Genetic variation in selenoprotein S influences inflammatory response

Chronic inflammation has a pathological role in many common diseases and is influenced by both genetic and environmental factors. Here we assess the role of genetic variation in selenoprotein S (SEPS1, also called SELS or SELENOS), a gene involved in stress response in the endoplasmic reticulum and inflammation control. After resequencing SEPS1, we genotyped 13 SNPs in 522 individuals from 92 families. As inflammation biomarkers, we measured plasma levels of IL-6, IL-1b and TNF-a. Bayesian quantitative trait nucleotide analysis identified associations between SEPS1 polymorphisms and all three proinflammatory
cytokines. One promoter variant, 105G-A, showed strong evidence for an association with each cytokine (multivariate P = 0.0000002). Functional analysis of this polymorphism showed that the A variant significantly impaired SEPS1 expression after exposure to endoplasmic reticulum stress agents (P = 0.00006). Furthermore, suppression of SEPS1 by short interfering RNA in macrophage cells increased the release of IL-6 and TNF-a. To investigate further the significance of the observed associations, we genotyped 105G-A in 419 Mexican American individuals from 23 families for replication. This analysis confirmed a significant
association with both TNF-a (P = 0.0049) and IL-1b (P = 0.0101). These results provide a direct mechanistic link between SEPS1 and the production of inflammatory cytokines and suggest that SEPS1 has a role in mediating inflammation.

Molecular characterization of Osh6p, an oxysterol binding protein homolog in the yeast Saccharomyces cerevisiae

Oxysterol binding protein (OSBP) and its homologs have been shown to regulate lipid metabolism and vesicular transport. However, the exact molecular function of individual OSBP homologs remains uncharacterized. Here we demonstrate that the yeast OSBP homolog, Osh6p, bound phosphatidic acid and phosphoinositides via its N-terminal half containing the conserved OSBP-related domain (ORD). Using a green fluorescent protein fusion chimera, Osh6p was found to localize to the cytosol and patch-like or punctate structures in the vicinity of the plasma membrane. Further examination by domain mapping demonstrated that the N-terminal half was associated with FM4-64 positive membrane compartments; however, the C-terminal half containing a putative coiled-coil was localized to the nucleoplasm. Functional analysis showed that the deletion of OSH6 led to a significant increase in total cellular ergosterols, whereas OSH6 overexpression caused both a significant decrease in ergosterol levels and resistance to nystatin. Oleate incorporation into sterol esters was affected in OSH6 overexpressing cells. However, Lucifer yellow internalization, and FM4-64 uptake and transport were unaffected in both OSH6 deletion and overexpressing cells. Furthermore, osh6Δ exhibited no defect in carboxypeptidase Y transport and maturation. Lastly, we demonstrated that both the conserved ORD and the putative coiled-coil motif were indispensable for the in vivo function of Osh6p. These data suggest that Osh6p plays a role primarily in regulating cellular sterol metabolism, possibly stero transport.

Mutation of the LUNATIC FRINGE gene in humans Causes spondylocostal dysostosis with a severe vertebral phenotype

The spondylocostal dysostoses (SCDs) are a heterogeneous group of vertebral malsegmentation disorders that arise during embryonic development by a disruption of somitogenesis. Previously, we had identified two genes that cause a subset of autosomal recessive forms of this disease: DLL3 (SCD1) and MESP2 (SCD2). These genes are important components of the Notch signaling pathway, which has multiple roles in development and disease. Here, we have used a candidate-gene approach to identify a mutation in a third Notch pathway gene, LUNATIC FRINGE (LFNG), in a family with autosomal recessive SCD. LFNG encodes a glycosyltransferase that modifies the Notch family of cell-surface receptors, a key step in the regulation of this signaling pathway. A missense mutation was identified in a highly conserved phenylalanine close to the active site of the enzyme. Functional analysis revealed that the mutant LFNG was not localized to the correct compartment of the cell, was unable to modulate Notch signaling in a cell-based assay, and was enzymatically inactive. This represents the first known mutation in the human LFNG gene and reinforces the hypothesis that proper regulation of the Notch signaling pathway is an absolute requirement for the correct patterning of the axial skeleton.

Two novel missense mutations in hairy-and-enhancer-of-split-7 in a family with spondylocostal dysostosis

Spondylocostal dysostosis (SCD) is an inherited disorder with abnormal vertebral segmentation that results in extensive hemivertebrae, truncal shortening and abnormally aligned ribs. It arises during embryonic development by a disruption of formation of somites (the precursor tissue of the vertebrae, ribs and associated tendons and muscles). Four genes causing a subset of autosomal recessive forms of this disease have been identified: DLL3 (SCDO1: MIM 277300), MESP2 (SCDO2: MIM 608681), LFNG (SCDO3: MIM609813) and HES7 (SCDO4). These genes are all essential components of the Notch signalling pathway, which has multiple roles in development and disease. Previously, only a single SCD-causative missense mutation was described in HES7. In this study, we have identified two new missense mutations in the HES7 gene in a single family, with only individuals carrying both mutant alleles being affected by SCD. In vitro functional analysis revealed that one of the mutant HES7 proteins was unable to repress gene expression by DNA binding or protein heterodimerization.

