Sequence analysis of Arthrospira maxima based on fosmid library

Arthrospira (Spirulina) (Setchell& Gardner) is an important cyanobacterium not only in its nutritional potential but in its special biological characteristics. An unbiased fosmid library of Arthrospira maxima FACHB438 that contains 4300 clones was constructed. The size distribution of insert fragments is from 15.5 to 48.9 kb and the average size is 37.6 kb. The recombination frequency is 100%. Therefore the library is 29.9 equivalents to the Arthrospira genome size of 5.4 Mb. A total of 719 sample clones were randomly chosen from the library and 602 available sequences, which consisted of 307,547 bases, covering 5.70% of the whole genome. The codon usage of A. maxima was not strongly biased. GC content at the first position of codons (46.9%) was higher than the second (39.8%) and the third (45.5%) positions. GC content of the genome was 43.6%. Of these sequences, 287 (47.7%) showed high similarities to known genes, 63 (10.5%) to hypothetical genes and the remaining 252 (41.8%) had no significant similarities. The assigned genes were classified into 22 categories with respect to different biological roles. Remarkably, the high presence of 25 sequences (4.2%) encoding reverse transcriptase indicates the RT gene may have multiple copies in the A. maxima genome and might play an important role in the evolutionary history and metabolic regulation. In addition, the sequences encoding the ATP-binding cassette transport system and the two-component signal transduction system were the second and third most frequent genes, respectively. These genomic features provide some clues as to the mechanisms by which this organism adapts to the high concentration of bicarbonate and to the high pH environment.

Construction and characterization of two bacterial artificial chromosome libraries of Zhikong scallop, Chlamys farreri Jones et Preston, and identification of BAC clones containing the genes involved in its innate immune system

Large-insert bacterial artificial chromosome (BAC) libraries are necessary for advanced genetics and genomics research. To facilitate gene cloning and characterization, genome analysis, and physical mapping of scallop, two BAC libraries were constructed from nuclear DNA of Zhikong scallop, Chlamys farreri Jones et Preston. The libraries were constructed in the BamHI and MboI sites of the vector pECBAC1, respectively. The BamHI library consists of 73,728 clones, and approximately 99% of the clones contain scallop nuclear DNA inserts with an average size of 110 kb, covering 8.0x haploid genome equivalents. Similarly, the MboI library consists of 7680 clones, with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1x. The combined libraries collectively contain a total of 81,408 BAC clones arrayed in 212 384-well microtiter plates, representing 9.1x haploid genome equivalents and having a probability of greater than 99% of discovering at least one positive clone with a single-copy sequence. High-density clone filters prepared from a subset of the two libraries were screened with nine pairs of Overgos designed from the cDNA or DNA sequences of six genes involved in the innate immune system of mollusks. Positive clones were identified for every gene, with an average of 5.3 BAC clones per gene probe. These results suggest that the two scallop BAC libraries provide useful tools for gene cloning, genome physical mapping, and large-scale sequencing in the species.

Characterization and mapping of expressed sequence tags isolated from a subtracted cDNA library of Litopenaeus vannamei injected with white spot syndrome virus

