Further field testing of the more heat-stable measles vaccines in Cameroon.

Two of the more heat-stable measles vaccines were field tested in Cameroon. Both maintained the minimum required infectivity titre and the ability to induce seroconversion after storage unreconstituted at 37 degrees C for 14 days. One of the vaccines, studied after reconstitution, maintained its ability to induce seroconversion after reconstitution and storage at 25 degrees C for 48 hours and at 37 degrees C for at least four hours. The increased heat stability of the studied vaccines will not eliminate the need for a well-monitored system of vaccine conservation and distribution but will ease the rigid cold-storage requirements of conventional measles vaccines.     

ASAP: automated sequence annotation pipeline for web-based updating of sequence information with a local dynamic database

The automated sequence annotation pipeline (ASAP) is designed to ease routine investigation of new functional annotations on unknown sequences, such as expressed sequence tags (ESTs), through querying of web-accessible resources and maintenance of a local database. The system allows easy use of the output from one search as the input for a new search, as well as the filtering of results. The database is used to store formats and parameters and information for parsing data from web sites. The database permits easy updating of format information should a site modify the format of a query or of a returned web page.     

Rapid RAPD screening of plant DNA using dot blot hybridization

Scoring of the results of RAPD analysis using gel electrophoresis imposes a constraint on throughput. To circumvent this barrier, dot-blot hybridization was substituted for electrophoresis. Arbitrarily amplified fragments from barley and wheat genomic DNA were labelled and used as probes for the identification of identical fragments in subsequent amplification reactions. None of the twelve fragments used as probes exhibited significant levels of croos-hybridization to other fragments amplified by the same arbitrary primer. The strength of the hybridization signal facilitates more accurate and more sensitive detection of diagnostic fragments than gel electrophoresis. In addition, the defined spatial orientation (microtitre dish format) of the ± results provide an excellent format for automated data collection. The use of dot blot hybridization to analyse PCR products well decrease the cost and time requirements of marker-assisted selection. This technique will also facilitate the rapid application of PCR-based maps.     

Abiotic Resource Depletion Different perceptions of the problem with mineral deposits

Goal, Scope and Background The goal of the present paper is to demonstrate how environmental product declarations (EPDs) are developed based on a set of product category rules (PCRs) in accordance with the requirements in the ISO 14025-standard. This is demonstrated by examples from the furniture industry in Norway, where several case models are evaluated. To ease the capability of developing EPDs in this industry, a database with specific environmental data for materials in furniture is developed. The database is used to produce the LCA for selected furniture models, and further, the database is the backbone of a data-assistance tool used to create the EPDs. Methods The LCA-data are produced based on traditional LCA-methodology. The PCR is based on a stakeholder analysis and the proposed methodology in the ISO 14025-standard. The EPDs developed so far, are results of close collaboration between companies and research centres in the Nordic countries. For the verification of the EPDs, auditing methodologies are used as a part of the audit of the companies' environmental management systems (EMS). Results and Conclusion Based on a hearing of a set of suggested PCRs, a consensus document for seating accommodation is developed. This is further the model for how to develop PCR-documents for all types of furniture, for example sleeping accommodations. Likewise, the database shall contain the most important data for the parts of a furniture model. Within the goal of the project period, EPDs will be developed for 80% of Norwegian furniture. The verification of the EPDs is done as a part of the certification procedures of EMS in accordance with the ISO 14001. Recommendation and Perspective The results presented in the paper are mainly for the pilot models in the project. However, the results will be further tested and the data-tool will be developed as a part of a product design tool where environmental requirements will be combined with quality requirements. The product design tool will be implemented in the furniture industry. Information on how to use EPDs in public purchasing will also be a part of future work.     

