New Shuttle Vector-Based Expression System To Generate Polyhistidine-Tagged Fusion Proteins in Staphylococcus aureus and Escherichia coli.

Four Staphylococcus aureus-Escherichia coli shuttle vectors were constructed for gene expression and production of tagged fusion proteins. Vectors pBUS1-HC and pTSSCm have no promoter upstream of the multiple cloning site (MCS), and this allows study of genes under the control of their native promoters, and pBUS1-Pcap-HC and pTSSCm-Pcap contain the strong constitutive promoter of S. aureus type 1 capsule gene 1A (Pcap) upstream of a novel MCS harboring codons for the peptide tag Arg-Gly-Ser-hexa-His (rgs-his6). All plasmids contained the backbone derived from pBUS1, including the E. coli origin ColE1, five copies of terminator rrnB T1, and tetracycline resistance marker tet(L) for S. aureus and E. coli. The minimum pAMα1 replicon from pBUS1 was improved through either complementation with the single-strand origin oriL from pUB110 (pBUS1-HC and pBUS1-Pcap-HC) or substitution with a pT181-family replicon (pTSSCm and pTSSCm-Pcap). The new constructs displayed increased plasmid yield and segregational stability in S. aureus. Furthermore, pBUS1-Pcap-HC and pTSSCm-Pcap offer the potential to generate C-terminal RGS-His6 translational fusions of cloned genes using simple molecular manipulation. BcgI-induced DNA excision followed by religation converts the TGA stop codon of the MCS into a TGC codon and links the rgs-his6 codons to the 3' end of the target gene. The generation of the rgs-his6 codon-fusion, gene expression, and protein purification were demonstrated in both S. aureus and E. coli using the macrolide-lincosamide-streptogramin B resistance gene erm(44) inserted downstream of Pcap. The new His tag expression system represents a helpful tool for the direct analysis of target gene function in staphylococcal cells.

An ultrasensitive NanoLuc-based luminescence system for monitoring Plasmodium berghei throughout its life cycle

BACKGROUND: Bioluminescence imaging is widely used for cell-based assays and animal imaging studies, both in biomedical research and drug development. Its main advantages include its high-throughput applicability, affordability, high sensitivity, operational simplicity, and quantitative outputs. In malaria research, bioluminescence has been used for drug discovery in vivo and in vitro, exploring host-pathogen interactions, and studying multiple aspects of Plasmodium biology. While the number of fluorescent proteins available for imaging has undergone a great expansion over the last two decades, enabling simultaneous visualization of multiple molecular and cellular events, expansion of available luciferases has lagged behind. The most widely used bioluminescent probe in malaria research is the Photinus pyralis firefly luciferase, followed by the more recently introduced Click-beetle and Renilla luciferases. Ultra-sensitive imaging of Plasmodium at low parasite densities has not been previously achieved. With the purpose of overcoming these challenges, a Plasmodium berghei line expressing the novel ultra-bright luciferase enzyme NanoLuc, called PbNLuc has been generated, and is presented in this work. RESULTS: NanoLuc shows at least 150 times brighter signal than firefly luciferase in vitro, allowing single parasite detection in mosquito, liver, and sexual and asexual blood stages. As a proof-of-concept, the PbNLuc parasites were used to image parasite development in the mosquito, liver and blood stages of infection, and to specifically explore parasite liver stage egress, and pre-patency period in vivo. CONCLUSIONS: PbNLuc is a suitable parasite line for sensitive imaging of the entire Plasmodium life cycle. Its sensitivity makes it a promising line to be used as a reference for drug candidate testing, as well as the characterization of mutant parasites to explore the function of parasite proteins, host-parasite interactions, and the better understanding of Plasmodium biology. Since the substrate requirements of NanoLuc are different from those of firefly luciferase, dual bioluminescence imaging for the simultaneous characterization of two lines, or two separate biological processes, is possible, as demonstrated in this work.

