High-salinity growth conditions promote tat-independent secretion of tat substrates in Bacillus subtilis

The Gram-positive bacterium Bacillus subtilis contains two Tat translocases, which can facilitate transport of folded proteins across the plasma membrane. Previous research has shown that Tat-dependent protein secretion in B. subtilis is a highly selective process and that heterologous proteins, such as the green fluorescent protein (GFP), are poor Tat substrates in this organism. Nevertheless, when expressed in Escherichia coli, both B. subtilis Tat translocases facilitated exclusively Tat-dependent export of folded GFP when the twin-arginine (RR) signal peptides of the E. coli AmiA, DmsA, or MdoD proteins were attached. Therefore, the present studies were aimed at determining whether the same RR signal peptide-GFP precursors would also be exported Tat dependently in B. subtilis. In addition, we investigated the secretion of GFP fused to the full-length YwbN protein, a strict Tat substrate in B. subtilis. Several investigated GFP fusion proteins were indeed secreted in B. subtilis, but this secretion was shown to be completely Tat independent. At high-salinity growth conditions, the Tat-independent secretion of GFP as directed by the RR signal peptides from the E. coli AmiA, DmsA, or MdoD proteins was significantly enhanced, and this effect was strongest in strains lacking the TatAy-TatCy translocase. This implies that high environmental salinity has a negative influence on the avoidance of Tat-independent secretion of AmiA-GFP, DmsA-GFP, and MdoD-GFP. We conclude that as-yet-unidentified control mechanisms reject the investigated GFP fusion proteins for translocation by the B. subtilis Tat machinery and, at the same time, set limits to their Tat-independent secretion, presumably via the Sec pathway

Effects of Added Vegetation on Sand Bar Stability and Stream Hydrodynamics

Vegetation was added to a fully developed sandy point bar in the meander of a constructed stream. Significant changes in the flow structure and bed topography were observed. As expected, the addition of vegetative resistance decreased the depth-averaged streamwise velocity over the bar and increased it in the open region. In addition, the secondary circulation increased in strength but became confined to the deepest section of the channel. Over the point bar, the secondary flow was entirely outward, i.e., toward the outer bank. The changes in flow led to changes in bar shape. Although the region of the bar closest to the inner bank accumulated sediment, erosion of the bar and the removal of plants by scouring were observed at the interface between the planted bar and the open channel.National Science Foundation (U.S.) (Grant No. EAR 0738352

Discharge and force distribution in a sinuous channel with vegetated floodplains during overbank flow

This is an Accepted Manuscript of an article published by Taylor & Francis Group in Journal of Hydraulic Research on 2019, available online at: http://www.tandfonline.com/10.1080/00221686.2019.1581667Overbank flow in a sinuous channel with roughened floodplains has been investigated, focusing on the effect of floodplain vegetation on overall flow resistance. The physical model of the Besòs River has allowed analysing the effect of flexible roughness elements which simulate the natural vegetation of rivers. The experimental measurements of horizontal velocities have been used to obtain zonal discharges and forces along a meander wavelength. The results illustrate that although mass transfer is the most important source of energy losses, in rivers with strongly vegetated floodplains the flow resistance increases considerably due to the strong apparent shear forces acting between the main channel and floodplains.Peer ReviewedPostprint (author's final draft

Detecting Lactococcus lactis prophages by Mitomycin C-mediated induction coupled to flow cytometry analysis

Most analyzed Lactococcus lactis strains are predicted to harbor one or more prophage genomes within their chromosome; however, the true extent of the inducibility and functionality of such prophages cannot easily be deduced from sequence analysis alone. Chemical treatment of lysogenic strains with Mitomycin C is known to cause induction of temperate phages, though it is not always easy to clearly identify a lysogenic strain or to measure the number of released phage particles. Here, we report the application of flow cytometry as a reliable tool for the detection and enumeration of released lactococcal prophages using the green dye SYTO-9

Staphylococcal PknB as the First Prokaryotic Representative of the Proline-Directed Kinases

