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1.
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.  相似文献   
2.
A novel method for fabricating a nano-Cu/Si3N4 ceramic substrate is proposed. The nano-Cu/Si3N4 ceramic substrate is first fabricated using spark plasma sintering (SPS) with the addition of nanoscale multilayer films (Ti/TiN/Ti/TiN/Ti) as transition layers. The microstructures of the nano-Cu metal layer and the interface between Cu and Si3N4 are investigated. The results show that a higher SPS temperature increases the grain size of the nano-Cu metal layer and affects the hardness. The microstructure of the transition layer evolves significantly after SPS. Ti in the transition layer can react with Si3N4 and with nano-Cu to form interfacial reaction layers of TiN and Ti–Cu, respectively; these ensure stronger bonding between nano-Cu and Si3N4. Higher SPS temperatures improve the diffusion ability of Ti and Cu, inducing the formation of Ti3Cu3O compounds in the nano-Cu metal layer and Ti2Cu in the transition layer. This study provides an important strategy for designing and constructing a new type of ceramic substrate.  相似文献   
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In this article we illustrate a methodology for building cross-language search engine. A synergistic approach between thesaurus-based approach and corpus-based approach is proposed. First, a bilingual ontology thesaurus is designed with respect to two languages: English and Spanish, where a simple bilingual listing of terms, phrases, concepts, and subconcepts is built. Second, term vector translation is used – a statistical multilingual text retrieval techniques that maps statistical information about term use between languages (Ontology co-learning). These techniques map sets of t f id f term weights from one language to another. We also applied a query translation method to retrieve multilingual documents with an expansion technique for phrasal translation. Finally, we present our findings.  相似文献   
6.
The Vehicle Routing Problem with Simultaneous Pickup and Delivery (VRPSPD) is an extension to the classical Vehicle Routing Problem (VRP), where customers may both receive and send goods simultaneously. The Vehicle Routing Problem with Mixed Pickup and Delivery (VRPMPD) differs from the VRPSPD in that the customers may have either pickup or delivery demand. However, the solution approaches proposed for the VRPSPD can be directly applied to the VRPMPD. In this study, an adaptive local search solution approach is developed for both the VRPSPD and the VRPMPD, which hybridizes a Simulated Annealing inspired algorithm with Variable Neighborhood Descent. The algorithm uses an adaptive threshold function that makes the algorithm self-tuning. The proposed approach is tested on well-known VRPSPD and VRPMPD benchmark instances derived from the literature. The computational results indicate that the proposed algorithm is effective in solving the problems in reasonable computation time.  相似文献   
7.
The identification of the Hammerstein–Wiener (H-W) systems based on the nonuniform input–output dataset remains a challenging problem. This article studies the identification problem of a periodically nonuniformly sampled-data H-W system. In addition, the product terms of the parameters in the H-W system are inevitable. In order to solve the problem, the key-term separation is applied and two algorithms are proposed. One is the key-term-based forgetting factor stochastic gradient (KT-FFSG) algorithm based on the gradient search. The other is the key-term-based hierarchical forgetting factor stochastic gradient (KT-HFFSG) algorithm. Compared with the KT-FFSG algorithm, the KT-HFFSG algorithm gives more accurate estimates. The simulation results indicate that the proposed algorithms are effective.  相似文献   
8.
The architectural choices underlying Linked Data have led to a compendium of data sources which contain both duplicated and fragmented information on a large number of domains. One way to enable non-experts users to access this data compendium is to provide keyword search frameworks that can capitalize on the inherent characteristics of Linked Data. Developing such systems is challenging for three main reasons. First, resources across different datasets or even within the same dataset can be homonyms. Second, different datasets employ heterogeneous schemas and each one may only contain a part of the answer for a certain user query. Finally, constructing a federated formal query from keywords across different datasets requires exploiting links between the different datasets on both the schema and instance levels. We present Sina, a scalable keyword search system that can answer user queries by transforming user-supplied keywords or natural-languages queries into conjunctive SPARQL queries over a set of interlinked data sources. Sina uses a hidden Markov model to determine the most suitable resources for a user-supplied query from different datasets. Moreover, our framework is able to construct federated queries by using the disambiguated resources and leveraging the link structure underlying the datasets to query. We evaluate Sina over three different datasets. We can answer 25 queries from the QALD-1 correctly. Moreover, we perform as well as the best question answering system from the QALD-3 competition by answering 32 questions correctly while also being able to answer queries on distributed sources. We study the runtime of SINA in its mono-core and parallel implementations and draw preliminary conclusions on the scalability of keyword search on Linked Data.  相似文献   
9.
在分析单一MU(Most Uncertainty)采样缺陷的基础上,提出一种"全局最优搜寻"方法 GOS(Global Optimal Search),并结合MU共同完成查询选择。GOS+MU方法中,GOS着眼全局寻找目标,在应用环境能提供的训练样本数量有限、分类器受训不充分时,该方法选择的对象学习价值高,能快速推进分类器学习进程;MU则能够在GOS采样失效情形下,利用分类器当前训练成果,选择查询不确定性最强的样本补充训练集。通过对网络商品的用户评论进行分类仿真,并比较其他采样学习方法的效果,证明了GOS+MU方法在压缩学习成本、提高训练效率方面的有效性。  相似文献   
10.
带关键字搜索的公钥加密(PEKS)是一种有用的加密原语,它允许用户将在加密数据上搜索的功能委托给不可信的第三方服务器,而不影响原始数据的安全性和隐私性。但是,由于缺乏对于数据的加密以及解密能力,PEKS方案不能单独进行使用,必须与标准的公钥加密方案(PKE)相结合。因此,Baek等人在2006年引入了一种新的加密原语,称为结合PKE和PEKS的加密方案(PKE+PEKS),它同时提供了PKE和PEKS的功能。目前,已有文献提出了几种PKE+PEKS方案。然而,他们都没有考虑关键字猜测攻击的问题。本文提出一个新的高效且能够抵抗关键字猜测攻击的PKE+PEKS方案,与已有方案相比,该方案在性能上有很大的提升,并且在生成关键字和数据密文时,不需要使用双线性对,极大地降低了计算和存储成本。安全性分析表明,本文中所提出的方案能够满足密文隐私安全性、陷门不可区分性和抗关键字猜测攻击的安全性。效率分析表明,本分提出的方案更加高效。  相似文献   
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