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91.
Dr. Emrah Kara Dr. Nis Valentin Nielsen Bergrun Eggertsdottir Dr. Bernd Thiede Dr. Sandip M. Kanse Dr. Geir Åge Løset 《Chembiochem : a European journal of chemical biology》2020,21(13):1875-1884
We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated. 相似文献
92.
Design and Fabrication of Concentration‐Gradient Generators with Two and Three Inlets in Microfluidic Chips 下载免费PDF全文
A simple and low‐cost method for designing and fabricating concentration‐gradient generators with two and three inlets is proposed which can generate different concentration gradients at varying flow velocities. The microchannel structure was designed in S‐shape and left‐right symmetry. The concentration‐gradient generator was simulated based on the finite element method. The microchannels were processed on a computer numerical control (CNC) engraving and milling machine on poly(methylmethacrylate) substrate, and then two concentration‐gradient generators were fabricated by hot bonding technology. The results of experiment and simulation were compared to prove the feasibility of the method. Flow velocity was an important factor for generating different concentration gradients. The concentration‐gradient profiles of the generators with two and three inlets present approximately linear and quadratic curves. 相似文献
93.
《Food Control》2015
A diverse range of genetic elements has been used to develop genetically modified organisms (GMOs) over the last 18 years. Screening methods that target few elements, such as the Cauliflower Mosaic Virus 35S promoter (P-35S) and Agrobacterium tumefaciens nopaline terminator (T-nos), are not sufficient to screen GMOs. In the present study, a multiplex PCR system for all globally commercialized GM soybean events was developed to easily trace the events. For this purpose, screening elements of 24 GM soybean events were investigated and 9 screening targets were selected and divided into three individual triplex PCR systems: P-35S, ribulose-1,5-bisphosphate carboxylase small subunit promoter of Arabidopsis thaliana, T-nos, T-35S, pea E9 terminator, open reading frame 23 terminator of A. tumefaciens, proteinase inhibitor II terminator of potato, acetohydroxy acid synthase large subunit terminator of A. thaliana, and the revealed 3′ flanking sequences of DP-305423-1. The specificity of the assays was confirmed using thirteen GM soybean events as the respective positive/negative controls. The limit of detection of each multiplex set, as determined using certified reference materials of specific GM events, ranged from 0.03 to 0.5%, depending upon target. Furthermore, 26 food samples that contained soybean ingredients, which were purchased from the USA, China, Japan, and Korea, were analyzed, 17 of which contained one or more GM soybean events. These results suggest that the developed screening method can be used to efficiently track and identify 24 GM soybean events in food and feed. 相似文献
94.
95.
The aim of the study was to identify the importance of individual food product attributes (the use of preservatives, processing method, shelf-life period and nutritional value) as well as their relation to the purchase behaviour. To achieve this, consumer preferences were decomposed in conditions of full access to information, and data was compared with actual consumer behaviour related to making purchasing decisions in front of the store shelf. Based on data from 338 respondents, conjoint analysis and repeated ANOVA measurements were carried out, allowing to eliminate individual behavioural patterns. The results showed a dissonance between the consumers attitudes towards the attributes and their shopping behaviour. The processing method was the most important declared attribute for consumers, meanwhile this information was the least searched for during purchase. At the same time, shelf-life period marked as the least important was the main information searched for by consumers when shopping. The results also show a clear dislike towards the use of microwaves as a preservation method, while traditional thermal preservation was marked as the most positive for consumers. In addition, consumers were classified into 3 heterogeneous groups, identifying differences in the preferences of food attributes. The obtained results have practical value relating not only to the preferences of individual attributes, but also to their various variants. The results can be a guide for companies to properly label food products. The indicated dissonance illustrates the educational gap and related insufficient motivation to get to know the product before buying it. This is extremely important in connection with technological development and new food processing methods. 相似文献
96.
97.
The fourth-order partial differential equations have good performance on noise smoothing and edge preservation without creating blocky effects on smooth regions. However, for low signal-to-noise ratio images, the discrimination between edges and noise is a challenging problem. A novel kernel-based fourth-order diffusion is proposed in this paper. It introduces a kernelized gradient operator in the fourth-order diffusion process, which leads to more effective noise removal capability. Experiment results show that this method outperforms several previous anisotropic diffusion methods for noise removal and edge preservation. 相似文献
98.
