3T3-L1 adipocytes display phenotypic characteristics of multiple adipocyte lineages

Differentiated 3T3-L1 adipocytes are a widely used in vitro model of white adipocytes. In addition to classical white and brown adipocytes that are derived from different cell lineages, beige adipocytes have also been identified, which have characteristics of both white and brown adipocytes. Here we show that 3T3-L1 adipocytes display features of multiple adipocytes lineages. While the gene expression profile and basal bioenergetics of 3T3-L1 adipocytes was typical of white adipocytes, they responded acutely to catecholamines by increasing oxygen consumption in an UCP1-dependent manner, and by increasing the expression of genes enriched in brown but not beige adipocytes. Chronic exposure to catecholamines exacerbated this phenotype. However, a beige adipocyte differentiation procedure did not induce a beige adipocyte phenotype in 3T3-L1 fibroblasts. These multiple lineage features should be considered when interpreting data from experiments utilizing 3T3-L1 adipocytes.

Fully automatic localization and segmentation of 3D vertebral bodies from CT/MR images via a learning-based method

In this paper, we address the problems of fully automatic localization and segmentation of 3D vertebral bodies from CT/MR images. We propose a learning-based, unified random forest regression and classification framework to tackle these two problems. More specifically, in the first stage, the localization of 3D vertebral bodies is solved with random forest regression where we aggregate the votes from a set of randomly sampled image patches to get a probability map of the center of a target vertebral body in a given image. The resultant probability map is then further regularized by Hidden Markov Model (HMM) to eliminate potential ambiguity caused by the neighboring vertebral bodies. The output from the first stage allows us to define a region of interest (ROI) for the segmentation step, where we use random forest classification to estimate the likelihood of a voxel in the ROI being foreground or background. The estimated likelihood is combined with the prior probability, which is learned from a set of training data, to get the posterior probability of the voxel. The segmentation of the target vertebral body is then done by a binary thresholding of the estimated probability. We evaluated the present approach on two openly available datasets: 1) 3D T2-weighted spine MR images from 23 patients and 2) 3D spine CT images from 10 patients. Taking manual segmentation as the ground truth (each MR image contains at least 7 vertebral bodies from T11 to L5 and each CT image contains 5 vertebral bodies from L1 to L5), we evaluated the present approach with leave-one-out experiments. Specifically, for the T2-weighted MR images, we achieved for localization a mean error of 1.6 mm, and for segmentation a mean Dice metric of 88.7% and a mean surface distance of 1.5 mm, respectively. For the CT images we achieved for localization a mean error of 1.9 mm, and for segmentation a mean Dice metric of 91.0% and a mean surface distance of 0.9 mm, respectively.

Stable feature selection with support vector machines

The support vector machine (SVM) is a popular method for classification, well known for finding the maximum-margin hyperplane. Combining SVM with l1-norm penalty further enables it to simultaneously perform feature selection and margin maximization within a single framework. However, l1-norm SVM shows instability in selecting features in presence of correlated features. We propose a new method to increase the stability of l1-norm SVM by encouraging similarities between feature weights based on feature correlations, which is captured via a feature covariance matrix. Our proposed method can capture both positive and negative correlations between features. We formulate the model as a convex optimization problem and propose a solution based on alternating minimization. Using both synthetic and real-world datasets, we show that our model achieves better stability and classification accuracy compared to several state-of-the-art regularized classification methods.

Stabilizing l1-norm prediction models by supervised feature grouping

Emerging Electronic Medical Records (EMRs) have reformed the modern healthcare. These records have great potential to be used for building clinical prediction models. However, a problem in using them is their high dimensionality. Since a lot of information may not be relevant for prediction, the underlying complexity of the prediction models may not be high. A popular way to deal with this problem is to employ feature selection. Lasso and l1-norm based feature selection methods have shown promising results. But, in presence of correlated features, these methods select features that change considerably with small changes in data. This prevents clinicians to obtain a stable feature set, which is crucial for clinical decision making. Grouping correlated variables together can improve the stability of feature selection, however, such grouping is usually not known and needs to be estimated for optimal performance. Addressing this problem, we propose a new model that can simultaneously learn the grouping of correlated features and perform stable feature selection. We formulate the model as a constrained optimization problem and provide an efficient solution with guaranteed convergence. Our experiments with both synthetic and real-world datasets show that the proposed model is significantly more stable than Lasso and many existing state-of-the-art shrinkage and classification methods. We further show that in terms of prediction performance, the proposed method consistently outperforms Lasso and other baselines. Our model can be used for selecting stable risk factors for a variety of healthcare problems, so it can assist clinicians toward accurate decision making.