The Role of Forest Soils in a Northern New England Effluent Management System

The Carrabassett Valley Sanitary District in Carrabassett Valley, Maine has utilized both a forest spray irrigation system and a Snowfluent™ system for the treatment of their wastewater effluent. This study was designed to evaluate potential changes in soil properties after approximately 20 years of treatment in the forested spray irrigation site and three years of treatment in the field Snowfluent™ site. In addition, grass yield and composition were evaluated on the field study sites. After treatment with effluent or Snowfluent™, soils showed an increase in soil exchangeable Ca, Mg, Na, and K, base saturation, and pH. While most constituents were higher in treated soils, available P was lower in treated soils compared to the controls. This difference was attributed to higher rates of P mineralization from soil organic matter due to an irrigation effect of the treatment, depleting available P pools despite the P addition with the treatment. Most of the differences due to treatment were greatest at the surface and diminished with depth. Depth patterns in soil properties mostly reflected the decreasing influence of organic matter and its decomposition products with depth as evidenced by significantly higher total C in the surface compared to lower horizons. There were decreasing concentrations of total N, and exchangeable or extractable Ca, Mg, Na, K, Mn, Zn, and P with depth. In addition, there was decreasing BS with depth, driven primarily by declining exchangeable Ca and Mg. Imgation with Snowfluent™ altered the chemical composition of the grass on the site. All element concentrations were significantly higher in the grass foliage except for Ca. The differences were attributed to the additional nutrients and moisture derived from the Snowfluent™. The use of forest spray imgation and Snowfluent™ as a wastewater treatment strategy appears to work well. The soil and vegetation were able to retain most of the applied nutrients, and do not appear to be moving toward saturation. Vegetation management may be a key tool for managing nutrient accumulation on the grass sites as the system ages.

A survey of the implementation status of environmental management systems in U.S. colleges and universities, and, An environmental management system implementation model for U.S. colleges and universities

A census of 925 U.S. colleges and universities offering masters and doctorate degrees was conducted in order to study the number of elements of an environmental management system as defined by ISO 14001 possessed by small, medium and large institutions. A 30% response rate was received with 273 responses included in the final data analysis. Overall, the number of ISO 14001 elements implemented among the 273 institutions ranged from 0 to 16, with a median of 12. There was no significant association between the number of elements implemented among institutions and the size of the institution (p = 0.18; Kruskal-Wallis test) or among USEPA regions (p = 0.12; Kruskal-Wallis test). The proportion of U.S. colleges and universities that reported having implemented a structured, comprehensive environmental management system, defined by answering yes to all 16 elements, was 10% (95% C.I. 6.6%–14.1%); however 38% (95% C.I. 32.0%–43.8%) reported that they had implemented a structured, comprehensive environmental management system, while 30.0% (95% C.I. 24.7%–35.9%) are planning to implement a comprehensive environmental management system within the next five years. Stratified analyses were performed by institution size, Carnegie Classification and job title. ^ The Osnabruck model, and another under development by the South Carolina Sustainable Universities Initiative, are the only two environmental management system models that have been proposed specifically for colleges and universities, although several guides are now available. The Environmental Management System Implementation Model for U.S. Colleges and Universities developed is an adaptation of the ISO 14001 standard and USEPA recommendations and has been tailored to U.S. colleges and universities for use in streamlining the implementation process. In using this implementation model created for the U.S. research and academic setting, it is hoped that these highly specialized institutions will be provided with a clearer and more cost-effective path towards the implementation of an EMS and greater compliance with local, state and federal environmental legislation. ^

Innovative self management system for guided cardiac rehabilitation

This paper describes the design and development of a system for cardio rehabilitation of patients that suffered a myocardial infarction. The proposed solution focuses on exercise prescriptions and the encouragement of healthy behaviors. The innovative strategy of the design takes into account health promotion models to provide safe, assistive exercise training sessions, personalized feedbacks, and educational contents.

