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Posters for AA/CE Reception

Kuether, Robert J.; Allensworth, Brooke M.; Backer, Adam B.; Chen, Elton Y.; Dingreville, Remi P.; Forrest, Eric C.; Knepper, Robert; Tappan, Alexander S.; Marquez, Michael P.; Vasiliauskas, Jonathan G.; Rupper, Stephen G.; Grant, Michael J.; Atencio, Lauren C.; Hipple, Tyler J.; Maes, Danae M.; Timlin, Jerilyn A.; Ma, Tian J.; Garcia, Rudy J.; Danford, Forest L.; Patrizi, Laura P.; Galasso, Jennifer G.; Draelos, Timothy J.; Gunda, Thushara G.; Venezuela, Otoniel V.; Brooks, Wesley A.; Anthony, Stephen M.; Carson, Bryan C.; Reeves, Michael J.; Roach, Matthew R.; Maines, Erin M.; Lavin, Judith M.; Whetten, Shaun R.; Swiler, Laura P.

Abstract not provided.

3D optical sectioning with a new hyperspectral confocal fluorescence imaging system

Haaland, David M.; Sinclair, Michael B.; Jones, Howland D.; Timlin, Jerilyn A.; Bachand, George B.; Sasaki, Darryl Y.; Davidson, George S.; Van Benthem, Mark V.

A novel hyperspectral fluorescence microscope for high-resolution 3D optical sectioning of cells and other structures has been designed, constructed, and used to investigate a number of different problems. We have significantly extended new multivariate curve resolution (MCR) data analysis methods to deconvolve the hyperspectral image data and to rapidly extract quantitative 3D concentration distribution maps of all emitting species. The imaging system has many advantages over current confocal imaging systems including simultaneous monitoring of numerous highly overlapped fluorophores, immunity to autofluorescence or impurity fluorescence, enhanced sensitivity, and dramatically improved accuracy, reliability, and dynamic range. Efficient data compression in the spectral dimension has allowed personal computers to perform quantitative analysis of hyperspectral images of large size without loss of image quality. We have also developed and tested software to perform analysis of time resolved hyperspectral images using trilinear multivariate analysis methods. The new imaging system is an enabling technology for numerous applications including (1) 3D composition mapping analysis of multicomponent processes occurring during host-pathogen interactions, (2) monitoring microfluidic processes, (3) imaging of molecular motors and (4) understanding photosynthetic processes in wild type and mutant Synechocystis cyanobacteria.

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Reverse engineering biological networks :applications in immune responses to bio-toxins

Faulon, Jean-Loup M.; Zhang, Zhaoduo Z.; Martino, Anthony M.; Timlin, Jerilyn A.; Haaland, David M.; Davidson, George S.; May, Elebeoba E.; Slepoy, Alexander S.

Our aim is to determine the network of events, or the regulatory network, that defines an immune response to a bio-toxin. As a model system, we are studying T cell regulatory network triggered through tyrosine kinase receptor activation using a combination of pathway stimulation and time-series microarray experiments. Our approach is composed of five steps (1) microarray experiments and data error analysis, (2) data clustering, (3) data smoothing and discretization, (4) network reverse engineering, and (5) network dynamics analysis and fingerprint identification. The technological outcome of this study is a suite of experimental protocols and computational tools that reverse engineer regulatory networks provided gene expression data. The practical biological outcome of this work is an immune response fingerprint in terms of gene expression levels. Inferring regulatory networks from microarray data is a new field of investigation that is no more than five years old. To the best of our knowledge, this work is the first attempt that integrates experiments, error analyses, data clustering, inference, and network analysis to solve a practical problem. Our systematic approach of counting, enumeration, and sampling networks matching experimental data is new to the field of network reverse engineering. The resulting mathematical analyses and computational tools lead to new results on their own and should be useful to others who analyze and infer networks.

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High throughput instruments, methods, and informatics for systems biology

Davidson, George S.; Sinclair, Michael B.; Thomas, Edward V.; Werner-Washburne, Margaret; Davidson, George S.; Boyack, Kevin W.; Wylie, Brian N.; Haaland, David M.; Timlin, Jerilyn A.; Keenan, Michael R.

