Advanced Programming Language Features for Executable Design Patterns "Better Patterns Through Reflection

The Design Patterns book [GOF95] presents 24 time-tested patterns that consistently appear in well-designed software systems. Each pattern is presented with a description of the design problem the pattern addresses, as well as sample implementation code and design considerations. This paper explores how the patterns from the "Gang of Four'', or "GOF'' book, as it is often called, appear when similar problems are addressed using a dynamic, higher-order, object-oriented programming language. Some of the patterns disappear -- that is, they are supported directly by language features, some patterns are simpler or have a different focus, and some are essentially unchanged.

Inferring 3D Structure with a Statistical Image-Based Shape Model

We present an image-based approach to infer 3D structure parameters using a probabilistic "shape+structure'' model. The 3D shape of a class of objects may be represented by sets of contours from silhouette views simultaneously observed from multiple calibrated cameras. Bayesian reconstructions of new shapes can then be estimated using a prior density constructed with a mixture model and probabilistic principal components analysis. We augment the shape model to incorporate structural features of interest; novel examples with missing structure parameters may then be reconstructed to obtain estimates of these parameters. Model matching and parameter inference are done entirely in the image domain and require no explicit 3D construction. Our shape model enables accurate estimation of structure despite segmentation errors or missing views in the input silhouettes, and works even with only a single input view. Using a dataset of thousands of pedestrian images generated from a synthetic model, we can perform accurate inference of the 3D locations of 19 joints on the body based on observed silhouette contours from real images.

Fast Pose Estimation with Parameter Sensitive Hashing

Example-based methods are effective for parameter estimation problems when the underlying system is simple or the dimensionality of the input is low. For complex and high-dimensional problems such as pose estimation, the number of required examples and the computational complexity rapidly becme prohibitively high. We introduce a new algorithm that learns a set of hashing functions that efficiently index examples relevant to a particular estimation task. Our algorithm extends a recently developed method for locality-sensitive hashing, which finds approximate neighbors in time sublinear in the number of examples. This method depends critically on the choice of hash functions; we show how to find the set of hash functions that are optimally relevant to a particular estimation problem. Experiments demonstrate that the resulting algorithm, which we call Parameter-Sensitive Hashing, can rapidly and accurately estimate the articulated pose of human figures from a large database of example images.

Virtual Visual Hulls: Example-Based 3D Shape Estimation from a Single Silhouette

Recovering a volumetric model of a person, car, or other object of interest from a single snapshot would be useful for many computer graphics applications. 3D model estimation in general is hard, and currently requires active sensors, multiple views, or integration over time. For a known object class, however, 3D shape can be successfully inferred from a single snapshot. We present a method for generating a ``virtual visual hull''-- an estimate of the 3D shape of an object from a known class, given a single silhouette observed from an unknown viewpoint. For a given class, a large database of multi-view silhouette examples from calibrated, though possibly varied, camera rigs are collected. To infer a novel single view input silhouette's virtual visual hull, we search for 3D shapes in the database which are most consistent with the observed contour. The input is matched to component single views of the multi-view training examples. A set of viewpoint-aligned virtual views are generated from the visual hulls corresponding to these examples. The 3D shape estimate for the input is then found by interpolating between the contours of these aligned views. When the underlying shape is ambiguous given a single view silhouette, we produce multiple visual hull hypotheses; if a sequence of input images is available, a dynamic programming approach is applied to find the maximum likelihood path through the feasible hypotheses over time. We show results of our algorithm on real and synthetic images of people.

Differential effects of insulin signaling on individual carbon fluxes for fatty acid synthesis in brown adipocytes