Literacy, technology and the economics of attention

This article is based on a project aimed at generating practical suggestions based on research findings about how new technologies might be used to enhance L1 literacy attainment in disadvantagedsettings. The project involved designing,implementing and researching an innovative approach to curriculum and pedagogy using new digital technologies in language and literacy education within classroom settings involving small groups of "disadvantaged'' learners. The paper reports activity and findings from one of four study sites. It focuses on four Grade 9 boys seen by their teachers as troublemakers and at risk of failing in English. The researchers draw on current conceptual and theoretical work associated with the emergence of an Attention Economy theory to design a collaborative activity around constructing a website, and to identify and analyse positive literacy learning outcomes associated with the pedagogical approach taken. The authors show how this new perspective on attention informs a critique of conventional approaches to school organization and classroom learning, and how it can be used to envisage alternative approaches to understanding and teaching students who display literacy learning difficulties at school.

The effect of insulin and exercise on c-Cbl protein abundance and phosphorylation in insulin-resistant skeletal muscle in lean and obese Zucker rats.

Aims/hypothesis: Recruitment of the protein c-Cbl to the insulin receptor (IR) and its tyrosine phosphorylation via a pathway that is independent from phosphatidylinositol 3prime-kinase is necessary for insulin-stimulated GLUT4 translocation in 3T3-L1 adipocytes. The activation of this pathway by insulin or exercise has yet to be reported in skeletal muscle. Methods: Lean and obese Zucker rats were randomly assigned to one of three treatment groups: (i) control, (ii) insulin-stimulated or (iii) acute, exhaustive exercise. Hind limb skeletal muscle was removed and the phosphorylation state of IR, Akt and c-Cbl measured.  Results:   Insulin receptor phosphorylation was increased 12-fold after insulin stimulation (p<0.0001) in lean rats and threefold in obese rats. Acute exercise had no effect on IR tyrosine phosphorylation. Similar results were found for serine phosphorylation of Akt. Exercise did not alter c-Cbl tyrosine phosphorylation in skeletal muscle of lean or obese rats. However, in contrast to previous studies in adipocytes, c-Cbl tyrosine phosphorylation was reduced after insulin treatment (p<0.001). Conclusions/interpretation: We also found that c-Cbl associating protein expression is relatively low in skeletal muscle of Zucker rats compared to 3T3-L1 adipocytes and this could account for the reduced c-Cbl tyrosine phosphorylation after insulin treatment. Interestingly, basal levels of c-Cbl tyrosine phosphorylation were higher in skeletal muscle from insulin-resistant Zucker rats (p<0.05), but the physiological relevance is not clear. We conclude that the regulation of c-Cbl phosphorylation in skeletal muscle differs from that previously reported in adipocytes.

Joint space-time trellis decoding and channel estimation in correlated fading channels

This letter addresses the issue of joint space-time trellis decoding and channel estimation in time-varying fading channels that are spatially and temporally correlated. A recursive space-time receiver which incorporates per-survivor processing (PSP) and Kalman filtering into the Viterbi algorithm is proposed. This approach generalizes existing work to the correlated fading channel case. The channel time-evolution is modeled by a multichannel autoregressive process, and a bank of Kalman filters is used to track the channel variations. Computer simulation results show that a performance close to the maximum likelihood receiver with perfect channel state information (CSI) can be obtained. The effects of the spatial correlation on the performance of a receiver that assumes independent fading channels are examined.

Long-term effects of calcium-vitamin-D3-fortified milk on bone geometry and strength in older men

The long-term effects of calcium and vitamin D supplementation on bone material and structural properties in older men are not known. The aim of this study was to examine the effects of high calcium (1000 mg/day)- and vitamin-D3 (800 IU/day)-fortified milk on cortical and trabecular volumetric BMD (vBMD) and bone geometry at the axial and appendicular skeleton in men aged over 50 years. One hundred and eleven men who were part of a larger 2-year randomized controlled trial had QCT scans of the mid-femur and lumbar spine (L1–L3) to assess vBMD, bone geometry and indices of bone strength [polar moment of inertia (Ipolar)]. After 2 years, there were no significant differences between the milk supplementation and control group for the change in any mid-femur or L1–L3 bone parameters for all men aged over 50 years. However, the mid-femur skeletal responses to the fortified milk varied according to age, with a split of ≤62 versus >62 years being the most significant for discriminating the changes between the two groups. Subsequent analysis revealed that, in the older men (>62 years), the expansion in mid-femur medullary area was 2.8% (P < 0.01) less in the milk supplementation compared to control group, which helped to preserve cortical area in the milk supplementation group (between group difference 1.1%, P < 0.01). Similarly, for mid-femur cortical vBMD and Ipolar, the net loss was 2.3 and 2.8% less in the milk supplementation compared to control group (P < 0.01 and <0.001, respectively). In conclusion, calcium–vitamin-D3-fortified milk may represent an effective strategy to maintain bone strength by preventing endocortical bone loss and slowing the loss in cortical vBMD in elderly men.

