Assessing German farmers’ attitudes regarding nature conservation set-aside in regions dominated by arable farming

This paper presents an analysis of the attitudes of farmers towards a policy approach that combines the instrument of set-aside of farmland with agri-environmental measures under the Common Agricultural Policy (CAP) for achieving nature goals. One of the stated objectives of current agricultural and environmental policies is to give greater consideration to issues of nature conservation in agricultural landscapes. To fulfil this objective, there is an urgent need to develop approaches that, on the one hand, are capable of delivering tangible improvements in the ecological situation and nature conservation in agriculture and, on the other hand, are structured in such a way that farmers are willing to put them into practice. In addition, such approaches need to be economical and affordable for society. The Leibniz-Centre for Agricultural Landscape Research (ZALF) has developed the concept of ‘nature-conservation set-aside’, especially for landscapes that are under predominantly arable use. Conservation set-aside refers to parts of arable farmland especially well suited to nature conservation, which are mandatorily withdrawn from agricultural production (for those who claim payment) and on which ecologically valuable habitats are created through specific management activities. To determine what kind of problems might arise for farmers in the course of implementing this concept as an agri-environmental measure, a survey was carried out in four largely arable regions of Germany. The results of this survey show the attitudes of German farmers regarding set-aside farmland for nature conservation in regions dominated by arable farming, and they demonstrate which factors influence the implementation of conservation set-aside.     

Discovery, SAR and X-ray structure of 1H-benzimidazole-5-carboxylic acid cyclohexyl-methyl-amides as inhibitors of inducible T-cell kinase (Itk)

A series of novel potent benzimidazole based inhibitors of interleukin-2 T-cell kinase (Itk) were prepared. In this report, we discuss the structure-activity relationship (SAR), selectivity, and cell-based activity for the series. We also discuss the SAR associated with an X-ray structure of one of the small-molecule inhibitors bound to ITK.     

Deoxyribonucleic Acid Repair in Bacillus subtilis: Development of Competent Cells into a Tester for Carcinogens

The development of competent transformed Bacillus subtilis into a tester system for carcinogens is described. Precocious or noninduced activation of SOS functions occur in competent cells. Thus, lower doses or concentrations of SOS inducing agents are needed to cause cell death due to indigenous prophage activation and lysis of bacteria. The two known defective prophages in B. subtilis enhance the sensitivity of competent cells to the carcinogens ultraviolet light, mitomycin C, and methyl methanesulfonate. However, these same cells have no enhanced sensitivity for the non-carcinogenic ethyl methanesulfonate or for nalidixic acid. Therefore, competent B. subtilis appear to be a sensitive tester for carcinogens.     

Tim-3+ T-bet+ tumor-specific Th1 cells colocalize with and inhibit development and growth of murine neoplasms

Although T cells infiltrate many types of murine and human neoplasms, in many instances tumor-specific cytotoxicity is not observed. Strategies to stimulate CTL-mediated antitumor immunity have included in vitro stimulation and/or genetic engineering of T cells, followed by adoptive transfer into tumor-bearing hosts. In this model of B cell lymphoma in SJL/J mice, we used Tim-3(+) T-bet(+) Th1 cells to facilitate the development of tumor-specific CTL. Tumor-specific Th1 cell lines were polarized with IL-12 during in vitro stimulation and long term maintenance. As few as 5 million Tim-3(+) T-bet(+) Th1 cells enabled recipients to resist growth of malignant transplantable cells. In addition, similar numbers of Th1 cells injected into 2- to 3-mo-old mice inhibited development of the spontaneous primary lymphomas, which normally arise in 90% of aging mice. CFSE(+) Th1 cells colocalized with injected tumor cells in vivo and formed conjugates with the tumor cells within follicles, whereas in nontumor-challenged recipients the CFSE(+) Th1 cells localized only within the T cell zones of the spleen. These results provide evidence that adoptive immunotherapy with Tim-3(+) T-bet(+) tumor-specific Th1 cells can be used to induce host cytotoxic responses that inhibit the development and growth of neoplastic cells.     

The IsdC protein from Staphylococcus aureus uses a flexible binding pocket to capture heme

Staphylococcus aureus scavenges heme-iron from host hemoproteins using iron-regulated surface determinant (Isd) proteins. IsdC is the central conduit through which heme is passed across the cell wall and binds this molecule using a NEAr Transporter (NEAT) domain. NMR spectroscopy was used to determine the structure of IsdC in complex with a heme analog, zinc-substituted protoporphyrin IX (ZnPPIX). The backbone coordinates of the ensemble of conformers representing the structure exhibit a root mean square deviation to the mean structure of 0.53 +/- 0.11 angstroms. IsdC partially buries protoporphyrin within a large hydrophobic pocket that is located at the end of its beta-barrel structure. The central metal ion of the analog adopts a pentacoordinate geometry in which a highly conserved tyrosine residue serves as a proximal ligand. Consistent with the structure and its role in heme transfer across the cell wall, we show that IsdC weakly binds heme (K(D) = 0.34 +/- 0.12 microm) and that ZnPPIX rapidly dissociates from the protein at a rate of 126 +/- 30 s(-1). NMR studies of the apo-form of IsdC reveal that a 3(10) helix within the binding pocket undergoes a flexible to rigid transition as heme is captured. This structural plasticity may increase the efficiency of heme transfer across the cell wall by facilitating protein-protein interactions between apoIsdC and upstream hemoproteins.     

