Identification and characterization of a serpin from Eimeria acervulina

Serpins are serine protease inhibitors that are widely distributed in metazoans but have not been previously characterized in Eimeria spp. A serpin from Eimeria acervulina was cloned, expressed and characterized. Random screening of an E.acervulina sporozoite cDNA library identified a single clone (D14) whose coding region shared high similarity to consensus structure of serpins. Clone D14 contained an entire open reading frame (ORF) consisting of 1,245 nts that encode a peptide 413 amino acids in length with a predicted molecular weight of 45.5 kDa and containing a signal peptide 28 residues in length. By Western blot analysis, polyclonal antiserum to the recombinant serpin (rbSp) recognized a major 55 kDa protein band in unsporulated oocysts and in oocysts sporulated up to 24 hr (fully sporulated). The anti-rbSp detected bands of 55 kDa and 48 kDa in sporozoites (SZ) and merozoites (MZ) respectively. Analysis of MZ secretion products revealed a single protein of 48 kDa which may correspond to secreted serpin. By immuno-staining the serpin was located in granules distributed throughout both the SZ and MZ but granules appeared to be concentrated in the parasite's anterior. Analysis of the structure predicts that the E. acervulina serpin should be an active inhibitor. However, rbSp was without inhibitory activity against common serine proteases. By Western blot analysis the endogenous serpin in MZ extracts did not form the expected high molecular weight complex when coincubated with either trypsin or subtilisin. The results demonstrate that E. acervulina contains a serpin gene and expresses a protein with structural properties similar to an active serine protease inhibitor. Although the function of the E. acervulina serpin remains unknown the results further suggest that serpin is secreted by the parasite where it may be involved in cell invasion and other basic developmental processes.     

A comparative metabolomic study of NHR-49 in Caenorhabditis elegans and PPAR-alpha in the mouse

Proton Nuclear Magnetic Resonance spectroscopy and Gas Chromatography Mass Spectrometry based metabolomics has been used in conjunction with multivariate statistics to examine the metabolic changes in Caenorhabditis elegans following the deletion of nuclear hormone receptor-49 (nhr-49). Deletion of the receptor produced profound changes in fatty acid metabolism, in particular an increase in the ratio of unsaturated to saturated fatty acids, a decrease in the concentration of glucose and increases in lactate and alanine. Given the proposed functional similarity between nhr-49 and the mammalian peroxisome proliferator-activated receptors (PPARs) these changes were compared with the metabolome of the PPAR-alpha null mouse. The metabolomic approach demonstrated a number of similarities including the regulation of lipid synthesis, beta-oxidation of fatty acids and changes in glycolysis/gluconeogenesis.     

Effects of landscape composition on farmland and red-listed birds in boreal agricultural-forest mosaics

Farmland birds have declined in large areas of western and northern Europe. This decline has been connected with changes in the agricultural landscape. We studied the effects of landscape composition on birds in a boreal agricultural-forest mosaic in SW Finland. This study was carried out with a grid-based approach: bird pairs were counted in 105 grid squares of 25 ha within an area of 26.25 km². The total density of farmland birds and density of red-listed species were related to the land cover variables using generalized linear modelling (GLM). Farmland birds consist of a variable group of species either breeding or feeding in agricultural land. The model explained a moderate proportion (49%) of the variation in the total density of farmland birds in the landscape. In a regression analysis cover of non-arable agricultural land (semi-natural grasslands, built-up areas) explained a much higher proportion (r²=0.49) of the variation in farmland bird density than that of arable land (cultivated fields and set-aside fields, r²=0.04). Semi-natural grasslands, which have drastically declined throughout NW Europe, and built-up areas (mainly farmyards) had the most significant positive effects on the density of red-listed species. The results emphasize the significance of semi-natural grasslands for the declining red-listed farmland bird species. Birds are usually not restricted to certain patches of habitat but use several patches in their home range. Thus, when studying bird-landscape relations for land use planning, we also recommend grid-based approaches covering the whole landscape variation.