) be considered for all aerial surveys; if g(0) is found to be very close to 1, estimation from subsequent surveys under the assumption that it is 1 may be reasonable, but without any estimation of g(0), the assumption that it is 1 is a matter of faith.     

An oral vaccine against NMDAR1 with efficacy in experimental stroke and epilepsy

The brain is generally considered immunoprivileged, although increasing examples of immunological responses to brain antigens, neuronal expression of major histocompatibility class I genes, and neurological autoimmunity have been recognized. An adeno-associated virus (AAV) vaccine generated autoantibodies that targeted a specific brain protein, the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor. After peroral administration of the AAV vaccine, transgene expression persisted for at least 5 months and was associated with a robust humoral response in the absence of a significant cell-mediated response. This single-dose vaccine was associated with strong anti-epileptic and neuroprotective activity in rats for both a kainate-induced seizure model and also a middle cerebral artery occlusion stroke model at 1 to 5 months following vaccination. Thus, a vaccination strategy targeting brain proteins is feasible and may have therapeutic potential for neurological disorders.