Electron capture dissociation as structural probe for noncovalent gas-phase protein assemblies

Electron capture dissociation (ECD) of proteins in Fourier transform ion cyclotron resonance mass spectrometry usually leads to charge reduction and backbone-bond cleavage, thereby mostly retaining labile, intramolecular noncovalent interactions. In this report, we evaluate ECD of the 84-kDa noncovalent heptameric gp31 complex and compare this with sustained off-resonance irradiation collisionally activated dissociation (SORI-CAD) of the same protein. Unexpectedly, the 21+ charge state of the gp31 oligomer exhibits a main ECD pathway resulting in a hexamer and monomer, disrupting labile, intermolecular noncovalent bonds and leaving the backbone intact. Unexpectedly, the charge separation over the two products is highly proportional to molecular weight. This indicates that a major charge redistribution over the subunits of the complex does not take place during ECD, in contrast to the behavior observed when using SORI-CAD. We speculate that the ejected monomer retains more of its original structure in ECD, when compared to SORI-CAD. ECD of lower charge states of gp31 does not lead to dissociation of noncovalent bonds. We hypothesize that the initial gas-phase structure of the 21+ charge state is significantly different from the lower charge states. These structural differences result in the different reaction pathways when using ECD.     

Detection of noroviruses in foods: a study on virus extraction procedures in foods implicated in outbreaks of human gastroenteritis

Disease outbreaks in which foods are epidemiologically implicated as the common source are frequently reported. Noroviruses and enteric hepatitis A viruses are among the most prevalent causative agents of foodborne diseases. However, the detection of these viruses in foods other than shellfish is often time-consuming and unsuccessful. In this study, three virus concentration methods were compared: polyethylene glycol (PEG) plus NaCl, ultracentrifugation, and ultrafiltration. Two RNA extraction methods, TRIzol and RNeasy Mini Kit (Qiagen), were compared for detection of viruses in whipped cream and lettuce (as representatives of the dairy and vegetable-fruit food groups, respectively). A seeding experiment with canine calicivirus was conducted to determine the efficiency of each virus extraction procedure. The PEG-NaCl-TRIzol method was most efficient for the detection of viruses in whipped cream and the ultracentrifugation-RNeasy-Mini Kit procedure was best for detection on lettuce. Based on the seeding experiments, food items implicated in norovirus-associated gastroenteritis outbreaks were subjected to the optimal procedure for a specific composition and matrix. No noroviruses were detected in the implicated food items, possibly because the concentration of virus on the food item was too low or because of the presence of inhibitory factors. For each food group, a specific procedure is optimal. Inhibitory factors should be controlled in these procedures because they influence virus detection in food.     

Axial beanspruchte Sandwichelemente in rahmenlosen Konstruktionen

Sandwichelemente werden üblicherweise als einachsig spannende Bauelemente verwendet. Diese werden durch auf sie einwirkende Querlasten auf Biegung beansprucht und leiten die Kräfte in eine Unterkonstruktion ab. Eine neuere Entwicklung ist es kleinere Gebäude aus Sandwichelementen auch ohne Unterkonstruktion herzustellen. Bei dieser Bauweise haben die Elemente nicht nur die Funktion des Raumabschlusses, sondern werden auch zur Lastabtragung und zur Aussteifung des Gebäudes herangezogen. Zusätzlich zu Biegemomenten und Querkräften wirken in den Elementen dann auch Normalkräfte. Ein Bemessungsvorschlag für axial beanspruchte Sandwichelemente, durch den auch die auftretenden Kriecherscheinungen berücksichtigt werden können, wird vorgestellt. Außerdem werden die maßgebenden Versagensmodi an der Lasteinleitungsstelle aufgezeigt und ein mechanisches Modell erläutert. Axially loaded sandwich panels in frameless buildings. Traditionally sandwich panels are used as unidirectional spanning flexural members transferring loads to a substructure. As a recent development, sandwich panels are used to design small buildings without substructure. In this application the panels are not only used as cladding element but also for load transfer and bracing of the building. A design model for axially loaded sandwich panels, which also considers creeping effects, is presented. Additionally the basic failure modes at the area of load application are illustrated and a mechanical model is introduced.     