Religious/spiritual well-being, coping styles and personality dimensions in people with substance use disorders

Religiosity and spirituality have been found to be negatively associated with a range of addictions. It has been suggested that religious/spiritual well-being might play an important role in the development, course and the recovery from addictive disorders. A sample of addiction in-patients (n=389) was assessed using the Multidimensional Inventory for Religious/Spiritual Well-Being (MI-RSWB) and compared with a matched group of non-addicted community controls (n=389). RSWB was found to be substantially lower in people with substance use disorders compared to the normal sample. Discriminate functional analysis showed that Experiences of Sense and Meaning, General Religiosity and Forgiveness were the dimensions of RSWB which strongly distinguished the groups. Within the group of people with substance use disorders, RSWB was strongly positively associated with the personality dimensions of Conscientiousness, Agreeableness and Openness as well as Sense of Coherence and positive Coping styles. The study suggests that therapeutic intervention programs focusing on building a positive and meaningful personal framework, akin to that of a religious/spiritual orientation, may contribute to positive outcomes in addiction treatment.

Monotreme lactation protein is highly expressed in monotreme milk and provides antimicrobial protection

Monotremes (platypus and echidna) are the descendants of the oldest ancestor of all extant mammals distinguished from other mammals by mode of reproduction. Monotremes lay eggs following a short gestation period and after an even briefer incubation period, altricial hatchlings are nourished over a long lactation period with milk secreted by nipple-less mammary patches located on the female's abdomen. Milk is the sole source of nutrition and immune protection for the developing young until weaning. Using transcriptome and mass spectrometry analysis of milk cells and milk proteins, respectively, a novel Monotreme Lactation Protein (MLP) was identified as a major secreted protein in milk. We show that platypus and short-beaked echidna MLP genes show significant homology and are unique to monotremes. The MLP transcript was shown to be expressed in a variety of tissues; however, highest expression was observed in milk cells and was expressed constitutively from early to late lactation. Analysis of recombinant MLP showed that it is an N-linked glycosylated protein and biophysical studies predicted that MLP is an amphipathic, α-helical protein, a typical feature of antimicrobial proteins. Functional analysis revealed MLP antibacterial activity against both opportunistic pathogenic Staphylococcus aureus and commensal Enterococcus faecalis bacteria but showed no effect on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Salmonella enterica. Our data suggest that MLP is an evolutionarily ancient component of milk-mediated innate immunity absent in other mammals. We propose that MLP evolved specifically in the monotreme lineage supporting the evolution of lactation in these species to provide bacterial protection, at a time when mammals lacked nipples.

Maternal creatine supplementation during pregnancy prevents long-term changes in diaphragm muscle structure and function after birth asphyxia

Using a model of birth asphyxia, we previously reported significant structural and functional deficits in the diaphragm muscle in spiny mice, deficits that are prevented by supplementing the maternal diet with 5% creatine from mid-pregnancy. The long-term effects of this exposure are unknown. Pregnant spiny mice were fed control or 5% creatine-supplemented diet for the second half of pregnancy, and fetuses were delivered by caesarean section with or without 7.5 min of in-utero asphyxia. Surviving pups were raised by a cross-foster dam until 33±2 days of age when they were euthanized to obtain the diaphragm muscle for ex-vivo study of twitch tension and muscle fatigue, and for structural and enzymatic analyses. Functional analysis of the diaphragm revealed no differences in single twitch contractile parameters between any groups. However, muscle fatigue, induced by stimulation of diaphragm strips with a train of pulses (330 ms train/sec, 40 Hz) for 300 sec, was significantly greater for asphyxia pups compared with controls (p<0.05), and this did not occur in diaphragms of creatine + asphyxia pups. Birth asphyxia resulted in a significant increase in the proportion of glycolytic, fast-twitch fibres and a reduction in oxidative capacity of Type I and IIb fibres in male offspring, as well as reduced cross-sectional area of all muscle fibre types (Type I, IIa, IIb/d) in both males and females at 33 days of age. None of these changes were observed in creatine + asphyxia animals. Thus, the changes in diaphragm fatigue and structure induced by birth asphyxia persist long-term but are prevented by maternal creatine supplementation.

