Enzymatic on-chip enhancement for high resolution genotyping DNA microarrays

Antibiotic resistance among pathogenic microorganisms is emerging as a major human healthcare concern. While there are a variety of resistance mechanisms, many can be related to single nucleotide polymorphisms and for which DNA microarrays have been widely deployed in bacterial genotyping. However, genotyping by means of allele-specific hybridization can suffer from the drawback that oligonucleotide probes with different nucleotide composition have varying thermodynamic parameters. This results in unpredictable hybridization behavior of mismatch probes. Consequently, the degree of discrimination between perfect match and mismatch probes is insufficient in some cases. We report here an on-chip enzymatic procedure to improve this discrimination in which false-positive hybrids are selectively digested. We find that the application of CEL1 Surveyor nuclease, a mismatch-specific endonuclease, significantly enhances the discrimination fidelity, as demonstrated here on a microarray for the identification of variants of carbapenem resistant Klebsiella pneumoniae carbapenemases and monitored by end point detection of fluorescence intensity. Further fundamental investigations applying total internal reflection fluorescence detection for kinetic real-time measurements confirmed the enzymatic enhancement for SNP discrimination.     

SVD-based MIMO Precoding and Equalization Schemes for Realistic Channel Knowledge: Design Criteria and Performance Evaluation

Multiple-input multiple-output (MIMO) based communication systems with precoding, bit-loading and equalization procedures are considered in this paper. Applications of precoding schemes, which are based on Singular Value Decomposition (SVD) of the channel matrix H assume almost always ideal channel knowledge at the transmitter and/or receiver site. This paper investigates an SVD based MIMO approach considering non ideal radio channel estimation results. In any case the MIMO channel matrix H is decomposed into Eigenmodes. In case of an ideal radio channel knowledge the SVD based precoding procedure, which is applied at the transmitter site, is going to consider all possible Eigenmodes which results in a perfect separation of all signals at the receive antenna output and into a minimum bit-error-rate (BER). It will be shown in this paper that in case of non ideal channel knowledge and a limited accuracy in the channel matrix H estimation a reduced number of Eigenmodes in the precoding process will become an optimum and will lead into an increased BER performance.