A large number of polymorphic simple sequence repeats (SSRs) or microsatellites are needed to develop a genetic map for shrimp. However, developing an SSR map is very time-consuming, expensive, and most SSRs are not specifically linked to gene loci of immediate interest. We report here on our strategy to develop polymorphic markers using expressed sequence tags (ESTs) by designing primers flanking single or multiple SSRs with three or more repeats. A subtracted cDNA library was prepared using RNA from specific pathogen-free (SPF) Litopenaeus vannamei juveniles (similar to 1 g) collected before (0) and after (48 h) inoculation with the China isolate of white spot syndrome virus (WSSV). A total of 224 clones were sequenced, 194 of which were useful for homology comparisons against annotated genes in NCBI nonredundant (nr) and protein databases, providing 179 sequences encoded by nuclear DNA, 4 mitochondrial DNA, and 11 were similar to portions of WSSV genome. The nuclear sequences clustered in 43 groups, 11 of which were homologous to various ESTs of unknown function, 4 had no homology to any sequence, and 28 showed similarities to known genes of invertebrates and vertebrates, representatives of cellular metabolic processes such as calcium ion balance, cytoskeleton mRNAs, and protein synthesis. A few sequences were homologous to immune system-related (allergens) genes and two were similar to motifs of the sex-lethal gene of Drosophila. A large number of EST sequences were similar to domains of the EF-hand superfamily (Ca2+ binding motif and FRQ protein domain of myosin light chains). Single or multiple SSRs with three or more repeats were found in approximately 61 % of the 179 nuclear sequences. Primer sets were designed from 28 sequences representing 19 known or putative genes and tested for polymorphism (EST-SSR marker) in a small test panel containing 16 individuals. Ten (53%) of the 19 putative or unknown function genes were polymorphic, 4 monomorphic, and 3 either failed to satisfactorily amplify genomic DNA or the allele amplification conditions need to be further optimized. Five polymorphic ESTs were genotyped with the entire reference mapping family, two of them (actin, accession #CX535973 and shrimp allergen arginine kinase, accession #CX535999) did not amplify with all offspring of the IRMF panel suggesting presence of null alleles, and three of them amplified in most of the IRM F offspring and were used for linkage analysis. EF-hand motif of myosin light chain (accession #CX535935) was placed in ShrimpMap's linkage group 7, whereas ribosomal protein S5 (accession #CX535957) and troponin I (accession #CX535976) remained unassigned. Results indicate that (a) a large number of ESTs isolated from this cDNA library are similar to cytoskeleton mRNAs and may reflect a normal pathway of the cellular response after im infection with WSSV, and (b) primers flanking single or multiple SSRs with three or more repeats from shrimp ESTs could be an efficient approach to develop polymorphic markers useful for linkage mapping. Work is underway to map additional SSR-containing ESTs from this and other cDNA libraries as a plausible strategy to increase marker density in ShrimpMap.

Automated Program Recognition

The key to understanding a program is recognizing familiar algorithmic fragments and data structures in it. Automating this recognition process will make it easier to perform many tasks which require program understanding, e.g., maintenance, modification, and debugging. This report describes a recognition system, called the Recognizer, which automatically identifies occurrences of stereotyped computational fragments and data structures in programs. The Recognizer is able to identify these familiar fragments and structures, even though they may be expressed in a wide range of syntactic forms. It does so systematically and efficiently by using a parsing technique. Two important advances have made this possible. The first is a language-independent graphical representation for programs and programming structures which canonicalizes many syntactic features of programs. The second is an efficient graph parsing algorithm.

Automated Assembly Using Feature Localization

Automated assembly of mechanical devices is studies by researching methods of operating assembly equipment in a variable manner; that is, systems which may be configured to perform many different assembly operations are studied. The general parts assembly operation involves the removal of alignment errors within some tolerance and without damaging the parts. Two methods for eliminating alignment errors are discussed: a priori suppression and measurement and removal. Both methods are studied with the more novel measurement and removal technique being studied in greater detail. During the study of this technique, a fast and accurate six degree-of-freedom position sensor based on a light-stripe vision technique was developed. Specifications for the sensor were derived from an assembly-system error analysis. Studies on extracting accurate information from the sensor by optimally reducing redundant information, filtering quantization noise, and careful calibration procedures were performed. Prototype assembly systems for both error elimination techniques were implemented and used to assemble several products. The assembly system based on the a priori suppression technique uses a number of mechanical assembly tools and software systems which extend the capabilities of industrial robots. The need for the tools was determined through an assembly task analysis of several consumer and automotive products. The assembly system based on the measurement and removal technique used the six degree-of-freedom position sensor to measure part misalignments. Robot commands for aligning the parts were automatically calculated based on the sensor data and executed.

