The Power of Regeneration: Lessons from a Degraded Grassland

To gain insight into mechanisms controlling ecosystem regeneration, we investigated a grassland that was heavily polluted by a phosphate fertilizer factory between 1960 and 1990. Abiotic and biotic filters that influenced species assemblage and succession were monitored and restoration approaches assessed. Studies included the investigation of soil parameters, impact of abiotic stress on microbes and plants, plant succession, recolonization mechanisms, functional group interactions, mycorrhizal diversity and function, and food web analysis. Results indicate that after cessation of pollution, the system had high potential for self‐recovery. About 15 years of research are now summarized by interrelating the results of all projects and fitting them into one conceptual model.     

A wide-range integrative yeast expression vector system based on <Emphasis Type="Italic">Arxula adeninivorans</Emphasis>-derived elements

An Arxula adeninivorans integration vector was applied to a range of alternative yeast species including Saccharomyces cerevisiae, Debaryomyces hansenii, Debaryomyces polymorphus, Hansenula polymorpha and Pichia pastoris. The vector harbours a conserved A. adeninivorans-derived 25S rDNA sequence for targeting, the A. adeninivorans-derived TEF1 promoter for expression control of the reporter sequence, and the Escherichia coli-derived hph gene conferring resistance against hygromycin B for selection of recombinants. Heterologous gene expression was assessed using a green fluorescent protein (GFP) reporter gene. The plasmid was found to be integrated into the genome of the various hosts tested; recombinant strains of all species exhibited heterologous gene expressions of a similar high level.     

Direct assignment of orosomucoid to human chromosome 9 and α2HS-glycoprotein to chromosome 3 using human fetal liver x rat hepatoma hybrids

Summary The production of plasma proteins has been monitored in somatic cell hybrids between a rat hepatoma cell line (7777) and human fetal liver cells. Production of 14 plasma proteins was assayed in concentrated serum-free culture supernatants by electroimmunoassay. 2HS-glycoprotein (AHSG) was produced by 10 of 19 hybrids; concordancy for presence or absence of protein production was 100% for human chromosome 3. Orosomucoid (ORM) was produced in 8 of 19 hybrids, with a concordancy for presence or absence of protein of 94.7% with human chromosome 9. The chromosome location for genes for these two proteins, previously assigned by linkage studies, is confirmed by direct assignment. These studies have also suggested possible chromosomal assignments for loci for ₁ and C1 esterase inhibitor. Other genes for proteins which could not be assigned to specific chromosomes using these hybrids were: complement C3, ceruloplasmin, hemopexin, inter--trypsin inhibitor, prealbumin, retinol-binding protein, transferrin and apolipoproteins CII, B, and sinking-pre-beta [Lp(a)].     

Sexual harassment in heterogeneous landscapes can mediate population regulation in a grasshopper

Population regulation has been related to differences in thequality among habitats, which mediate differences in vital ratessuch that in poor habitats reproductive rates are lower thanthose in high-quality habitats. The spatial distribution ofanimals in such habitats depends on their preferences and thedegree to which individuals have a free choice of a particularhabitat. The identified mechanisms that lead to a particularspatial distribution and eventually to regulation mainly includeforaging-related interference, for example, ideal free distribution,or simple selection of available high-quality habitats, thatis, site-dependent habitat selection. However, in insect speciesthese mechanisms might not be applicable, but density-dependenthabitat selection still occurs. We therefore suggest a mechanismthat refers to the nearly universal observation that matingsalso bear fitness costs. Although these costs have been investigatedon the individual level in many insect species, their consequencesfor population dynamics have not yet been addressed. In thegrasshopper species Stenobothrus lineatus, females in a nonreceptivemating status escape sexually approaching males by undirectedjumps. By including such avoidance behavior in a spatially-explicitsimulation model, we investigated its potential to result inprogressive use of low-quality habitats at increasing populationdensities. In particular, we show that (1) such behavior changeshabitat selection, (2) altered habitat selection results inpopulation regulation, and (3) the degree of habitat heterogeneityinfluences regulation such that (4) heterogeneous habitats showfine-tuned regulation and homogeneous habitats tend to supportlarge fluctuations.     

