Actin Re-Organization Induced by Chlamydia trachomatis Serovar D - Evidence for a Critical Role of the Effector Protein CT166 Targeting Rac

The intracellular bacterium Chlamydia trachomatis causes infections of urogenital tract, eyes or lungs. Alignment reveals homology of CT166, a putative effector protein of urogenital C. trachomatis serovars, with the N-terminal glucosyltransferase domain of clostridial glucosylating toxins (CGTs). CGTs contain an essential DXD-motif and mono-glucosylate GTP-binding proteins of the Rho/Ras families, the master regulators of the actin cytoskeleton. CT166 is preformed in elementary bodies of C. trachomatis D and is detected in the host-cell shortly after infection. Infection with high MOI of C. trachomatis serovar D containing the CT166 ORF induces actin re-organization resulting in cell rounding and a decreased cell diameter. A comparable phenotype was observed in HeLa cells treated with the Rho-GTPase-glucosylating Toxin B from Clostridium difficile (TcdB) or HeLa cells ectopically expressing CT166. CT166 with a mutated DXD-motif (CT166-mut) exhibited almost unchanged actin dynamics, suggesting that CT166-induced actin re-organization depends on the glucosyltransferase motif of CT166. The cytotoxic necrotizing factor 1 (CNF1) from E. coli deamidates and thereby activates Rho-GTPases and transiently protects them against TcdB-induced glucosylation. CNF1-treated cells were found to be protected from TcdB- and CT166-induced actin re-organization. CNF1 treatment as well as ectopic expression of non-glucosylable Rac1-G12V, but not RhoA-G14A, reverted CT166-induced actin re-organization, suggesting that CT166-induced actin re-organization depends on the glucosylation of Rac1. In accordance, over-expression of CT166-mut diminished TcdB induced cell rounding, suggesting shared substrates. Cell rounding induced by high MOI infection with C. trachomatis D was reduced in cells expressing CT166-mut or Rac1-G12V, and in CNF1 treated cells. These observations indicate that the cytopathic effect of C. trachomatis D is mediated by CT166 induced Rac1 glucosylation. Finally, chlamydial uptake was impaired in CT166 over-expressing cells. Our data strongly suggest CT166's participation as an effector protein during host-cell entry, ensuring a balanced uptake into host-cells by interfering with Rac-dependent cytoskeletal changes

Nitrogen transport and retention dynamics across central European catchments using large-sample data

Human activities, especially agricultural practices, have significantly altered the Earth’s landscape and the global cycle of nitrogen. In Europe, diffuse nitrogen (N) input from agriculture has been identified as a major driver of marine eutrophication. Despite a long history of measures, little improvement in groundwater and surface water quality has been observed. Recent studies have attempted to provide insights into nitrogen dynamics at the catchment scale, helping to explain the causes and effects of persistent water quality problems. However, there is a lack of large-scale, long-term studies that provide insights into both biogeochemical and hydrological N legacies under different landscape settings. Here using data of more than 100 German catchments of the last seven decades, we synthesis the nitrogen transport and retention dynamics, as well as their dominant (landscape and climate) controls in a large-sample setting. To this end, we adapted the mHM-SAS model (Nguyen et al., 2021) to reflect regional-scale biogeochemical and hydrological N legacies, taking into account the historical development of both diffuse and point sources. The underlying parameterizations were constrained using instream N concentrations. We found high heterogeneity in catchment responses to N inputs. The fractions of N surplus that were stored in the soil, removed by denitrification, stored in the subsurface, and finally exported to the stream vary over a wide range. Our analysis of the long-term (1950-2014) average N balances from all catchments suggests that a majority (mean = 57%) of N surplus was removed by denitrification, followed by stream N export (27%) and the rest was stored in the catchment (16%). Despite the reduction in N surplus after 1990s, biogeochemical legacy reflected in the soil N build-up showed an increasing trend over the analyzed period (1950-2014) across a majority of the study catchments. As for the hydrologic legacy, we found a varying range of mean transit times of discharge between 3.5 years and 13.1 years (95% confidence interval) among the analyzed catchments. Overall, our large-sample analysis provides a detailed overview of biogeochemical and hydrological N legacies across Germany; and thus provides useful insights for an improvement of agricultural practices and water quality management in Central European landscapes

Inhaled nitric oxide improves transpulmonary blood flow and clinical outcomes after prolonged cardiac arrest: a large animal study

Introduction: The probability to achieve a return of spontaneous circulation (ROSC) after cardiac arrest can be improved by optimizing circulation during cardiopulomonary resuscitation using a percutaneous left ventricular assist device (iCPR). Inhaled nitric oxide may facilitate transpulmonary blood flow during iCPR and may therefore improve organ perfusion and outcome. Methods: Ventricular fibrillation was electrically induced in 20 anesthetized male pigs. Animals were left untreated for 10 minutes before iCPR was attempted. Subjects received either 20 ppm of inhaled nitric oxide (iNO, n = 10) or 0 ppm iNO (Control, n = 10), simultaneously started with iCPR until 5 hours following ROSC. Animals were weaned from the respirator and followed up for five days using overall performance categories (OPC) and a spatial memory task. On day six, all animals were anesthetized again, and brains were harvested for neurohistopathologic evaluation. Results: All animals in both groups achieved ROSC. Administration of iNO markedly increased iCPR flow during CPR (iNO: 1.81 ± 0.30 vs Control: 1.64 ± 0.51 L/min, p < 0.001), leading to significantly higher coronary perfusion pressure (CPP) during the 6 minutes of CPR (25 ± 13 vs 16 ± 6 mmHg, p = 0.002). iNO-treated animals showed significantly lower S-100 serum levels thirty minutes post ROSC (0.26 ± 0.09 vs 0.38 ± 0.15 ng/mL, p = 0.048), as well as lower blood glucose levels 120–360 minutes following ROSC. Lower S-100 serum levels were reflected by superior clinical outcome of iNO-treated animals as estimated with OPC (3 ± 2 vs. 5 ± 1, p = 0.036 on days 3 to 5). Three out of ten iNO-treated, but none of the Control animals were able to successfully participate in the spatial memory task. Neurohistopathological examination of vulnerable cerebral structures revealed a trend towards less cerebral lesions in neocortex, archicortex, and striatum in iNO-treated animals compared to Controls. Conclusions: In pigs resuscitated with mechanically-assisted CPR from prolonged cardiac arrest, the administration of 20 ppm iNO during and following iCPR improved transpulmonary blood flow, leading to improved clinical neurological outcomes

