RAB5A and TRAPPC6B are novel targets for Shiga toxin 2a inactivation in kidney epithelial cells

The cardinal virulence factor of human-pathogenic enterohaemorrhagic Escherichia coli (EHEC) is Shiga toxin (Stx), which causes severe extraintestinal complications including kidney failure by damaging renal endothelial cells. In EHEC pathogenesis, the disturbance of the kidney epithelium by Stx becomes increasingly recognised, but how this exactly occurs is unknown. To explore this molecularly, we investigated the Stx receptor content and transcriptomic profile of two human renal epithelial cell lines: highly Stx-sensitive ACHN cells and largely Stx-insensitive Caki-2 cells. Though both lines exhibited the Stx receptor globotriaosylceramide, RNAseq revealed strikingly different transcriptomic responses to an Stx challenge. Using RNAi to silence factors involved in ACHN cells’ Stx response, the greatest protection occurred when silencing RAB5A and TRAPPC6B, two host factors that we newly link to Stx trafficking. Silencing these factors alongside YKT6 fully prevented the cytotoxic Stx effect. Overall, our approach reveals novel subcellular targets for potential therapies against Stx-mediated kidney failure.publishedVersio

The PyCBC search for gravitational waves from compact binary coalescence

We describe the PyCBC search for gravitational waves from compact-object binary coalescences in advanced gravitational-wave detector data. The search was used in the first Advanced LIGO observing run and unambiguously identified two black hole binary mergers, GW150914 and GW151226. At its core, the PyCBC search performs a matched-filter search for binary merger signals using a bank of gravitational-wave template waveforms. We provide a complete description of the search pipeline including the steps used to mitigate the effects of noise transients in the data, identify candidate events and measure their statistical significance. The analysis is able to measure false-alarm rates as low as one per million years, required for confident detection of signals. Using data from initial LIGO's sixth science run, we show that the new analysis reduces the background noise in the search, giving a 30% increase in sensitive volume for binary neutron star systems over previous searches.Comment: 29 pages, 7 figures, accepted by Classical and Quantum Gravit

Croup Is Associated with the Novel Coronavirus NL63

BACKGROUND: The clinical relevance of infections with the novel human coronavirus NL63 (HCoV-NL63) has not been investigated systematically. We therefore determined its association with disease in young children with lower respiratory tract infection (LRTI). METHODS AND FINDINGS: Nine hundred forty-nine samples of nasopharyngeal secretions from children under 3 y of age with LRTIs were analysed by a quantitative HCoV-NL63-specific real-time PCR. The samples had been collected from hospitalised patients and outpatients from December 1999 to October 2001 in four different regions in Germany as part of the prospective population-based PRI.DE study and analysed for RNA from respiratory viruses. Forty-nine samples (5.2%), mainly derived from the winter season, were positive for HCoV-NL63 RNA. The viral RNA was more prevalent in samples from outpatients (7.9%) than from hospitalised patients (3.2%, p = 0.003), and co-infection with either respiratory syncytial virus or parainfluenza virus 3 was observed frequently. Samples in which only HCoV-NL63 RNA could be detected had a significantly higher viral load than samples containing additional respiratory viruses (median 2.1 × 10(6) versus 2.7 × 10(2) copies/ml, p = 0.0006). A strong association with croup was apparent: 43% of the HCoV-NL63-positive patients with high HCoV-NL63 load and absence of co-infection suffered from croup, compared to 6% in the HCoV-NL63-negative group, p < 0.0001. A significantly higher fraction (17.4%) of samples from croup patients than from non-croup patients (4.2%) contained HCoV-NL63 RNA. CONCLUSION: HCoV-NL63 infections occur frequently in young children with LRTI and show a strong association with croup, suggesting a causal relationship

Modeling Native EHEC Outer Membrane Vesicles by Creating Synthetic Surrogates

Enterohemorrhagic Escherichia coli (EHEC) is a zoonotic pathogen responsible for life-threating diseases such as hemolytic uremic syndrome. While its major virulence factor, the Shiga toxin (Stx), is known to exert its cytotoxic effect on various endothelial and epithelial cells when in its free, soluble form, Stx was also recently found to be associated with EHEC outer membrane vesicles (OMVs). However, depending on the strain background, other toxins can also be associated with native OMVs (nOMVs), and nOMVs are also made up of immunomodulatory agents such as lipopolysaccharides and flagellin. Thus, it is difficult to determine to which extent a single virulence factor in nOMVs, such as Stx, contributes to the molecular pathogenesis of EHEC. To reduce this complexity, we successfully developed a protocol for the preparation of synthetic OMVs (sOMVs) with a defined lipid composition resembling the E. coli outer membrane and loaded with specific proteins, i.e., bovine serum albumin (BSA) as a proxy for functional Stx2a. Using BSA for parameter evaluation, we found that (1) functional sOMVs can be prepared at room temperature instead of potentially detrimental higher temperatures (e.g., 45 &deg;C), (2) a 1:10 ratio of protein to lipid, i.e., 100 &micro;g protein with 1 mg of lipid mixture, yields homogenously sized sOMVs, and (3) long-term storage for up to one year at 4 &deg;C is possible without losing structural integrity. Accordingly, we reproducibly generated Stx2a-loaded sOMVs with an average diameter of 132.4 &plusmn; 9.6 nm that preserve Stx2a&rsquo;s injuring activity, as determined by cytotoxicity assays with Vero cells. Overall, we successfully created sOMVs and loaded them with an EHEC toxin, which opens the door for future studies on the degree of virulence associated with individual toxins from EHEC and other bacterial pathogens

