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Bacterial cells alter gene expression in response to changes in population density in a process called quorum sensing (QS). In Vibrio harveyi, LuxO, a low cell density activator of sigma factor-54 (RpoN), is required for transcription of five non-coding regulatory sRNAs, Qrr1-Qrr5, which each repress translation of the master QS regulator LuxR. Vibrio parahaemolyticus, the leading cause of bacterial seafood-borne gastroenteritis, also contains five Qrr sRNAs that control OpaR (the LuxR homolog), controlling capsule polysaccharide (CPS), motility, and metabolism. We show that in a Δ luxO deletion mutant, opaR was de-repressed and CPS and biofilm were produced. However, in a Δ rpoN mutant, opaR was repressed, no CPS was produced, and less biofilm production was observed compared to wild type. To determine why opaR was repressed, expression analysis in Δ luxO showed all five qrr genes were repressed, while in Δ rpoN the qrr2 gene was significantly de-repressed. Reporter assays and mutant analysis showed Qrr2 sRNA can act alone to control OpaR. Bioinformatics analysis identified a sigma-70 (RpoD) -35 -10 promoter overlapping the canonical sigma-54 (RpoN) -24 -12 promoter in the qrr2 regulatory region. The qrr2 sigma-70 promoter element was also present in additional Vibrio species indicating it is widespread. Mutagenesis of the sigma-70 -10 promoter site in the Δ rpoN mutant background, resulted in repression of qrr2. Analysis of qrr quadruple deletion mutants, in which only a single qrr gene is present, showed that only Qrr2 sRNA can act independently to regulate opaR . Mutant and expression data also demonstrated that RpoN and the global regulator, Fis, act additively to repress qrr2 . Our data has uncovered a new mechanism of qrr expression and shows that Qrr2 sRNA is sufficient for OpaR regulation. Importance The quorum sensing non-coding sRNAs are present in all Vibrio species but vary in number and regulatory roles among species. In the Harveyi clade, all species contain five qrr genes, and in V. harveyi these are transcribed by sigma-54 and are additive in function. In the Cholerae clade, four qrr genes are present, and in V. cholerae the qrr genes are redundant in function. In V. parahaemolyticus , qrr2 is controlled by two overlapping promoters. In an rpoN mutant, qrr2 is transcribed from a sigma-70 promoter that is present in all V. parahaemolyticus strains and in other species of the Harveyi clade suggesting a conserved mechanism of regulation. Qrr2 sRNA can function as the sole Qrr sRNA to control OpaR. 

A multiline ring anchor (MRA) system has been developed as a cost-effective alternative for securing arrays of floating offshore wind turbines (FOWTs) to the seabed. Multiline attachments can improve the economically competitiveness of FOWTs by reducing the capital cost of the support system for the floating structures. FOWTs can be subjected to severe wind and wave conditions resulting in extreme loads to the anchor system. Thus, the reliable design of the anchor system requires proper determination of the extreme mooring line loads acting on the anchor needed to secure FOWTs to the seabed. Previous studies showed the MRA in soft clay has clear advantages over existing anchors under the extreme horizontal loading conditions imposed by catenary moorings; however, its performance relative to conventional anchors under extreme vertical loading imposed by taut mooring systems requires further investigation. This study presents predictions of extreme loads on floating structures secured by taut mooring systems and evaluates the potential for developing an economical anchor for resisting these extreme loads. 

Photosynthetic O 2 evolution is catalyzed by the Mn 4 CaO 5 cluster of the water oxidation complex of the photosystem II (PSII) complex. The photooxidative self-assembly of the Mn 4 CaO 5 cluster, termed photoactivation, utilizes the same highly oxidizing species that drive the water oxidation in order to drive the incorporation of Mn 2+ into the high-valence Mn 4 CaO 5 cluster. This multistep process proceeds with low quantum efficiency, involves a molecular rearrangement between light-activated steps, and is prone to photoinactivation and misassembly. A sensitive polarographic technique was used to track the assembly process under flash illumination as a function of the constituent Mn 2+ and Ca 2+ ions in genetically engineered membranes of the cyanobacterium Synechocystis sp. PCC6803 to elucidate the action of Ca 2+ and peripheral proteins. We show that the protein scaffolding organizing this process is allosterically modulated by the assembly protein Psb27, which together with Ca 2+ stabilizes the intermediates of photoactivation, a feature especially evident at long intervals between photoactivating flashes. The results indicate three critical metal-binding sites: two Mn and one Ca, with occupation of the Ca site by Ca 2+ critical for the suppression of photoinactivation. The long-observed competition between Mn 2+ and Ca 2+ occurs at the second Mn site, and its occupation by competing Ca 2+ slows the rearrangement. The relatively low overall quantum efficiency of photoactivation is explained by the requirement of correct occupancy of these metal-binding sites coupled to a slow restructuring of the protein ligation environment, which are jointly necessary for the photooxidative trapping of the first stable assembly intermediate. 

The trend of offshore wind energy in deeper water that is expected to shift from fixed to floating platforms requires a cost-effective anchor solution for floating offshore wind turbines (FOWTs). Multiline ring anchor (MRA) has been developed as a cost-effective solution for FOWTs due to its capability of anchoring multiple mooring lines, its high efficiency, and its availability to a wide range of soils and loading conditions. While previous preliminary studies on the anchor performance provide useful insights on how the potential advantages of the MRA can improve load capacity, these studies are limited to focusing on optimizing the anchor design in certain soil and loading conditions. By contrast, the MRA will be installed in seabeds under more complex conditions that depend on geological location, water depth of at-place, and environmental conditions, of which wind, current, and wave are major components. These may result in additional substantial extra capital costs, delays in the projects, and safety issues, when the complex conditions are not properly considered. Specifically, the installation time and expenses of the offshore anchor are very susceptible to anchor types, installation methods, and environmental conditions. For this reason, this paper compares two existing offshore anchor installation methods and different anchor types on the basis of their performance under the same severe environmental condition. In evaluating the installability of the MRA, this paper conducts a comparative scenario study. The results show that the anchor installations and anchor handling vessel (AHV) operations are sensitive to weather conditions and AHV sizes. In view of total weather standby, the results show that anchor types or installation methods have little effect on it due to their relatively shorter duration than other work sequences. However, the MRA can benefit in substantially reducing transport time and costs due to its compact size. The MRA can be more efficient and cost-effective than other alternatives under complex and severe weather conditions. 

A search for pair production of squarks or gluinos decaying via sleptons or weak bosons is reported. The search targets a final state with exactly two leptons with same-sign electric charge or at least three leptons without any charge requirement. The analysed data set corresponds to an integrated luminosity of 139 fb⁻¹of proton-proton collisions collected at a centre-of-mass energy of 13 TeV with the ATLAS detector at the LHC. Multiple signal regions are defined, targeting several SUSY simplified models yielding the desired final states. A single control region is used to constrain the normalisation of theWZ+ jets background. No significant excess of events over the Standard Model expectation is observed. The results are interpreted in the context of several supersymmetric models featuring R-parity conservation or R-parity violation, yielding exclusion limits surpassing those from previous searches. In models considering gluino (squark) pair production, gluino (squark) masses up to 2.2 (1.7) TeV are excluded at 95% confidence level.