Hybrid Approach Combining Machine Learning and a Rule-Based Expert System for Text Categorization

This paper discusses a novel hybrid approach for text categorization that combines a machine learning algorithm, which provides a base model trained with a labeled corpus, with a rule-based expert system, which is used to improve the results provided by the previous classifier, by filtering false positives and dealing with false negatives. The main advantage is that the system can be easily fine-tuned by adding specific rules for those noisy or conflicting categories that have not been successfully trained. We also describe an implementation based on k-Nearest Neighbor and a simple rule language to express lists of positive, negative and relevant (multiword) terms appearing in the input text. The system is evaluated in several scenarios, including the popular Reuters-21578 news corpus for comparison to other approaches, and categorization using IPTC metadata, EUROVOC thesaurus and others. Results show that this approach achieves a precision that is comparable to top ranked methods, with the added value that it does not require a demanding human expert workload to train

Is Europe ready to provide a pan-European Identity Management System?

European public administrations must manage citizens' digital identities, particularly considering interoperability among different countries. Owing to the diversity of electronic identity management (eIDM) systems, when users of one such system seek to communicate with governments using a different system, both systems must be linked and understand each other. To achieve this, the European Union is working on an interoperability framework. This article provides an overview of eIDM systems' current state at a pan-European level. It identifies and analyzes issues on which agreement exists, as well as those that aren't yet resolved and are preventing the adoption of a large-scale model.

Sustainable Air Traffic Management System Development Methodology

As it is defined in ATM 2000+ Strategy (Eurocontrol 2001), the mission of the Air Traffic Management (ATM) System is: “For all the phases of a flight, the ATM system should facilitate a safe, efficient, and expedite traffic flow, through the provision of adaptable ATM services that can be dimensioned in relation to the requirements of all the users and areas of the European air space. The ATM services should comply with the demand, be compatible, operate under uniform principles, respect the environment and satisfy the national security requirements.” The objective of this paper is to present a methodology designed to evaluate the status of the ATM system in terms of the relationship between the offered capacity and traffic demand, identifying weakness areas and proposing solutions. The first part of the methodology relates to the characterization and evaluation of the current system, while a second part proposes an approach to analyze the possible development limit. As part of the work, general criteria are established to define the framework in which the analysis and diagnostic methodology presented is placed. They are: the use of Air Traffic Control (ATC) sectors as analysis unit, the presence of network effects, the tactical focus, the relative character of the analysis, objectivity and a high level assessment that allows assumptions on the human and Communications, Navigation and Surveillance (CNS) elements, considered as the typical high density air traffic resources. The steps followed by the methodology start with the definition of indicators and metrics, like the nominal criticality or the nominal efficiency of a sector; scenario characterization where the necessary data is collected; network effects analysis to study the relations among the constitutive elements of the ATC system; diagnostic by means of the “System Status Diagram”; analytical study of the ATC system development limit; and finally, formulation of conclusions and proposal for improvement. This methodology was employed by Aena (Spanish Airports Manager and Air Navigation Service Provider) and INECO (Spanish Transport Engineering Company) in the analysis of the Spanish ATM System in the frame of the Spanish airspace capacity sustainability program, although it could be applied elsewhere.

A Multingual Web-based Educational System for Professional Musicians

This paper presents the main results of the eContent HARMOS project. The project has developed a webbased educational system for professional musicians. The main idea of the project consists of recording master classes taught by highly recognised maestros and annotate this multimedia material using an educational musical taxonomy and automatic annotation tools. Users of the system access a multi-criteria search engine that allows them to find and play video segments according to a combination of criteria, which include instrument, teacher, composer, composition, movement and pedagogical concept. In order to preserve teachers and students rights, a DRM and protection system has been developed. The system is being publicly exploited. This model preserves musical heritage, since these valuable master classes are usually not recorded and it also provides a sustainable model for musical institutions.

