Structural and functional characterization of the aryl hydrocarbon receptor ligand binding domain by homology modeling and mutational analysis

The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that is activated by a structurally diverse array of synthetic and natural chemicals, including toxic halogenated aromatic hydrocarbons such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Analysis of the occurring in the AhR ligand binding and activation processes requires structural information on the AhR Per-Arnt-Sim (PAS) B-containing ligand binding domain, for which no experimentally determined structure has been reported. With the availability of extensive structural information on homologous PAS-containing proteins, a reliable model of the mouse AhR PAS B domain was developed by comparative modeling techniques. The PAS domain structures of the functionally related hypoxia-inducible factor 2α (HIF-2α) and AhR nuclear translocator (ARNT) proteins, which exhibit the highest degree of sequence identity and similarity with AhR, were chosen to develop a two-template model. To confirm the features of the modeled domain, the effects of point mutations in selected residue positions on both TCDD binding to the AhR and TCDD-dependent transformation and DNA binding were analyzed. Mutagenesis and functional analysis results are consistent with the proposed model and confirm that the cavity modeled in the interior of the domain is indeed involved in ligand binding. Moreover, the physicochemical characteristics of some residues and of their mutants, along with the effects of mutagenesis on TCDD and DNA binding, also suggest some key features that are required for ligand binding and activation of mAhR at a molecular level, thus providing a framework for further studies. © 2007 American Chemical Society

Detection of the TCDD binding-fingerprint within the Ah receptor ligand binding domain by structurally driven mutagenesis and functional analysis

The aryl hydrocarbon receptor (AhR) is a ligand-dependent, basic helix-loop-helix Per-Arnt-Sim (PAS)-containing transcription factor that can bind and be activated by structurally diverse chemicals, including the toxic environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Our previous three-dimensional homology model of the mouse AhR (mAhR) PAS B ligand binding domain allowed identification of the binding site and its experimental validation. We have extended this analysis by conducting comparative structural modeling studies of the ligand binding domains of six additional highaffinity mammalian AhRs. These results, coupled with site-directed mutagenesis and AhR functional analysis, have allowed detection of the "TCDD binding-fingerprint" of conserved residues within the ligand binding cavity necessary for high-affinity TCDD binding and TCDD-dependent AhR transformation DNA binding. The essential role of selected residues was further evaluated using molecular docking simulations of TCDD with both wild-type and mutant mAhRs. Taken together, our results dramatically improve our understanding of the molecular determinants of TCDD binding and provide a basis for future studies directed toward rationalizing the observed species differences in AhR sensitivity to TCDD and understanding the mechanistic basis for the dramatic diversity in AhR ligand structure. © 2009 American Chemical Society

Three-dimensional image-based numerical homogenisation using octree meshes

The determination of effective material properties of composites based on a three-dimensional representative volume element (RVE) is considered in this paper. The material variation in the RVE is defined based on the colour intensity in each voxel of an image which can be obtained from imaging techniques such as X-ray computed tomography (XCT) scans. The RVE is converted into a numerical model using hierarchical meshing based on octree decompositions. Each octree cell in the mesh is modelled as a scaled boundary polyhedral element, which only requires a surface discretisation on the polyhedron's boundary. The problem of hanging (incompatible) nodes – typically encountered when using the finite element method in conjunction with octree meshes – is circumvented by employing special transition elements. Two different types of boundary conditions (BCs) are used to obtain the homogenised material properties of various samples. The numerical results confirm that periodic BCs provide a better agreement with previously published results. The reason is attributed to the fact that the model based on the periodic BCs is not over-constrained as is the case for uniform displacement BCs

Realization of GHZ States and the GHZ Test via Cavity QED

In this article we discuss the realization of atomic GHZ states involving three-level atoms and we show explicitly how to use this state to perform the GHZ test in which it is possible to decide between local realism theories and quantum mechanics. The experimental realizations proposed makes use of the interaction of Rydberg atoms with a cavity prepared in a coherent state.Comment: 16 pages and 3 figures. submitted to J. Mod. Op

Dynamic clamp with StdpC software

Dynamic clamp is a powerful method that allows the introduction of artificial electrical components into target cells to simulate ionic conductances and synaptic inputs. This method is based on a fast cycle of measuring the membrane potential of a cell, calculating the current of a desired simulated component using an appropriate model and injecting this current into the cell. Here we present a dynamic clamp protocol using free, fully integrated, open-source software (StdpC, for spike timing-dependent plasticity clamp). Use of this protocol does not require specialist hardware, costly commercial software, experience in real-time operating systems or a strong programming background. The software enables the configuration and operation of a wide range of complex and fully automated dynamic clamp experiments through an intuitive and powerful interface with a minimal initial lead time of a few hours. After initial configuration, experimental results can be generated within minutes of establishing cell recording

Facilitators' influence on student PBL small group session online information resource use: a survey

