Retrotransposon expression profiling: Unveiling the hidden SINE transcriptome through Next-Generation Sequencing data analysis

Of the ~1.7 million SINE elements in the human genome, only a tiny number are estimated to be active in transcription by RNA polymerase (Pol) III. Tracing the individual loci from which SINE transcripts originate is complicated by their highly repetitive nature. By exploiting RNA-Seq datasets and unique SINE DNA sequences, we devised a bioinformatic pipeline allowing us to identify Pol III-dependent transcripts of individual SINE elements. When applied to ENCODE transcriptomes of seven human cell lines, this search strategy identified ~1300 Alu loci and ~1100 MIR loci corresponding to detectable transcripts, with ~120 and ~60 respectively Alu and MIR loci expressed in at least three cell lines. In vitro transcription of selected SINEs did not reflect their in vivo expression properties, and required the native 5’-flanking region in addition to internal promoter. We also identified a cluster of expressed AluYa5-derived transcription units, juxtaposed to snaR genes on chromosome 19, formed by a promoter-containing left monomer fused to an Alu-unrelated downstream moiety. Autonomous Pol III transcription was also revealed for SINEs nested within Pol II-transcribed genes raising the possibility of an underlying mechanism for Pol II gene regulation by SINE transcriptional units. Moreover the application of our bioinformatic pipeline to both RNA-seq data of cells subjected to an in vitro pro-oncogenic stimulus and of in vivo matched tumor and non-tumor samples allowed us to detect increased Alu RNA expression as well as the source loci of such deregulation. The ability to investigate SINE transcriptomes at single-locus resolution will facilitate both the identification of novel biologically relevant SINE RNAs and the assessment of SINE expression alteration under pathological conditions.

Data visualization during the early stages of drug discovery

Multidimensional compound optimization is a new paradigm in the drug discovery process, yielding efficiencies during early stages and reducing attrition in the later stages of drug development. The success of this strategy relies heavily on understanding this multidimensional data and extracting useful information from it. This paper demonstrates how principled visualization algorithms can be used to understand and explore a large data set created in the early stages of drug discovery. The experiments presented are performed on a real-world data set comprising biological activity data and some whole-molecular physicochemical properties. Data visualization is a popular way of presenting complex data in a simpler form. We have applied powerful principled visualization methods, such as generative topographic mapping (GTM) and hierarchical GTM (HGTM), to help the domain experts (screening scientists, chemists, biologists, etc.) understand and draw meaningful decisions. We also benchmark these principled methods against relatively better known visualization approaches, principal component analysis (PCA), Sammon's mapping, and self-organizing maps (SOMs), to demonstrate their enhanced power to help the user visualize the large multidimensional data sets one has to deal with during the early stages of the drug discovery process. The results reported clearly show that the GTM and HGTM algorithms allow the user to cluster active compounds for different targets and understand them better than the benchmarks. An interactive software tool supporting these visualization algorithms was provided to the domain experts. The tool facilitates the domain experts by exploration of the projection obtained from the visualization algorithms providing facilities such as parallel coordinate plots, magnification factors, directional curvatures, and integration with industry standard software. © 2006 American Chemical Society.

Discovery of active proteins directly from combinatorial randomized protein libraries without display, purification or sequencing:identification of novel zinc finger proteins.

We have successfully linked protein library screening directly with the identification of active proteins, without the need for individual purification, display technologies or physical linkage between the protein and its encoding sequence. By using 'MAX' randomization we have rapidly constructed 60 overlapping gene libraries that encode zinc finger proteins, randomized variously at the three principal DNA-contacting residues. Expression and screening of the libraries against five possible target DNA sequences generated data points covering a potential 40,000 individual interactions. Comparative analysis of the resulting data enabled direct identification of active proteins. Accuracy of this library analysis methodology was confirmed by both in vitro and in vivo analyses of identified proteins to yield novel zinc finger proteins that bind to their target sequences with high affinity, as indicated by low nanomolar apparent dissociation constants.

Towards an operations strategy for product-centric servitization

Purpose – This paper aims to present a framework that will help manufacturing firms to configure their internal production and support operations to enable effective and efficient delivery of products and their closely associated services. Design/methodology/approach – First, the key definitions and literature sources directly associated with servitization of manufacturing are established. Then, a theoretical framework that categorises the key characteristics of a manufacturer's operations strategy is developed and this is populated using both evidence from the extant literature and empirical data. Findings – The framework captures a set of operations principles, structures and processes that can guide a manufacturer in the delivery of product-centric servitized offering. These are illustrated and contrasted against operations that deliver purely product (production operations) and those which deliver purely services (services operations). Research limitations/implications – The work is based on a review of the literature supported by data collected from an exploratory case study. Whilst it provides an essential platform, further research will be needed to validate the framework. Originality/value – The principal contribution of this paper is a framework that captures the key characteristics of operations for product-centric servitized manufacture.

