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Combining Whispering‐Gallery Mode Optical Biosensors with Microfluidics for Real‐Time Detection of Protein Secretion from Living Cells in Complex Media 下载免费PDF全文
Ying‐Jen Chen Chung‐Hsuan Huang Frank Vollmer 《Small (Weinheim an der Bergstrasse, Germany)》2018,14(22)
The noninvasive monitoring of protein secretion of cells responding to drug treatment is an effective and essential tool in latest drug development and for cytotoxicity assays. In this work, a surface functionalization method is demonstrated for specific detection of protein released from cells and a platform that integrates highly sensitive optical devices, called whispering‐gallery mode biosensors, with precise microfluidics control to achieve label‐free and real‐time detection. Cell biomarker release is measured in real time and with nanomolar sensitivity. The surface functionalization method allows for antibodies to be immobilized on the surface for specific detection, while the microfluidics system enables detection in a continuous flow with a negligible compromise between sensitivity and flow control over stabilization and mixing. Cytochrome c detection is used to illustrate the merits of the system. Jurkat cells are treated with the toxin staurosporine to trigger cell apoptosis and cytochrome c released into the cell culture medium is monitored via the newly invented optical microfluidic platform. 相似文献
3.
Dr. Raysa Khan Tareque Dr. Storm Hassell-Hart Dr. Tobias Krojer Dr. Anthony Bradley Dr. Srikannathasan Velupillai Dr. Romain Talon Dr. Michael Fairhead Dr. Iain J. Day Kamlesh Bala Dr. Robert Felix Dr. Paul D. Kemmitt Prof. Paul Brennan Prof. Frank von Delft Dr. Laura Díaz Sáez Prof. Kilian Huber Prof. John Spencer 《ChemMedChem》2020,15(24):2513-2520
Combined photochemical arylation, “nuisance effect” (SNAr) reaction sequences have been employed in the design of small arrays for immediate deployment in medium-throughput X-ray protein–ligand structure determination. Reactions were deliberately allowed to run “out of control” in terms of selectivity; for example the ortho-arylation of 2-phenylpyridine gave five products resulting from mono- and bisarylations combined with SNAr processes. As a result, a number of crystallographic hits against NUDT7, a key peroxisomal CoA ester hydrolase, have been identified. 相似文献
4.
《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
5.
In recent years, various heuristic optimization methods have been developed. Many of these methods are inspired by swarm behaviors in nature, such as particle swarm optimization (PSO), firefly algorithm (FA) and cuckoo optimization algorithm (COA). Recently introduced COA, has proven its excellent capabilities, such as faster convergence and better global minimum achievement. In this paper a new approach for solving graph coloring problem based on COA was presented. Since COA at first was presented for solving continuous optimization problems, in this paper we use the COA for the graph coloring problem, we need a discrete COA. Hence, to apply COA to discrete search space, the standard arithmetic operators such as addition, subtraction and multiplication existent in COA migration operator based on the distance's theory needs to be redefined in the discrete space. Redefinition of the concept of the difference between the two habitats as the list of differential movements, COA is equipped with a means of solving the discrete nature of the non-permutation. A set of graph coloring benchmark problems are solved and its performance is compared with some well-known heuristic search methods. The obtained results confirm the high performance of the proposed method. 相似文献
6.
The paper presents a novel methodology for the rapid identification of the water diffusion coefficients of composite materials. The methodology consists in employing a numerical parametric Proper Generalized Decomposition (PGD) method allowing incorporating the diffusion coefficients among the number of degrees of freedom. Compared to classical identification schemes, often based on Finite Element Method (FEM) iterations, the proposed method allows achieving consistent CPU time gain. The method is general and can be applied when diffusion anomalies take place or when diffusion–reaction coupling must be taken into account, moreover can deal with anisotropic materials. However, for the scope of illustration, in the present case, it is applied to the simple case of “classical diffusion” (Fick’s model with constant boundary conditions) and concerns isotropic materials. 相似文献
7.
《Food Control》2016
Crop-specific GMO matrices of 199 genetically modified (GM) events, comprising 143 GM maize events with 75 genetic elements and 56 cotton events with 45 genetic elements, were developed to screen globally approved GM maize and cotton events. As per the compiled information in the matrix, frequently present genetic elements were identified using GMOSeek algorithm: eight genetic elements ([P-35S] [r-act] [T-35S] [T-nos] [pinII] [pat] [aad-1] [cp4 epsps]) in maize and four ([P-35S] [T-nos] [pat] [nptII]) in cotton. Based on the cost-efficiency, feasibility of plexing and coverage of GM events, maize-specific tetraplex PCR, targeting Cauliflower Mosaic Virus 35S promoter (P-35S), Agrobacterium tumefaciens nos terminator (T-nos), Cauliflower Mosaic Virus 35S terminator (T-35S) and endogenous alcohol dehydrogenase (Adh1) gene, with limit of detection (LOD) up to 0.5% was developed. For screening of GM cotton events, pentaplex PCR, targeting P-35S, T-nos, neomycin phosphotransferase II (nptII), phosphinothricin acetyltransferase (pat) and endogenous stearoyl-ACP desaturase (Sad1) gene, with LOD up to 0.25% was developed. Practical applicability of multiplex PCR assays was confirmed with six maize samples of proficiency testing and eight spiked cotton samples. The reported tetraplex and pentaplex PCR assays could efficiently screen 94% of maize and 93% of cotton events approved globally. The developed GMO matrices in combination with multiplex PCR could facilitate checking the GM status of seed samples or food and feed products, and monitoring for presence of authorized GMOs in food and supply chain. This approach can be easily employed by low resource GM testing laboratories in the developing countries, as the multiplex PCR assays are easy to operate with less time and cost inputs. The GMO matrices being presented herein are based on the current information of approved GM events of maize and cotton, which can be further upgraded by including new approved GM events. As most of the newly approved GM events are stacked versions of already commercialized GM events, the developed multiplex PCR assays could also be employed to screen for their presence. 相似文献
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9.
《Journal of Computer and System Sciences》2016,82(5):690-711
Counting independent sets on bipartite graphs (#BIS) is considered a canonical counting problem of intermediate approximation complexity. It is conjectured that #BIS neither has an FPRAS nor is as hard as #Sat to approximate. We study #BIS in the general framework of two-state spin systems on bipartite graphs. We define two notions, nearly-independent phase-correlated spins and unary symmetry breaking. We prove that it is #BIS-hard to approximate the partition function of any 2-spin system on bipartite graphs supporting these two notions. Consequently, we classify the complexity of approximating the partition function of antiferromagnetic 2-spin systems on bounded-degree bipartite graphs. 相似文献
10.
《Journal of Computer and System Sciences》2016,82(5):739-755
The partner units problem (PUP) is an acknowledged hard benchmark problem for the Logic Programming community with various industrial application fields like surveillance, electrical engineering, computer networks or railway safety systems. Although it is already known for a while that the PUP is NP-complete in its general form, complexity for subproblems reflecting the real problems in industrial fields remained widely unclear so far. In this article we provide all missing complexity results. For the subclass of the PUP that belongs to the complexity class P we present a polynomial-time algorithm and give in-depth algorithmic complexity results. 相似文献