An architecture for a heterogeneous private IaaS management system

Cloud computing and, more particularly, private IaaS, is seen as a mature technology with a myriad solutions tochoose from. However, this disparity of solutions and products has instilled in potential adopters the fear of vendor and data lock-in. Several competing and incompatible interfaces and management styles have given even more voice to these fears. On top of this, cloud users might want to work with several solutions at the same time, an integration that is difficult to achieve in practice. In this paper, we propose a management architecture that tries to tackle these problems; it offers a common way of managing several cloud solutions, and an interface that can be tailored to the needs of the user. This management architecture is designed in a modular way, and using a generic information model. We have validated our approach through the implementation of the components needed for this architecture to support a sample private IaaS solution: OpenStack

Development of a management system for virtual machines on private clouds

Cloud computing and, more particularly, private IaaS, is seen as a mature technol- ogy with a myriad solutions to choose from. However, this disparity of solutions and products has instilled in potential adopters the fear of vendor and data lock- in. Several competing and incompatible interfaces and management styles have increased even more these fears. On top of this, cloud users might want to work with several solutions at the same time, an integration that is difficult to achieve in practice. In this Master Thesis I propose a management architecture that tries to solve these problems; it provides a generalized control mechanism for several cloud infrastructures, and an interface that can meet the requirements of the users. This management architecture is designed in a modular way, and using a generic infor- mation model. I have validated the approach through the implementation of the components needed for this architecture to support a sample private IaaS solution: OpenStack.

A proposal for an Internet of Things-based monitoring system composed by low capability, open source and open hardware devices

The Internet of Things makes use of a huge disparity of technologies at very different levels that help one to the other to accomplish goals that were previously regarded as unthinkable in terms of ubiquity or scalability. If the Internet of Things is expected to interconnect every day devices or appliances and enable communications between them, a broad range of new services, applications and products can be foreseen. For example, monitoring is a process where sensors have widespread use for measuring environmental parameters (temperature, light, chemical agents, etc.) but obtaining readings at the exact physical point they want to be obtained from, or about the exact wanted parameter can be a clumsy, time-consuming task that is not easily adaptable to new requirements. In order to tackle this challenge, a proposal on a system used to monitor any conceivable environment, which additionally is able to monitor the status of its own components and heal some of the most usual issues of a Wireless Sensor Network, is presented here in detail, covering all the layers that give it shape in terms of devices, communications or services.

Resource allocation in GMD and SVD-based MIMO System

Singular-value decomposition (SVD)-based multiple-input multiple output (MIMO) systems, where the whole MIMO channel is decomposed into a number of unequally weighted single-input single-output (SISO) channels, have attracted a lot of attention in the wireless community. The unequal weighting of the SISO channels has led to intensive research on bit- and power allocation even in MIMO channel situation with poor scattering conditions identified as the antennas correlation effect. In this situation, the unequal weighting of the SISO channels becomes even much stronger. In comparison to the SVD-assisted MIMO transmission, geometric mean decomposition (GMD)-based MIMO systems are able to compensate the drawback of weighted SISO channels when using SVD, where the decomposition result is nearly independent of the antennas correlation effect. The remaining interferences after the GMD-based signal processing can be easily removed by using dirty paper precoding as demonstrated in this work. Our results show that GMD-based MIMO transmission has the potential to significantly simplify the bit and power loading processes and outperforms the SVD-based MIMO transmission as long as the same QAM-constellation size is used on all equally-weighted SISO channels.

RESSIST: a research object-based recommender system

The scientific method is a methodological approach to the process of inquiry { in which empirically grounded theory of nature is constructed and verified [14]. It is a hard, exhaustive and dedicated multi-stage procedure that a researcher must perform to achieve valuable knowledge. Trying to help researchers during this process, a recommender system, intended as a researcher assistant, is designed to provide them useful tools and information for each stage of the procedure. A new similarity measure between research objects and a representational model, based on domain spaces, to handle them in dif ferent levels are created as well as a system to build them from OAI-PMH (and RSS) resources. It tries to represents a sound balance between scientific insight into individual scientific creative processes and technical implementation using innovative technologies in information extraction, document summarization and semantic analysis at a large scale.