Active vibration control at machinery feet

The unmitigated transmission of undesirable vibration can result in problems by way of causing human discomfort, machinery and equipment failure, and affecting the quality of a manufacturing process. When identifiable transmission paths are discernible, vibrations from the source can be isolated from the rest of the system and this prevents or minimises the problems. The approach proposed here for vibration isolation is active force cancellation at points close to the vibration source. It uses force feedback for multiple-input and multiple-output control at the mounting locations. This is particularly attractive for rigid mounting of machine on relative flexible base where machine alignment and motions are to be restricted. The force transfer function matrix is used as a disturbance rejection performance specification for the design of MIMO controllers. For machine soft-mounted via flexible isolators, a model for this matrix has been derived. Under certain conditions, a simple multiplicative uncertainty model is obtained that shows the amount of perturbation a flexible base has on the machine-isolator-rigid base transmissibility matrix. Such a model is very suitable for use with robust control design paradigm. A different model is derived for the machine on hard-mounts without the flexible isolators. With this model, the level of force transmitted from a machine to a final mounting structure using the measurements for the machine running on another mounting structure can be determined. The two mounting structures have dissimilar dynamic characteristics. Experiments have verified the usefulness of the expression. The model compares well with other methods in the literature. The disadvantage lies with the large amount of data that has to be collected. Active force cancellation is demonstrated on an experimental rig using an AC industrial motor hard-mounted onto a relative flexible structure. The force transfer function matrix, determined from measurements, is used to design H and Static Output Feedback controllers. Both types of controllers are stable and robust to modelling errors within the identified frequency range. They reduce the RMS of transmitted force by between 30?80% at all mounting locations for machine running at 1340 rpm. At the rated speed of 1440 rpm only the static gain controller is able to provide 30?55% reduction at all locations. The H controllers on the other hand could only give a small reduction at one mount location. This is due in part to the deficient of the model used in the design. Higher frequency dynamics has been ignored in the model. This can be resolved by the use of a higher order model that can result in a high order controller. A low order static gain controller, with some tuning, performs better. But it lacks the analytical framework for analysis and design.

Asymmetric reactions of organosilicon compounds and properties of novel optically active organosilanes

Partial reduction of racemic methoxysilanes by 1:1 complexes of lithium aluminium hydride with optically active cinchona and ephedra alkaloids give optically active silanes and methoxysilanes. Optical yields depend on the groups attached to silicon and the alkaloid used but in some cases approach 50%, The method has been used to prepare novel optically active organosilanes, possessing an asymmetric silicon centre, which are either inaccessible by any of the other available routes or would require a time consuming preparation. Such compounds are of use in the study of the mechanism of substitutions at silicon. Attempts have been made to rationalize the results of the asymmetric reductions in terms of differences in sterio and electronic interactions in diastereoisomeric transition states. Circular dichroism and optical rotatory dispersion spectra have been obtained for the optically active products in an attempt to elucidate the absolute configurations of the novel asymmetric organosilanes. The results from these studies provide a useful addition to the data so far accumulated for asymmetrically perturbed aromatic chromophores. Nuclear magnetic resonanoe studies of diastereoisomaric (-)-menthoxysilanes show that these compounds possess resonances extremely useful in the determination of optical purities for asymmetric organosilanes which possess an aromatic group. The effect of variable temperature on the spectra has revealed evidence for the conformational preferences in these compounds. Other diastereoisomeric alkoxysilanes have been prepared and their n.m.r.spectra studied in the hope of establishing trends. Exploratory studies for other asymmetric reactions proceeding at silicon have proved unfruitful.

Towards Active Vision in the DUAL Cognitive Architecture

The paper describes an extension of the cognitive architecture DUAL with a model of visual attention and perception. The goal of this attempt is to account for the construction and the categorization of object and scene representations derived from visual stimuli in the TextWorld microdomain. Low-level parallel computations are combined with an active serial deployment of visual attention enabling the construction of abstract symbolic representations. A limited-capacity short-term visual store holding information across attention shifts forms the core of the model interfacing between the low-level representation of the stimulus and DUAL’s semantic memory. The model is validated by comparing the results of a simulation with real data from an eye movement experiment with human subjects.