From a Bodily-based Format of Knowledge to Symbols. The Evolution of Human Language

Although ontogeny cannot recapitulate phylogeny, a two-level model of the acquisition of language will be here proposed and its implication for the evolution of the faculty of language will be discussed. It is here proposed that the identification of the cognitive requirements of language during ontogeny could help us in the task of identifying the phylogenetic achievements that concurred, at some point, to the acquisition of language during phylogeny. In this model speaking will be considered as a complex ability that arises in two different steps. The first step of competence widely relies on a bodily-based format of knowledge. The second step relies on more abstract meta-representations and implies high-level socio-cognitive skills. It is hypothesized that in order to reach the second level of language competence, symbolic communication and interaction with a cultural community are needed. Hence, the origins of species-specific human complex language and cognition are in both the brain and culture. Moreover, in this model, data from the embodied language research will be discussed in the light of a usage-based account of language.     

Mesofluidic devices for DNA-programmed combinatorial chemistry

Hybrid combinatorial chemistry strategies that use DNA as an information-carrying medium are proving to be powerful tools for molecular discovery. In order to extend these efforts, we present a highly parallel format for DNA-programmed chemical library synthesis. The new format uses a standard microwell plate footprint and is compatible with commercially available automation technology. It can accommodate a wide variety of combinatorial synthetic schemes with up to 384 different building blocks per chemical step. We demonstrate that fluidic routing of DNA populations in the highly parallel format occurs with excellent specificity, and that chemistry on DNA arrayed into 384 well plates proceeds robustly, two requirements for the high-fidelity translation and efficient in vitro evolution of small molecules.     

The Open AUC Project

Progress in analytical ultracentrifugation (AUC) has been hindered by obstructions to hardware innovation and by software incompatibility. In this paper, we announce and outline the Open AUC Project. The goals of the Open AUC Project are to stimulate AUC innovation by improving instrumentation, detectors, acquisition and analysis software, and collaborative tools. These improvements are needed for the next generation of AUC-based research. The Open AUC Project combines on-going work from several different groups. A new base instrument is described, one that is designed from the ground up to be an analytical ultracentrifuge. This machine offers an open architecture, hardware standards, and application programming interfaces for detector developers. All software will use the GNU Public License to assure that intellectual property is available in open source format. The Open AUC strategy facilitates collaborations, encourages sharing, and eliminates the chronic impediments that have plagued AUC innovation for the last 20 years. This ultracentrifuge will be equipped with multiple and interchangeable optical tracks so that state-of-the-art electronics and improved detectors will be available for a variety of optical systems. The instrument will be complemented by a new rotor, enhanced data acquisition and analysis software, as well as collaboration software. Described here are the instrument, the modular software components, and a standardized database that will encourage and ease integration of data analysis and interpretation software.     

Single framework recombinant antibody fragments designed for protein chip applications

High-throughput proteomics, based on the microarray platform, requires stable, highly functional components that will yield a highly sensitive read-out of low abundance proteins. Although antibodies are the best characterized binding molecules for this purpose, only a fraction of them appear to behave satisfactorily in the chip format. Therefore, high demands need to be placed on their molecular design. In the present study, we have focused on recombinant antibody design based on a single framework for protein chip applications, aiming at defining crucial molecular probe parameters. Our results show that engineered human recombinant scFv antibody fragments that displayed appropriate biophysical properties (molecular [functional] stability in particular) can be generated, making them prime candidates for high-density antibody arrays. In fact, a superior framework that displays both multifaceted adsorption properties and very high functional stability over several months on chips (stored in a dried-out state) was identified. Taken together, designed scFv fragments based on a single molecular scaffold, readily accessible in large phage display libraries, can undoubtedly meet the requirements of probe content in antibody microarrays, particularly for global proteome analysis.     

Lifting Industrial Ecology Modeling to a New Level of Quality and Transparency: A Call for More Transparent Publications and a Collaborative Open Source Software Framework

Industrial ecology (IE) is a maturing scientific discipline. The field is becoming more data and computation intensive, which requires IE researchers to develop scientific software to tackle novel research questions. We review the current state of software programming and use in our field and find challenges regarding transparency, reproducibility, reusability, and ease of collaboration. Our response to that problem is fourfold: First, we propose how existing general principles for the development of good scientific software could be implemented in IE and related fields. Second, we argue that collaborating on open source software could make IE research more productive and increase its quality, and we present guidelines for the development and distribution of such software. Third, we call for stricter requirements regarding general access to the source code used to produce research results and scientific claims published in the IE literature. Fourth, we describe a set of open source modules for standard IE modeling tasks that represent our first attempt at turning our recommendations into practice. We introduce a Python toolbox for IE that includes the life cycle assessment (LCA) framework Brightway2, the ecospold2matrix module that parses unallocated data in ecospold format, the pySUT and pymrio modules for building and analyzing multiregion input‐output models and supply and use tables, and the dynamic_stock_model class for dynamic stock modeling. Widespread use of open access software can, at the same time, increase quality, transparency, and reproducibility of IE research.     