Inspection Methods in Programming

The work reported here lies in the area of overlap between artificial intelligence software engineering. As research in artificial intelligence, it is a step towards a model of problem solving in the domain of programming. In particular, this work focuses on the routine aspects of programming which involve the application of previous experience with similar programs. I call this programming by inspection. Programming is viewed here as a kind of engineering activity. Analysis and synthesis by inspection area prominent part of expert problem solving in many other engineering disciplines, such as electrical and mechanical engineering. The notion of inspections methods in programming developed in this work is motivated by similar notions in other areas of engineering. This work is also motivated by current practical concerns in the area of software engineering. The inadequacy of current programming technology is universally recognized. Part of the solution to this problem will be to increase the level of automation in programming. I believe that the next major step in the evolution of more automated programming will be interactive systems which provide a mixture of partially automated program analysis, synthesis and verification. One such system being developed at MIT, called the programmer's apprentice, is the immediate intended application of this work. This report concentrates on the knowledge are of the programmer's apprentice, which is the form of a taxonomy of commonly used algorithms and data structures. To the extent that a programmer is able to construct and manipulate programs in terms of the forms in such a taxonomy, he may relieve himself of many details and generally raise the conceptual level of his interaction with the system, as compared with present day programming environments. Also, since it is practical to expand a great deal of effort pre-analyzing the entries in a library, the difficulty of verifying the correctness of programs constructed this way is correspondingly reduced. The feasibility of this approach is demonstrated by the design of an initial library of common techniques for manipulating symbolic data. This document also reports on the further development of a formalism called the plan calculus for specifying computations in a programming language independent manner. This formalism combines both data and control abstraction in a uniform framework that has facilities for representing multiple points of view and side effects.

Early library management systems in the UK

A 5-minute clip from a 1975 video on Bath University Library has been incorporated as slide 21 of this PowerPoint. The video was produced by the author to show library and information science students details of a library with a computer-based cataloguing and circulation system. To make sure that the video clip functions correctly within the presentation, please make sure that you have downloaded both the presentation (.ppt file) and the video (.wmv file) into the same directory. To activate the video move the cursor to the middle of Slide 21 (i.e. the screen that says Bath University) and then click the mouse.

Library management systems:current issues and developments

Tedd, L.A. (2006). Library management systems: current issues and developments. Umbrella 2007 conference of the Chartered Institute of Library and Information Professionals in Hatfield, July 2007.

Automated cross-modal mapping in robotic eye/hand systems using plastic radial basis function networks

Q. Meng and M. H Lee, Automated cross-modal mapping in robotic eye/hand systems using plastic radial basis function networks, Connection Science, 19(1), pp 25-52, 2007.

Automated Placement of Cameras in a Floorplan to Satisfy Task-Specific Constraints

In many multi-camera vision systems the effect of camera locations on the task-specific quality of service is ignored. Researchers in Computational Geometry have proposed elegant solutions for some sensor location problem classes. Unfortunately, these solutions utilize unrealistic assumptions about the cameras' capabilities that make these algorithms unsuitable for many real-world computer vision applications: unlimited field of view, infinite depth of field, and/or infinite servo precision and speed. In this paper, the general camera placement problem is first defined with assumptions that are more consistent with the capabilities of real-world cameras. The region to be observed by cameras may be volumetric, static or dynamic, and may include holes that are caused, for instance, by columns or furniture in a room that can occlude potential camera views. A subclass of this general problem can be formulated in terms of planar regions that are typical of building floorplans. Given a floorplan to be observed, the problem is then to efficiently compute a camera layout such that certain task-specific constraints are met. A solution to this problem is obtained via binary optimization over a discrete problem space. In preliminary experiments the performance of the resulting system is demonstrated with different real floorplans.

Automated Camera Layout to Satisfy Task-Specific and Floorplan-Specific Coverage Requirements

In many multi-camera vision systems the effect of camera locations on the task-specific quality of service is ignored. Researchers in Computational Geometry have proposed elegant solutions for some sensor location problem classes. Unfortunately, these solutions utilize unrealistic assumptions about the cameras' capabilities that make these algorithms unsuitable for many real-world computer vision applications: unlimited field of view, infinite depth of field, and/or infinite servo precision and speed. In this paper, the general camera placement problem is first defined with assumptions that are more consistent with the capabilities of real-world cameras. The region to be observed by cameras may be volumetric, static or dynamic, and may include holes that are caused, for instance, by columns or furniture in a room that can occlude potential camera views. A subclass of this general problem can be formulated in terms of planar regions that are typical of building floorplans. Given a floorplan to be observed, the problem is then to efficiently compute a camera layout such that certain task-specific constraints are met. A solution to this problem is obtained via binary optimization over a discrete problem space. In experiments the performance of the resulting system is demonstrated with different real floorplans.

