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.

Cost analysis of opt-in consent procedures for the Texas statewide immunization information system, ImmTrac

Background. Childhood immunization programs have dramatically reduced the morbidity and mortality associated with vaccine-preventable diseases. Proper documentation of immunizations that have been administered is essential to prevent duplicate immunization of children. To help improve documentation, immunization information systems (IISs) have been developed. IISs are comprehensive repositories of immunization information for children residing within a geographic region. The two models for participation in an IIS are voluntary inclusion, or "opt-in," and voluntary exclusion, or "opt-out." In an opt-in system, consent must be obtained for each participant, conversely, in an opt-out IIS, all children are included unless procedures to exclude the child are completed. Consent requirements for participation vary by state; the Texas IIS, ImmTrac, is an opt-in system.^ Objectives. The specific objectives are to: (1) Evaluate the variance among the time and costs associated with collecting ImmTrac consent at public and private birthing hospitals in the Greater Houston area; (2) Estimate the total costs associated with collecting ImmTrac consent at selected public and private birthing hospitals in the Greater Houston area; (3) Describe the alternative opt-out process for collecting ImmTrac consent at birth and discuss the associated cost savings relative to an opt-in system.^ Methods. Existing time-motion studies (n=281) conducted between October, 2006 and August, 2007 at 8 birthing hospitals in the Greater Houston area were used to assess the time and costs associated with obtaining ImmTrac consent at birth. All data analyzed are deidentified and contain no personal information. Variations in time and costs at each location were assessed and total costs per child and costs per year were estimated. The cost of an alternative opt-out system was also calculated.^ Results. The median time required by birth registrars to complete consent procedures varied from 72-285 seconds per child. The annual costs associated with obtaining consent for 388,285 newborns in ImmTrac's opt-in consent process were estimated at $702,000. The corresponding costs of the proposed opt-out system were estimated to total $194,000 per year. ^ Conclusions. Substantial variation in the time and costs associated with completion of ImmTrac consent procedures were observed. Changing to an opt-out system for participation could represent significant cost savings. ^

Characterisation factors for life cycle impact assessment of sound emissions

abstract to be added by authors

Effects of ocean acidification on different life-cycle stages of the kelp Laminaria hyperborea (Phaeophyceae), link to supplementary data

Our objective for this study was to evaluate the influence of preindustrial and expected future atmospheric CO2 concentrations (280 µatm and 700 µatm pCO2, respectively) on different life-cycle stages of the kelp Laminaria hyperborea from Helgoland (Germany, North Sea). Zoospore germination, gametogenesis, vegetative growth, sorus formation and photosynthetic performance of vegetative and fertile tissue were examined. The contribution of external carbonic anhydrase (exCA) to C-supply for net-photosynthesis (net-PS) and the Chla- and phlorotannin content were investigated. Female gametogenesis and vegetative growth of sporophytes were significantly enhanced under the expected future pCO2. rETR(max) and net-PS of young vegetative sporophytes tended to increase performance at higher pCO2. The trend towards elevated net-PS vanished after inhibition of exCA. In vegetative sporophytes, phlorotannin content and Chla content were not significantly affected by pCO2.

Ocean Acidification Affects Redox-Balance and Ion-Homeostasis in the Life-Cycle Stages of Emiliania huxleyi

Ocean Acidification (OA) has been shown to affect photosynthesis and calcification in the coccolithophore Emiliania huxleyi, a cosmopolitan calcifier that significantly contributes to the regulation of the biological carbon pumps. Its non-calcifying, haploid life-cycle stage was found to be relatively unaffected by OA with respect to biomass production. Deeper insights into physiological key processes and their dependence on environmental factors are lacking, but are required to understand and possibly estimate the dynamics of carbon cycling in present and future oceans. Therefore, calcifying diploid and non-calcifying haploid cells were acclimated to present and future CO2 partial pressures (pCO2; 38.5 Pa vs. 101.3 Pa CO2) under low and high light (50 vs. 300 µmol photons/m**2 /s). Comparative microarray-based transcriptome profiling was used to screen for the underlying cellular processes and allowed to follow up interpretations derived from physiological data. In the diplont, the observed increases in biomass production under OA are likely caused by stimulated production of glycoconjugates and lipids. The observed lowered calcification under OA can be attributed to impaired signal-transduction and ion-transport. The haplont utilizes distinct genes and metabolic pathways, reflecting the stage-specific usage of certain portions of the genome. With respect to functionality and energy-dependence, however, the transcriptomic OA-responses resemble those of the diplont. In both life-cycle stages, OA affects the cellular redox-state as a master regulator and thereby causes a metabolic shift from oxidative towards reductive pathways, which involves a reconstellation of carbon flux networks within and across compartments. Whereas signal transduction and ion-homeostasis appear equally OA-sensitive under both light intensities, the effects on carbon metabolism and light physiology are clearly modulated by light availability. These interactive effects can be attributed to the influence of OA and light on the redox equilibria of NAD and NADP, which function as major sensors for energization and stress. This generic mode of action of OA may therefore provoke similar cell-physiological responses in other protists.