Dual analyte detection using tandem flash luminescence

A heterogeneous, dual analyte-binding assay which makes use of the flash luminescence from both aequorin and an acridinium-9-carboxamide label is presented. The signal generating species were triggered both differentially and sequentially using Ca(2+) followed by basic peroxide. Both signals were resolved readily using a single photomultiplier tube without the need for multiwavelength detection. To demonstrate the tandem luminescence concept in a model assay system, dose-response curves for two analytes, biotinylated BSA and myoglobin, were generated using a competitive binding format. Because of the relatively short assay time and the well-resolved signals, this format will be useful in the development of dual analyte high-throughput assays.     

The pronghorn (Antilocapra americana): A review as a model for development of a “Species Management Plan” format,Part I

A format is suggested for a “Species Management Plan”; the model for this presentation is the pronghorn (Antilocapra americana). By using the Species Management Plan format, the captive maintenance of pronghorn is analyzed in relation to habitat and microhabitat needs, behavioral requirements, and nutritional needs. A strategy for captive maintenance of pronghorn in relatively wet climates is presented. It is suggested that development of a Species Management Plan is necessary before any new species is acquired for captive maintenance and reproduction.     

Structure-radical scavenging activity relationships of flavonoids

The objective of this work is to establish the structural requirements of flavonoids for appreciable radical-scavenging activity (RSA) and elucidate a comprehensive mechanism that can explain their activity. To this end, the RSA of 52 flavonoids against 2,2-diphenyl-1-picrylhydrazyl was determined. The relative change in energy (DeltaH(f)) associated with the formation of various flavonoidal and other phenolic radicals and also the spin distribution in these radicals were determined using computational programmes. By correlating experimental data with DeltaH(f), structural features that affect activity have been identified and considered in perspective. It was shown with compelling evidences that the RSA of flavonoids could be mapped to one of their ring systems, making it possible to study their RSA by dissecting their structures and designing representative simpler models. Consequently, hydroxytoluene units were demonstrated to successfully account for the RSA of flavonoids due to ring B and also to satisfactorily do so for activities due to ring A. Further, a comprehensive model for the radical scavenging reactions of flavonoids (and in general, phenolic compounds), which could account for hydrogen atom donation and the termination of aroxyl radicals, was proposed. Finally, prediction of structural features that could endow flavonoids with appreciable radical scavenging capability was made by considering the stability data and the ease of termination. In conclusion, the underlying molecular phenomena of the RSA of flavonoids could be explained by the ease of hydrogen atom abstraction and the ease of the termination of the flavonoidal aroxyl radicals.     

Release of bovine serum albumin from a hydrogel-cored biodegradable polymer fiber

We have developed a novel biodegradable, polymeric fiber construct that is coextruded using a wet-spinning process into a core-sheath format with a polysaccharide pre-hydrogel solution as the core fluid and poly(L-lactic acid) (PLLA) as the sheath. The biodegradable, biocompatible fibers were extruded from polymeric emulsions comprised of solutions of various molecular weights of PLLA dissolved in chloroform and containing dispersed, protein-free aqueous phases comprising up to 10% of the emulsion volume. Biologically sensitive agents can be loaded via a dispersed aqueous phase in the polymer, and/or directly into the polysaccharide. We show that this core-sheath fiber format will load a model protein that can be delivered for extended periods in vitro. Bovine serum albumin (BSA) was loaded into the fiber core as a model protein. We have shown that the greater the volume of the protein-free aqueous phase dispersed into the polymeric continuous-phase emulsion, the greater the total release of BSA encapsulated by a core gel comprised of 1% sodium alginate solution. We conclude this fiber format provides a promising vehicle for in vivo delivery of biological molecules. Its biocompatibility and biodegradability also allow for its use as a possible substrate for tissue engineering applications.     