Mammalian cell cultures as a model system for the determination of cytotoxicity induced by cholesterol oxidation products

Oxysterols are products of cholesterol oxidation, which may be produced endogenously or may be absorbed from the diet where they are commonly found in foods of animal origin. Oxysterols are known to be cyctotoxic to cells in culture and mode of toxicity has been identified as apoptosis in certain cell lines. The cytotoxicity of the oxysterols 25-hydroxycholesterol (25-OH) and 7β-hydroxycholesterol (7β-OH) was examined in two human cell lines, HepG2, a hepatoma cell line, and U937, a monocytic cell line. Both 25-OH and 7β-OH were cytotoxic to the HepG2 cell line but apoptotic cells were not detected and it was concluded that cells underwent necrosis. 25-OH was not cytotoxic to the U937 cell line but it was found to have a cytostatic effect. 7β-OH was shown to induce apoptosis in the U937 line. The mechanism of oxysterol-induced apoptosis has not yet been fully elucidated, however the generation of an oxidative stress and the depletion of glutathione have been associated with the initial stages of the apoptotic process. The concentration of cellular antioxidant enzyme, superoxide dismutase (SOD) was increased in association with 7β-OH induced apoptosis in the U937 cell line. There was no change in the glutathione concentration or the SOD activity of HepG2 cells, which underwent necrosis in the presence of 7β-OH. Many apoptotic pathways center on the activation of caspase-3, which is the key executioner protease of apoptosis. Caspase-3 activity was also shown to increase in association with 7β-OH-induced apoptosis in U937 cells but there was no significant increase in caspase-3 activity in HepG2 cells. DNA fragmentation is regarded as the biochemical hallmark of apoptosis, therefore the comet assay as a measure of DNA fragmentation was assessed as a measure of apoptosis. The level of DNA fragmentation induced by 7β-OH, as measured using the comet assay, was similar for both cell lines. Therefore, it was concluded that the comet assay could not be used to distinguish between 7β-OH-induced apoptosis in U937 cells and 7β-OH-induced necrosis in HepG2 cells. The cytotoxicity and apoptotic potency of oxysterols 25-OH, 7β-OH, cholesterol- 5a,6a-epoxide (a-epoxide), cholesterol-5β,6β-epoxide (β-epoxide), 19-hydroxy-cholesterol (19-OH), and 7-ketocholesterol (7-keto) was compared in the U937 cell line. 7 β-OH, β-epoxide and 7-keto were found to induce apoptosis in U937 cells. 7β-OH-induced apoptosis was associated with a decrease in the cellular glutathione concentration and an increase in SOD activity, 7-keto and β-epoxide did not affect the glutathione concentration or the SOD activity of the cells.a-Epoxide, 19-OH and 25-OH were not cytotoxic to the U937 cell line.

The development of a computer model to optimize the performance of a 50kWp photovoltaic system

A computer model has been developed to optimize the performance of a 50kWp photovoltaic system which supplies electrical energy to a dairy farm at Fota Island in Cork Harbour. Optimization of the system involves maximising the efficiency and increasing the performance and reliability of each hardware unit. The model accepts horizontal insolation, ambient temperature, wind speed, wind direction and load demand as inputs. An optimization program uses the computer model to simulate the optimum operating conditions. From this analysis, criteria are established which are used to improve the photovoltaic system operation. This thesis describes the model concepts, the model implementation and the model verification procedures used during development. It also describes the techniques which are used during system optimization. The software, which is written in FORTRAN, is structured in modular units to provide logical and efficient programming. These modular units may also be used in the modelling and optimization of other photovoltaic systems.

An entirely cell-based system to generate single-chain antibodies against cell surface receptors.

