A shortcut to identifying small molecule signals that regulate behavior and development in Caenorhabditis elegans

Small molecule metabolites play important roles in Caenorhabditis elegans biology, but effective approaches for identifying their chemical structures are lacking. Recent studies revealed that a family of glycosides, the ascarosides, differentially regulate C. elegans development and behavior. Low concentrations of ascarosides attract males and thus appear to be part of the C. elegans sex pheromone, whereas higher concentrations induce developmental arrest at the dauer stage, an alternative, nonaging larval stage. The ascarosides act synergistically, which presented challenges for their identification via traditional activity-guided fractionation. As a result the chemical characterization of the dauer and male attracting pheromones remained incomplete. Here, we describe the identification of several additional pheromone components by using a recently developed NMR-spectroscopic approach, differential analysis by 2D NMR spectroscopy (DANS), which simplifies linking small molecule metabolites with their biological function. DANS-based comparison of wild-type C. elegans and a signaling-deficient mutant, daf-22, enabled identification of 3 known and 4 previously undescribed ascarosides, including a compound that features a p-aminobenzoic acid subunit. Biological testing of synthetic samples of these compounds revealed additional evidence for synergy and provided insights into structure–activity relationships. Using a combination of the three most active ascarosides allowed full reconstitution of the male-attracting activity of wild-type pheromone extract. Our results highlight the efficacy of DANS as a method for identifying small-molecule metabolites and placing them within a specific genetic context. This study further supports the hypothesis that ascarosides represent a structurally diverse set of nematode signaling molecules regulating major life history traits

Anthracimycin activity against contemporary methicillin-resistant Staphylococcus aureus.

Anthracimycin is a recently discovered novel marine-derived compound with activity against Bacillus anthracis. We tested anthracimycin against an expanded panel of Staphylococcus aureus strains in vitro and in vivo. All strains of S. aureus tested, including methicillin-susceptible, methicillin-resistant (MRSA) and vancomycin-resistant strains of S. aureus, were susceptible to anthracimycin at MIC values of ⩽0.25 mg l(-1). Although its postantibiotic effects were minimal, anthracimycin exhibited potent and rapid bactericidal activity, with a >4-log kill of USA300 MRSA within 3 h at five times its MIC. At concentrations significantly below the MIC, anthracimycin slowed MRSA growth and potentiated the bactericidal activity of the human cathelicidin, LL-37. The bactericidal activity of anthracimycin was somewhat mitigated in the presence of 20% human serum, and the compound was minimally toxic to human cells, with an IC50 (inhibitory concentration 50)=70 mg l(-1) against human carcinoma cells. At concentrations near the MIC, anthracimycin inhibited S. aureus nucleic acid synthesis as determined by optimized macromolecular synthesis methodology, with inhibition of DNA and RNA synthesis occurring in the absence of DNA intercalation. Anthracimycin at a single dose of 1 or 10 mg kg(-1) was able to protect mice from MRSA-induced mortality in a murine peritonitis model of infection. Anthracimycin provides an interesting new scaffold for future development of a novel MRSA antibiotic

Bestimmungsgründe von Arbeitszeitkonten bei Betrieben und Beschäftigten

Arbeitszeitkonten ermöglichen es Arbeitszeiten zu variieren und den Arbeitseinsatz an das Arbeitsaufkommen anzupassen. Mit logistischen Regressionsmodellen und Daten des IAB-Betriebspanels wird in einem ersten Schritt analysiert, welche Faktoren Arbeitszeitkonten in Betrieben bestimmen. Die industriellen Beziehungen und betriebliche Beschäftigungsstruktur sind wesentliche Determinanten. In einem zweiten Schritt wird mit Daten des Sozio-oekonomischen Panels analysiert, welche Faktoren den Zugang von Beschäftigten zu einem Arbeitszeitkonto bestimmen. Beschäftigte mit einem höheren Maß an Humankapital haben überdurchschnittlich häufig Zugang zu einem Arbeitszeitkonto, während geringfügig beschäftigte Frauen und befristet beschäftigte Männer unterdurchschnittlich häufig Arbeitszeitkonten haben

"Kultur" als Form symbolischer Gewalt: Grenzziehungsprozesse im Kontext von Migration am Beispiel der Schweiz

