Tuning a Protein‐Labeling Reaction to Achieve Highly Site Selective Lysine Conjugation

Activated esters are widely used to label proteins at lysine side chains and N termini. These reagents are useful for labeling virtually any protein, but robust reactivity toward primary amines generally precludes site‐selective modification. In a unique case, fluorophenyl esters are shown to preferentially label human kappa antibodies at a single lysine (Lys188) within the light‐chain constant domain. Neighboring residues His189 and Asp151 contribute to the accelerated rate of labeling at Lys188 relative to the ≈40 other lysine sites. Enriched Lys188 labeling can be enhanced from 50–70 % to >95 % by any of these approaches: lowering reaction temperature, applying flow chemistry, or mutagenesis of specific residues in the surrounding protein environment. Our results demonstrated that activated esters with fluoro‐substituted aromatic leaving groups, including a fluoronaphthyl ester, can be generally useful reagents for site‐selective lysine labeling of antibodies and other immunoglobulin‐type proteins.     

Acceptance sampling plans for photovoltaic modules with two‐sided specification limits

The quality control of photovoltaic modules in terms of the output power to satisfy the technical specification is of great importance for producers as well as consumers and also represents a major issue of certification procedures. Previous work focused on one‐sided specification limits to reject underperforming samples (lots) of photovoltaic modules or solar cells. In the present paper, we generalize the classic acceptance sampling methodology and derive sampling plans on the basis of two‐sided specification limits. Those sampling plans can be constructed for arbitrary output power distributions by making use of flash data tables. For the out‐of‐spec setting, the sampling plans are solutions of rather involved nonlinear equations. Explicit formulas, which resemble known sampling plans, can only be obtained under symmetry assumptions. Further, the solution depends on the ratio of overperforming modules to underperforming modules. We investigate by numerical studies to which extent the required sample size depends on that ratio and the shape of the underlying output power distribution. The application to real examples indicates that in practice, the new approach often results in substantially smaller control samples than classic approaches. Copyright © 2012 John Wiley & Sons, Ltd.     

Monitoring a reaction at submillisecond resolution in picoliter volumes

Well-established rapid mixing techniques such as stopped-flow have been used to push the dead time for kinetic experiments down to a few milliseconds. However, very fast reactions are difficult to resolve below this limit. We now outline an approach that provides access to ultrafast kinetics but does not rely on active mixing at all. Here, the reagents are compartmentalized into water-in-oil emulsion microdroplets (diameter ～50 μm) that are statically arrayed in pairs, resting side-by-side in a well feature of a poly(dimethylsiloxane) (PDMS) device. A reaction between the contents of two droplets arrayed in such a holding trap is initiated by droplet fusion that is brought about by electrocoalescence and known to occur on a time scale of about 100 μs. A reaction between the reactants (Fe(3+) and SCN(-)) is monitored by image analysis measuring the product formation in the newly merged drop in both space and time, by use of a fast camera. A comparison of the concentration field of the reaction product with the output of a reaction-diffusion system of equations yields a rate constant k ～ 3 × 10(4) M(-3)·s(-1). Since reaction and diffusion are formally included in the mathematical model, measurements can proceed immediately after the drop fusion, removing the need to allow time for mixing. This approach is different from existing methodologies, for example, experiments in a conventional stopped-flow apparatus but also electrofusion of moving droplets where contents are mixed by chaotic advection before a reaction is monitored. Our analysis makes kinetics accessible that are several times faster than techniques that have to allow time for mixing.     

Is there semantic interference in delayed naming?

The semantic interference effect in the picture–word interference task is interpreted as an index of lexical competition in prominent speech production models. Janssen, Schirm, Mahon, and Caramazza (2008) challenged this interpretation on the basis of experiments with a novel version of this task, which introduced a task-switching component. Participants either named the picture or read the word, depending on the word's color. Janssen et al. reported semantic interference in picture naming, regardless of whether the word appeared simultaneously with the picture (immediate naming) or 1,000 ms after the picture (delayed naming). Because picture name retrieval is completed in less than 1,000 ms, the finding in delayed naming was taken as evidence against the lexical competition account. In 3 sets of experiments conducted in German and English, we tested for semantic effects in Janssen et al.'s task-switching version and in the standard picture–word interference task. Using identical materials, we obtained sizeable interference effects in the standard task (Experiments 2, 4, and 6) but no effects in the task-switching version (Experiments 1, 3, and 5). When the word reading trials of the task-switching version were replaced with no-go trials (Experiment 7), semantic interference reemerged in immediate naming but was still absent in delayed naming. The experiments question the reliability of Janssen et al.'s critical finding and suggest that theoretical inferences about the origin of semantic effects in the standard picture–word interference task based on results from the task-switching version used by Janssen et al. are difficult to draw. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Qualifications in the German construction industry: stocks,flows and comparisons with the British construction sector

