Detection of constitutive heterodimerization of the integrin Mac-1 subunits by fluorescence resonance energy transfer in living cells

Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to beta(2) subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasma membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we suggested that the conformation of Mac-1 subunits is not affected by the fusion of CFP or YFP and inferred that Mac-1 subunits take different conformation when expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293T cells, respectively. (c) 2006 Elsevier Inc. All rights reserved.

Detection of constitutive homomeric associations of the integrins Mac-1 subunits by fluorescence resonance energy transfer in living cells

Integrins alpha(M)beta(2) plays important role on leukocytes, such as adhesion, migration, phagocytosis, and apoptosis. It was hypothesized that homomeric associations of integrin subunits provide a driving force for integrins activation, and simultaneously inducing the formation of integrins clusters. However, experimental reports on homomeric associations between integrin subunits are still controversial. Here, we proved the homomeric associations of the isolated Mac-1 subunits in living cells using three-channel fluorescence resonance energy transfer (FRET) microscopy and FRET spectra methods. We found that the extent of homomeric associations between beta(2) subunits is higher than alpha(M) subunits. Furthermore, FRET imaging indicated that the extent of homomeric associations of the Mac-1 subunits is higher along the plasma membrane than in the cytoplasm. Finally, we suggested that homomeric associations of the transmernbrane domains or/and cytoplasmic domains may provide the driving force for the formation of constitutive homomeric associations between alpha(M) or beta(2) subunits. (c) 2006 Elsevier Inc. All rights reserved.

工业无线网络化控制系统结构及调度问题研究

无线通信技术在工业网络化控制系统中的应用已成为当今工业领域中的研究热点。然而无线通信技术在恶劣的工业环境中通信质量严重下降，使得现有无线通信技术在工业网络化控制系统中的应用受阻。为此在不断提高和改进无线信号传输性能的同时，更要在控制系统级层面上充分考虑无线通信技术的特点，进行相关系统结构和实时调度机制等方面的改进，以达到使系统产生的通信任务在应用层上得到可靠、实时调度的目的。同时也要从无线网络技术层面上入手，解决上层下达的通信任务在数据链路层中的实时调度问题。要设计适合工业无线网络化控制系统应用特点的MAC协议来有效调度网络所有节点的信息收发，减少高层下达的通信任务在网络中的平均时延，提高吞吐量，进而保证工业无线网络化控制系统的实时性。 因此，本文从工业无线网络化控制系统中的通信任务在数据链路层和应用层上的调度角度出发，以解决、改善工业无线网络化控制系统的实时性和可靠性等性能指标为目的。对工业无线网络化控制系统的结构、应用层调度策略和优化以及数据链路层中的MAC协议进行了系统的研究。主要工作和贡献有以下几点： （1）对工业网络化控制系统的基本问题和引入无线通信技术后产生的问题进行了探讨。在定性分析了工业环境对无线通信的影响及要求的同时，又推导出了无线通信的链路预算模型，并利用Matlab对该预算模型进行了建模，计算出了在工业级噪声指数和工业要求的误码率下，IEEE802.11b和IEEE802.15.4两大主流无线通信技术的链路预算情况及具体的量化数据。 （2）提出了工业无线网络化控制系统的双频二级拓扑结构和多路径传输机制。利用了无线信道特点和路径冗余优势，在系统级上改进了目前无线通信技术在工业网络化控制系统中的应用模式，部分缓解了无线通信技术在工业恶劣环境中的实时性和可靠性问题。为工业无线网络化控制系统结构的设计做了有益的尝试。同时，首次将DS-UWB技术应用于工业网络化控制系统中，设计出了基于该结构的UWB和FF及PROFIBUS的集成方案，为该结构的快速应用提供了有效的途径。 （3）提出了极大容忍无线通信时延的调度算法(GTD)。有效解决了工业无线网络化控制系统中周期性通信任务的实时性问题,部分弥补了因无线网络时延而导致周期性通信任务超时的缺陷。并针对GTD调度算法产生的通信窗口碎片问题进行了优化处理，又提出了面向非周期信息吞吐量最大化的优化调度算法，该算法提高了非周期性通信任务的吞吐量。同时，给出了基于这两种调度算法的非线性规划模型的调度时间表求解方法。 （4）提出了根据通信任务调度时间表进行分配无竞争期的混合CAP和CFP的MAC协议（HAF-MAC）。解决了工业无线网络化控制系统上层下达的周期性通信任务和非周期性通信任务在数据链路层中的实时调度问题，并有效解决了本文提出的应用层调度算法生成的预留时间可能存在浪费的问题。同时，在理论分析了HAF-MAC协议时隙利用率的基础上，提出了基于时隙的HAF-MAC（TSHAF-MAC）协议。TSHAF-MAC协议比HAF-MAC协议提高了时隙利用率，使吞吐量和平均延时得到了进一步改善，进而提高了非周期性通信任务的实时性。最后，对这两个协议进行了仿真建模与验证，进而较系统地解决了工业无线网络化控制系统中的通信任务的实时调度问题。 （5）分别将论文中的第三章和第四章的部分研究成果,在本人博士研究阶段研发的无线智能照明控制系统这一物理实验平台上进行了一定的验证。