Camera traps are an effective tool for monitoring insect–plant interactions

Insect and pollinator populations are vitally important to the health of ecosystems, food production, and economic stability, but are declining worldwide. New, cheap, and simple monitoring methods are necessary to inform management actions and should be available to researchers around the world. Here, we evaluate the efficacy of a commercially available, close‐focus automated camera trap to monitor insect–plant interactions and insect behavior. We compared two video settings—scheduled and motion‐activated—to a traditional human observation method. Our results show that camera traps with scheduled video settings detected more insects overall than humans, but relative performance varied by insect order. Scheduled cameras significantly outperformed motion‐activated cameras, detecting more insects of all orders and size classes. We conclude that scheduled camera traps are an effective and relatively inexpensive tool for monitoring interactions between plants and insects of all size classes, and their ease of accessibility and set‐up allows for the potential of widespread use. The digital format of video also offers the benefits of recording, sharing, and verifying observations.     

In vitro culture conditions to study keratinocyte differentiation using the HaCaT cell line

In vitro models to study the process of keratinocyte differentiation have been hindered by the stringent culture requirements and limitations imposed by the inherent properties of the cells. Primary keratinocytes only have a finite life span, while transformed cell lines exhibit many phenotypic features not found in normal cells. The spontaneously immortalized HaCaT cell line has been a widely employed keratinocyte model due to its ease of propagation and near normal phenotype, but protocols for differentiation and gene delivery into HaCaT cells vary widely in the literature. Here we report culture conditions for maintaining HaCaT cells in a basal-like state, for efficient differentiation of these cells, and for delivery of transgenes by transfection or adenoviral infection. This technological report will provide guidance to a large audience of scientists interested in investigating mechanisms of differentiation and skin morphogenesis.     

Production,purification and diagnostic application of filarial recombinant protein WbSXP-1 expressed in salt inducible <Emphasis Type="Italic">Escherichia coli</Emphasis>

Wuchereria bancrofti protein WbSXP-1 was identified and established as a potential candidate for the diagnosis of lymphatic filariasis. For the economic production of rWbSXP-1, osmotically (salt) inducible Escherichia coli GJ1158 was preferred. Cultivation and expression was optimized in 3 L airlift bioreactor (ALB) and was successfully extended to 30 L ALB. Purification of rWbSXP-1 his-tag protein was optimized in technical scale using FPLC and the maximal recovery of rWbSXP-1 with significant level of purity was achieved using the combination of IMAC and gel filtration. Quality criteria for immuno-reactivity of purified rWbSXP-1 were established for diagnostic applications. Enhancement of sensitivity in rapid diagnostic format was optimized to effectively detect weak to strong antibody reactivity in individuals exposed to lymphatic filariasis. Performance of the rapid format during field evaluation was successful. The accelerated stability assessment of the rapid format satisfied the requirements of WHO-cGMP norms. This investigation presents a successful technical scale production and purification of rWbSXP-1 considering the future industrial application and an enhanced rapid flow through antibody assay for the diagnosis of human lymphatic filariasis.     

Neutron, proton, and photonuclear cross-sections for radiation therapy and radiation protection

I review recent work at Los Alamos undertaken to evaluate neutron, proton, and photonuclear cross-sections up to 150 MeV (to 250 MeV for protons), based on experimental data and nuclear model calculations. These data are represented in the ENDF format and can be used in computer codes to simulate radiation transport. They permit calculations of absorbed dose in the body from therapy beams, and through use of kerma coefficients allow absorbed dose to be estimated for a given neutron energy distribution. In radiation protection, these data can be used to determine shielding requirements in accelerator environments and to calculate neutron, proton, gamma-ray, and radionuclide production. Illustrative comparisons of the evaluated cross-section and kerma coefficient data with measurements are given.