Analysis of perinatal gene expression: hormone response elements mediate activation of a lacZ reporter gene in liver of transgenic mice.

The transcription of genes encoding gluconeogenic enzymes is tightly regulated during the perinatal period. These genes are induced by glucagon (cAMP) and glucocorticoids and repressed by insulin. To address the role of cAMP and glucocorticoids in the physiological activation of genes encoding gluconeogenic enzymes in the perinatal period, transgenic mice have been generated with chimeric constructs containing the reporter gene lacZ under the control of hormone response elements. The activity of the transgene is restricted to the liver by the presence of the enhancers from the alpha-fetoprotein gene and its transcription is driven by a promoter that contains a TATA box linked to either cAMP response elements (CREs) or glucocorticoid response elements (GREs). We demonstrate cAMP and glucocorticoid regulation, liver-specific expression, and perinatal activation of the reporter gene. These data indicate that the CRE and GRE are, independently, necessary and sufficient to mediate perinatal gene activation. Perinatal activation was not impaired when a CRE reporter transgene was assayed in mice that contain a targeted mutation of the CRE-binding protein (CREB) gene, providing further evidence for functional redundancy among the members of the CREB/ATF gene family.

Reducing task’s redundancy – the strategy of Faculty of Pharmacy Library to centralize all scientific research

Introduction – Based on a previous project of University of Lisbon (UL) – a Bibliometric Benchmarking Analysis of University of Lisbon, for the period of 2000-2009 – a database was created to support research information (ULSR). However this system was not integrated with other existing systems at University, as the UL Libraries Integrated System (SIBUL) and the Repository of University of Lisbon (Repositório.UL). Since libraries were called to be part of the process, the Faculty of Pharmacy Library’ team felt that it was very important to get all systems connected or, at least, to use that data in the library systems. Objectives – The main goals were to centralize all the scientific research produced at Faculty of Pharmacy, made it available to the entire Faculty, involve researchers and library team, capitalize and reinforce team work with the integration of several distinct projects and reducing tasks’ redundancy. Methods – Our basis was the imported data collection from the ISI Web of Science (WoS), for the period of 2000-2009, into ULSR. All the researchers and indexed publications at WoS, were identified. A first validation to identify all the researchers and their affiliation (university, faculty, department and unit) was done. The final validation was done by each researcher. In a second round, concerning the same period, all Pharmacy Faculty researchers identified their published scientific work in other databases/resources (NOT WoS). To our strategy, it was important to get all the references and essential/critical to relate them with the correspondent digital objects. To each researcher previously identified, was requested to register all their references of the ‘NOT WoS’ published works, at ULSR. At the same time, they should submit all PDF files (for both WoS and NOT WoS works) in a personal area of the Web server. This effort enabled us to do a more reliable validation and prepare the data and metadata to be imported to Repository and to Library Catalogue. Results – 558 documents related with 122 researchers, were added into ULSR. 1378 bibliographic records (WoS + NOT WoS) were converted into UNIMARC and Dublin Core formats. All records were integrated in the catalogue and repository. Conclusions – Although different strategies could be adopted, according to each library team, we intend to share this experience and give some tips of what could be done and how Faculty of Pharmacy created and implemented her strategy.

Removing the redundancy from randomised gene libraries

Amino acid substitution plays a vital role in both the molecular engineering of proteins and analysis of structure-activity relationships. High-throughput substitution is achieved by codon randomisation, which generates a library of mutants (a randomised gene library) in a single experiment. For full randomisation, key codons are typically replaced with NNN (64 sequences) or NNG CorT (32 sequences). This obligates cloning of redundant codons alongside those required to encode the 20 amino acids. As the number of randomised codons increases, there is therefore a progressive loss of randomisation efficiency; the number of genes required per protein rises exponentially. The redundant codons cause amino acids to be represented unevenly; for example, methionine is encoded just once within NNN, whilst arginine is encoded six times. Finally, the organisation of the genetic code makes it impossible to encode functional subsets of amino acids (e.g. polar residues only) in a single experiment. Here, we present a novel solution to randomisation where genetic redundancy is eliminated; the number of different genes equals the number of encoded proteins, regardless of codon number. There is no inherent amino acid bias and any required subset of amino acids may be encoded in one experiment. This generic approach should be widely applicable in studies involving randomisation of proteins. © 2003 Elsevier Ltd. All rights reserved.