Chemerin is a novel adipokine associated with obesity and metabolic syndrome

Soluble protein hormones are key regulators of a number of metabolic processes, including food intake and insulin sensitivity. We have used a signal sequence trap to identify genes that encode secreted or membrane-bound proteins in Psammomys obesus, an animal model of obesity and type 2 diabetes (T2D). Using this signal sequence trap, we identified the chemokine chemerin as being a novel adipokine. Gene expression of chemerin and its receptor, chemokine-like receptor 1 (CMKLR1), was significantly higher in adipose tissue of obese and type 2 diabetic P. obesus compared with lean, normoglycemic P. obesus. Fractionation of P. obesus adipose tissue confirmed that chemerin was predominantly expressed in adipocytes, whereas CMKLR1 was expressed in both adipocytes and stromal-vascular cells of adipose tissue. In 3T3-L1 adipocytes, chemerin was markedly induced during differentiation, whereas CMKLR1 was down-regulated during differentiation. Serum chemerin levels were measured by ELISA in human plasma samples from 114 subjects with T2D and 142 normal glucose tolerant controls. Plasma chemerin levels were not significantly different between subjects with T2D and normal controls. However, in normal glucose tolerant subjects, plasma chemerin levels were significantly associated with body mass index, circulating triglycerides, and blood pressure. Here we report, for the first time, that chemerin is an adipokine, and circulating levels of chemerin are associated with several key aspects of metabolic syndrome.

Three-dimensional Finite Element modelling of laser shock peening process

Laser shock peening (LSP) is an innovative surface treatment technique for metal alloys, with the great improvement of their fatigue, corrosion and wear resistance performance. Finite element method has been widely applied to simulate the LSP to provide the theoretically predictive assessment and optimally parametric design. In the current work, 3-D numerical modelling approaches, combining the explicit dynamic analysis, static equilibrium analysis algorithms and different plasticity models for the high strain rate exceeding 106s-1, are further developed. To verify the proposed methods, 3-D static and dynamic FEA of AA7075-T7351 rods subject to two-sided laser shock peening are performed using the FEA package–ABAQUS. The dynamic and residual stress fields, shock wave propagation and surface deformation of the treated metal from different material modelling approaches have a good agreement.

The effect of laser power density on the fatigue life of laser-shock-peened 7050 aluminium alloy

Laser shock peening (LSP) is an innovative surface treatment method that can result in significant improvement in the fatigue life of many metallic components. The process produces very little or no surface profile modification while producing a considerably deeper compressive residual stress layer than traditional shot peening operations. The work discussed here was designed to: (a) quantify the fatigue life improvement achieved by LSP in a typical high strength aircraft aluminium alloy and (b) identify any technological risks associated with its use. It is shown that when LSP conditions are optimal for the material and specimen configuration, a —three to four times increase in fatigue life over the as-machined specimens could be achieved for a representative fighter aircraft loading spectrum when applied at a representative load level. However, if the process parameters are not optimal for the material investigated here, fatigue lives of LSP treated specimens may be reduced instead of increased due to the occurrence of internal cracking. This paper details the effect of laser power density on fatigue life of 7050-T7451 aluminium alloy by experimental and numerical analysis.

Writing, digital culture and English curriculum

In their out-of-school lives, young people are immersed in rich and complex digital worlds, characterised by image and multimodality. Computer games in particular present young people with specific narrative genres and textual forms: contexts in which meaning is constructed interactively and drawing explicitly on a wide range of design elements including sound, image, gesture, symbol, colour and so on. As English curriculum seeks to address the changing nature of literacy, challenges are raised, particularly with respect to the ways in which multimodal texts might be incorporated alongside print based forms of literacy. Questions focus both on the ways in which such texts might be created, studied and assessed, and on the implications of the introduction of such texts for print based literacies. This paper explores intersections between writing and computer games within the English classroom, from a number of junior secondary examples. In particular it considers tensions that arise when young people use writing to recreate or respond to multimodal forms. It explores ways in which writing is stretched and challenged by enterprises such as these, ways in which students utilise and adapt print based modes to represent multimodal forms of narrative, and how teachers and curriculum might respond. Consideration is given to the challenges posed to teaching and assessment by bringing writing to bear as the medium of analysis of, and response to, multimodal texts.