The paper provides an overview of qualifications in the German construction workforce and draws comparisons with the situation in the British construction industry. The German system of general education and vocational training is outlined. Data from a variety of sources are used to describe both the stocks and the flows of qualifications in the German construction workforce. Particular attention is paid to vocational qualifications, such as apprenticeships and the Meister qualification. Key findings are that the majority of German construction workers are qualified up to apprenticeship level, and that most of these employees have achieved a general schooling certificate from at least a lower secondary school prior to their vocational training. These factors are taken as signs of strength of the German construction workforce. Comparisons between the stocks of qualifications in the construction industry in the two countries suggest that intermediate qualifications of the apprenticeship type are far less prevalent in the British construction workforce.     

Grundlagen und Grenzgebiete der Lebensmittelwissenschaft

European Food Research and Technology -     

Beitrag zur Kenntnis der antimikrobiellen Wirkung der schwefligen S?ureIII. Mitteilung Aufhebung der antimikrobiellen Wirkung durch Bildung von Sulfonaten

Zusammenfassung Es wurde versucht, den Einfluß von Carbonylen auf die antimikrobielle Wirkung der schwefligen Säuren zu charakterisieren. Die Arbeiten wurden mitSaccharomyces cerevisiae durchgeführt.Durch die Bildung eines rel. beständigen Sulfonats hebt Acetaldehyd die antimikrobielle Wirkung der schwefligen Säure - wenn der Aldehyd bis zu äquimolaren Konzentrationen vorliegt - in bestimmten Bereichen vollständig auf. Auch Benzaldehyd mit einer rel. starken Eigenhemmung vermindert die Giftwirkung von schwefliger Säure bedeutend, da das Sulfonat ebenfalls ziemlich beständig ist. Ist dieses Sulfonat unbeständig, so wird die antimikrobielle Wirkung der schwefligen Säure wesentlich weniger vermindert, wie z. B. mit Salicylaldehyd, Vanillin, Glucose, Fructose und Arabinose. Die drei Ketone, Aceton, Na-Pyruvat und -Ketoglutarsäure vermindern die Wirkung der schwefligen Säure rel. stark, obwohl die Konzentrationen der Abbindung stark differieren.Es ergaben sich bei den Untersuchungen deutliche Hinweise auf eine eigene antimikrobielle Wirkung der gebildeten Sulfonate.Die Arbeit wurde mit Unterstützung der Deutschen Forschungsgemeinschaft durchgeführt, der wir hierfür unseren Dank sagen. Frau E.Niebäumer danken wir für die sehr sorgfältige technische Unterstützung.     

The Role of Silver in Mitigation of Whisker Formation on Thin Tin Films

The mitigating effect of alloying Sn thin films with Ag on the formation of Sn whiskers was investigated by time-resolved investigations employing x-ray diffraction for phase and stress analyses and focused ion beam microscopy for morphological characterization of the surfaces and cross-sections of the specimens. The investigated Sn-6 wt.%Ag thin films were prepared by galvanic co-deposition. The results are compared with those obtained from investigation of pure Sn films and discussed with regard to current whisker-growth models. The simultaneous deposition of Sn and Ag leads to a fine-grained microstructure consisting of columnar and equiaxed grains, i.e. an imperfect columnar Sn film microstructure. Isolated Ag3Sn grains are present at the Sn grain boundaries in the as-deposited films. Pronounced grain growth was observed during aging at room temperature, which provides a global stress relaxation mechanism that prevents Sn whisker growth.     

The High Resolution NMR Structure of Parvulustat (Z‐2685) from Streptomyces parvulus FH‐1641: Comparison with Tendamistat from Streptomyces tendae 4158

The protein parvulustat (Z‐2685) from Streptomyces parvulus comprises 78 amino acids and functions as a highly efficient α‐amylase inhibitor. Parvulustat shares 29.6 % overall amino acid sequence identity to the well‐known α‐amylase inhibitor tendamistat. Among the conserved residues are the two disulfide bridges (C9–C25, C43–C70) and the active‐site motif (W16, R17, Y18). Here, we report the high‐resolution NMR structure of parvulustat based on NOEs, J couplings, chemical shifts and hydrogen‐exchange data. In addition, we studied the dynamical properties of parvulustat by heteronuclear relaxation measurements. We compare the structure of parvulustat with the structure of tendamistat in terms of secondary structure elements, charges and hydrophobicity. The overall structural composition is very similar, but there are distinct differences including the active‐site region. These structural and dynamical differences indicate that for parvulustat an induced‐fit mechanism for binding to α‐amylase might take place, since the structure of tendamistat does not change upon binding to α‐amylase.