Copper complexes as a source of redox active MRI contrast agents

The study reports an advance in designing copper-based redox sensing MRI contrast agents. Although the data demonstrate that copper(II) complexes are not able to compete with lanthanoids species in terms of contrast, the redox-dependent switch between diamagnetic copper(I) and paramagnetic copper(II) yields a novel redox-sensitive contrast moiety with potential for reversibility.

A comparison of breathing patterns of normal full term infants in supine versus right side-lying during quiet and active behavioral sleep states

This study investigated the effect of sleep position on breathing patterns of normal full term infants during quiet and active behavioral sleep states. Tidal volume, percent contribution of rib cage to tidal volume, and respiration rate were measured via respiratory inductive plethysmography (RIP) and pneumotachograph (PNT) in ten infants sleeping in supine versus right side-lying. Data was collected immediately following two consecutive feedings. Paired t tests and ANOVA comparisons showed no significant differences between the two postures (p $<$.05) in mean tidal volume (supine, M = 19.16, right side, M = 22.45), percent contribution of rib cage to tidal volume (supine, M = 30.55, right side M = 33.20), or respiration rate (supine, M = 49.13, right side, M = 49.37) during quiet sleep. Comparisons also showed no significant differences between the two postures (p $<$.05) in mean tidal volume (supine, M = 18.89, right side, M = 20.12), percent contribution of rib cage to tidal volume (supine, M = 6.43, right side, M = 6.97) or respiration rate (supine, M = 62.18, right side, M = 61.04) during active sleep. Therefore, no differences were found in the three respiratory variables measured between the supine and right side-lying positions. These findings suggest that infants may be positioned in either sleep position without detriment to respiratory function. This information may benefit occupational therapists and other health professionals involved in the education of parents on infant positioning and their respective advantages. ^

Dynamic data retrieval on the World Wide Web

Methods for accessing data on the Web have been the focus of active research over the past few years. In this thesis we propose a method for representing Web sites as data sources. We designed a Data Extractor data retrieval solution that allows us to define queries to Web sites and process resulting data sets. Data Extractor is being integrated into the MSemODB heterogeneous database management system. With its help database queries can be distributed over both local and Web data sources within MSemODB framework. ^ Data Extractor treats Web sites as data sources, controlling query execution and data retrieval. It works as an intermediary between the applications and the sites. Data Extractor utilizes a twofold “custom wrapper” approach for information retrieval. Wrappers for the majority of sites are easily built using a powerful and expressive scripting language, while complex cases are processed using Java-based wrappers that utilize specially designed library of data retrieval, parsing and Web access routines. In addition to wrapper development we thoroughly investigate issues associated with Web site selection, analysis and processing. ^ Data Extractor is designed to act as a data retrieval server, as well as an embedded data retrieval solution. We also use it to create mobile agents that are shipped over the Internet to the client's computer to perform data retrieval on behalf of the user. This approach allows Data Extractor to distribute and scale well. ^ This study confirms feasibility of building custom wrappers for Web sites. This approach provides accuracy of data retrieval, and power and flexibility in handling of complex cases. ^

Abstractions to support dynamic adaptation of communication frameworks for user-centric communication

The convergence of data, audio and video on IP networks is changing the way individuals, groups and organizations communicate. This diversity of communication media presents opportunities for creating synergistic collaborative communications. This form of collaborative communication is however not without its challenges. The increasing number of communication service providers coupled with a combinatorial mix of offered services, varying Quality-of-Service and oscillating pricing of services increases the complexity for the user to manage and maintain ‘always best’ priced or performance services. Consumers have to manually manage and adapt their communication in line with differences in services across devices, networks and media while ensuring that the usage remain consistent with their intended goals. This dissertation proposes a novel user-centric approach to address this problem. The proposed approach aims to reduce the aforementioned complexity to the user by (1) providing high-level abstractions and a policy based methodology for automated selection of the communication services guided by high-level user policies and (2) providing services through the seamless integration of multiple communication service providers and providing an extensible framework to support the integration of multiple communication service providers. The approach was implemented in the Communication Virtual Machine (CVM), a model-driven technology for realizing communication applications. The CVM includes the Network Communication Broker, the layer responsible for providing a network-independent API to the upper layers of CVM. The initial prototype for the NCB supported only a single communication framework which limited the number, quality and types of services available. Experimental evaluation of the approach show the additional overhead of the approach is minimal compared to the individual communication services frameworks. Additionally the automated approach proposed out performed the individual communication services frameworks for cross framework switching.