Die Schweiz gilt international als Modell eines gelungenen Multikulturalismus, dann nämlich wenn es das Zusammenleben der vier Sprachgruppen (Romands, DeutschschweizerInnen, TessinerInnen, RäteromanInnen) betrifft. Ein sprachlicher wie auch religiöser Pluralismus ist und war stets ein Grundbaustein des Selbstverständnisses der „Willensnation“ Schweiz. Geht es aber um MigrantInnen präsentiert sich die Geschichte anders, denn in diesem Falle erscheinen religiöse und ethnisch-kulturelle Pluralität vorwiegend als problematisch. MigrantInnen gehören entsprechend den öffentlichen und politischen Diskursen nicht zum multikulturellen Staat, vielmehr sind Prozesse kollektiver Grenzziehungen und damit Schließungsmechanismen zu beobachten, in denen Ethnizität, Religion und Kultur zu den wichtigsten Differenzierungsmerkmale werden, wie Gemeinsamkeiten gegen innen (SchweizerInnen) und Barrieren gegen außen (Ausländer, Migranten, Muslims, etc.) hergestellt werden. Ich argumentiere in diesem Kapitel, dass sich dieser „Kulturdiskurs“ im letzten Jahrzehnt verstärkt hat und gleichzeitig semantischen Verschiebungen unterworfen war. Mittels der Grenzziehungsperspektive wird historisch nachvollzogen, wie Zuwanderung und Integration in politischen Debatten und Gesetz zunehmend kulturalisiert und ethnisiert wurden. Ein Fallbeispiel aus der Forschung dient mir anschließend der Veranschaulichung dieser theoretischen Perspektive und dieses „neuen“ Essentialismus

mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein

The nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus plays important roles in both viral replication and modulation of host cell processes. New ligands that target the N protein may thus provide tools to track the protein inside cells, detect interaction hot spots on the protein surface, and discover sites that could be used to develop new anti-SARS therapies. Using mRNA display selection and directed evolution, we designed novel antibody-like protein affinity reagents that target SARS N protein with high affinity and selectivity. Our libraries were based on an 88-residue variant of the 10th fibronectin type III domain from human fibronectin (10Fn3). This selection resulted in eight independent 10Fn3 intrabodies, two that require the N-terminal domain for binding and six that recognize the C terminus, one with K_d = 1.7 nM. 10Fn3 intrabodies are well expressed in mammalian cells and are relocalized by N in SARS-infected cells. Seven of the selected intrabodies tested do not perturb cellular function when expressed singly in vivo and inhibit virus replication from 11- to 5900-fold when expressed in cells prior to infection. Targeting two sites on SARS-N simultaneously using two distinct 10Fn3s results in synergistic inhibition of virus replication

Paraklooramfetamiini mõju psühhomotoorsele aktiivsusele ja serotoniini ja dopamiini tasemetele septumis ja prefrontaalkorteksis erineva püsisotsiaalsusega loomadel

Social behaviour plays important role in many psychopathologies. Sociability has been shown to be a stable trait in animals and individual differences in sociability are related to 5-HT transmission in septum and prefrontal cortex. The aim of the current study was to investigate whether animals with different levels of sociability are differently affected by p-chloroamphetamine treatment. 5-HT, 5-HIAA and DA levels were measured in septum and PFC and locomotor activity was recorded. Although there was no difference in monoamine levels or locomotor activity between LS- and HS-animals after PCA treatment, our findings confirm previous research on the different neurodegenerative effect of PCA in different brain regions. PCA had stronger neurodegenerative effect in PFC as 5-HT and 5-HIAA levels there were lower than in septum

A role for jasmonates in the release of dormancy by cold stratification in wheat

Hydration at low temperatures, commonly referred to as cold stratification, is widely used for releasing dormancy and triggering germination in a wide range of species including wheat. However, the molecular mechanism that underlies its effect on germination has largely remained unknown. Our previous studies showed that methyl-jasmonate, a derivative of jasmonic acid (JA), promotes dormancy release in wheat. In this study, we found that cold-stimulated germination of dormant grains correlated with a transient increase in JA content and expression of JA biosynthesis genes in the dormant embryos after transfer to 20 (o)C. The induction of JA production was dependent on the extent of cold imbibition and precedes germination. Blocking JA biosynthesis with acetylsalicylic acid (ASA) inhibited the cold-stimulated germination in a dose-dependent manner. In addition, we have explored the relationship between JA and abscisic acid (ABA), a well-known dormancy promoter, in cold regulation of dormancy. We found an inverse relationship between JA and ABA content in dormant wheat embryos following stratification. ABA content decreased rapidly in response to stratification, and the decrease was reversed by addition of ASA. Our results indicate that the action of JA on cold-stratified grains is mediated by suppression of two key ABA biosynthesis genes, TaNCED1 and TaNCED2.This project was funded by a CSIRO Office of the Chief Executive PDF scheme