A simplified energy consumption model for fiber-based Next Generation Access Networks

ICTs account nowadays for 2% of total carbon emissions. However, in a time when strict measures to reduce energyconsumption in all the industrial and services sectors are required, the ICT sector faces an increase in services and bandwidth demand. The deployment of NextGenerationNetworks (NGN) will be the answer to this new demand and specifically, the NextGenerationAccessNetworks (NGANs) will provide higher bandwidth access to users. Several policy and cost analysis are being carried out to understand the risks and opportunities of new deployments, though the question of which is the role of energyconsumption in NGANs seems off the table. Thus, this paper proposes amodel to analyze the energyconsumption of the main fiber-based NGAN architectures, i.e. Fiber To The House (FTTH) in both Passive Optical Network (PON) and Point-to-Point (PtP) variations, and FTTx/VDSL. The aim of this analysis is to provide deeper insight on the impact of new deployments on the energyconsumption of the ICT sector and the effects of energyconsumption on the life-cycle cost of NGANs. The paper presents also an energyconsumption comparison of the presented architectures, particularized in the specific geographic and demographic distribution of users of Spain, but easily extendable to other countries.

The Road from Community Ideas to Organisational Innovation: A Life Cycle Survey of Idea Management Systems

This paper introduces a new emerging software component, the idea management system, which helps to gather, organise, select and manage the innovative ideas provided by the communities gathered around organisations or enterprises. We define the notion of the idea life cycle, which provides a framework for characterising tools and techniques that drive the evolution of community submitted data inside idea management systems. Furthermore, we show the dependencies between the community-created information and the enterprise processes that are a result of using idea management systems and point out the possible benefits.

New Constructive Solutions to Improve Energy Efficiency in Existing Buildings and their Economic Viability

The improvement of energy efficiency in existing buildings is always a challenge due to their particular, and sometimes protected, constructive solutions. New constructive regulations in Spain leave a big undefined gap when a restoration is considered because they were developed for new buildings. However, rehabilitation is considered as an opportunity for many properties because it allows owners to obtain benefits from the use of the buildings. The current financial and housing crisis has turned society point of view to existing buildings and making them more efficient is one of the Spanish government’s aims. The economic viability of a rehabilitation action should take all factors into account: both construction costs and the future operative costs of the building must be considered. Nevertheless, the application of these regulations in Spain is left to the designer’s opinion and always under a subjective point of view. With the research work described in this paper and with the help of some case-studies, the cost of adapting an existing building to the new constructive regulations will be studied and Energetic Efficiency will be evaluated depending on how the investment is recovered. The interest of the research is based on showing how new constructive solutions can achieve higher levels of efficiency in terms of energy, construction and economy and it will demonstrate that Life Cycle Costing analysis can be a mechanism to find the advantages and disadvantages of using these new constructive solutions. Therefore, this paper has the following objectives: analysing constructive solutions in existing buildings - to establish a process for assessing total life cycle costs (LCC) during the planning stages with consideration of future operating costs - to select the most advantageous operating system – To determine the return on investment in terms of construction costs based on new techniques, the achieved energy savings and investment payback periods.