The generation of recombinant antibodies (Abs) using phage display is a proven method to obtain a large variety of Abs that bind with high affinity to a given antigen. Traditionally, the generation of single-chain Abs depends on the use of recombinant proteins in several stages of the procedure. This can be a problem, especially in the case of cell-surface receptors, because Abs generated and selected against recombinant proteins may not bind the same protein expressed on a cell surface in its native form and because the expression of some receptors as recombinant proteins is problematic. To overcome these difficulties, we developed a strategy to generate single-chain Abs that does not require the use of recombinant protein at any stage of the procedure. In this strategy, stably transfected cells are used for the immunization of mice, measuring Ab responses to immunization, panning the phage library, high-throughput screening of arrayed phage clones, and characterization of recombinant single-chain variable regions. This strategy was used to generate a panel of single-chain Abs specific for the innate immunity receptor Toll-like receptor 2. Once generated, individual single-chain variable regions were subcloned into an expression vector allowing the production of recombinant Abs in insect cells, thus avoiding the contamination of recombinant Abs with microbial products. This cell-based system efficiently generates Abs that bind to native molecules on the cell surface, bypasses the requirement of recombinant protein production, and avoids risks of microbial component contamination.

Real-Time and Data-Driven Operation Optimization and Knowledge Discovery for an Enterprise Information System

An enterprise information system (EIS) is an integrated data-applications platform characterized by diverse, heterogeneous, and distributed data sources. For many enterprises, a number of business processes still depend heavily on static rule-based methods and extensive human expertise. Enterprises are faced with the need for optimizing operation scheduling, improving resource utilization, discovering useful knowledge, and making data-driven decisions.

This thesis research is focused on real-time optimization and knowledge discovery that addresses workflow optimization, resource allocation, as well as data-driven predictions of process-execution times, order fulfillment, and enterprise service-level performance. In contrast to prior work on data analytics techniques for enterprise performance optimization, the emphasis here is on realizing scalable and real-time enterprise intelligence based on a combination of heterogeneous system simulation, combinatorial optimization, machine-learning algorithms, and statistical methods.

On-demand digital-print service is a representative enterprise requiring a powerful EIS.We use real-life data from Reischling Press, Inc. (RPI), a digit-print-service provider (PSP), to evaluate our optimization algorithms.

In order to handle the increase in volume and diversity of demands, we first present a high-performance, scalable, and real-time production scheduling algorithm for production automation based on an incremental genetic algorithm (IGA). The objective of this algorithm is to optimize the order dispatching sequence and balance resource utilization. Compared to prior work, this solution is scalable for a high volume of orders and it provides fast scheduling solutions for orders that require complex fulfillment procedures. Experimental results highlight its potential benefit in reducing production inefficiencies and enhancing the productivity of an enterprise.

We next discuss analysis and prediction of different attributes involved in hierarchical components of an enterprise. We start from a study of the fundamental processes related to real-time prediction. Our process-execution time and process status prediction models integrate statistical methods with machine-learning algorithms. In addition to improved prediction accuracy compared to stand-alone machine-learning algorithms, it also performs a probabilistic estimation of the predicted status. An order generally consists of multiple series and parallel processes. We next introduce an order-fulfillment prediction model that combines advantages of multiple classification models by incorporating flexible decision-integration mechanisms. Experimental results show that adopting due dates recommended by the model can significantly reduce enterprise late-delivery ratio. Finally, we investigate service-level attributes that reflect the overall performance of an enterprise. We analyze and decompose time-series data into different components according to their hierarchical periodic nature, perform correlation analysis,

and develop univariate prediction models for each component as well as multivariate models for correlated components. Predictions for the original time series are aggregated from the predictions of its components. In addition to a significant increase in mid-term prediction accuracy, this distributed modeling strategy also improves short-term time-series prediction accuracy.

In summary, this thesis research has led to a set of characterization, optimization, and prediction tools for an EIS to derive insightful knowledge from data and use them as guidance for production management. It is expected to provide solutions for enterprises to increase reconfigurability, accomplish more automated procedures, and obtain data-driven recommendations or effective decisions.