Minority enterprise in the clothing industry: an analysis of Asian jeans manufacturers in Birmingham

This thesis discusses and assesses the resources available to Asian entrepreneurs in the West Midlands' clothing industry and how they are used by these small businessmen in order to address opportunities in the market economy within the constraints imposed. The fashion industry is volatile and is dependent upon flexible firms which can respond quickly to shortrun production schedules. Small firms are best able to respond to this market environment. Production of jeans presents an interesting departure from the mainstream fashion industry. It is traditionally gared towards longrun production schedules where multinational enterprises have artificially diversified the market, promoting the 'right' brand name and have established control of the upper end of the market, whilst imports from Newly Developing Countries have catered for cheap copies at the lower end of the market. In recent years, a fashion element to jeans has emerged, thus opening a market gap for U.K. manufacturers to respond in the same way as for other fashion articles. A large immigrant population, previously serving the now declining factories and foundries of the West Midlands but, through redundancy, no longer a part of this employment sector, has ~5ponded to economic constraints and market opportunities by drawing on ethnic network resources for competitive access to labour, finance and contacts, to attack the emergent market gap. Two models of these Asian entrepreneurs are developed. One being somecne who has professionally and actively tackled the market gap and become established. These entrepreneurs are usually educated and have personal experience in business and were amongst the first to perceive opportunities to enter the industry, actively utilising their ethnicity as a resource upon which to draw for favorable access to cheap, flexible labour and capital. The second model is composed of later entrants to jeans manufacturing. They have less formal education and experience and have been pushed into self-employment by constraints of unemployment. Their ethnicity is passively used as a resource. They are more likely confined to the marginal activity of 'cut make and trim' and have little opportunity to increase profit margins, become estalished or expand.

Second life battery energy storage systems:converter topology and redundancy selection

Battery energy storage systems have traditionally been manufactured using new batteries with a good reliability. The high cost of such a system has led to investigations of using second life transportation batteries to provide an alternative energy storage capability. However, the reliability and performance of these batteries is unclear and multi-modular power electronics with redundancy have been suggested as a means of helping with this issue. This paper reviews work already undertaken on battery failure rate to suggest suitable figures for use in reliability calculations. The paper then uses reliability analysis and a numerical example to investigate six different multi-modular topologies and suggests how the number of series battery strings and power electronic module redundancy should be determined for the lowest hardware cost using a numerical example. The results reveal that the cascaded dc-side modular with single inverter is the lowest cost solution for a range of battery failure rates.

Petri Net Siphon Analysis and Network Centrality Measures for Identifying Combination Therapies in Signaling Pathways

Aberrant behavior of biological signaling pathways has been implicated in diseases such as cancers. Therapies have been developed to target proteins in these networks in the hope of curing the illness or bringing about remission. However, identifying targets for drug inhibition that exhibit good therapeutic index has proven to be challenging since signaling pathways have a large number of components and many interconnections such as feedback, crosstalk, and divergence. Unfortunately, some characteristics of these pathways such as redundancy, feedback, and drug resistance reduce the efficacy of single drug target therapy and necessitate the employment of more than one drug to target multiple nodes in the system. However, choosing multiple targets with high therapeutic index poses more challenges since the combinatorial search space could be huge. To cope with the complexity of these systems, computational tools such as ordinary differential equations have been used to successfully model some of these pathways. Regrettably, for building these models, experimentally-measured initial concentrations of the components and rates of reactions are needed which are difficult to obtain, and in very large networks, they may not be available at the moment. Fortunately, there exist other modeling tools, though not as powerful as ordinary differential equations, which do not need the rates and initial conditions to model signaling pathways. Petri net and graph theory are among these tools. In this thesis, we introduce a methodology based on Petri net siphon analysis and graph network centrality measures for identifying prospective targets for single and multiple drug therapies. In this methodology, first, potential targets are identified in the Petri net model of a signaling pathway using siphon analysis. Then, the graph-theoretic centrality measures are employed to prioritize the candidate targets. Also, an algorithm is developed to check whether the candidate targets are able to disable the intended outputs in the graph model of the system or not. We implement structural and dynamical models of ErbB1-Ras-MAPK pathways and use them to assess and evaluate this methodology. The identified drug-targets, single and multiple, correspond to clinically relevant drugs. Overall, the results suggest that this methodology, using siphons and centrality measures, shows promise in identifying and ranking drugs. Since this methodology only uses the structural information of the signaling pathways and does not need initial conditions and dynamical rates, it can be utilized in larger networks.

