Sphingosine-1-phosphate mediates proliferation maintaining the multipotency of human adult bone marrow and adipose tissue-derived stem cells

High renewal and maintenance of multipotency of human adult stem cells (hSCs), are a prerequisite for experimental analysis as well as for potential clinical usages. The most widely used strategy for hSC culture and proliferation is using serum. However, serum is poorly defined and has a considerable degree of inter-batch variation, which makes it difficult for large-scale mesenchymal stem cells (MSCs) expansion in homogeneous culture conditions. Moreover, it is often observed that cells grown in serum-containing media spontaneously differentiate into unknown and/or undesired phenotypes. Another way of maintaining hSC development is using cytokines and/or tissue-specific growth factors; this is a very expensive approach and can lead to early unwanted differentiation. In order to circumvent these issues, we investigated the role of sphingosine-1-phosphate (S1P), in the growth and multipotency maintenance of human bone marrow and adipose tissue-derived MSCs. We show that S1P induces growth, and in combination with reduced serum, or with the growth factors FGF and platelet-derived growth factor-AB, S1P has an enhancing effect on growth. We also show that the MSCs cultured in S1P-supplemented media are able to maintain their differentiation potential for at least as long as that for cells grown in the usual serum-containing media. This is shown by the ability of cells grown in S1P-containing media to be able to undergo osteogenic as well as adipogenic differentiation. This is of interest, since S1P is a relatively inexpensive natural product, which can be obtained in homogeneous high-purity batches: this will minimize costs and potentially reduce the unwanted side effects observed with serum. Taken together, S1P is able to induce proliferation while maintaining the multipotency of different human stem cells, suggesting a potential for S1P in developing serum-free or serum-reduced defined medium for adult stem cell cultures.

In vitro cultivation of adult human chondrocytes : importance of culture system, oxygen and zonal differences

Articular cartilage exhibits limited intrinsic regenerative capacity and focal tissue defects can lead to the development of osteoarthritis (OA), a painful and debilitating loss of cartilage tissue. In Australia, 1.4 million people are affected by OA and its prevalence is increasing in line with current demographics. As treatment options are limited, new therapeutic approaches are being investigated including biological resurfacing of joints with tissue-engineered cartilage. Despite some progress in the field, major challenges remain to be addressed for large scale clinical success. For example, large numbers of chondrogenic cells are required for cartilage formation, but chondrocytes lose their chondrogenic phenotype (dedifferentiate) during in vitro propagation. Additionally, the zonal organization of articular cartilage is critical for normal cartilage function, but development of zonal structure has been largely neglected in cartilage repair strategies. Therefore, we hypothesised that culture conditions for freshly isolated human articular chondrocytes from non-OA and OA sources can be improved by employing microcarrier cultures and a reduced oxygen environment and that oxygen is a critical factor in the maintenance of the zonal chondrocyte phenotype. Microcarriers have successfully been used to cultivate bovine chondrocytes, and offer a potential alternative for clinical expansion of human chondrocytes. We hypothesised that improved yields can be achieved by propagating human chondrocytes on microcarriers. We found that cells on microcarriers acquired a flattened, polygonal morphology and initially proliferated faster than monolayercultivated cells. However, microcarrier cultivation over four weeks did not improve growth rates or the chondrogenic potential of non-OA and OA human articular chondrocytes over conventional monolayer cultivation. Based on these observations, we aimed to optimise culture conditions by modifying oxygen tension, to more closely reflect the in vivo environment. We found that propagation at 5% oxygen tension (moderate hypoxia) did not improve proliferation or redifferentiation capacity of human osteoarthritic chondrocytes. Moderate hypoxia increased the expression of chondrogenic markers during redifferentiation. However, osteoarthritic chondrocytes cultivated on microcarriers exhibited lower expression levels of chondrogenic surface marker proteins and had at best equivalent redifferentiation capacities compared to monolayer-cultured cells. This suggests that monolayer culture with multiple passaging potentially selects for a subpopulation of cells with higher differentiation capacity, which are otherwise rare in osteoarthritic, aged cartilage. However, fibroblastic proteins were found to be highly expressed in all cultures of human osteoarthritic chondrocytes indicating the presence of a high proportion of dedifferentiated, senescent cells with a chondrocytic phenotype that was not rescued by moderate hypoxia. The different zones of cartilage support chondrocyte subpopulations, which exhibit characteristic protein expression and experience varying oxygen tensions. We, therefore, hypothesised that oxygen tension affects the zonal marker expression of human articular chondrocytes isolated from the different cartilage layers. We found that zonal chondrocytes maintained these phenotypic differences during in vitro cultivation. Low oxygen environments favoured the expression of the zonal marker proteoglycan 4 in superficial cells, most likely through the promotion of chondrogenesis. The putative zonal markers clusterin and cartilage intermediate layer protein were found to be expressed by all subpopulations of human osteoarthritic chondrocytes ex vivo and, thus, may not be reliable predictors of in vitro stratification using these clinically relevant cells. The findings in this thesis underline the importance of considering low oxygen conditions and zonal stratification when creating native-like cartilaginous constructs. We have not yet found the right cues to successfully cultivate clinically-relevant human osteoarthritic chondrocytes in vitro. A more thorough understanding of chondrocyte biology and the processes of chondrogenesis are required to ensure the clinical success of cartilage tissue engineering.

