Fluorescence Polarization as a Functional Parameter in Monitoring Living Cells: Theory and Practice

The use of fluorescence polarization as a functional parameter in monitoring cellular activation calls for the reliable and accurate measurement of the fluorescence intensity and polarization (FI and FP) of microscopic objects. The relevant experimental parameters that enter such measurements are thoroughly discussed. The possibility of executing FP measurements properly by flow-through systems is compared with that of static cytometry. Remarks on the effects of high-power excitation on markers and cells conclude the paper.     

Real-time quantification of protein expression and translocation at individual cell resolution using imaging-dish-based live cell array

Cell populations represent intrinsically heterogeneous systems with a high level of spatiotemporal complexity. Monitoring and understanding cell-to-cell diversity is essential for the research and application of intra- and interpopulation variations. Optical analysis of live cells is challenging since both adherent and nonadherent cells change their spatial location. However, most currently available single-cell techniques do not facilitate treatment and monitoring of the same live cells over time throughout multistep experiments. An imaging-dish-based live cell array (ID-LCA) has been developed and produced for cell handling, culturing, and imaging of numerous live cells. The dish is composed of an array of pico scale cavities—pico wells (PWs) embossed on its glass bottom. Cells are seeded, cultured, treated, and spatiotemporally measured on the ID-LCA, while each cell or small group of cells are locally constrained in the PWs. Finally, predefined cells can be retrieved for further evaluation. Various types of ID-LCAs were used in this proof-of-principle work, to demonstrate on-ID-LCA transfection of fluorescently tagged chimeric proteins, as well as the detection and kinetic analysis of their induced translocation. High variability was evident within cell populations with regard to protein expression levels as well as the extent and dynamics of protein redistribution. The association of these parameters with cell morphology and functional parameters was examined. Both the new methodology and the device facilitate research of the translocation process at individual cell resolution within large populations and thus, can potentially be used in high-throughput fashion. Graphical Abstract

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Translation-invariant and positive-homogeneous risk measures and optimal portfolio management in the presence of a riskless component

Risk portfolio optimization, with translation-invariant and positive-homogeneous risk measures, leads to the problem of minimizing a combination of a linear functional and a square root of a quadratic functional for the case of elliptical multivariate underlying distributions.This problem was recently treated by the authors for the case when the portfolio does not contain a riskless component. When it does, however, the initial covariance matrix Σ becomes singular and the problem becomes more complicated. In the paper we focus on this case and provide an explicit closed-form solution of the minimization problem, and the condition under which this solution exists. The results are illustrated using data of 10 stocks from the NASDAQ Computer Index.     

单畴Ni球观测及NicoreNiOshell的铁磁/反铁磁耦合研究

 使用微波辅助聚合方法制备了单分散单畴Ni纳米球，由MFM发现，尺度分布在100～180 nm的Ni球的一个相关特征是条型磁畴结构。用XRD、TEM、XPS以及EDAX测量了由Ni球进一步制备的Ni_coreNiO_shell高度球型纳米结构。用VSM 和SQUID进一步讨论了其铁磁/反铁磁界面耦合效应，估算了交换耦合场与粒子尺寸的关系。     

Inhibition of Cysteine Proteases by 6,6′-Dihydroxythiobinupharidine (DTBN) from Nuphar lutea

The specificity of inhibition by 6,6′-dihydroxythiobinupharidine (DTBN) on cysteine proteases was demonstrated in this work. There were differences in the extent of inhibition, reflecting active site structural-steric and biochemical differences. Cathepsin S (IC₅₀ = 3.2 μM) was most sensitive to inhibition by DTBN compared to Cathepsin B, L and papain (IC₅₀ = 1359.4, 13.2 and 70.4 μM respectively). DTBN is inactive for the inhibition of M^pro of SARS-CoV-2. Docking simulations suggested a mechanism of interaction that was further supported by the biochemical results. In the docking results, it was shown that the cysteine sulphur of Cathepsin S, L and B was in close proximity to the DTBN thiaspirane ring, potentially forming the necessary conditions for a nucleophilic attack to form a disulfide bond. Covalent docking and molecular dynamic simulations were performed to validate disulfide bond formation and to determine the stability of Cathepsins-DTBN complexes, respectively. The lack of reactivity of DTBN against SARS-CoV-2 M^pro was attributed to a mismatch of the binding conformation of DTBN to the catalytic binding site of M^pro. Thus, gradations in reactivity among the tested Cathepsins may be conducive for a mechanism-based search for derivatives of nupharidine against COVID-19. This could be an alternative strategy to the large-scale screening of electrophilic inhibitors.     

A self-powered, one-step chip for rapid, quantitative and multiplexed detection of proteins from pinpricks of whole blood

We describe an automated, self-powered chip based on lateral flow immunoassay for rapid, quantitative, and multiplex protein detection from pinpricks of whole blood. The device incorporates on-chip purification of blood plasma by employing inertial forces to focus blood cells away from the assay surface, where plasma proteins are captured and detected on antibody "barcode" arrays. Power is supplied from the capillary action of a piece of adsorbent paper, and sequentially drives, over a 40 minute period, the four steps required to capture serum proteins and then develop a multiplex immunoassay. An 11 protein panel is assayed from whole blood, with high sensitivity and high reproducibility. This inexpensive, self-contained, and easy to operate chip provides a useful platform for point-of-care diagnoses, particularly in resource-limited settings.     

Graphical balanced allocations and the (1 + β)‐choice process

Suppose m balls are sequentially thrown into n bins where each ball goes into a random bin. It is well‐known that the gap between the load of the most loaded bin and the average is , for large m. If each ball goes to the lesser loaded of two random bins, this gap dramatically reduces to independent of m. Consider a constrained setting where not all pairs of bins can be sampled. We are given a graph where each node corresponds to a bin. The process sequentially samples an edge from the graph and places a ball in the lesser loaded of its endpoints. We show the gap is at most where σ is the edge expansion of the graph. Our results extend naturally to the hypergraph version of this question. Our technique involves a tight analysis of what we call the “‐choice” process for some parameter : each ball goes to a random bin with probability and the lesser loaded of two random bins with probability β. For this process we show that the gap is , irrespective of m. Moreover the gap stays at in the weighted case for a large class of weight distributions. No non‐trivial bounds were previously known in the weighted case, even for the 2‐choice case. © 2014 Wiley Periodicals, Inc. Random Struct. Alg., 47, 760–775, 2015