The mosaic of habitats of the Seine estuary: Insights from food-web modelling and network analysis

Ecological network analysis was applied in the Seine estuary ecosystem, northern France, integrating ecological data from the years 1996 to 2002. The Ecopath with Ecosim (EwE) approach was used to model the trophic flows in 6 spatial compartments leading to 6 distinct EwE models: the navigation channel and the two channel flanks in the estuary proper, and 3 marine habitats in the eastern Seine Bay. Each model included 12 consumer groups, 2 primary producers, and one detritus group. Ecological network analysis was performed, including a set of indices, keystoneness, and trophic spectrum analysis to describe the contribution of the 6 habitats to the Seine estuary ecosystem functioning. Results showed that the two habitats with a functioning most related to a stressed state were the northern and central navigation channels, where building works and constant maritime traffic are considered major anthropogenic stressors. The strong top-down control highlighted in the other 4 habitats was not present in the central channel, showing instead (i) a change in keystone roles in the ecosystem towards sediment-based, lower trophic levels, and (ii) a higher system omnivory. The southern channel evidenced the highest system activity (total system throughput), the higher trophic specialisation (low system omnivory), and the lowest indication of stress (low cycling and relative redundancy). Marine habitats showed higher fish biomass proportions and higher transfer efficiencies per trophic levels than the estuarine habitats, with a transition area between the two that presented intermediate ecosystem structure. The modelling of separate habitats permitted disclosing each one's response to the different pressures, based on their a priori knowledge. Network indices, although non-monotonously, responded to these differences and seem a promising operational tool to define the ecological status of transitional water ecosystems.

Genome-scale analysis of the non-cultivable Treponema pallidum reveals extensive within-patient genetic variation

Insights into the genomic adaptive traits of Treponema pallidum, the causative bacterium of syphilis, have long been hampered due to the absence of in vitro culture models and the constraints associated with its propagation in rabbits. Here, we have bypassed the culture bottleneck by means of a targeted strategy never applied to uncultivable bacterial human pathogens to directly capture whole-genome T. pallidum data in the context of human infection. This strategy has unveiled a scenario of discreet T. pallidum interstrain single-nucleotide-polymorphism-based microevolution, contrasting with a rampant within-patient genetic heterogeneity mainly targeting multiple phase-variable loci and a major antigen-coding gene (tprK). TprK demonstrated remarkable variability and redundancy, intra- and interpatient, suggesting ongoing parallel adaptive diversification during human infection. Some bacterial functions (for example, flagella- and chemotaxis-associated) were systematically targeted by both inter- and intrastrain single nucleotide polymorphisms, as well as by ongoing within-patient phase variation events. Finally, patient-derived genomes possess mutations targeting a penicillin-binding protein coding gene (mrcA) that had never been reported, unveiling it as a candidate target to investigate the impact on the susceptibility to penicillin. Our findings decode the major genetic mechanisms by which T. pallidum promotes immune evasion and survival, and demonstrate the exceptional power of characterizing evolving pathogen subpopulations during human infection.

Raman microscopy of formamide-intercalated kaolinites treated by controlled-rate thermal analysis technology

Raman spectroscopy of formamide-intercalated kaolinites treated using controlled-rate thermal analysis technology (CRTA), allowing the separation of adsorbed formamide from intercalated formamide in formamide-intercalated kaolinites, is reported. The Raman spectra of the CRTA-treated formamide-intercalated kaolinites are significantly different from those of the intercalated kaolinites, which display a combination of both intercalated and adsorbed formamide. An intense band is observed at 3629 cm-1, attributed to the inner surface hydroxyls hydrogen bonded to the formamide. Broad bands are observed at 3600 and 3639 cm-1, assigned to the inner surface hydroxyls, which are hydrogen bonded to the adsorbed water molecules. The hydroxyl-stretching band of the inner hydroxyl is observed at 3621 cm-1 in the Raman spectra of the CRTA-treated formamide-intercalated kaolinites. The results of thermal analysis show that the amount of intercalated formamide between the kaolinite layers is independent of the presence of water. Significant differences are observed in the CO stretching region between the adsorbed and intercalated formamide.

Detailed analysis of a conservative difference approximation for the time fractional diffusion equation

Diffusion equations that use time fractional derivatives are attractive because they describe a wealth of problems involving non-Markovian Random walks. The time fractional diffusion equation (TFDE) is obtained from the standard diffusion equation by replacing the first-order time derivative with a fractional derivative of order α ∈ (0, 1). Developing numerical methods for solving fractional partial differential equations is a new research field and the theoretical analysis of the numerical methods associated with them is not fully developed. In this paper an explicit conservative difference approximation (ECDA) for TFDE is proposed. We give a detailed analysis for this ECDA and generate discrete models of random walk suitable for simulating random variables whose spatial probability density evolves in time according to this fractional diffusion equation. The stability and convergence of the ECDA for TFDE in a bounded domain are discussed. Finally, some numerical examples are presented to show the application of the present technique.

Determination of preventive maintenance lead time using hybrid analysis

The time for conducting Preventive Maintenance (PM) on an asset is often determined using a predefined alarm limit based on trends of a hazard function. In this paper, the authors propose using both hazard and reliability functions to improve the accuracy of the prediction particularly when the failure characteristic of the asset whole life is modelled using different failure distributions for the different stages of the life of the asset. The proposed method is validated using simulations and case studies.