Large-scale functional expression of WT and truncated human adenosine A(2A) receptor in Pichia pastoris bioreactor cultures

Background: The large-scale production of G-protein coupled receptors (GPCRs) for functional and structural studies remains a challenge. Recent successes have been made in the expression of a range of GPCRs using Pichia pastoris as an expression host. P. pastoris has a number of advantages over other expression systems including ability to post-translationally modify expressed proteins, relative low cost for production and ability to grow to very high cell densities. Several previous studies have described the expression of GPCRs in P. pastoris using shaker flasks, which allow culturing of small volumes (500 ml) with moderate cell densities (OD600 similar to 15). The use of bioreactors, which allow straightforward culturing of large volumes, together with optimal control of growth parameters including pH and dissolved oxygen to maximise cell densities and expression of the target receptors, are an attractive alternative. The aim of this study was to compare the levels of expression of the human Adenosine 2A receptor (A(2A)R) in P. pastoris under control of a methanol-inducible promoter in both flask and bioreactor cultures. Results: Bioreactor cultures yielded an approximately five times increase in cell density (OD600 similar to 75) compared to flask cultures prior to induction and a doubling in functional expression level per mg of membrane protein, representing a significant optimisation. Furthermore, analysis of a C-terminally truncated A2AR, terminating at residue V334 yielded the highest levels (200 pmol/mg) so far reported for expression of this receptor in P. pastoris. This truncated form of the receptor was also revealed to be resistant to C-terminal degradation in contrast to the WT A(2A)R, and therefore more suitable for further functional and structural studies. Conclusion: Large-scale expression of the A(2A)R in P. pastoris bioreactor cultures results in significant increases in functional expression compared to traditional flask cultures.

Planning rigid body motions using elastic curves

This paper tackles the problem of computing smooth, optimal trajectories on the Euclidean group of motions SE(3). The problem is formulated as an optimal control problem where the cost function to be minimized is equal to the integral of the classical curvature squared. This problem is analogous to the elastic problem from differential geometry and thus the resulting rigid body motions will trace elastic curves. An application of the Maximum Principle to this optimal control problem shifts the emphasis to the language of symplectic geometry and to the associated Hamiltonian formalism. This results in a system of first order differential equations that yield coordinate free necessary conditions for optimality for these curves. From these necessary conditions we identify an integrable case and these particular set of curves are solved analytically. These analytic solutions provide interpolating curves between an initial given position and orientation and a desired position and orientation that would be useful in motion planning for systems such as robotic manipulators and autonomous-oriented vehicles.

Integrable quadratic Hamiltonians on the Euclidean group of motions

In this paper, we discuss the problem of globally computing sub-Riemannian curves on the Euclidean group of motions SE(3). In particular, we derive a global result for special sub-Riemannian curves whose Hamiltonian satisfies a particular condition. In this paper, sub-Riemannian curves are defined in the context of a constrained optimal control problem. The maximum principle is then applied to this problem to yield an appropriate left-invariant quadratic Hamiltonian. A number of integrable quadratic Hamiltonians are identified. We then proceed to derive convenient expressions for sub-Riemannian curves in SE(3) that correspond to particular extremal curves. These equations are then used to compute sub-Riemannian curves that could potentially be used for motion planning of underwater vehicles.

An implicit pole-assignment self-tuning regulator

A new self-tuning implicit pole-assignment algorithm is presented which, through the use of a pole compression factor and different RLS model and control structures, overcomes stability and convergence problems encountered in previously available algorithms. Computational requirements of the technique are much reduced when compared to explicit pole-assignment schemes, whereas the inherent robustness of the strategy is retained.

Auto-weighted minimum-variance self-tuning controller

A self-tuning controller which automatically assigns weightings to control and set-point following is introduced. This discrete-time single-input single-output controller is based on a generalized minimum-variance control strategy. The automatic on-line selection of weightings is very convenient, especially when the system parameters are unknown or slowly varying with respect to time, which is generally considered to be the type of systems for which self-tuning control is useful. This feature also enables the controller to overcome difficulties with non-minimum phase systems.