A furostanol glycoside from Helleborus macranthus

From the roots and rhizomes of Helleborus macranthus a new glycoside, macranthoside I, has been isolated and shown to have the structure 25(27)-dehydro-5β-furostan-3β,22,26-triol-3-O-β-d-glucopyranosyl(1 → 6)-β-d-glucopyranoside, 26-O-β-d-glucopyranoside.     

KLP-18, a Klp2 kinesin,is required for assembly of acentrosomal meiotic spindles in Caenorhabditis elegans

The proper segregation of chromosomes during meiosis or mitosis requires the assembly of well organized spindles. In many organisms, meiotic spindles lack centrosomes. The formation of such acentrosomal spindles seems to involve first assembly or capture of microtubules (MTs) in a random pattern around the meiotic chromosomes and then parallel bundling and bipolar organization by the action of MT motors and other proteins. Here, we describe the structure, distribution, and function of KLP-18, a Caenorhabditis elegans Klp2 kinesin. Previous reports of Klp2 kinesins agree that it concentrates in spindles, but do not provide a clear view of its function. During prometaphase, metaphase, and anaphase, KLP-18 concentrates toward the poles in both meiotic and mitotic spindles. Depletion of KLP-18 by RNA-mediated interference prevents parallel bundling/bipolar organization of the MTs that accumulate around female meiotic chromosomes. Hence, meiotic chromosome segregation fails, leading to haploid or aneuploid embryos. Subsequent assembly and function of centrosomal mitotic spindles is normal except when aberrant maternal chromatin is present. This suggests that although KLP-18 is critical for organizing chromosome-derived MTs into a parallel bipolar spindle, the order inherent in centrosome-derived astral MT arrays greatly reduces or eliminates the need for KLP-18 organizing activity in mitotic spindles.     

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich structures suggest alterations in synaptic integrity and connectivity. Here we provide a detailed protocol for culturing primary rat cortical neurons, Phalloidin staining for F-actin puncta, and subsequent quantification techniques. First, the frontal cortex of E18 rat embryos are dissociated into low-density cell culture, then the neurons grown in vitro for at least 12-14 days. Following experimental treatment, the cortical neurons are stained with AlexaFluor 488 Phalloidin (to label the dendritic F-actin puncta) and microtubule-associated protein 2 (MAP2; to validate the neuronal cells and dendritic integrity). Finally, specialized software is used to analyze and quantify randomly selected neuronal dendrites. F-actin rich structures are identified on second order dendritic branches (length range 25-75 µm) with continuous MAP2 immunofluorescence. The protocol presented here will be a useful method for investigating changes in dendritic synapse structures subsequent to experimental treatments.     

Age‐dependent dispersal and habitat choice in black‐tailed godwits Limosa limosa limosa across a mosaic of traditional and modern grassland habitats

Whether to disperse, and where to, are two of the most prominent decisions in an individual's life, with major consequences for reproductive success. We studied natal and breeding dispersal in the monogamous black‐tailed godwit Limosa limosa limosa in the Netherlands, where they breed in agricultural grasslands. The majority of these grasslands recently changed from wet herb‐rich meadows into well‐drained grassland monocultures, on which godwits have a lower reproductive success. Here we examine habitat selection with a multistate mark–recapture analysis. Habitat transition probabilities between meadows and monocultures were estimated on the basis of 1810 marked chicks and 531 adults during seven years in a 8500 ha study area. Young and adult godwits may differ in habitat selection because: 1) adults may have gained experience from previous nest success where to settle, 2) younger individuals may find it harder to compete for the best territories. Both young and adults moved at a higher rate from the predominant monocultures to meadows than the other way around, thus actively selecting the habitat with better quality. However, dispersal distance of adults was not affected by previous nest success. The average dispersal distance from place of birth of godwits breeding for the first time was ten times larger than that of adult godwits. That godwits breeding in their second calendar year arrived and laid at similar dates and were equally able to select territories in areas with high breeding densities, suggests that young birds were not competitively inferior to adults. Although on monocultures reproduction is insufficient to maintain constant populations, birds sometimes moved from meadows to monocultures. This explains why even after 30 years of land‐use intensification, godwits still breed in low‐quality habitat. The adjustment to changing habitat conditions at the population level appears to be a slow process.