High throughput instruments and analysis techniques are required in order to make good use of the genomic sequences that have recently become available for many species, including humans. These instruments and methods must work with tens of thousands of genes simultaneously, and must be able to identify the small subsets of those genes that are implicated in the observed phenotypes, or, for instance, in responses to therapies. Microarrays represent one such high throughput method, which continue to find increasingly broad application. This project has improved microarray technology in several important areas. First, we developed the hyperspectral scanner, which has discovered and diagnosed numerous flaws in techniques broadly employed by microarray researchers. Second, we used a series of statistically designed experiments to identify and correct errors in our microarray data to dramatically improve the accuracy, precision, and repeatability of the microarray gene expression data. Third, our research developed new informatics techniques to identify genes with significantly different expression levels. Finally, natural language processing techniques were applied to improve our ability to make use of online literature annotating the important genes. In combination, this research has improved the reliability and precision of laboratory methods and instruments, while also enabling substantially faster analysis and discovery.

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Carbon sequestration in Synechococcus Sp.: from molecular machines to hierarchical modeling

Proposed for publication in OMICS: A Journal of Integrative Biology, Vol. 6, No.4, 2002.

Heffelfinger, Grant S.; Faulon, Jean-Loup M.; Frink, Laura J.; Haaland, David M.; Hart, William E.; Lane, Todd L.; Heffelfinger, Grant S.; Plimpton, Steven J.; Roe, Diana C.; Timlin, Jerilyn A.; Martino, Anthony M.; Rintoul, Mark D.; Davidson, George S.

The U.S. Department of Energy recently announced the first five grants for the Genomes to Life (GTL) Program. The goal of this program is to ''achieve the most far-reaching of all biological goals: a fundamental, comprehensive, and systematic understanding of life.'' While more information about the program can be found at the GTL website (, this paper provides an overview of one of the five GTL projects funded, ''Carbon Sequestration in Synechococcus Sp.: From Molecular Machines to Hierarchical Modeling.'' This project is a combined experimental and computational effort emphasizing developing, prototyping, and applying new computational tools and methods to elucidate the biochemical mechanisms of the carbon sequestration of Synechococcus Sp., an abundant marine cyanobacteria known to play an important role in the global carbon cycle. Understanding, predicting, and perhaps manipulating carbon fixation in the oceans has long been a major focus of biological oceanography and has more recently been of interest to a broader audience of scientists and policy makers. It is clear that the oceanic sinks and sources of CO(2) are important terms in the global environmental response to anthropogenic atmospheric inputs of CO(2) and that oceanic microorganisms play a key role in this response. However, the relationship between this global phenomenon and the biochemical mechanisms of carbon fixation in these microorganisms is poorly understood. The project includes five subprojects: an experimental investigation, three computational biology efforts, and a fifth which deals with addressing computational infrastructure challenges of relevance to this project and the Genomes to Life program as a whole. Our experimental effort is designed to provide biology and data to drive the computational efforts and includes significant investment in developing new experimental methods for uncovering protein partners, characterizing protein complexes, identifying new binding domains. We will also develop and apply new data measurement and statistical methods for analyzing microarray experiments. Our computational efforts include coupling molecular simulation methods with knowledge discovery from diverse biological data sets for high-throughput discovery and characterization of protein-protein complexes and developing a set of novel capabilities for inference of regulatory pathways in microbial genomes across multiple sources of information through the integration of computational and experimental technologies. These capabilities will be applied to Synechococcus regulatory pathways to characterize their interaction map and identify component proteins in these pathways. We will also investigate methods for combining experimental and computational results with visualization and natural language tools to accelerate discovery of regulatory pathways. Furthermore, given that the ultimate goal of this effort is to develop a systems-level of understanding of how the Synechococcus genome affects carbon fixation at the global scale, we will develop and apply a set of tools for capturing the carbon fixation behavior of complex of Synechococcus at different levels of resolution. Finally, because the explosion of data being produced by high-throughput experiments requires data analysis and models which are more computationally complex, more heterogeneous, and require coupling to ever increasing amounts of experimentally obtained data in varying formats, we have also established a companion computational infrastructure to support this effort as well as the Genomes to Life program as a whole.

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6 Results
6 Results