SYSTOMONAS — an integrated database for systems biology analysis of Pseudomonas

To provide an integrated bioinformatics platform for a systems biology approach to the biology of pseudomonads in infection and biotechnology the database SYSTOMONAS (SYSTems biology of pseudOMONAS) was established. Besides our own experimental metabolome, proteome and transcriptome data, various additional predictions of cellular processes, such as gene-regulatory networks were stored. Reconstruction of metabolic networks in SYSTOMONAS was achieved via comparative genomics. Broad data integration is realized using SOAP interfaces for the well established databases BRENDA, KEGG and PRODORIC. Several tools for the analysis of stored data and for the visualization of the corresponding results are provided, enabling a quick understanding of metabolic pathways, genomic arrangements or promoter structures of interest. The focus of SYSTOMONAS is on pseudomonads and in particular Pseudomonas aeruginosa, an opportunistic human pathogen. With this database we would like to encourage the Pseudomonas community to elucidate cellular processes of interest using an integrated systems biology strategy. The database is accessible at

QUADICA: Water quality, discharge and catchment attributes for large-sample studies in Germany

Environmental data are the key to define and address water quality and quantity challenges at catchment scale. Here, we present the first large-sample water quality data set for 1386 German catchments covering a large range of hydroclimatic, topographic, geologic, land use and anthropogenic settings. QUADICA (water QUAlity, DIscharge and Catchment Attributes for large-sample studies in Germany) combines water quality with water quantity data, meteorological and nutrient forcing data, and catchment attributes. The data set comprises time series of riverine macronutrient concentrations (species of nitrogen, phosphorus and organic carbon) and diffuse nitrogen forcing data at catchment scale (nitrogen surplus, atmospheric deposition and fixation). Time series are generally aggregated to an annual basis; however, for 140 stations with long-term water quality and quantity data (more than 20 years), we additionally present monthly median discharge and nutrient concentrations, flow-normalized concentrations and corresponding mean fluxes as outputs from weighted regressions on time, discharge, and season (WRTDS). The catchment attributes include catchment nutrient inputs from point and diffuse sources and characteristics from topography, climate, land cover, lithology and soils. This comprehensive, freely available data collection can facilitate large-sample data-driven water quality assessments at catchment scale as well as mechanistic modeling studies

QUADICA: A large-sample data set of water quality, discharge and catchment attributes for Germany

Environmental data are critical for understanding and managing ecosystems, including mitigation of degraded water quality. Therefore, we provide the first large-sample water quality data set of riverine water quality combined with water quantity, meteorological and nutrient forcing data, and catchment attributes for Germany in a preprocessed and structured form. The QUADICA data set (water QUAlity, DIscharge and Catchment Attributes for large-sample studies in Germany) covers 1386 German and transboundary catchments with a large range of hydroclimatic, topographic, geologic, land use and anthropogenic settings. The data set comprises time series of riverine macronutrient concentrations (species of nitrogen, phosphorus and organic carbon), discharge, meteorological and diffuse nitrogen forcing data (nitrogen surplus, atmospheric deposition and fixation). The time series are generally aggregated to an annual basis; however, for 140 stations with long-term water quality and quantity data (more than 20 years), we additionally provide monthly median discharge and nutrient concentrations, flow-normalized concentrations and corresponding mean fluxes as outputs from weighted regressions on time, discharge, and season (WRTDS). The catchment attributes include catchment nutrient inputs from point and diffuse sources and characteristics from topography, hydroclimate, land cover, lithology and soils. QUADICA is a comprehensive, freely available, ready-to-use data set that facilitates large-sample data-driven water quality assessments at catchment scale as well as mechanistic modeling studies. We hope to stimulate the hydrological and water quality communities to provide similar data sets to create novel research opportunities, increase our understanding of catchment functioning, and ultimately improve water quality management

Upper limits on the strength of periodic gravitational waves from PSR J1939+2134

The first science run of the LIGO and GEO gravitational wave detectors presented the opportunity to test methods of searching for gravitational waves from known pulsars. Here we present new direct upper limits on the strength of waves from the pulsar PSR J1939+2134 using two independent analysis methods, one in the frequency domain using frequentist statistics and one in the time domain using Bayesian inference. Both methods show that the strain amplitude at Earth from this pulsar is less than a few times $10^{-22}$.Comment: 7 pages, 1 figure, to appear in the Proceedings of the 5th Edoardo Amaldi Conference on Gravitational Waves, Tirrenia, Pisa, Italy, 6-11 July 200