Deficiencies of the T and natural killer cell system in major depressive disorder

Background: In a previous study, we found an up-regulated inflammatory monocyte gene expression profile in major depressive disorder (MDD) patients aged >= 28 years and a down-regulated inflammatory gene expression profile in MDD patients aged = 28 years additionally exhibited decreased percentages of CD4(+)CD25(high)FoxP3(+) T regulatory cells, next to signs of the above described partial T cell defects. Natural T regulatory cells were inversely associated with the pro-inflammatory state of the monocytes (r = -.311; p = .034) that characterized this patient subgroup. Conclusions: Deficiencies of the NK and T (regulatory) cell system and inflammatory monocyte immune activation co-occur as partly interrelated phenomena within the same MDD patients. (C) 2015 Elsevier Inc. All rights reserved

The CRESST dark matter search

The CRESST (Cryogenic Rare Event Search with Superconducting Thermometers) experiment at the Gran Sasso Laboratory will search for dark matter WIMPs using cryogenic detectors. In the first stage we will use four 262 g sapphire crystals with thresholds of about 0.5 keV. This low threshold gives us sensitivity to WIMP masses below 10 GeV, making CRESST complementary to other dark matter searches. The main installation in Gran Sasso is now complete and we report on the first detector tests

Global Food Security Governance: Civil Society Engagement in the Reformed Committee on World Food Security

In 2007/8 world food prices spiked and global economic crisis set in, leaving hundreds of millions of people unable to access adequate food. The international reaction was swift. In a bid for leadership, the 123 member countries of the United Nations’ Committee on World Food Security (CFS) adopted a series of reforms with the aim of becoming the foremost international, inclusive and intergovernmental platform for food security. Central to the reform was the inclusion of participants (including civil society and the private sector) across all activities of the Committee. Drawing on data collected from policy documents, interviews and participant observation, this book examines the re-organization and functioning of a UN Committee that is coming to be known as a best practice in global governance. Framed by key challenges that plague global governance, the impact and implication of increased civil society engagement are examined by tracing policy negotiations within the CFS, in particular, policy roundtables on smallholder sensitive investment and food price volatility and negotiations on the Voluntary Guidelines on the Responsible Governance of Tenure of Land, Fisheries and Forests in the Context of National Food Security, and the Global Strategic Framework for Food Security and Nutrition. The author shows that through their participation in the Committee, civil society actors are influencing policy outcomes. Yet analysis also reveals that the CFS is being undermined by other actors seeking to gain and maintain influence at the global level. By way of this analysis, this book provides empirically-informed insights into increased participation in global governance processes

Subcellular localization of proteins in the anaerobic sulfate reducer Desulfovibrio vulgaris via SNAP-tag labeling and photoconversion

Systems Biology studies the temporal and spatial 3D distribution of macromolecular complexes with the aim that such knowledge will allow more accurate modeling of biological function and will allow mathematical prediction of cellular behavior. However, in order to accomplish accurate modeling precise knowledge of spatial 3D organization and distribution inside cells is necessary. And while a number of macromolecular complexes may be identified by its 3D structure and molecular characteristics alone, the overwhelming number of proteins will need to be localized using a reporter tag. GFP and its derivatives (XFPs) have been traditionally employed for subcelllar localization using photoconversion approaches, but this approach cannot be taken for obligate anaerobic bacteria, where the intolerance towards oxygen prevents XFP approaches. As part of the GTL-funded PCAP project (now ENIGMA) genetic tools have been developed for the anaerobe sulfate reducer Desulfovibrio vulgaris that allow the high-throughput generation of tagged-protein mutant strains, with a focus on the commercially available SNAP-tag cell system (New England Biolabs, Ipswich, MA), which is based on a modified O6-alkylguanine-DNA alkyltransferase (AGT) tag, that has a dead-end reaction with a modified O6-benzylguanine (BG) derivative and has been shown to function under anaerobic conditions. After initial challenges with respect to variability, robustness and specificity of the labeling signal we have optimized the labeling. Over the last year, as a result of the optimized labeling protocol, we now obtain robust labeling of 20 out of 31 SNAP strains. Labeling for 13 strains were confirmed at least five times. We have also successfully performed photoconversion on 5 of these 13 strains, with distinct labeling patterns for different strains. For example, DsrC robustly localizes to the periplasmic portion of the inner membrane, where as a DNA-binding protein localizes to the center of the cell, where the chromosome is located. Two other proteins - Thiosulfate reductase and ATP binding protein were found to be cytoplasmically distributed, whereas a molybdenum transporter was found to locate to the cell periphery. We judge labeling outcome by (1) SDS gel electrophoresis, followed by direct fluorescence imaging of the gel to address specificity of labeling/confirm expected molecular weight, and subsequent Coomassie analysis to ensure comparable protein levels (2) fluorescence intensity of culture by plate reader for statistical sampling (after adjustment for respective cell numbers) and (3) fluorescence microscopy for addressing cell-to-cell signal variation and potential localization patterns. All three assays were usually found to be consistent with one another. While we have been able to improve the efficacy of photoconversion by drastically reducing (eliminating) non-specific binding with our altered labeling protocol, we are currently working on reducing non-specific photoconversion reaction arising occasionally in non-labeled cells. In addition, we have confirmed the presence of SNAP tagged constructs in three recently cloned E.coli strains under promotor control, and are in the process of utilizing them for evaluating the sensitivity of the photoconversion protocol. Fluorescent Activated Cell Sorting was successfully applied to labeled E.coli cells containing SNAP tagged AtpA protein. Different batches of sorted cells, representing low and high labeling intensity, were re-grown and re-labeled and displayed a labeling efficiency similar to the starter culture, supporting the notion that cell-to-cell differences in labeling reflect difference in protein expression, rather then genetic differences