Berücksichtigung der Fließgeschwindigkeit bei Hochwasser‐Schadensmodellen

Die Quantifizierung der Schadenspotentiale infolge Hochwasser setzt realistische Zusammenhänge zwischen Einwirkungs‐ und Verlustkenngrößen voraus. Da sich herkömmliche Schadensfunktionen auf den Zusammenhag zwischen Fluthöhe und den versicherungsseitig nachvollziehbaren Kosten für eine bestimmte Nutzungsklasse (wie. z. B. private Wohngebäude) beschränken, fehlt zum einen die notwendige Differenzierung nach den für die Widerstandsseite relevanten Merkmalen, zum anderen bleiben weitere Einwirkungsgrößen wie die Fließgeschwindigkeit unberücksichtigt. Auf der Grundlage einer einheitlichen Datenerhebung wurde ein System entwickelt, mit dem die strukturelle Schädigung eines Bauwerks oder eines betroffenen Bauwerksbestands für konkrete Hochwasserszenarien bestimmt werden kann. Dieses Instrumentarium wird erweitert, so dass der Zusammenhang zwischen den Einwirkungskenngrößen Überflutungshöhe und Fließgeschwindigkeit und dem strukturellen Schaden hergestellt werden kann. Die Studie stützt sich auf dabei zwei Datensätze, die nach dem Hochwasser 2002 erhoben wurden: – Datensatz 1 bezieht sich auf eine schriftliche Befragung, welche zwischen 2002 und 2004 in Sachsen durchgeführt wurde. – Datensatz 2 stellt dabei das Ergebnis einer Auswertung von Schadensgutachten im Auftrag der Landestalsperrenverwaltung Sachsen dar. Im Ergebnis kann ein neuartiger Typ von Verletzbarkeitsfunktionen bereitgestellt werden, der den Zusammenhang zwischen Überflutungshöhe, Fließgeschwindigkeit und den Schadensgraden D_i in Abhängigkeit von der konkreten Bauwerksverletzbarkeit beschreibt. Mit Hilfe ebenfalls neu entwickelter Schadensfunktionen werden die so berechneten Schadensgrade in konkrete Verlustaussagen überführt. Wie an drei Testgebieten im Freistaat Sachsen nachgewiesen werden kann, gelingt es mit diesen Funktionen, die gemeldeten Verluste infolge des Hochwassers 2002 in guter Übereinstimmung nachzuvollziehen. Im Weiteren werden erste Untersuchungen zum Einfluss der städtebaulichen Einbindung und der Anströmrichtung auf die strukturellen Schäden vorgestellt. Consideration of flow velocity in flood damage models. Reliable prognoses of building damage caused by flood impact require realistic relationships between action and damage or loss describing parameters. Due to the fact that commonly applied damage functions are related to the different usage classes (i.e. private housing), the required differentiation according to the parameters on the resistance side is still missing and further impact parameters like flow velocity remain unconsidered. On the basis of the August 2002 Saxony flood data base, a method to determine the structural damage for any given flood scenario was developed. This method is improved to consider the correlation between inundation level and flow velocity and structural damage. The study is based on two different datasets collected after the flood 2002 in Saxony: – Dataset 1 is related to the outcome of a questionnaire survey performed by the authors between 2002 and 2004. – Dataset 2 is the outcome of an evaluation of damage expertises by order of the “Landestalsperrenverwaltung Sachsen”. A set of new Specific Vulnerability Functions (SVF) for calculation of damage grades is derived considering the inundation level and the flow velocity depending on the vulnerability of the predominant building types. Based on new developed Specific Damage Functions (SDF) the calculated damage grades are transferred into loss estimates. The developed tools are applied to the reinterpretation of the August 2002 flood damage. Results of the approach are presented for three different study areas in the free state of Saxony. In all cases a remarkable good agreement between the predicted and the reported loss can be stated. First investigations with respect to the location of buildings, the direction of inflow and the structural damage are represented.     

A dose-controlled system for air-liquid interface cell exposure and application to zinc oxide nanoparticles

Background  

Engineered nanoparticles are becoming increasingly ubiquitous and their toxicological effects on human health, as well as on the ecosystem, have become a concern. Since initial contact with nanoparticles occurs at the epithelium in the lungs (or skin, or eyes), in vitro cell studies with nanoparticles require dose-controlled systems for delivery of nanoparticles to epithelial cells cultured at the air-liquid interface.     

Early colonization of stone by freshwater lichens of restored habitats: A case study in northern Italy

This study evaluated the effectiveness and life-strategies of freshwater lichens in colonizing newly constructed stone structures in low-elevation streams in a small nature reserve in northern Italy. Species richness, size of thalli, morphological and ontogenetic traits of the species were related to the age of restored habitats. Lichen colonization was surprisingly rapid, indicating the high potential of these organisms in colonizing restored habitats. However, the species pool found in the restored habitats was different than that found in natural sites in the same study area. The age of newly constructed habitats influenced both species richness and thallus size of the two most frequent Verrucaria species. Verrucaria aquatilis was a rapid colonizer invading the substrate by several small-sized and thin thalli which soon supported a large number of small perithecia whose development began in the earlier phase of thallus formation. V. elaeomelaena, on the contrary, developed according to a different strategy, establishing a thick thallus on which relatively large perithecia were formed much later than in V. aquatilis. As these taxa are important photoautotrophic components of freshwater ecosystems more ecological knowledge is needed to evaluate the effectiveness of different measures of river restoration on lichen communities. The main practical implication of our study is related to the value of small stone structures, such as riffles and ramps, for enhancing the establishment of pioneer freshwater lichens to rapidly colonize newly available substrata.     