User-centric music information retrieval

The rapid growth of the Internet and the advancements of the Web technologies have made it possible for users to have access to large amounts of on-line music data, including music acoustic signals, lyrics, style/mood labels, and user-assigned tags. The progress has made music listening more fun, but has raised an issue of how to organize this data, and more generally, how computer programs can assist users in their music experience. An important subject in computer-aided music listening is music retrieval, i.e., the issue of efficiently helping users in locating the music they are looking for. Traditionally, songs were organized in a hierarchical structure such as genre->artist->album->track, to facilitate the users’ navigation. However, the intentions of the users are often hard to be captured in such a simply organized structure. The users may want to listen to music of a particular mood, style or topic; and/or any songs similar to some given music samples. This motivated us to work on user-centric music retrieval system to improve users’ satisfaction with the system. The traditional music information retrieval research was mainly concerned with classification, clustering, identification, and similarity search of acoustic data of music by way of feature extraction algorithms and machine learning techniques. More recently the music information retrieval research has focused on utilizing other types of data, such as lyrics, user-access patterns, and user-defined tags, and on targeting non-genre categories for classification, such as mood labels and styles. This dissertation focused on investigating and developing effective data mining techniques for (1) organizing and annotating music data with styles, moods and user-assigned tags; (2) performing effective analysis of music data with features from diverse information sources; and (3) recommending music songs to the users utilizing both content features and user access patterns.

An Analytical Workflow for Metagenomic Data and its Application to the Study of Chronic Obstructive Pulmonary Disease (COPD)

Metagenomics is the culture-independent study of genetic material obtained directly from environmental samples. It has become a realistic approach to understanding microbial communities thanks to advances in high-throughput DNA sequencing technologies over the past decade. Current research has shown that different sites of the human body house varied bacterial communities. There is a strong correlation between an individual’s microbial community profile at a given site and disease. Metagenomics is being applied more often as a means of comparing microbial profiles in biomedical studies. The analysis of the data collected using metagenomics can be quite challenging and there exist a plethora of tools for interpreting the results. An automatic analytical workflow for metagenomic analyses has been implemented and tested using synthetic datasets of varying quality. It is able to accurately classify bacteria by taxa and correctly estimate the richness and diversity of each set. The workflow was then applied to the study of the airways microbiome in Chronic Obstructive Pulmonary Disease (COPD). COPD is a progressive lung disease resulting in narrowing of the airways and restricted airflow. Despite being the third leading cause of death in the United States, little is known about the differences in the lung microbial community profiles of healthy individuals and COPD patients. Bronchoalveolar lavage (BAL) samples were collected from COPD patients, active or ex-smokers, and never smokers and sequenced by 454 pyrosequencing. A total of 56 individuals were recruited for the study. Substantial colonization of the lungs was found in all subjects and differentially abundant genera in each group were identified. These discoveries are promising and may further our understanding of how the structure of the lung microbiome is modified as COPD progresses. It is also anticipated that the results will eventually lead to improved treatments for COPD.

What factors influence persistence rates in active older adult group exercise programs?

The purpose of this research is to determine factors that influence the persistence rates of active older adults participating in group exercise classes. During the study, group participants and instructors in a Florida facility were asked to respond to surveys ascertaining motivational factors regarding participation in active older adult classes. Utilizing participant data, instructors were given a professional development course as part of the treatment. Data indicates participants prefer working in group settings to prevent health related illness, for socialization, and for instructors. Research demonstrates that the instructor plays a critical role in advancing exercise understanding and providing motivation that encourages participants to attend classes. Instructor response included a heightened awareness of motivational factors and a determination to improve standards.

Dynamic data retrieval on the world wide web

Methods for accessing data on the Web have been the focus of active research over the past few years. In this thesis we propose a method for representing Web sites as data sources. We designed a Data Extractor data retrieval solution that allows us to define queries to Web sites and process resulting data sets. Data Extractor is being integrated into the MSemODB heterogeneous database management system. With its help database queries can be distributed over both local and Web data sources within MSemODB framework. Data Extractor treats Web sites as data sources, controlling query execution and data retrieval. It works as an intermediary between the applications and the sites. Data Extractor utilizes a two-fold "custom wrapper" approach for information retrieval. Wrappers for the majority of sites are easily built using a powerful and expressive scripting language, while complex cases are processed using Java-based wrappers that utilize specially designed library of data retrieval, parsing and Web access routines. In addition to wrapper development we thoroughly investigate issues associated with Web site selection, analysis and processing. Data Extractor is designed to act as a data retrieval server, as well as an embedded data retrieval solution. We also use it to create mobile agents that are shipped over the Internet to the client's computer to perform data retrieval on behalf of the user. This approach allows Data Extractor to distribute and scale well. This study confirms feasibility of building custom wrappers for Web sites. This approach provides accuracy of data retrieval, and power and flexibility in handling of complex cases.