Reverse logistics systems for waste generated throughout vehicles life-cycle

Waste produced during the service life of automobiles has received much less attention than end-of-life vehicles themselves. In this paper, we deal with the set up of a reverse logistics system for the collection and treatment of use-phase residues. First, the type of waste arising during vehicles? service life is characterized. Data were collected in collaboration with SIGRAUTO, the product stewardship organization in charge of vehicles? recovery in Spain. Next, three organizational models are proposed. The three alternatives are benchmarked and assessed from a double organizational and operational perspective for the particular case of the Madrid region in Spain

Termination and cost analysis: Complexity and precision issues

The research in this thesis is related to static cost and termination analysis. Cost analysis aims at estimating the amount of resources that a given program consumes during the execution, and termination analysis aims at proving that the execution of a given program will eventually terminate. These analyses are strongly related, indeed cost analysis techniques heavily rely on techniques developed for termination analysis. Precision, scalability, and applicability are essential in static analysis in general. Precision is related to the quality of the inferred results, scalability to the size of programs that can be analyzed, and applicability to the class of programs that can be handled by the analysis (independently from precision and scalability issues). This thesis addresses these aspects in the context of cost and termination analysis, from both practical and theoretical perspectives. For cost analysis, we concentrate on the problem of solving cost relations (a form of recurrence relations) into closed-form upper and lower bounds, which is the heart of most modern cost analyzers, and also where most of the precision and applicability limitations can be found. We develop tools, and their underlying theoretical foundations, for solving cost relations that overcome the limitations of existing approaches, and demonstrate superiority in both precision and applicability. A unique feature of our techniques is the ability to smoothly handle both lower and upper bounds, by reversing the corresponding notions in the underlying theory. For termination analysis, we study the hardness of the problem of deciding termination for a speci�c form of simple loops that arise in the context of cost analysis. This study gives a better understanding of the (theoretical) limits of scalability and applicability for both termination and cost analysis.

Quantity of CO2 Emissions in the Life Cycle of a Highway Infrastructure

There is strong evidence to indicate that carbon dioxide and other greenhouse gases are accumulating at unprecedented concentrations in out atmosphere contributing to global climate change. Evidence is equally strong that human activities, mainly the burning of fossil fuels, are driving force in this process (IPCC 2007). While different industries contribute varying amounts to total anthropogenic greenhouse gases, it is incumbent upon each to understand its contribution and search for sensible ways to reduce overall greenhouse gas production. The aim of this paper is the development of a methodology to determine the amount of CO2 emissions of a highway, allowing providing solutions that can improve the energy footprint and reduce its emissions

A Simplified Energy Consumption Model for Fibre-Based Next Generation Access Networks

Presentación realizada en el PhD Seminar del ITS 2011 en Budapest. ICTs (Information and Communication Technologies) currently account for 2% of total carbon emissions. However, although modern standards require strict measures to reduce energy consumption across all industrial and services sectors, the ICT sector also faces an increase in services and bandwidth demand. The deployment of Next Generation Networks (NGN) will be the answer to this new demand; more specifically, Next Generation Access Networks (NGANs) will provide higher bandwidth access to users. Several policy and cost analyses are being carried out to understand the risks and opportunities of new deployments, but the question of what role energy consumption plays in NGANs seems off the table. Thus, this paper proposes a model to analyse the energy consumption of the main fibre-based NGAN architectures: Fibre To The House (FTTH), in both Passive Optical Network (PON) and Point-to-Point (PtP) variations, and FTTx/VDSL. The aim of this analysis is to provide deeper insight on the impact of new deployments on the energy consumption of the ICT sector and the effects of energy consumption on the life-cycle cost of NGANs. The paper also presents an energy consumption comparison of the presented architectures, particularised to the specific geographic and demographic distribution of users of Spain but easily extendable to other countries.

Análisis de ciclo de vida de una nueva solución arquitectónica que mejora el rendimiento térmico de la envolvente del edificio: fachada natural aljibe. Life Cycle Assessment of a new architectural solution that improves the thermal performance of the building envelope: Natural Green Tank Façade

Análisis de ciclo de vida de una nueva solución arquitectónica que mejora el rendimiento térmico de la envolvente del edificio: fachada natural aljibe.

Quantity of CO2 Emissions in the Life Cycle of a Highway Infrastructure.

The complexity of climate change and its evolution during the last few years has a positive impact on new developments and approaches to reduce the emissions of CO2. Looking for a methodology to evaluate the sustainability of a roadway, a tool has been developed. Life Cycle Assessment (LCA) is being accepted by the road industry to measure and evaluate the environmental impacts of an infrastructure, as the energy consumption and carbon footprint. This paper describes the methodology to calculate the CO2 emissions associated with the energy embodied on a roadway along its life cycle, including construction, operations and demolition. It will assist to find solutions to improve the energy footprint and reduce the amount of CO2 emissions. Details are provided of both, the methodology and the data acquisition. This paper is an application of the methodology to the Spanish highways, using a local database. Two case studies and a practical example are studied to show the model as a decision support for sustainable construction in the road industry.