Regulation of HSL serine phosphorylation in skeletal muscle and adipose tissue

Hormone-sensitive lipase (HSL) is important for the degradation of triacylglycerol in adipose and muscle tissue, but the tissue-specific regulation of this enzyme is not fully understood. We investigated the effects of adrenergic stimulation and AMPK activation in vitro and in circumstances where AMPK activity and catecholamines are physiologically elevated in humans in vivo (during physical exercise) on HSL activity and phosphorylation at Ser⁵⁶³ and Ser⁶⁶⁰, the PKA regulatory sites, and Ser⁵⁶⁵, the AMPK regulatory site. In human experiments, skeletal muscle, subcutaneous adipose and venous blood samples were obtained before, at 15 and 90 min during, and 120 min after exercise. Skeletal muscle HSL activity was increased by ~80% at 15 min compared with rest and returned to resting rates at the cessation of and 120 min after exercise. Consistent with changes in plasma epinephrine, skeletal muscle HSL Ser⁵⁶³ and Ser⁶⁶⁰ phosphorylation were increased by 27% at 15 min (P < 0.05), remained elevated at 90 min, and returned to preexercise values postexercise. Skeletal muscle HSL Ser⁵⁶⁵ phosphorylation and AMPK signaling were increased at 90 min during, and after, exercise. Phosphorylation of adipose tissue HSL paralleled changes in skeletal muscle in vivo, except HSL Ser⁶⁶⁰ was elevated 80% in adipose compared with 35% in skeletal muscle during exercise. Studies in L6 myotubes and 3T3-L1 adipocytes revealed important tissue differences in the regulation of HSL. AMPK inhibited epinephrine-induced HSL activity in L6 myotubes and was associated with reduced HSL Ser660 but not Ser563 phosphorylation. HSL activity was reduced in L6 myotubes expressing constitutively active AMPK, confirming the inhibitory effects of AMPK on HSL activity. Conversely, in 3T3-L1 adipocytes, AMPK activation after epinephrine stimulation did not prevent HSL activity or glycerol release, which coincided with maintenance of HSL Ser660 phosphorylation. Taken together, these data indicate that HSL activity is maintained in the face of AMPK activation as a result of elevated HSL Ser660 phosphorylation in adipose tissue but not skeletal muscle.

The subcellular fractionation properties and function of insulin receptor substrate-1 (IRS-1) are independent of cytoskeletal integrity

Efficient insulin action requires spatial and temporal coordination of signaling cascades. The prototypical insulin receptor substrate, IRS-1 plays a central role in insulin signaling. By subcellular fractionation IRS-1 is enriched in a particulate fraction, termed the high speed pellet (HSP), and its redistribution from this fraction is associated with signal attenuation and insulin resistance. Anecdotal evidence suggests the cytoskeleton may underpin the localization of IRS-1 to the HSP. In the present study we have taken a systematic approach to examine whether the cytoskeleton contributes to the subcellular fractionation properties and function of IRS-1. By standard microscopy or immunoprecipitation we were unable to detect evidence to support a specific interaction between IRS-1 and the major cytoskeletal components actin (microfilaments), vimentin (intermediate filaments), and tubulin (microtubules) in 3T3-L1 adipocytes or in CHO.IR.IRS-1 cells. Pharmacological disruption of microfilaments and microtubules, individually or in combination, was without effect on the subcellular distribution of IRS-1 or insulin-stimulated tyrosine phosphorylation in either cell type. Phosphorylation of Akt was modestly reduced (20–35%) in 3T3-L1 adipocytes but not in CHO.IR.IRS-1 cells. In cells lacking intermediate filaments (Vim^−/−) IRS-1 expression, distribution and insulin-stimulated phosphorylation appeared normal. Even after depolymerisation of microfilaments and microtubules, insulin-stimulated phosphorylation of IRS-1 and Akt were maintained in Vim^−/− cells. Taken together these data indicate that the characteristic subcellular fractionation properties and function of IRS-1 are unlikely to be mediated by cytoskeletal networks and that proximal insulin signaling does not require an intact cytoskeleton.

Geometrical effects on residual stresses in 7050-T7451 aluminum alloy rods subject to laser shock peening

Laser shock peening (LSP) is an emerging surface treatment technology for metallic materials, which appears to produce more significant compressive residual stresses than those from the conventional shot peening (SP) for fatigue, corrosion and wear resistance, etc. The finite element method has been applied to simulate the laser shock peening treatment to provide the overall numerical assessment of the characteristic physical processes and transformations. However, the previous researchers mostly focused on metallic specimens with simple geometry, e.g. flat surface. The current work investigates geometrical effects of metallic specimens with curved surface on the residual stress fields produced by LSP process using three-dimensional finite element (3-D FEM) analysis and aluminium alloy rods with a middle scalloped section subject to two-sided laser shock peening. Specimens were numerically studied to determine dynamic and residual stress fields with varying laser parameters and geometrical parameters, e.g. laser power intensity and radius of the middle scalloped section. The results showed that the geometrical effects of the curved target surface greatly influenced residual stress fields.