Syn‐ and post‐eruptive gully formation near the Laacher See volcano

The Laacher See volcano (LSV) is located at the western margin of the Neuwied Basin, the central part of the Middle Rhine Basin of Germany. Its paroxysmal Plinian eruption c. 13 ka ago (Laacher See event; LSE) deposited a complex tephra sequence in the Neuwied Basin, whilst the distal ashes became one of the most important chronostratigraphic markers in Central Europe. However, some other impacts on landscape formation have thus far been largely neglected, such as buried gully structures in the proximity of the LSV. In this contribution, we map and discuss the spatial extent of these landforms at the site Lungenkärchen c. 4 km south of the LSV based on geophysical prospection as well as contrasting pedo‐sedimentary characteristics of the gully infill (particle‐size distribution, bulk‐sediment density, thin‐section analysis, saturated hydraulic conductivity) and the surrounding soils and tephra layers. These data are combined with a luminescence‐ and carbon‐14 (14C)‐based age model that relates them to the LSE. It is demonstrated how these gullies seem to have been formed and rapidly infilled by rainfall and surface discharge both during and subsequent to the eruptive phase, with modern analog processes documented for the 1980 Mount St Helens eruption (Washington State, USA). Given the density of the gullies at the site and their deviating pedo‐sedimentary properties compared to the surrounding soils, we propose a significant influence on agricultural production in the proximity of the LSV, which remains to be tested in future studies. Finally, in contrast, gullies of similar lateral and vertical dimensions identified in post‐LSE reworked loess and tephra deposits of the Wingertsbergwand (close to the main study site and proximal to the LSV) have shown to be unrelated to the LSE and can either be attributed to periglacial processes at the Younger Dryas‐Preboreal transition or to linear incision during the early Holocene.Linear subsurface gullies were identified close to the Laacher See volcano in magnetometer and ground‐penetrating radar prospection. Optically stimulated luminescence data indicate they incised during or shortly after the Laacher See event 13 ka ago. Pedo‐sedimentary characteristics of the gully infill differs from the surrounding regosols and brown earths, possibly influencing regional agricultural land use. imageUniversity of Cologne http://dx.doi.org/10.13039/501100008001Deutsche Forschungsgemeinschaft http://dx.doi.org/10.13039/50110000165

RAB5A and TRAPPC6B are novel targets for Shiga toxin 2a inactivation in kidney epithelial cells

The cardinal virulence factor of human-pathogenic enterohaemorrhagic Escherichia coli (EHEC) is Shiga toxin (Stx), which causes severe extraintestinal complications including kidney failure by damaging renal endothelial cells. In EHEC pathogenesis, the disturbance of the kidney epithelium by Stx becomes increasingly recognised, but how this exactly occurs is unknown. To explore this molecularly, we investigated the Stx receptor content and transcriptomic profile of two human renal epithelial cell lines: highly Stx-sensitive ACHN cells and largely Stx-insensitive Caki-2 cells. Though both lines exhibited the Stx receptor globotriaosylceramide, RNAseq revealed strikingly different transcriptomic responses to an Stx challenge. Using RNAi to silence factors involved in ACHN cells’ Stx response, the greatest protection occurred when silencing RAB5A and TRAPPC6B, two host factors that we newly link to Stx trafficking. Silencing these factors alongside YKT6 fully prevented the cytotoxic Stx effect. Overall, our approach reveals novel subcellular targets for potential therapies against Stx-mediated kidney failure

Syn- and post-eruptive gully formation near the Laacher See volcano

The Laacher See volcano (LSV) is located at the western margin of the Neuwied Basin, the central part of the Middle Rhine Basin of Germany. Its paroxysmal Plinian eruption c. 13 ka ago (Laacher See event; LSE) deposited a complex tephra sequence in the Neuwied Basin, whilst the distal ashes became one of the most important chronostratigraphic markers in Central Europe. However, some other impacts on landscape formation have thus far been largely neglected, such as buried gully structures in the proximity of the LSV. In this contribution, we map and discuss the spatial extent of these landforms at the site Lungenkarchen c. 4 km south of the LSV based on geophysical prospection as well as contrasting pedo-sedimentary characteristics of the gully infill (particle-size distribution, bulk-sediment density, thin-section analysis, saturated hydraulic conductivity) and the surrounding soils and tephra layers. These data are combined with a luminescence- and carbon-14 (C-14)-based age model that relates them to the LSE. It is demonstrated how these gullies seem to have been formed and rapidly infilled by rainfall and surface discharge both during and subsequent to the eruptive phase, with modern analog processes documented for the 1980 Mount St Helens eruption (Washington State, USA). Given the density of the gullies at the site and their deviating pedo-sedimentary properties compared to the surrounding soils, we propose a significant influence on agricultural production in the proximity of the LSV, which remains to be tested in future studies. Finally, in contrast, gullies of similar lateral and vertical dimensions identified in post-LSE reworked loess and tephra deposits of the Wingertsbergwand (close to the main study site and proximal to the LSV) have shown to be unrelated to the LSE and can either be attributed to periglacial processes at the Younger Dryas-Preboreal transition or to linear incision during the early Holocene