Empowerment and role stress in the human interface between the firm and its markets

Frontline employees constitute one of the key interfaces that service organisations have with their markets. Many strategies to enhance the ability of these employees to satisfy the needs of customers have been proposed. Amongst these, empowering employees has been suggested to enhance the customer orientation of the firm and consequently its effectiveness in serving the market. However, the impact of empowerment in service organisations remains somewhat contentious. This paper examines the role of empowerments an organisational service strategy and identifies its consequences for role stress, job satisfaction and the willingness of service employees to serve their customers.

Liking vs. wanting food : importance for human appetite control and weight regulation

Current train of thought in appetite research is favouring an interest in non-homeostatic or hedonic (reward) mechanisms in relation to overconsumption and energy balance. This tendency is supported by advances in neurobiology that precede the emergence of a new conceptual approach to reward where affect and motivation (liking and wanting) can be seen as the major force in guiding human eating behaviour. In this review, current progress in applying processes of liking and wanting to the study of human appetite are examined by discussing the following issues: How can these concepts be operationalised for use in human research to reflect the neural mechanisms by which they may be influenced? Do liking and wanting operate independently to produce functionally significant changes in behaviour? Can liking and wanting be truly experimentally separated or will an expression of one inevitably contain elements of the other? The review contains a re-examination of selected human appetite research before exploring more recent methodological approaches to the study of liking and wanting in appetite control. In addition, some theoretical developments are described in four diverse models that may enhance current understanding of the role of these processes in guiding ingestive behaviour. Finally, the implications of a dual process modulation of food reward for weight gain and obesity are discussed. The review concludes that processes of liking and wanting are likely to have independent roles in characterising susceptibility to weight gain. Further research into the dissociation of liking and wanting through implicit and explicit levels of processing would help to disclose the relative importance of these components of reward for appetite control and weight regulation.

Appetite sensations and satiety quotient : predictors of energy intake and weight loss

PURPOSE: The aim of this study was to further evaluate the validity and clinical meaningfulness of appetite sensations to predict overall energy intake as well as body weight loss. METHODS: Men (n=176) and women (n=139) involved in six weight loss studies were selected to participate in this study. Visual analogue scales were used to measure appetite sensations before and after a fixed test meal. Fasting appetite sensations, 1 h post-prandial area under the curve (AUC) and the satiety quotient (SQ) were used as predictors of energy intake and body weight loss. Two separate measures of energy intake were used: a buffet style ad libitum test lunch and a three-day self-report dietary record. RESULTS: One-hour post-prandial AUC for all appetite sensations represented the strongest predictors of ad libitum test lunch energy intake (p0.001). These associations were more consistent and pronounced for women than men. Only SQ for fullness was associated with ad libitum test lunch energy intake in women. Similar but weaker relationships were found between appetite sensations and the 3-day self-reported energy intake. Weight loss was associated with changes in appetite sensations (p0.01) and the best predictors of body weight loss were fasting desire to eat; hunger; and PFC (p0.01). CONCLUSIONS: These results demonstrate that appetite sensations are relatively useful predictors of spontaneous energy intake, free-living total energy intake and body weight loss. They also confirm that SQ for fullness predicts energy intake, at least in women.

Spherical image-based visual servo and structure estimation

This paper presents a formulation of image-based visual servoing (IBVS) for a spherical camera where coordinates are parameterized in terms of colatitude and longitude: IBVSSph. The image Jacobian is derived and simulation results are presented for canonical rotational, translational as well as general motion. Problems with large rotations that affect the planar perspective form of IBVS are not present on the sphere, whereas the desirable robustness properties of IBVS are shown to be retained. We also describe a structure from motion (SfM) system based on camera-centric spherical coordinates and show how a recursive estimator can be used to recover structure. The spherical formulations for IBVS and SfM are particularly suitable for platforms, such as aerial and underwater robots, that move in SE(3).