Yeast pyruvate dehydrogenase complex is regulated by a concerted activity of two kinases and two phosphatases

The activity of yeast pyruvate dehydrogenase complex is regulated by reversible phosphorylation. Recently we identified two enzymes that are involved in the phosphorylation (Pkp1p) and dephosphorylation (Ppp1p) of Pda1p, the alpha-subunit of the pyruvate dehydrogenase complex. Here we provide evidence that two additional mitochondrial proteins, Pkp2p (Ygl059wp) and Ppp2p (Ycr079wp), are engaged in the regulation of this complex by affecting the phosphorylation state of Pda1p. Our data indicate complementary activities of the kinases and a redundant function for the phosphatases. Both proteins are associated with the complex. We propose a model for the role of the regulatory enzymes and the phosphorylation state of Pda1p in the assembly process of the pyruvate dehydrogenase complex.     

Maize mutants affected at distinct stages of the arbuscular mycorrhizal symbiosis

Maize mutants affected in the symbiotic interaction with the arbuscular mycorrhizal fungus Glomus mosseae have been found by a visual, macroscopic screen in a Mutator-tagged population of maize. Seven mutants have been identified, falling into three phenotypic classes. For each class one mutant has been characterized in more detail. The nope1 (no perception 1) mutant does not support appressoria formation of G. mosseae, suggesting the absence of a plant-encoded function necessary for early recognition prior to contact. The phenotype segregated as a monogenic recessive trait, indicating that a mutation in a single locus abolished compatibility of maize to G. mosseae. On a second mutant termed taci1 (taciturn 1), appressoria form at reduced frequency but their morphology is normal and leads to penetration of the rhizodermis. However, intraradically, the majority of hyphae are septate, resulting in terminated fungal spreading. This phenotype suggests that the mutation of taci1 has an effect on recognition and on cortex invasion. Segregation analysis indicates taci1 to carry a recessive mutation. In contrast, wild-type fungal morphology has been recorded in the Pram1 (Precocious arbuscular mycorrhiza 1) mutant, which displays enhanced and earlier fungal invasion. This trait segregates in a dominant fashion indicative of a gain-of-function mutation affecting the plant's control over restricting fungal colonization.     

Murine CD146 is widely expressed on endothelial cells and is recognized by the monoclonal antibody ME-9F1

The endothelium plays an important role in the exchange of molecules, but also of immune cells between blood and the underlying tissue. The endothelial molecule S-Endo 1 antigen (CD146) is preferentially located at endothelial junctions and has been claimed to support endothelial integrity. In this study we show that the monoclonal antibody ME-9F1 recognizes the extracellular portion of murine CD146. Making use of ME-9F1 we found CD146 highly expressed and widely spread on endothelial cells in the analyzed murine tissues. In contrast to humans that express CD146 also on T cells or follicular dendritic cells, murine CD146 albeit at low levels was only found on a subset of NK1.1⁺ cells. The antibody against murine CD146 is useful for immunomagnetic sorting of primary endothelial cells not only from the liver but from various other organs. In vitro, no evidence was seen that the formation and integrity of endothelial monolayers or the transendothelial migration of T cells was affected by antibody binding to CD146 or by crosslinking of the antigen. This makes the antibody ME-9F1 an excellent tool especially for the ex vivo isolation of murine endothelial cells intended to be used in functional studies.     

Purification and characterization of natural Ara h 8, the Bet v 1 homologous allergen from peanut, provides a novel isoform

The peanut allergen Ara h 8 is an important allergen for birch pollen allergic patients because of the cross-reactivity to the homologous Bet v 1. As the existence of Ara h 8 has been shown at the cDNA level so far (AY328088) and the allergen has indirectly been detected as natural protein, it was the aim of our study to identify natural Ara h 8 in peanut extract and to develop a purification strategy. This was achieved using a unique combination of purification steps, including optimized extraction conditions, size exclusion and ion exchange chromatography and treatment of the interfering contaminants with iodoacetic acid. A characterization of the protein by microsequencing showed discrepancies to the deduced amino acid sequence of AY328088. For this reason, we cloned and expressed a new Ara h 8 isoform from cDNA (EU046325). This IgE-reactive protein corresponds to the results of microsequencing, ESI-FTICR-MS and trypsin fingerprinting analysis of the authentic and purified nAra h 8. Apart from the ultimate use of recombinant allergens for diagnostic procedures, there is also a scientific need for the natural counterpart, as it represents an excellent reference point by which to compare protein characteristics and to standardize diagnostic and therapeutic allergens.