Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain

In response to light, as part of a two-component system, the Brucella blue light-activated histidine kinase (LOV-HK) increases its autophosphorylation, modulating the virulence of this microorganism. The Brucella histidine kinase (HK) domain belongs to the HWE family, for which there is no structural information. The HWE family is exclusively present in proteobacteria and usually coupled to a wide diversity of light sensor domains. This work reports the crystal structure of the Brucella HK domain, which presents two different dimeric assemblies in the asymmetric unit: one similar to the already described canonical parallel homodimers (C) and the other, an antiparallel non-canonical (NC) dimer, each with distinct relative subdomain orientations and dimerization interfaces. Contrary to these crystallographic structures and unlike other HKs, in solution, the Brucella HK domain is monomeric and still active, showing an astonishing instability of the dimeric interface. Despite this instability, using cross-linking experiments, we show that the C dimer is the functionally relevant species. Mutational analysis demonstrates that the autophosphorylation activity occurs in cis. The different relative subdomain orientations observed for the NC and C states highlight the large conformational flexibility of the HK domain. Through the analysis of these alternative conformations by means of molecular dynamics simulations, we also propose a catalytic mechanism for Brucella LOV-HK.Fil: Rinaldi, Jimena Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Arrar, Mehrnoosh. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; ArgentinaFil: Sycz, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Cerutti, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Plataforma Argentina de Biología Estructural y Metabolómica PLABEM; ArgentinaFil: Berguer, Paula Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Paris, Gastón. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Estrin, Dario Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; ArgentinaFil: Marti, Marcelo Adrian. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Klinke, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Plataforma Argentina de Biología Estructural y Metabolómica PLABEM; ArgentinaFil: Goldbaum, Fernando Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Plataforma Argentina de Biología Estructural y Metabolómica PLABEM; Argentin

Crystal structure of Hop2-Mnd1 and mechanistic insights into its role in meiotic recombination

In meiotic DNA recombination, the Hop2-Mnd1 complex promotes Dmc1-mediated single-stranded DNA (ssDNA) invasion into homologous chromosomes to form a synaptic complex by a yet-unclear mechanism. Here, the crystal structure of Hop2-Mnd1 reveals that it forms a curved rod-like structure consisting of three leucine zippers and two kinked junctions. One end of the rod is linked to two juxtaposed winged-helix domains, and the other end is capped by extra ?-helices to form a helical bundle-like structure. Deletion analysis shows that the helical bundle-like structure is sufficient for interacting with the Dmc1-ssDNA nucleofilament, and molecular modeling suggests that the curved rod could be accommodated into the helical groove of the nucleofilament. Remarkably, the winged-helix domains are juxtaposed at fixed relative orientation, and their binding to DNA is likely to perturb the base pairing according to molecular simulations. These findings allow us to propose a model explaining how Hop2-Mnd1 juxtaposes Dmc1-bound ssDNA with distorted recipient double-stranded DNA and thus facilitates strand invasion

Predictive Tools in Protein Chromatography: Extension and Optimization of a Molecular Modeling Approach

The aim of this study was to increase the understanding of the binding behavior of biomolecules on different adsorbents. This ought to be used for prediction and problem analysis. The approach was to develop a molecular dynamics simulations tool, which represents both: the biomolecule and the ligand surface at the atomic level and realistically maps the interaction processes between them. Results were validated by corresponding laboratory experiments

Improved reception of in-body signals by means of a wearable multi-antenna system

High data-rate wireless communication for in-body human implants is mainly performed in the 402-405 MHz Medical Implant Communication System band and the 2.45 GHz Industrial, Scientific and Medical band. The latter band offers larger bandwidth, enabling high-resolution live video transmission. Although in-body signal attenuation is larger, at least 29 dB more power may be transmitted in this band and the antenna efficiency for compact antennas at 2.45 GHz is also up to 10 times higher. Moreover, at the receive side, one can exploit the large surface provided by a garment by deploying multiple compact highly efficient wearable antennas, capturing the signals transmitted by the implant directly at the body surface, yielding stronger signals and reducing interference. In this paper, we implement a reliable 3.5 Mbps wearable textile multi-antenna system suitable for integration into a jacket worn by a patient, and evaluate its potential to improve the In-to-Out Body wireless link reliability by means of spatial receive diversity in a standardized measurement setup. We derive the optimal distribution and the minimum number of on-body antennas required to ensure signal levels that are large enough for real-time wireless endoscopy-capsule applications, at varying positions and orientations of the implant in the human body

Probing white-matter microstructure with higher-order diffusion tensors and susceptibility tensor MRI.