Wide-angle visual feature matching for outdoor localization

Wide-angle images exhibit significant distortion for which existing scale-space detectors such as the scale-invariant feature transform (SIFT) are inappropriate. The required scale-space images for feature detection are correctly obtained through the convolution of the image, mapped to the sphere, with the spherical Gaussian. A new visual key-point detector, based on this principle, is developed and several computational approaches to the convolution are investigated in both the spatial and frequency domain. In particular, a close approximation is developed that has comparable computation time to conventional SIFT but with improved matching performance. Results are presented for monocular wide-angle outdoor image sequences obtained using fisheye and equiangular catadioptric cameras. We evaluate the overall matching performance (recall versus 1-precision) of these methods compared to conventional SIFT. We also demonstrate the use of the technique for variable frame-rate visual odometry and its application to place recognition.

Generic decoupled image-based visual servoing for cameras obeying the unified projection model

In this paper a generic decoupled imaged-based control scheme for calibrated cameras obeying the unified projection model is proposed. The proposed decoupled scheme is based on the surface of object projections onto the unit sphere. Such features are invariant to rotational motions. This allows the control of translational motion independently from the rotational motion. Finally, the proposed results are validated with experiments using a classical perspective camera as well as a fisheye camera mounted on a 6 dofs robot platform.

Mobile robot navigation as a planar visual servoing problem

This paper demonstrates some interesting connections between the hitherto disparate fields of mobile robot navigation and image-based visual servoing. A planar formulation of the well-known image-based visual servoing method leads to a bearing-only navigation system that requires no explicit localization and directly yields desired velocity. The well known benefits of image-based visual servoing such as robustness apply also to the planar case. Simulation results are presented.

An introduction to inertial and visual sensing

In this paper we present a tutorial introduction to two important senses for biological and robotic systems — inertial and visual perception. We discuss the fundamentals of these two sensing modalities from a biological and an engineering perspective. Digital camera chips and micro-machined accelerometers and gyroscopes are now commodities, and when combined with today's available computing can provide robust estimates of self-motion as well 3D scene structure, without external infrastructure. We discuss the complementarity of these sensors, describe some fundamental approaches to fusing their outputs and survey the field.

Dynamic Image-based visual servo control using centroid and optic flow features

This paper considers the question of designing a fully image-based visual servo control for a class of dynamic systems. The work is motivated by the ongoing development of image-based visual servo control of small aerial robotic vehicles. The kinematics and dynamics of a rigid-body dynamical system (such as a vehicle airframe) maneuvering over a flat target plane with observable features are expressed in terms of an unnormalized spherical centroid and an optic flow measurement. The image-plane dynamics with respect to force input are dependent on the height of the camera above the target plane. This dependence is compensated by introducing virtual height dynamics and adaptive estimation in the proposed control. A fully nonlinear adaptive control design is provided that ensures asymptotic stability of the closed-loop system for all feasible initial conditions. The choice of control gains is based on an analysis of the asymptotic dynamics of the system. Results from a realistic simulation are presented that demonstrate the performance of the closed-loop system. To the author's knowledge, this paper documents the first time that an image-based visual servo control has been proposed for a dynamic system using vision measurement for both position and velocity.

Visual motion estimation for an autonomous underwater reef monitoring robot

Performing reliable localisation and navigation within highly unstructured underwater coral reef environments is a difficult task at the best of times. Typical research and commercial underwater vehicles use expensive acoustic positioning and sonar systems which require significant external infrastructure to operate effectively. This paper is focused on the development of a robust vision-based motion estimation technique using low-cost sensors for performing real-time autonomous and untethered environmental monitoring tasks in the Great Barrier Reef without the use of acoustic positioning. The technique is experimentally shown to provide accurate odometry and terrain profile information suitable for input into the vehicle controller to perform a range of environmental monitoring tasks.

Visual motion estimation for an autonomous underwater reef monitoring robot

Performing reliable localisation and navigation within highly unstructured underwater coral reef environments is a difficult task at the best of times. Typical research and commercial underwater vehicles use expensive acoustic positioning and sonar systems which require significant external infrastructure to operate effectively. This paper is focused on the development of a robust vision-based motion estimation technique using low-cost sensors for performing real-time autonomous and untethered environmental monitoring tasks in the Great Barrier Reef without the use of acoustic positioning. The technique is experimentally shown to provide accurate odometry and terrain profile information suitable for input into the vehicle controller to perform a range of environmental monitoring tasks.

Sensing for visual homing

In this paper, we outline the sensing system used for the visual pose control of our experimental car-like vehicle, the Autonomous Tractor. The sensing system consists of a magnetic compass, an omnidirectional camera and a low-resolution odometry system. In this work, information from these sensors is fused using complementary filters. Complementary filters provide a means of fusing information from sensors with different characteristics in order to produce a more reliable estimate of the desired variable. Here, the range and bearing of landmarks observed by the vision system are fused with odometry information and a vehicle model, providing a more reliable estimate of these states. We also present a method of combining a compass sensor with odometry and a vehicle model to improve the heading estimate.