Enzymatic degradation of hyperbranched polyesters

In this work, the enzyme‐catalyzed degradation of hyperbranched polyesters (HBPEs) was investigated. Enzymatic degradation experiments were performed in a phosphate buffer in the presence of the lipases Candida cylindracea, Pseudomonas cepacia, Novozym 388, Amano CE, Lipomod 34P, and Cal‐B, whereas control experiments were performed in the same system without lipases. The extent of polymer degradation was determined by quantification of the released free fatty acids by gas chromatography. The influence of the alkane chain length and the number of alkane chain end groups on the lipase‐catalyzed hydrolysis of esterified HBPEs was investigated systematically. It was found that the increase in the alkane chain length of the end groups diminished the enzymatic degradation of the polymer, whereas the number of end groups had no influence on the degradation rate. The effect of temperature on the rate of degradation was also described. Surface morphological changes that occurred during the degradation were assessed with reflected electron microscopy. The changes in the crystallinity of the polymers after they were subjected to degradation were qualitatively determined with differential scanning calorimetry through the quantification of the enthalpy of melting. The enthalpy of melting of one HBPE sample increased from 79 to 90 and 94 J/g with and without the action of Lipomod 34P, respectively, in 7 days, showing the changes in the crystallinity of the polymer. The results prove that modified HBPEs are an important new class of biodegradable materials with a predictable degradation mechanism, and the degradation can be adjusted on the basis of the molecular engineering. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Das Allergie-Internetportal im Aktionsplan gegen Allergien des Bundesministeriums für Ernährung, Landwirtschaft und Verbraucherschutz

Ohne Zusammenfassung Eingegangen: 7. Januar 2009     

A near real-time dual-band-spatial approach to determine the source of increased radiance from closely spaced active volcanoes in coarse resolution satellite data

Bezymianny and Kliuchevskoi volcanoes (Kamchatka) present a danger as both inject ash into North Pacific air routes. Current automated monitoring algorithms do not distinguish them in real time due to their mutual proximity (10 km) and poor geolocation accuracy of Advanced Very High Resolution Radiometer (AVHRR) data. Contrasting mid- and thermal infrared volcanic radiances are influenced by (1) differences in temperature and eruptive style of Bezymianny's andesite and Kliuchevskoi's basalt and (2) different temperatures of the non-volcanic portion of pixels located over their summits, due to different elevations. Data from 571 AVHRR images show the latter is more significant. Discriminant function analysis using summit and regional band 4 pixel-integrated radiant temperatures (pirT) correctly identifies the source volcano of a thermal anomaly in 89% of cases. Weather permitting, a spatial component can be added, leading to improved accuracy. The approach used here can also be applied at other closely spaced volcanoes with substantially different summit elevations.     

Facilitating peripheral interaction: design and evaluation of peripheral interaction for a gesture-based lighting control with multimodal feedback

Most interactions with today’s interfaces require a person’s full and focused attention. To alleviate the potential clutter of focal information, we investigated how interactions could be designed to take place in the background or periphery of attention. This paper explores whether gestural, multimodal interaction styles of an interactive light system allow for this. A study compared the performance of interactions with the light system in two conditions: the central condition in which participants interacted only with the light system, and the peripheral condition in which they interacted with the system while performing a high-attentional task simultaneously. Our study furthermore compared different feedback styles (visual, auditory, haptic, and a combination). Results indicated that especially for the combination feedback style, the interaction could take place without participants’ full visual attention, and performance did not significantly decrease in the peripheral condition. This seems to indicate that these interactions at least partly took place in their periphery of attention and that the multimodal feedback style aided this process.