New developments in batch process input profile optimisation

A novel algorithm for solving nonlinear discrete time optimal control problems with model-reality differences is presented. The technique uses dynamic integrated system optimisation and parameter estimation (DISOPE) which achieves the correct optimal solution in spite of deficiencies in the mathematical model employed in the optimisation procedure. A new method for approximating some Jacobian trajectories required by the algorithm is introduced. It is shown that the iterative procedure associated with the algorithm naturally suits applications to batch chemical processes.

Predicting random effects with an expanded finite population mixed model

Prediction of random effects is an important problem with expanding applications. In the simplest context, the problem corresponds to prediction of the latent value (the mean) of a realized cluster selected via two-stage sampling. Recently, Stanek and Singer [Predicting random effects from finite population clustered samples with response error. J. Amer. Statist. Assoc. 99, 119-130] developed best linear unbiased predictors (BLUP) under a finite population mixed model that outperform BLUPs from mixed models and superpopulation models. Their setup, however, does not allow for unequally sized clusters. To overcome this drawback, we consider an expanded finite population mixed model based on a larger set of random variables that span a higher dimensional space than those typically applied to such problems. We show that BLUPs for linear combinations of the realized cluster means derived under such a model have considerably smaller mean squared error (MSE) than those obtained from mixed models, superpopulation models, and finite population mixed models. We motivate our general approach by an example developed for two-stage cluster sampling and show that it faithfully captures the stochastic aspects of sampling in the problem. We also consider simulation studies to illustrate the increased accuracy of the BLUP obtained under the expanded finite population mixed model. (C) 2007 Elsevier B.V. All rights reserved.

Early impact of the BEST intervention for parents stressed by adolescent substance abuse.

Youth involvement in substance abuse can be a source of considerable distress for their parents. Unilateral family interventions have been advocated as one means by which concerned family members can be supported to assist substance-abusing family members. To date there has been little research examining the impact of unilateral family interventions on the directly participating family members. In this study the early impact of an 8-week parent-group programme known as Behavioural Exchange Systems Training (BEST) was evaluated using a quasi-experimental, waiting list control design. The professionally led programme had been developed to support and assist parents in their efforts to cope with adolescent substance abuse. Subjects were 66 parents (48 families) accepted for entry into the programme between 1997 and 1998. Comparison was made between 46 parents offered immediate entry into the programme and 20 parents whose entry to the programme was delayed by an 8-week waiting list. At the first assessment 87% of parents showed elevated mental health symptoms on the General Health Questionnaire. Evidence suggested exposure to the intervention had a positive impact on parents. Compared to parents on the waiting list, parents entered immediately into the intervention demonstrated greater reductions in mental health symptoms, increased parental satisfaction, and increased use of assertive parenting behaviours.

Path planning for sensor data collecting mobile robot

A mobile robot employed for data collection is faced with the problem of travelling from an initial location to a final location while maintaining as close a distance as possible to all the sensors at a given time in the journey. Here we employ optimal control ideas in forming the necessary control commands for such a robot resulting not only the necessary acceleration commands for the underlying robot, but also the resulting trajectory. This approach can also be easily extended for the case of producing the optimal trajectory for an ariel vehicle used for data collection from indiscriminately scattered ad-hoc sensors located on the ground. We demonstrate the implementation of our algorithm using a Pioneer 3-AT robot.

Sedentary lifestyle as a risk factor for low back pain : a systematic review

Objectives To review systematically studies examining the association between sedentary lifestyle and low back pain (LBP) using a comprehensive definition of sedentary behaviour including prolonged sitting both at work and during leisure time.

Methods Journal articles published between 1998 and 2006 were obtained by searching computerized bibliographical databases. Quality assessment of studies employing a cohort or case–control design was performed to assess the strength of the evidence.

