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Acinetobacter baumannii is an opportunistic pathogen with a high mortality rate due to multi-drug-resistant strains. The synthesis and uptake of the iron-chelating siderophores acinetobactin (Acb) and preacinetobactin (pre-Acb) have been shown to be essential for virulence. Here, we report the kinetic and structural characterization of BauF, a flavin-dependent siderophore-interacting protein (SIP) required for the reduction of Fe(III) bound to Acb/pre-Acb and release of Fe(II). Stopped-flow spectrophotometric studies of the reductive half-reaction show that BauF forms a stable neutral flavin semiquinone intermediate. Reduction with NAD(P)H is very slow (k obs, 0.001 s-1) and commensurate with the rate of reduction by photobleaching, suggesting that NAD(P)H are not the physiological partners of BauF. The reduced BauF was oxidized by Acb-Fe (k obs, 0.02 s-1) and oxazole pre-Acb-Fe (ox-pre-Acb-Fe) (k obs, 0.08 s-1), a rigid analogue of pre-Acb, at a rate 3-11 times faster than that with molecular oxygen alone. The structure of FAD-bound BauF was solved at 2.85 Å and was found to share a similarity to Shewanella SIPs. The biochemical and structural data presented here validate the role of BauF in A. baumannii iron assimilation and provide information important for drug design.

Acinetobacter baumannii is an opportunistic pathogen with a high mortality rate due to multi-drug-resistant strains. The synthesis and uptake of the iron-chelating siderophores acinetobactin (Acb) and preacinetobactin (pre-Acb) have been shown to be essential for virulence. Here, we report the kinetic and structural characterization of BauF, a flavin-dependent siderophore-interacting protein (SIP) required for the reduction of Fe(III) bound to Acb/pre-Acb and release of Fe(II). Stopped-flow spectrophotometric studies of the reductive half-reaction show that BauF forms a stable neutral flavin semiquinone intermediate. Reduction with NAD(P)H is very slow (k obs, 0.001 s-1) and commensurate with the rate of reduction by photobleaching, suggesting that NAD(P)H are not the physiological partners of BauF. The reduced BauF was oxidized by Acb-Fe (k obs, 0.02 s-1) and oxazole pre-Acb-Fe (ox-pre-Acb-Fe) (k obs, 0.08 s-1), a rigid analogue of pre-Acb, at a rate 3-11 times faster than that with molecular oxygen alone. The structure of FAD-bound BauF was solved at 2.85 Å and was found to share a similarity to Shewanella SIPs. The biochemical and structural data presented here validate the role of BauF in A. baumannii iron assimilation and provide information important for drug design.

© The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in St. Laurent, L., Ijichi, T., Merrifield, S. T., Shapiro, J., & Simmons, H. L. Turbulence and vorticity in the Wake of Palau. Oceanography, 32(4), (2019): 102-109, doi: 10.5670/oceanog.2019.416., The interaction of flow with steep island and ridge topography at the Palau island chain leads to rich vorticity fields that generate a cascade of motions. The energy transfer to small scales removes energy from the large-scale mean flow of the equatorial current systems and feeds energy to the fine and microstructure scales where instability mechanisms lead to turbulence and dissipation. Until now, direct assessments of the turbulence associated with island wakes have received only minimal attention. Here, we examine data collected from an ocean glider equipped with microstructure sensors that flew in the island wake of Palau. We use a combination of submesoscale modeling and direct observation to quantify the relationship between vorticity and turbulence levels. We find that direct wind-driven mixing only accounts for about 10% of the observed turbulence levels, suggesting that most of the energy for mixing is extracted from the shear associated with the vorticity field in the island’s wake. Below the surface layer, enhanced turbulence correlates with the phase and magnitude of the relative vorticity and strain levels of the mesoscale flow., We thank the Palau National Government for permission to carry out the research in Palau. We also thank the US Office of Naval Research for supporting this work. We especially thank Pat and Lori Colin of the Coral Reef Research Foundation and their team for accommodating our research team in Koror, Palau, and running vessel operations in support of glider deployments and recoveries. Sean Whelan of the Woods Hole Oceanographic Institution and Lance Braasch of Scripps Institution of Oceanography provided technical support in the field. Funding for the development of HYCOM has been provided by the National Ocean Partnership Program and the Office of Naval Research. Data assimilative products using HYCOM are funded by the US Navy. Computer time was made available by the Department of Defense High Performance Computing Modernization Program. The output is publicly available at https://www.hycom.org/.

This dissertation examines the relationships between the physical environment and the history of public housing in Washington, D.C. from the 1940s to the 1960s. The environmental features of public housing complexes, as well as those of the landscape around them, significantly shaped the outcomes of the National Capital Housing Authority’s (NCHA) projects. The scale of public housing construction during that period entailed sweeping and dramatic transformations in the landscape. At the same time, the NCHA found itself constrained by material and financial pressures coming from a variety of bureaucratic and institutional sources. Those pressures limited the NCHA’s ability to respond to environmental stresses at various public housing sites. In the absence of adequate responses from the NCHA, the environment played a significant role in determining the outcomes of the District of Columbia’s public housing program. The physical nature of the NCHA’s choice of sites, as well as the materials that it used, turned public housing complexes into sites of environmental injustice rather than the decent, safe, and sanitary housing that the Authority envisioned.