In eukaryotic cell types, virtually all cellular processes are under control of proline-directed kinases and especially MAP kinases. Serine/threonine kinases in general were originally considered as a eukaryote-specific enzyme family. However, recent studies have revealed that orthologues of eukaryotic serine/threonine kinases exist in bacteria. Moreover, various pathogenic species, such as Yersinia and Mycobacterium, require serine/threonine kinases for successful invasion of human host cells. The substrates targeted by bacterial serine/threonine kinases have remained largely unknown. Here we report that the serine/threonine kinase PknB from the important pathogen Staphylococcus aureus is released into the external milieu, which opens up the possibility that PknB does not only phosphorylate bacterial proteins but also proteins of the human host. To identify possible human targets of purified PknB, we studied in vitro phosphorylation of peptide microarrays and detected 68 possible human targets for phosphorylation. These results show that PknB is a proline-directed kinase with MAP kinase-like enzymatic activity. As the potential cellular targets for PknB are involved in apoptosis, immune responses, transport, and metabolism, PknB secretion may help the bacterium to evade intracellular killing and facilitate its growth. In apparent agreement with this notion, phosphorylation of the host-cell response coordinating transcription factor ATF-2 by PknB was confirmed by mass spectrometry. Taken together, our results identify PknB as the first prokaryotic representative of the proline-directed kinase/MAP kinase family of enzymes

Retention time and dispersion associated with submerged aquatic canopies

Author Posting. © American Geophysical Union, 2007. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Water Resources Research 43 (2007): W04422, doi:10.1029/2006WR005362.The shear layer at the top of a submerged canopy generates coherent vortices that control exchange between the canopy and the overflowing water. Unlike free shear layers, the vortices in a canopy shear layer do not grow continuously downstream but reach and maintain a finite scale determined by a balance between shear production and canopy dissipation. This balance defines the length scale of vortex penetration into the canopy, δ e , and the region of rapid exchange between the canopy and overflow. Deeper within the canopy, transport is constrained by smaller turbulence scales. A two-box canopy model is proposed on the basis of the length scale δ e . Using diffusivity and exchange rates defined in previous studies, the model predicts the timescale required to flush the canopy through vertical exchange over a range of canopy density and height. The predicted canopy retention times, which range from minutes to an hour, are consistent with canopy retention inferred from tracer observations in the field and comparable to retention times for some hyporheic regions. The timescale for vertical exchange, along with the in-canopy velocity, determines the minimum canopy length for which vertical exchange dominates water renewal. Shorter canopies renew interior water through longitudinal advection. Finally, canopy water retention influences longitudinal dispersion through a transient storage process. When vertical exchange controls canopy retention, the transient storage dispersion increases with canopy height. When longitudinal advection controls water renewal, dispersion increases with canopy patch length.This material is based upon work supported by the National Science Foundation under grant EAR0309188

Avalanche amplification of a single exciton in a semiconductor nanowire

Interfacing single photons and electrons is a crucial ingredient for sharing quantum information between remote solid-state qubits. Semiconductor nanowires offer the unique possibility to combine optical quantum dots with avalanche photodiodes, thus enabling the conversion of an incoming single photon into a macroscopic current for efficient electrical detection. Currently, millions of excitation events are required to perform electrical read-out of an exciton qubit state. Here we demonstrate multiplication of carriers from only a single exciton generated in a quantum dot after tunneling into a nanowire avalanche photodiode. Due to the large amplification of both electrons and holes (> 10^4), we reduce by four orders of magnitude the number of excitation events required to electrically detect a single exciton generated in a quantum dot. This work represents a significant step towards single-shot electrical read-out and offers a new functionality for on-chip quantum information circuits

Detectability of colorectal neoplasia with fluorine-18-2-fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography (FDG-PET/CT)

The purpose of this study was to analyze the detectability of colorectal neoplasia with fluorine-18-2-fluoro-2-deoxy-d-glucose positron emission tomography/computed tomography (FDG-PET/CT). Data for a total of 492 patients who had undergone both PET/CT and colonoscopy were analyzed. After the findings of PET/CT and colonoscopy were determined independently, the results were compared in each of the six colonic sites examined in all patients. The efficacy of PET/CT was determined using colonoscopic examination as the gold standard. In all, 270 colorectal lesions 5 mm or more in size, including 70 pathologically confirmed malignant lesions, were found in 172 patients by colonoscopy. The sensitivity and specificity of PET/CT for detecting any of the colorectal lesions were 36 and 98%, respectively. For detecting lesions 11 mm or larger, the sensitivity was increased to 85%, with the specificity remaining consistent (97%). Moreover, the sensitivity for tumors 21 mm or larger was 96% (48/50). Tumors with malignant or high-grade pathology were likely to be positive with PET/CT. A size of 10 mm or smaller [odds ratio (OR) 44.14, 95% confidence interval (95% CI) 11.44-221.67] and flat morphology (OR 7.78, 95% CI 1.79-36.25) were significant factors that were associated with false-negative cases on PET/CT. The sensitivity of PET/CT for detecting colorectal lesions is acceptable, showing size- and pathology-dependence, suggesting, for the most part, that clinically relevant lesions are detectable with PET/CT. However, when considering PET/CT for screening purposes caution must be exercised because there are cases of false-negative results