Trypsiligase-Catalyzed Labeling of Proteins on Living Cells

Fluorescent fusion proteins are powerful tools for studying biological processes in living cells, but universal application is limited due to the voluminous size of those tags, which might have an impact on the folding, localization or even the biological function of the target protein. The designed biocatalyst trypsiligase enables site-directed linkage of small-sized fluorescence dyes on the N terminus of integral target proteins located in the outer membrane of living cells through a stable native peptide bond. The function of the approach was tested by using the examples of covalent derivatization of the transmembrane proteins CD147 as well as the EGF receptor, both presented on human HeLa cells. Specific trypsiligase recognition of the site of linkage was mediated by the dipeptide sequence Arg-His added to the proteins’ native N termini, pointing outside the cell membrane. The labeling procedure takes only about 5 minutes, as demonstrated for couplings of the fluorescence dye tetramethyl rhodamine and the affinity label biotin as well.     

Cell-Free Synthesis of Selenoproteins in High Yield and Purity for Selective Protein Tagging

The selenol group of selenocysteine is much more nucleophilic than the thiol group of cysteine. Selenocysteine residues in proteins thus offer reactive points for rapid post-translational modification. Herein, we show that selenoproteins can be expressed in high yield and purity by cell-free protein synthesis by global substitution of cysteine by selenocysteine. Complete alkylation of solvent-exposed selenocysteine residues was achieved in 10 minutes with 4-chloromethylene dipicolinic acid (4Cl-MDPA) under conditions that left cysteine residues unchanged even after overnight incubation. Gd^III−Gd^III distances measured by double electron–electron resonance (DEER) experiments of maltose binding protein (MBP) containing two selenocysteine residues tagged with 4Cl-MDPA-Gd^III were indistinguishable from Gd^III−Gd^III distances measured of MBP containing cysteine reacted with 4Br-MDPA tags.     

Selective Analysis of Sulfur-Containing Species in a Heavy Crude Oil by Deuterium Labeling Reactions and Ultrahigh Resolution Mass Spectrometry

A heavy crude oil has been treated with deuterated alkylating reagents (CD₃I and C₂D₅I) and directly analyzed without any prior fractionation and chromatographic separation by high-field Orbitrap Fourier Transform Mass Spectrometry (FTMS) and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS) using electrospray ionization (ESI). The reaction of a polycyclic aromatic sulfur heterocycles (PASHs) dibenzothiophene (DBT), in the presence of silver tetrafluoroborate (AgBF₄) with ethyl iodide (C₂H₅I) in anhydrous dichloroethane (DCE) was optimized as a sample reaction to study heavy crude oil mixtures, and the reaction yield was monitored and determined by proton nuclear magnetic resonance spectroscopy (¹H-NMR). The obtained conditions were then applied to a mixture of standard aromatic CH-, N-, O- and S-containing compounds and then a heavy crude oil, and only sulfur-containing compounds were selectively alkylated. The deuterium labeled alkylating reagents, iodomethane-d₃ (CD₃I) and iodoethane-d₅ (C₂D₅I), were employed to the alkylation of heavy crude oil to selectively differentiate the tagged sulfur species from the original crude oil.     

Overcoming the Taste Stigma of Soy

ABSTRACT: Can labels unknowingly influence a person's taste perceptions? Using a “phantom ingredient” taste test, the presence or absence of a labeled ingredient (soy) was shown to influence sensory evaluations of a food. In particular, a nutrition bar wrongly purporting to contain soy generated negative ratings of taste, aftertaste, and attitude, yet generated favorable ratings of its nutritiousness. Because there was actually no soy in the product, these differences represent biases caused by ingredient labeling. Given the appropriate segment of consumers (health‐oriented), ingredient labeling may have more favorable consequences. This power of suggestion might also be expanded to other ingredients or processes, such as those involving biotechnology or organic production.     

Compact and localized distributed data structures

This survey concerns the role of data structures for compactly storing and representing various types of information in a localized and distributed fashion. Traditional approaches to data representation are based on global data structures, which require access to the entire structure even if the sought information involves only a small and local set of entities. In contrast, localized data representation schemes are based on breaking the information into small local pieces, or labels , selected in a way that allows one to infer information regarding a small set of entities directly from their labels, without using any additional (global) information. The survey concentrates mainly on combinatorial and algorithmic techniques, such as adjacency and distance labeling schemes and interval schemes for routing, and covers complexity results on various applications, focusing on compact localized schemes for message routing in communication networks.Received: August 2001, Accepted: May 2002, Supported in part by a grant from the Israel Science Foundation.     

Do 9-month-old infants expect distinct words to refer to kinds?

In 3 experiments, 9-month-old infants' expectations for what distinct count noun labels refer to were investigated. In Experiment 1, a box was opened to reveal 2 objects inside during familiarization: either 2 identical objects or 2 different objects. Test trials followed the same procedure, except before the box was opened, the contents were described using 2 distinct labels ("I see a wug! I see a dak!") or the same label twice ("I see a zav! I see a zav!"). Infants who heard a label repeated twice looked longer at 2 different objects versus 2 identical objects, whereas infants who heard 2 distinct labels showed a different pattern of looking. Experiments 2 and 3 presented infants with object pairs that only differed in shape or color, and it was found that infants expected the different-shaped (but not the different-colored) objects to be labeled by distinct count nouns. Because the property of shape is a cue to kind membership and the property of color is not, these results suggest that even at the beginning of word learning, infants may expect distinct labels to refer to distinct kinds of objects. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

基于中值滤波和梯度锐化的边缘检测

实现一种基于中值滤波和梯度锐化的边缘检测方法。首先采用既能过滤噪声又能保护边缘信息的中值滤波对图像进行平滑处理。然后采用梯度锐化加强边缘像素强度,通过简单的二值化获得图像的初始边缘,系统采用连通区域标记的方法去除位于图像封闭边界内的残留黑块。最后通过实验结果与人工提取的结果比较,进行误差分析,找出本文算法的不足之处,以便对该算法进行改进。实践证明,本文的边缘检测方法对含有噪声点且灰度分布比较均匀的图像有很好的效果。