Hai Huang Haichao An Haibo Ma Shenyan Chen 《International journal for numerical methods in engineering》2019,117(3):291-315
This work presents an engineering method for optimizing structures made of bars, beams, plates, or a combination of those components. Corresponding problems involve both continuous (size) and discrete (topology) variables. Using a branched multipoint approximate function, which involves such mixed variables, a series of sequential approximate problems are constructed to make the primal problem explicit. To solve the approximate problems, genetic algorithm (GA) is utilized to optimize discrete variables, and when calculating individual fitness values in GA, a second-level approximate problem only involving retained continuous variables is built to optimize continuous variables. The solution to the second-level approximate problem can be easily obtained with dual methods. Structural analyses are only needed before improving the branched approximate functions in the iteration cycles. The method aims at optimal design of discrete structures consisting of bars, beams, plates, or other components. Numerical examples are given to illustrate its effectiveness, including frame topology optimization, layout optimization of stiffeners modeled with beams or shells, concurrent layout optimization of beam and shell components, and an application in a microsatellite structure. Optimization results show that the number of structural analyses is dramatically decreased when compared with pure GA while even comparable to pure sizing optimization. 相似文献
99.
为解决采空区突发涌水给矿山安全生产带来的安全威胁,以齐大山铁矿为研究对象,以高密度电阻率法含水构造带理论模型的正演模拟研究为基础,运用高密度电阻率法对采场南帮含水构造带的异常范围进行了圈定;运用瞬变电磁法进行了二次探测,并对高密度电阻率法所圈定的异常范围进行了验证。研究表明:含水构造带显示为低阻异常特征,在高密度电阻率法探测成果图中显示的两处低阻异常与含水构造带理论模型的正演模拟结果一致;同时,在瞬变电磁法探测成果图中显示的低阻区域均在高密度电阻率法所圈定的低阻异常范围内,二者相互验证,这两处低阻异常均为含水构造带的地球物理异常特征的显示,表明采用高密度电阻率法和瞬变电磁法组合识别技术可实现对隐伏含水构造带的先导性探测。 相似文献
100.
讨论了3种制样方法对直流辉光放电质谱法(dc-GDMS)检测氮化硼中的Na、Mg、Al、Si等27种杂质元素的影响。3种制样方法分别如下所示:方法1,直接把氮化硼压在铟薄片上;方法2,把氮化硼压在铟薄片上后,再盖上一层铟罩;方法3,把压碎后的氮化硼放在针状钽勺上。在优化的辉光放电参数下对比了3种不同制样方法对基体信号强度的影响。试验表明:在方法1中,当氮化硼尺寸约为3mm×3mm,厚度小于1mm时,基体11B的信号可达1.8×107 cps;在方法2中,选择铟孔大小合适的铟罩,基体11B的信号可达1.0×107 cps;方法3获得基体信号强度比方法1、方法2高一个数量级。大部分元素在中分辨率下可获得较好的结果,而对于在高分辨率下也较难分离的元素,可选择丰度较低的同位素在中分辨率下进行测定,如Ge选择70Ge+,Se选择82Se+,Cd选择111Cd+,Sn选择119Sn+,Ag选择109Ag+,Pt选择194Pt+。氮化硼中的杂质元素含量可通过样品片中待测元素含量减去来自于铟薄片或钽勺中该元素贡献的含量来计算获得。将样品平行测定5次,相对标准偏差均在20%以内。对于Al、Si、Ti等元素的测定,3种制样方法的测定结果基本一致;方法1、方法2中检测到的In含量较大,使得铟中的Ni、Cu对氮化硼的测定值影响较大;方法3由于钽中Fe、Cu的贡献导致氮化硼中Fe、Cu的检测值较大,但方法3获得的基体信号强度大,可降低部分元素的检出限,如Cr、Mn、Ga、Ge等。综上所述,方法3为优选方法。 相似文献