Diffusion MRI has become an invaluable tool for studying white matter microstructure and brain connectivity. The emergence of quantitative susceptibility mapping and susceptibility tensor imaging (STI) has provided another unique tool for assessing the structure of white matter. In the highly ordered white matter structure, diffusion MRI measures hindered water mobility induced by various tissue and cell membranes, while susceptibility sensitizes to the molecular composition and axonal arrangement. Integrating these two methods may produce new insights into the complex physiology of white matter. In this study, we investigated the relationship between diffusion and magnetic susceptibility in the white matter. Experiments were conducted on phantoms and human brains in vivo. Diffusion properties were quantified with the diffusion tensor model and also with the higher order tensor model based on the cumulant expansion. Frequency shift and susceptibility tensor were measured with quantitative susceptibility mapping and susceptibility tensor imaging. These diffusion and susceptibility quantities were compared and correlated in regions of single fiber bundles and regions of multiple fiber orientations. Relationships were established with similarities and differences identified. It is believed that diffusion MRI and susceptibility MRI provide complementary information of the microstructure of white matter. Together, they allow a more complete assessment of healthy and diseased brains

Research on Nursing Unit Layouts: An Integrative Review

This is the author's accepted manuscript. This article is (c) Emerald Group Publishing and permission has been granted for this version to appear in KU ScholarWorks (kuscholarworks.ku.edu). Emerald does not grant permission for this article to be further copied/distributed or hosted elsewhere without the express permission from Emerald Group Publishing Limited. - See more at: http://www.emeraldgrouppublishing.com/authors/writing/author_rights.htm#sthash.J18UbKQW.dpufPURPOSE: To present an integrative review of the research studies on nursing unit layouts. DESIGN/METHODOLOGY/APPROACH: Studies selected for review were published between 1956 and 2014. For the purpose of this review, a framework for integrative review was developed using research orientations. The three primary dimensions–technical, psychological, and social– of the designed environment and various combinations of these dimensions were used to define the research orientations of these studies. FINDINGS: Of all the publications reviewed for the paper, 21 presented technical orientations, 16 psychological orientations, 3 social orientations, 20 psychotechnical orientations, 10 sociotechnical orientations, 2 psychosocial orientations, and 13 presented psychosociotechnical orientations. With only a few exceptions, several issues related to nursing unit layouts were investigated no more than one time in any one category of research orientations. Several other seemingly important issues including patient and family behavior and perception, health outcomes, and social and psychosocial factors in relation to unit layouts have not been studied adequately. RESEARCH LIMITATIONS/IMPLICATIONS: Future studies on nursing unit layouts will need to focus on patient and family behavior and perception, health outcomes, and social and psychosocial factors in different units. They will also need to focus on developing theories concerning the effects of layouts on the technical, psychological, and social dimensions of nursing units. ORIGINALITY/VALUE: Despite a long history of research on nursing unit layouts, an integrative review of these studies is still missing in the literature. This review fills in the gap using a novel framework for integrative review developed based on research orientations

Quantifying Cross-scatter Contamination in Biplane Fluoroscopy Motion Analysis Systems

Biplane fluoroscopy is used for dynamic in vivo three-dimensional motion analysis of various joints of the body. Cross-scatter between the two fluoroscopy systems may limit tracking accuracy. This study measured the magnitude and effects of cross-scatter in biplane fluoroscopic images. Four cylindrical phantoms of 4-, 6-, 8-, and 10-in. diameter were imaged at varying kVp levels to determine the cross-scatter fraction and contrast-to-noise ratio (CNR). Monte Carlo simulations quantified the effect of the gantry angle on the cross-scatter fraction. A cadaver foot with implanted beads was also imaged. The effect of cross-scatter on marker-based tracking accuracy was investigated. Results demonstrated that the cross-scatter fraction varied from 0.15 for the 4-in. cylinder to 0.89 for the 10-in. cylinder when averaged across kVp. The average change in CNR due to cross-scatter ranged from 5% to 36% CNR decreases for the 4- and 10-in. cylinders, respectively. In simulations, the cross-scatter fraction increased with the gantry angle for the 8- and 10-in. cylinders. Cross-scatter significantly increased static-tracking error by 15%, 25%, and 38% for the 6-, 8-, and 10-in. phantoms, respectively, with no significant effect for the foot specimen. The results demonstrated submillimeter marker-based tracking for a range of phantom sizes, despite cross-scatter degradation

Functional dynamics of the folded ankyrin repeats of I kappa B alpha revealed by nuclear magnetic resonance.

Inhibition of nuclear factor kappaB (NF-kappaB) is mainly accomplished by IkappaB alpha, which consists of a signal response sequence at the N-terminus, a six-ankyrin repeat domain (ARD) that binds NF-kappaB, and a C-terminal PEST sequence. Previous studies with the ARD revealed that the fifth and sixth repeats are only partially folded in the absence of NF-kappaB. Here we report NMR studies of a truncated version of IkappaB alpha, containing only the first four ankyrin repeats, IkappaB alpha(67-206). This four-repeat segment is well-structured in the free state, enabling full resonance assignments to be made. H-D exchange, backbone dynamics, and residual dipolar coupling (RDC) experiments reveal regions of flexibility. In addition, regions consistent with the presence of micro- to millisecond motions occur periodically throughout the repeat structure. Comparison of the RDCs with the crystal structure gave only moderate agreement, but an ensemble of structures generated by accelerated molecular dynamics gave much better agreement with the measured RDCs. The regions showing flexibility correspond to those implicated in entropic compensation for the loss of flexibility in ankyrin repeats 5 and 6 upon binding to NF-kappaB. The regions showing micro- to millisecond motions in the free protein are the ends of the beta-hairpins that directly interact with NF-kappaB in the complex