Evaluation of numerical procedures to determine seismic response of structures under influence of soil-structure interaction

In this study, the accuracy and reliability of fully nonlinear method against equivalent linear method for dynamic analysis of soil-structure interaction is investigated comparing the predicted results of both numerical procedures with the results of experimental shaking table tests. An enhanced numerical soil-structure model has been developed which treats the behaviour of the soil and the structure with equal rigour. The soil-structural model comprises a 15 storey structural model resting on a soft soil inside a laminar soil container. The structural model was analysed under three different conditions: (i) fixed base model performing conventional time history dynamic analysis, (ii) flexible base model (considering full soil-structure interaction) conducting equivalent linear dynamic analysis, and (iii) flexible base model performing fully nonlinear dynamic analysis. The results of the above mentioned three cases in terms of lateral storey deflections and inter-storey drifts are determined and compared with the experimental results of shaking table tests. Comparing the experimental results with the numerical analysis predictions, it is noted that equivalent linear method of dynamic analysis underestimates the inelastic seismic response of mid-rise moment resisting building frames resting on soft soils in comparison to the fully nonlinear dynamic analysis method. Thus, inelastic design procedure, using equivalent linear method, cannot adequately guarantee the structural safety for mid-rise building frames resting on soft soils. However, results obtained from the fully nonlinear method of analysis fit the experimental results reasonably well. Therefore, this method is recommended to be used by practicing engineers.

Nonlinear large deformation dynamic analysis of electroactive polymer actuators

Electroactive polymers have attracted considerable attention in recent years due to their sensing and actuating properties which make them a material of choice for a wide range of applications including sensors, biomimetic robots, and biomedical micro devices. This paper presents an effective modeling strategy for nonlinear large deformation (small strains and moderate rotations) dynamic analysis of polymer actuators. Considering that the complicated electro-chemo-mechanical dynamics of these actuators is a drawback for their application in functional devices, establishing a mathematical model which can effectively predict the actuator's dynamic behavior can be of paramount importance. To effectively predict the actuator's dynamic behavior, a comprehensive mathematical model is proposed correlating the input voltage and the output bending displacement of polymer actuators. The proposed model, which is based on the rigid finite element (RFE) method, consists of two parts, namely electrical and mechanical models. The former is comprised of a ladder network of discrete resistive-capacitive components similar to the network used to model transmission lines, while the latter describes the actuator as a system of rigid links connected by spring-damping elements (sdes). Both electrical and mechanical components are validated through experimental results.

A family of simple and robust finite elements for linear and geometrically nonlinear analysis of laminated composite plates

A family of simple, displacement-based and shear-flexible triangular and quadrilateral flat plate/shell elements for linear and geometrically nonlinear analysis of thin to moderately thick laminate composite plates are introduced and summarized in this paper.

The developed elements are based on the first-order shear deformation theory (FSDT) and von-Karman’s large deflection theory, and total Lagrangian approach is employed to formulate the element for geometrically nonlinear analysis. The deflection and rotation functions of the element boundary are obtained from Timoshenko’s laminated composite beam functions, thus convergence can be ensured theoretically for very thin laminates and shear-locking problem is avoided naturally.

The flat triangular plate/shell element is of 3-node, 18-degree-of-freedom, and the plane displacement interpolation functions of the Allman’s triangular membrane element with drilling degrees of freedom are taken as the in-plane displacements of the element. The flat quadrilateral plate/shell element is of 4-node, 24-degree-of-freedom, and the linear displacement interpolation functions of a quadrilateral plane element with drilling degrees of freedom are taken as the in-plane displacements.

The developed elements are simple in formulation, free from shear-locking, and include conventional engineering degrees of freedom. Numerical examples demonstrate that the elements are convergent, not sensitive to mesh distortion, accurate and efficient for linear and geometric nonlinear analysis of thin to moderately thick laminates.

A marginalised Markov Chain Monte Carlo approach for model based analysis of EEG data

The work presented in this paper focuses on fitting of a neural mass model to EEG data. Neurophysiology inspired mathematical models were developed for simulating brain's electrical activity imaged through Electroencephalography (EEG) more than three decades ago. At the present well informative models which even describe the functional integration of cortical regions also exists. However, a very limited amount of work is reported in literature on the subject of model fitting to actual EEG data. Here, we present a Bayesian approach for parameter estimation of the EEG model via a marginalized Markov Chain Monte Carlo (MCMC) approach.

Identification of nonlinear fMRI models using Auxiliary Particle Filter and kernel smoothing method

Hemodynamic models have a high potential in application to understanding the functional differences of the brain. However, full system identification with respect to model fitting to actual functional magnetic resonance imaging (fMRI) data is practically difficult and is still an active area of research. We present a simulation based Bayesian approach for nonlinear model based analysis of the fMRI data. The idea is to do a joint state and parameter estimation within a general filtering framework. One advantage of using Bayesian methods is that they provide a complete description of the posterior distribution, not just a single point estimate. We use an Auxiliary Particle Filter adjoined with a kernel smoothing approach to address this joint estimation problem.