Results Using pre-determined keywords, we identified 1,778 titles of which 1,391 were considered irrelevant. Then, 20 of the remaining 387 publications were scrutinized for full review after an examination of all the 387 abstracts. Finally, 15 studies (10 prospective cohorts and 5 case–controls) were included in the methodological quality assessment, of which 8 (6 cohorts and 2 case–controls; 53%) were classified as high-quality studies. One high-quality cohort study reported a positive association, between LBP and sitting at work only; all other studies reported no significant associations. Hence, there was limited evidence to demonstrate that sedentary behaviour is a risk factor for developing LBP.

Conclusions The present review confirms that sedentary lifestyle by itself is not associated with LBP.

Dynamic advertising under vertical product differentiation

We investigate a dynamic advertising model where product quality is endogenous. In the differential game between single-product firms, there exists a parameter range where the low-quality firm uses a more efficient advertising technology and earns higher profits than the rival. Moreover, we show that equilibrium qualities are the same under duopoly, multiproduct monopoly, and social planning, the only distortion being concerned with the output levels.

Joint velocity redistribution for fault tolerant manipulators

If the end-effector of a robotic manipulator moves on a specified trajectory, then for the fault tolerant operation, it is required that the end-effector continues the trajectory with a minimum velocity jump when a fault occurs within a joint. This problem is addressed in the paper. A way to tolerate the fault is to find new joint velocities for the faulty manipulator in which results into the same end-effector velocity provided by the healthy manipulator. The aim of this study is to find a strategy which optimally redistributes the joint velocities for the remained healthy joints of the manipulators. The optimality is defined by the minimum end-effector velocity jump. A solution of the problem is presented and it is applied to a robotics manipulator. Then through a case study and a simulation study it is validated. The paper shows that if would be possible the joint velocity redistribution results into a zero velocity jump.

Association between abnormal psychosocial situations in childhood, generalised anxiety disorder and oppositional defiant disorder

Objective: Psychosocial stressors are important in the pathogenesis of most mental disorders. However, little is known about the way psychosocial stressors uniquely combine to create risk for different expressions of child and adolescent psychopathology. The purpose of this study was to determine whether core dimensions of stressful psychosocial situations are differentially associated with childhood generalized anxiety disorder and oppositional defi ant disorder.

Method: A case-control design conducted in Trondheim (Norway) from 2002 to 2004 comparing exposure to ICD-10-defi ned abnormal psychosocial situations (Z-codes) among 21 children with oppositional defi ant disorder (ODD) and 22 children with generalized anxiety disorder (GAD) recruited from a university outpatient clinic with 42 non-patient school controls.

Results: Multigroup discriminant analysis extracted two signifi cant dimensions within the psychosocial variables assessed. Function 1 was characterized by overprotection, parental pressures and acute life events and was associated with GAD. Function 2 was characterized by parental abuse/hostility and interpersonal stress and was associated with ODD. Both dimensions were able to correctly classify 89.7% of the cases, compared to 35.9% by chance.

Conclusions: The results indicate that specifi c psychosocial dimensions are differentially related to childhood GAD and ODD. This may be useful in targeting at-risk populations for preventive intervention as well as informing more accurate alignment of psychosocial resources for treatment.

Image reconstruction using Loop subdivision

It is an ultimate objective to reconstruct an image with high quality using a compact representation, which is the basic step in image-manipulation fields. We propose an effective vectorization based approach to reconstruct an image using a triangular mesh associated with Loop subdivision scheme in the present paper. With an initial control mesh obtained by simplifying a dense mesh from a quality-preserved triangulation, we produce the final optimal control mesh by optimizing a mesh over topologies and colors to approximate the given image. The main advantages of the approach include: (1) the reconstruction of an image is not restricted to be aligned with image coordinate axes; (2) a high order continuous function is defined over a triangle instead of a bilinear interpolation; (3) it is a compact and vector-based representation easy to edit and transmit. Experimental results are presented to confirm the effectiveness of the method. Comparisons with the bi-cubic spline and the mesh simplification methods demonstrate the merits of our method in reconstruction quality and representation size.