Alternatively Activated Macrophages Are Host Cells for Chlamydia trachomatis and Reverse Anti-chlamydial Classically Activated Macrophages

The obligate intracellular pathogen Chlamydia trachomatis (Ctr) is the causative agent of the most common form of sexually transmitted disease in the United States. Genital infections with C. trachomatis can lead to inflammatory tissue damage followed by scarring and tissue remodeling during wound healing. Extensive scarring can lead to ectopic pregnancy or infertility. Classically activated macrophages (CA mϕ), with their anti-microbial effector mechanisms, are known to be involved in acute inflammatory processes during the course of infection. In contrast, alternatively activated macrophages (AA mϕ) contribute to tissue repair at sites of wound healing, and have reduced bactericidal functions. They are present during infection, and thus potentially can provide a growth niche for C. trachomatis during a course of infection. To address this question, macrophages derived from CD14-positive monocytes magnetically isolated from peripheral blood mononuclear cells (PBMC) were treated with interferon-γ or interleukin-4 to produce CA mϕ or AA mϕ, respectively. Confocal microscopy of chlamydial inclusions and quantification of infectious yields revealed better pathogen growth and development in AA mϕ than CA mϕ, which correlated with the reduced expression of indoleamine 2,3-dioxygenase, a known anti-chlamydial effector of the host. Furthermore, AA mϕ stained strongly for transferrin receptor and secreted higher amounts of anti-inflammatory interleukin-10 compared to CA mϕ, characteristics that indicate its suitability as host to C. trachomatis. CA, AA, and resting mϕ were infected with Ctr serovar L2. The data suggest that IL-10 produced by infected AA mϕ attenuated the anti-chlamydial function of CA mϕ with growth recovery observed in infected CA mϕ in the presence of infected, but not mock-infected AA mϕ. This could be related to our observation that IL-10 treatment of infected CA mϕ promoted better chlamydial growth. Thus, in addition to serving as an additional niche, AA mϕ might also serve as a means to modulate the immediate environment by attenuating the anti-chlamydial functions of nearby CA mϕ in a manner that could involve IL-10 produced by infected AA mϕ

Daytime naps improve motor imagery learning

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Determinants of GBP Recruitment to Toxoplasma gondii Vacuoles and the Parasitic Factors That Control It

IFN-γ is a major cytokine that mediates resistance against the intracellular parasite Toxoplasma gondii. The p65 guanylate-binding proteins (GBPs) are strongly induced by IFN-γ. We studied the behavior of murine GBP1 (mGBP1) upon infection with T. gondii in vitro and confirmed that IFN-γ-dependent re-localization of mGBP1 to the parasitophorous vacuole (PV) correlates with the virulence type of the parasite. We identified three parasitic factors, ROP16, ROP18, and GRA15 that determine strain-specific accumulation of mGBP1 on the PV. These highly polymorphic proteins are held responsible for a large part of the strain-specific differences in virulence. Therefore, our data suggest that virulence of T. gondii in animals may rely in part on recognition by GBPs. However, phagosomes or vacuoles containing Trypanosoma cruzi did not recruit mGBP1. Co-immunoprecipitation revealed mGBP2, mGBP4, and mGBP5 as binding partners of mGBP1. Indeed, mGBP2 and mGBP5 co-localize with mGBP1 in T. gondii-infected cells. T. gondii thus elicits a cell-autonomous immune response in mice with GBPs involved. Three parasitic virulence factors and unknown IFN-γ-dependent host factors regulate this complex process. Depending on the virulence of the strains involved, numerous GBPs are brought to the PV as part of a large, multimeric structure to combat T. gondii.National Institutes of Health (U.S.)Massachusetts Life Sciences Center (New Investigator Award)National Institute of General Medical Sciences (U.S.) (Pre-Doctoral Grant in the Biological Sciences (5-T32-GM007287-33))Studienstiftung des deutschen VolkesCancer Research Institute (New York, N.Y.)Cleo and Paul Schimmel FoundationBayer HealthcareHuman Frontier Science Program (Strasbourg, France

Intracellular Trafficking of Guanylate-Binding Proteins Is Regulated by Heterodimerization in a Hierarchical Manner

