Multiplex SNP genotyping using Locked Nucleic Acid and microfluidics

Locked Nucleic Acid's or LNA are a new class of bicyclic DNA analogues that have a high affinity and specificity towards complementary nucleic acids. LNA containing oligonucleotides were used to develop a multiplex SNP genotyping assay based entirely on hybridization between capture probe and target. The approach incorporates a polymer microarray platform, photochemistry for immobilization of oligonucleotides onto microarrays, and a dedicated software tool to aid primer and capture probe design for highly multiplex genotyping. Furthermore, these technologies are combined in an integrated microfluidics platform for simple, highly multiplex and robust SNP genotyping.     

Slow Technology – Designing for Reflection

As computers are increasingly woven into the fabric of everyday life, interaction design may have to change – from creating only fast and efficient tools to be used during a limited time in specific situations, to creating technology that surrounds us and therefore is a part of our activities for long periods of time. We present slow technology: a design agenda for technology aimed at reflection and moments of mental rest rather than efficiency in performance. The aim of this paper is to develop a design philosophy for slow technology, to discuss general design principles and to revisit some basic issues in interaction design from a more philosophical point of view. We discuss examples of soniture and informative art as instances of slow technology and as examples of how the design principles can be applied in practice.     

Development of an optimized refolding process for recombinant Ala-Glu-IGF-1

Denatured and reduced N-terminal extended insulin-like growthfactor-1 (AE-IGF-1) was purified from Escherichia coli extractsand subjected to in vitro folding. The renaturation processwas shown to be a function of the redox potential of the solution.Folding by different methods had no significant effect on therenaturation. A maximal yield of 60% (w/w)was obtained. Thefolded AE-IGF-1 was enzymatically converted to IGF-1. The majorby-product (20% w/w) was identified as scrambled IGF-1. Enzymaticdigestion at alkaline and acidic pH suggested two possible disulphidebond arrangements: (i) Cys⁶–Cys⁴⁷, Cys¹⁸–Cys⁶¹,Cys⁴⁸–Cys⁵²; or (if) Cys⁶–Cys⁵², Cys^l8–Cys⁶¹,Cys⁴⁷ and Cys⁴⁸ being in their reduced forms. Energy minimizationand molecular modelling suggested that the scrambled IGF-1,having reduced cysteines at positions 47 and 48, was the energeticallymost stable conformation of the two.     

Phthalates,nonylphenols and LAS in an alternately operated wastewater treatment plant--fate modelling based on measured concentrations in wastewater and sludge

The performance of an alternately operated activated sludge wastewater treatment plant (WWTP) has been investigated with respect to six phthalates, nonylphenol (NP) and nonylphenol diethoxylate (NPDE) and linear alkylbenzene sulphonates (LAS). Samples of raw sewage, primary and secondary sludge and treated water were collected during an 8-day period in May 1999 and analysed for dissolved and sorbed substances. To evaluate the system performance with respect to substance removal through biodegradation and sorption to sludge the measured data were applied in a model describing the different bioreactors as one single reactor, corresponding to the concepts of, e.g. SimpleTreat. The most abundant of the investigated phthalates was di-(2-ethylhexyl)-phthalate (DEHP) with a measured mean inlet flow of 240g/day. Two percent of this amount was found in the treated water, 70% was biodegraded and 28% was found in the sludge. For LAS the mean inlet flow was 20,300g/day, of which less than 1% was found in the treated water, 84% was biodegraded and 15% was found in the sludge. The mean inlet flow of NP and NPDE was 44 and 590g/day, of which 4% and 2% was found in the treated water, 80% was biodegraded for both substances, and 16% and 18% was found in the sludge, respectively. The WWTP removal of the investigated substances was thus high compared to other studies of conventional activated sludge WWTPs. The simple model set-up presents a strong tool for predicting substance removal and system sensitivity related to changes in the inlet conditions, such as concentrations and flow. Furthermore, it allows the inclusion of complex alternately operated WWTPs in risk assessment tools such as e.g. SimpleTreat.     

Health-Related Evaluation of Building Products based on Climate Chamber Tests

A procedure is developed for assessing the health effects of the emissions from building products. The procedure is based on:

• ? emission rates measured in environment test chambers
• ? a fixed standard room with fixed standard conditions
• ? a maximum acceptable concentration in the indoor air of each of the chemical compounds emitted, I_v a maximum permissible contribution to I_v from building products.

The procedure has two elements: evaluation of the emitted compound alone and evaluation of the compound together with other compounds and indoor air factors. The evaluation includes odour and health effects of the compounds. The procedure has been applied to the emission test results for two building products: a rubber floor covering, and a water-borne acrylic watt paint. Maximum acceptable indoor air concentrations are listed fin-selected chemical compounds emitted by these two materials.     

Reconstruction of thickness and composition of stratified materials by means of 3D micro X-ray fluorescence spectroscopy

The recently developed 3D micro X-ray fluorescence spectroscopy (3D Micro-XRF) enables three-dimensional resolved, nondestructive investigation of elemental distribution in samples in the micrometer regime. Establishing a reliable quantification procedure is the precondition to render this spectroscopic method into a true analytical tool. One prominent field of application is the investigation of stratified material. A procedure for the quantitative reconstruction of the composition of stratified material by means of 3D Micro-XRF is proposed and validated. With the procedure, it is now possible to determine nondestructively the chemical composition and the thickness of layers. As no adequate stratified reference samples were available for validation, stratified reference material has been developed that is appropriate for 3D Micro-XRF or other depth-sensitive X-ray techniques.     