Considering the major role of insulin signaling on fatty acid synthesis via stimulation of lipogenic enzymes, differential effects of insulin signaling on individual carbon fluxes for fatty acid synthesis have been investigated by comparing the individual lipogenic fluxes in WT and IRS-1 knockout (IRS-1 KO) brown adipocytes. Results from experiments on WT and IRS-1 KO cells incubated with [5-¹³C] glutamine were consistent with the existence of reductive carboxylation pathway. Analysis of isotopomer distribution of nine metabolites related to the lipogenic routes from glucose and glutamine in IRS-1 KO cells using [U-¹³C] glutamine as compared to that in WT cells indicated that flux through reductive carboxylation pathway was diminished while flux through conventional TCA cycle was stimulated due to absence of insulin signaling in IRS-1 KO cells. This observation was confirmed by quantitative estimation of individual lipogenic fluxes in IRS-1 KO cells and their comparison with fluxes in WT cells. Thus, these results suggest that glutamine’s substantial contribution to fatty acid synthesis can be directly manipulated by controlling the flux through reductive carboxylation of alpha-ketoglutarate to citrate using hormone (insulin).

Inverse Metabolic Engineering of Synechocystis PCC 6803 for Improved Growth Rate and Poly-3-hydroxybutyrate Production

Synechocystis PCC 6803 is a photosynthetic bacterium that has the potential to make bioproducts from carbon dioxide and light. Biochemical production from photosynthetic organisms is attractive because it replaces the typical bioprocessing steps of crop growth, milling, and fermentation, with a one-step photosynthetic process. However, low yields and slow growth rates limit the economic potential of such endeavors. Rational metabolic engineering methods are hindered by limited cellular knowledge and inadequate models of Synechocystis. Instead, inverse metabolic engineering, a scheme based on combinatorial gene searches which does not require detailed cellular models, but can exploit sequence data and existing molecular biological techniques, was used to find genes that (1) improve the production of the biopolymer poly-3-hydroxybutyrate (PHB) and (2) increase the growth rate. A fluorescence activated cell sorting assay was developed to screen for high PHB producing clones. Separately, serial sub-culturing was used to select clones that improve growth rate. Novel gene knock-outs were identified that increase PHB production and others that increase the specific growth rate. These improvements make this system more attractive for industrial use and demonstrate the power of inverse metabolic engineering to identify novel phenotype-associated genes in poorly understood systems.

Protein Microarray: "Theory" to "Real Practice"

Fueled by ever-growing genomic information and rapid developments of proteomics–the large scale analysis of proteins and mapping its functional role has become one of the most important disciplines for characterizing complex cell function. For building functional linkages between the biomolecules, and for providing insight into the mechanisms of biological processes, last decade witnessed the exploration of combinatorial and chip technology for the detection of bimolecules in a high throughput and spatially addressable fashion. Among the various techniques developed, the protein chip technology has been rapid. Recently we demonstrated a new platform called “Spacially addressable protein array” (SAPA) to profile the ligand receptor interactions. To optimize the platform, the present study investigated various parameters such as the surface chemistry and role of additives for achieving high density and high-throughput detection with minimal nonspecific protein adsorption. In summary the present poster will address some of the critical challenges in protein micro array technology and the process of fine tuning to achieve the optimum system for solving real biological problems.

SCAPA (Spacially Addressable Protein Array) – a Novel Protein Array for the Differential Profiling of Ligand-receptor Induced Signalling

While protein microarray technology has been successful in demonstrating its usefulness for large scale high-throughput proteome profiling, performance of antibody/antigen microarrays has been only moderately productive. Immobilization of either the capture antibodies or the protein samples on solid supports has severe drawbacks. Denaturation of the immobilized proteins as well as inconsistent orientation of antibodies/ligands on the arrays can lead to erroneous results. This has prompted a number of studies to address these challenges by immobilizing proteins on biocompatible surfaces, which has met with limited success. Our strategy relates to a multiplexed, sensitive and high-throughput method for the screening quantification of intracellular signalling proteins from a complex mixture of proteins. Each signalling protein to be monitored has its capture moiety linked to a specific oligo ‘tag’. The array involves the oligonucleotide hybridization-directed localization and identification of different signalling proteins simultaneously, in a rapid and easy manner. Antibodies have been used as the capture moieties for specific identification of each signaling protein. The method involves covalently partnering each antibody/protein molecule with a unique DNA or DNA derivatives oligonucleotide tag that directs the antibody to a unique site on the microarray due to specific hybridization with a complementary tag-probe on the array. Particular surface modifications and optimal conditions allowed high signal to noise ratio which is essential to the success of this approach.