Guanylate-binding proteins (GBPs) belong to the dynamin family of large GTPases and represent the major IFN-γ-induced proteins. Here we systematically investigated the mechanisms regulating the subcellular localization of GBPs. Three GBPs (GBP-1, GBP-2 and GBP-5) carry a C-terminal CaaX-prenylation signal, which is typical for small GTPases of the Ras family, and increases the membrane affinity of proteins. In this study, we demonstrated that GBP-1, GBP-2 and GBP-5 are prenylated in vivo and that prenylation is required for the membrane association of GBP-1, GBP-2 and GBP-5. Using co-immunoprecipitation, yeast-two-hybrid analysis and fluorescence complementation assays, we showed for the first time that GBPs are able to homodimerize in vivo and that the membrane association of GBPs is regulated by dimerization similarly to dynamin. Interestingly, GBPs could also heterodimerize. This resulted in hierarchical positioning effects on the intracellular localization of the proteins. Specifically, GBP-1 recruited GBP-5 and GBP-2 into its own cellular compartment and GBP-5 repositioned GBP-2. In addition, GBP-1, GBP-2 and GBP-5 were able to redirect non-prenylated GBPs to their compartment in a prenylation-dependent manner. Overall, these findings prove in vivo the ability of GBPs to dimerize, indicate that heterodimerization regulates sub-cellular localization of GBPs and underscore putative membrane-associated functions of this family of proteins

Investigation of the Suppression of the Narrow Channel Effect in Deep Sub-Micron EXTIGATE Transistors

A suppression of the narrow channel effect for deep sub-micron CMOS transistors when using a novel device architecture called EXTIGATE has recently been shown. This work compares the narrow channel effect in the EXTIGATE and in conventional shallow trench isolated MOS transistors by means of three-dimensional coupled process and device simulation. The 3D simulation analysis shows that the bird's beak shaped thickening of the gate oxide and the sharply terminated gate electrode at the edge of the active area which are typical for the EXTIGATE technology play an important role in the suppression of the narrow channel effect

A Computationally Efficient Method for Three-Dimensional Simulation of Ion Implantation

Abstract The high accuracy which is necessary for modern process simulation often requires the use of Monte-Carlo ion implantation simulation methods, with the disadvantage of very long simulation times especially for three-dimensional applications. In this work a new method for an accurate and CPU time efficient three-dimensional simulation of ion implantation is suggested. The approach is based on a combination of the algorithmic capabilities of fast analytical and the Monte-Carlo simulation methods

Treffgenauigkeit, Rationalität und Streuung von Konjunkturprognosen für Deutschland

Konjunkturprognostiker und deren prognostische Fähigkeiten standen schon oft im Kreuzfeuer der öffentlichen Kritik. Seit einiger Zeit mehren sich die Fragen über die Tauglichkeit von Konjunkturprognosen. Da war vom "Blindflug der Forscher" zu lesen (vgl. Der Spiegel 2005), Feuilletonisten forderten "Entlasst die Experten" (vgl. FAZ 2005a) und auch Altbundeskanzler Gerhard Schröder hegte wenig Sympathie für "diese Art der Wissenschaft, die ihn an Meteorologie erinnere" (vgl. FAZ 2005b). Dass viele Wirtschaftsforschungsinstitute mit öffentlichen Geldern gefördert werden, steigert den Legitimationsdruck zusätzlich. Umso wichtiger ist es, die Debatte zu versachlichen und sich Grenzen, Möglichkeiten und Chancen der Konjunkturprognosen unter Verwendung nachprüfbarer Kriterien vor Augen zu führen.1 Der folgende Beitrag soll dazu beitragen. Dazu wird zunächst im Abschnitt 2 der Frage nachgegangen, welche erkenntnistheoretischen Probleme es grundsätzlich bei Prognosen gibt und was diese leisten können. Abschnitt 3 beschreibt den für die folgenden empirischen Untersuchungen verwendeten Datensatz. Im Abschnitt 4 geht es um die Treffgenauigkeit und Rationalität von Prognosen und im Abschnitt 5 soll geklärt werden, warum Prognosen differieren und welche Bestimmungsgründe es für Phasen hoher Divergenz bei den Prognosen geben könnte. Abschnitt 6 fasst die Ergebnisse zusammen

ProteoPlex: Stability optimization of macromolecular complexes by sparse-matrix screening of chemical space.

Molecular machines or macromolecular complexes are supramolecular assemblies of biomolecules with a variety of functions. Structure determination of these complexes in a purified state is often tedious owing to their compositional complexity and the associated relative structural instability. To improve the stability of macromolecular complexes in vitro, we present a generic method that optimizes the stability, homogeneity and solubility of macromolecular complexes by sparse-matrix screening of their thermal unfolding behavior in the presence of various buffers and small molecules. The method includes the automated analysis of thermal unfolding curves based on a biophysical unfolding model for complexes. We found that under stabilizing conditions, even large multicomponent complexes reveal an almost ideal two-state unfolding behavior. We envisage an improved biochemical understanding of purified macromolecules as well as a substantial boost in successful macromolecular complex structure determination by both X-ray crystallography and cryo-electron microscopy