Use of alkyl ketene dimer (AKD) for surface modification of particleboard chips

The study objective was to impart urea-formaldehyde (UF) bonded particleboards higher and longer-lasting hydrophobicity than that resulting from conventionally used paraffin. Alkyl ketene dimer (AKD) is a paper sizing agent that can theoretically esterify wood compounds and result in a surface modification. Particles were 1) impregnated with an aqueous AKD-solution and cured at 130 °C prior to gluing, or 2) sprayed with a mixture of AKD-solution and UF-resin in a single step. Boards with and without paraffin wax were used as controls. Thickness swelling after 2, 24, and 48 h immersion was decreased by 90, 62, and 59% when the chips were impregnated with AKD in comparison to untreated control boards. Water uptake after 2, 24, and 48 h was reduced by 91, 75, and 60%. AKD-impregnation with subsequent curing decreased the internal bond strength by 53%, indicating that AKD impedes the adhesion. The mixture of AKD and UF-glue did not result in considerable hydrophobicity. Increased methyl/methylene and carbonyl bands in FTIR-spectra after toluene-extraction suggest that AKD partially formed ester bonds at the wood surface.     

The efficacy of commercial silicones against blue stain and mould fungi in wood

Scots pine (Pinus sylvestris L.) samples were vacuum pressure impregnated with quaternary ammonium (quat)-silicone micro-emulsion (<40 nm particle size), amino-silicone macro-emulsion (110 nm) and silicone macro-emulsion with alkyl modified side groups (740 nm). Quat-silicone micro-emulsion caused highest cell wall bulking (4.8%) and anti-swelling efficiency (21.8%) in wood when treated with 30% concentration of silicone in the treatment solution. All three formulations made wood hydrophobic which was evident from a capillary (longitudinal, tangential and radial) water uptake test. Amino-silicone macro-emulsion (10% silicone in treatment solution) resulted in strong resistance to blue stain attack with both pre-weathered and not pre-weathered wood samples. Signs of blue staining were more evident on the pre-weathered samples. Treatment with amino-silicone macro-emulsion (10% silicone in treatment solution) resulted in a certain resistance to mould growth on wood surfaces, whereas wood treated with the same concentration of quat-silicone micro-emulsion and alkyl-modified silicone macro-emulsion exhibited comparatively lower resistance.     

Procedural influence on the properties of particleboards made from AKD modified chips

Wood chips were treated with alkyl ketene dimer (AKD) using three different processes to impart water resistance to particleboards. In the first process, AKD was blended with UF resin. Thickness swelling and water uptake after a 24 h immersion period (20 and 69%) were lower than in the control boards (28 and 81%) but were higher than in the paraffin references (10 and 22%). In process 2, AKD and UF resin were sprayed separately on the chips resulting in a greater reduction of thickness swelling (15%) and water uptake (49%) than in process 1. Paraffin references revealed a thickness swelling and water uptake of 7 and 25%, respectively. An extension of the pressing time in processes 1 and 2 did not increase water repellence. In process 3, particleboards were made from AKD-treated chips that were cured at 130 °C (24 h) prior to gluing. They showed a thickness swelling of 7% and a water uptake of 25%, whereas particleboards with paraffin exhibited levels of 8 and 29%. The thickness swelling and water uptake of boards with AKD increased when the curing time was reduced from 24 to 12 to 6 h (130 °C). Changing the curing temperature from 130 to 100 °C (12 h) had no effect on board properties. The IB of boards made from pre-cured chips with AKD (24 h/130 °C) was 44% lower than in controls and 35% lower than in paraffin references. This indicates that AKD impedes the adhesion.     

Modellierung einer Einscheibentrockenkupplung mithilfe des C&C2-Ansatzes zur Identifikation von kritischen Gestalt-Funktion-Zusammenhängen bezüglich des Schwingungsphänomens zwangserregtes Kupplungsrupfen

Forschung im Ingenieurwesen - In diesem Beitrag wird ein systematisches Vorgehen zur Identifikation von kritischen Gestalt-Funktion-Zusammenhängen mit Einfluss auf zwangserregtes...