Improving the quality of phase maps in phase object digital holographic interferometry by finding the right reconstruction distance

Improved quality of phase maps in pulsed digital holographic interferometry is demonstrated by finding the right reconstruction distance. The objective is to improve the optical phase information when the object under study is a phase object and when it is out of focus, leading to low contrast fringes in the phase map. A numerical refocusing is performed by introducing an ideal lens as a multiplication by a phase field in the Fourier domain, and then a region of maximum speckle correlation is found by comparing undisturbed and disturbed subimages in different refocused imaging planes. After finding the right reconstruction distance, a phase map of high visibility is constructed. By this technique a 30% reduction of the phase error for a flow of helium gas and a 50% reduction of the phase error for a weak thin lens were obtained, which resulted in a significant improvement of the visual appearance of the phase maps.     

Recovery from groundwater extraction in a small catchment area with crystalline bedrock and thin soil cover in Sweden

An experiment has been in progress since 1997 in a small catchment area (28,000 m(2)) with crystalline bedrock and thin soil cover to study the conceivable impact on groundwater conditions of tunneling and the use of groundwater. The impact on hydrology and hydrochemistry from intensive extraction of groundwater at a depth of 50 m in the bedrock has been studied at Lake G?rdsj?n in Sweden. The catchment area was first monitored under pristine conditions, followed by four and a half years of extraction and then a recovery phase. The geological conditions result in a low buffer capacity and high sensitivity to acidification. During the period of extraction, the surface runoff decreased by approximately 50% compared to a nearby reference area. The groundwater extraction caused increased fluctuation in groundwater levels in a wetland, which in turn caused oxidation of reduced sulfur to sulfate. The sulfate concentrations increased almost 100-fold in some instances, causing a lowering of the pH by one unit in shallow groundwater. Since extraction of the groundwater was discontinued, the pH has gradually risen and the sulfate concentrations have decreased. However, the concentration of sulfate in groundwater in the wetland has remained stable at approximately double the pre-experiment levels. Magnesium concentrations were lower after the experiment, caused by exhaustion of the magnesium pool in the wetland through acidification. The extraction of water from the bedrock shortened the retention times and increased the recharge of groundwater in the bedrock. After extraction was terminated, the groundwater levels in the boreholes recovered within a month to levels similar to those before extraction. The hydrochemistry of the bedrock groundwater, which was strongly affected by the hydrochemistry of shallow groundwater during the experiment, has also gradually begun to regain its pre-extraction signature. However, the surface runoff has remained low during the first 2 years of recovery, at about 60% of the volume compared to the unaffected catchment area. This could be explained by delayed recovery in resaturation of the shallow rock that was unsaturated during the experiment.     

Exposure to ambient concentrations of particulate air pollution does not influence vascular function or inflammatory pathways in young healthy individuals

Background  

Particulate air pollution is associated with increased risk of cardiovascular events although the involved mechanisms are poorly understood. The objective of the present study was to investigate the effects of controlled exposure to ambient air fine and ultrafine particles on microvascular function and biomarkers related to inflammation, haemostasis and lipid and protein oxidation.     

Phthalates and nonylphenols in profiles of differently dressed soils

The concentrations of nonylphenols and phthalates in depth profiles of eight differently dressed, fertilised and cultured fields were investigated. The fields were typical for Danish agriculture and comprised an uncultured location, two manured fields, an artificially fertilised field and three fields amended with different amounts of sewage sludge. In addition, a location receiving run-off from a sewage sludge storage facility were investigated. At each location, two 50 cm vertical soil cores were taken, divided into sections of 10 cm each and analysed for nonylphenols and phthalates by high-resolution mass spectrometry. Di-(2-ethylhexyl)-phthalate (DEHP) was the most abundant phthalate in all samples whereas Di-(n-butyl)-phthalate (DBP) played a minor role. Nonylphenols occurred in significant concentrations only in soil samples exposed to high amounts of sludge and at the run-off location. A close relationship was found between the concentrations of contaminants in the soil samples and the method of dressing. The concentrations were low at comparable levels in the soil samples from the artificially fertilised field and in the fields amended with low amounts of sludge, as well as in the manured fields. Remarkably, these concentrations did not differ significantly from the level in an uncultured recreational preserved area, which was used as a reference. In contrast, much higher levels of contaminants were found in the soil samples from fields exposed to high amounts of sludge. We must conclude that sludge amendment below a certain limit does not lead to elevated levels of nonylphenols and phthalates in the soils, whereas heavy sludge amendment leads to the accumulation of these contaminants. For the vertical distribution of substances in the soils, an influence of soil characteristics on the concentration profile was noted. Thus, in soils with visible clay in the upper layers, a DEHP maximum occurred at a depth of 10-20 cm, whereas in most sandy soils no such maximum was observed. A 2-year time trend study of the highly sludge amended soil showed no measurable reduction in the substances during this period. The time study further suggested a downward movement of the DEHP maximum of approximately 10 cm per year.