BACKGROUND: In problem-based learning curricular research has focused on the characteristics of good facilitators and how they influence student performance and satisfaction. Far less frequently addressed has been the question of how PBL facilitators influence the small group session activity of students. We investigated the impact that facilitators' encouragement of use or non-use of the Internet would have on the students' use of online information resources. METHODS: Survey of student and facilitator perceptions of facilitator behavior and student use of online information resources. RESULTS: Students who used online information resources rated their facilitators' behavior as more encouraging, while students in groups who didn't use online information resources during problem-based learning small group sessions rated their facilitators' behavior as less encouraging. This result was statistically significant. CONCLUSIONS: Our study supports the role of the facilitator as an influence on medical students in small groups, particularly with respect to facilitator verbal behavior encouraging or discouraging student use of information technology in the problem-based learning small group session

Natural killer cells attenuate cytomegalovirus-induced hearing loss in mice

<div><p>Congenital cytomegalovirus (CMV) infection is the most common non-hereditary cause of sensorineural hearing loss (SNHL) yet the mechanisms of hearing loss remain obscure. Natural Killer (NK) cells play a critical role in regulating murine CMV infection via NK cell recognition of the Ly49H cell surface receptor of the viral-encoded m157 ligand expressed at the infected cell surface. This Ly49H NK receptor/m157 ligand interaction has been found to mediate host resistance to CMV in the spleen, and lung, but is much less effective in the liver, so it is not known if this interaction is important in the context of SNHL. Using a murine model for CMV-induced labyrinthitis, we have demonstrated that the Ly49H/m157 interaction mediates host resistance in the temporal bone. BALB/c mice, which lack functional Ly49H, inoculated with mCMV at post-natal day 3 developed profound hearing loss and significant outer hair cell loss by 28 days of life. In contrast, C57BL/6 mice, competent for the Ly49H/m157 interaction, had minimal hearing loss and attenuated outer hair cell loss with the same mCMV dose. Administration of Ly49H blocking antibody or inoculation with a mCMV viral strain deleted for the m157 gene rendered the previously resistant C57BL/6 mouse strain susceptible to hearing loss to a similar extent as the BALB/c mouse strain indicating a direct role of the Ly49H/m157 interaction in mCMV-dependent hearing loss. Additionally, NK cell recruitment to sites of infection was evident in the temporal bone of inoculated susceptible mouse strains. These results demonstrate participation of NK cells in protection from CMV-induced labyrinthitis and SNHL in mice.</p></div

Social Determinants of Community Health Services Utilization among the Users in China: A 4-Year Cross-Sectional Study

Background To identify social factors determining the frequency of community health service (CHS) utilization among CHS users in China. Methods Nationwide cross-sectional surveys were conducted in 2008, 2009, 2010, and 2011. A total of 86,116 CHS visitors selected from 35 cities were interviewed. Descriptive analysis and multinomial logistic regression analysis were employed to analyze characteristics of CHS users, frequency of CHS utilization, and the socio-demographic and socio-economic factors influencing frequency of CHS utilization. Results Female and senior CHS clients were more likely to make 3–5 and ≥6 CHS visits (as opposed to 1–2 visits) than male and young clients, respectively. CHS clients with higher education were less frequent users than individuals with primary education or less in 2008 and 2009; in later surveys, CHS clients with higher education were the more frequent users. The association between frequent CHS visits and family income has changed significantly between 2008 and 2011. In 2011, income status did not have a discernible effect on the likelihood of making ≥6 CHS visits, and it only had a slight effect on making 3–5 CHS visits. Conclusion CHS may play an important role in providing primary health care to meet the demands of vulnerable populations in China. Over time, individuals with higher education are increasingly likely to make frequent CHS visits than individuals with primary school education or below. The gap in frequency of CHS utilization among different economic income groups decreased from 2008 to 2011

Tomography of quantum dots in a non-hermitian photonic chip

© 2019 IEEE. Quantum optical information systems offer the potential for secure communication and fast quantum computation. To fully characterise a quantum optical system one has to use quantum tomography [1]. Integration of quantum optics onto photonic chips provides advantages such as miniaturisation and stability, and also significantly improves quantum tomography using both re-configurable [2], and more recently, simpler static designs [3,4]. These on-chip designs have, so far, only used probabilistic single photon sources. Here we are working towards quantum tomography using a true deterministic source - a quantum dot. The scheme of the proposed experiment is shown in Fig. 1A. So far we have fabricated and characterised the performance of an InGaAs quantum dot monolithically integrated into a microlens [5], and completed the design, fabrication and classical characterisation of a photonic chip for quantum tomography

On the classical equivalence of monodromy matrices in squashed sigma model

We proceed to study the hybrid integrable structure in two-dimensional non-linear sigma models with target space three-dimensional squashed spheres. A quantum affine algebra and a pair of Yangian algebras are realized in the sigma models and, according to them, there are two descriptions to describe the classical dynamics 1) the trigonometric description and 2) the rational description, respectively. For every description, a Lax pair is constructed and the associated monodromy matrix is also constructed. In this paper we show the gauge-equivalence of the monodromy matrices in the trigonometric and rational description under a certain relation between spectral parameters and the rescalings of sl(2) generators.Comment: 32pages, 3figures, references added, introduction and discussion sections revise