双聚焦偏移速度估计与可分表示叠前弹性反演

In exploration seismology, the geologic target of oil and gas reservoir in complex medium request the high accuracy image of the structure and lithology of the medium. So the study of the prestack image and the elastic inversion of seismic wave in the complex medium come to the leading edge. The seismic response measured at the surface carries two fundamental pieces of information: the propagation effects of the medium and the reflections from the different layer boundaries in the medium. The propagation represent the low-wavenumber component of the medium, it is so-called the trend or macro layering, whereas the reflections represent the high-wavenumber component of the medium, it is called the detailed or fine layering. The result of migration velocity analysis is the resolution of the low-wavenumber component of the medium, but the prestack elastic inversion provided the resolution of the high-wavvenumber component the medium. In the dissertation, the two aspects about the migration velocity estimation and the elastic inversion have been studied.Firstly, any migration velocity analysis methods must include two basic elements: the criterion that tell us how to know whether the model parameters are correct and the updating that tell us how to update the model parameters when they are incorrect, which are effected on the properties and efficiency of the velocity estimation method. In the dissertation, a migration velocity analysis method based on the CFP technology has been presented in which the strategy of the top-down layer stripping approach are adapted to avoid the difficult of the selecting reduce .The proposed method has a advantage that the travel time errors obtained from the DTS panel are defined directly in time which is the difference with the method based on common image gather in which the residual curvature measured in depth should be converted to travel time errors.In the proposed migration velocity analysis method, the four aspects have been improved as follow:? The new parameterization of velocity model is provided in which the boundaries of layers are interpolated with the cubic spline of the control location and the velocity with a layer may change along with lateral position but the value is calculated as a segmented linear function of the velocity of the lateral control points. The proposed parameterization is suitable to updating procedure.? The analytical formulas to represent the travel time errors and the model parameters updates in the t-p domain are derived under local lateral homogeneous. The velocity estimations are iteratively computed as parametric inversion. The zero differential time shift in the DTS panel for each layer show the convergence of the velocity estimation.? The method of building initial model using the priori information is provided to improve the efficiency of velocity analysis. In the proposed method, Picking interesting events in the stacked section to define the boundaries of the layers and the results of conventional velocity analysis are used to define the velocity value of the layers? An interactive integrate software environment with the migration velocity analysis and prestack migration is built.The proposed method is firstly used to the synthetic data. The results of velocity estimation show both properties and efficiency of the velocity estimation are very good.The proposed method is also used to the field data which is the marine data set. In this example, the prestack and poststack depth migration of the data are completed using the different velocity models built with different method. The comparison between them shows that the model from the proposed method is better and improves obviously the quality of migration.In terms of the theoretical method of expressing a multi-variable function by products of single-variable functions which is suggested by Song Jian (2001), the separable expression of one-way wave operator has been studied. A optimization approximation with separable expression of the one-way wave operator is presented which easily deal with the lateral change of velocity in space and wave number domain respectively and has good approach accuracy. A new prestack depth migration algorithm based on the optimization approximation separable expression is developed and used to testing the results of velocity estimation.Secondly, according to the theory of the seismic wave reflection and transmission, the change of the amplitude via the incident angle is related to the elasticity of medium in the subsurface two-side. In the conventional inversion with poststack datum, only the information of the reflection operator at the zero incident angles can be used. If the more robust resolutions are requested, the amplitudes of all incident angles should be used.A natural separable expression of the reflection/transmission operator is represented, which is the sum of the products of two group functions. One group function vary with phase space whereas other group function is related to elastic parameters of the medium and geological structure.By employing the natural separable expression of the reflection/transmission operator, the method of seismic wave modeling with the one-way wave equation is developed to model the primary reflected waves, it is adapt to a certain extent heterogeneous media and confirms the accuracy of AVA of the reflections when the incident angle is less than 45'. The computational efficiency of the scheme is greatly high.The natural separable expression of the reflection/transmission operator is also used to construct prestack elastic inversion algorithm. Being different from the AVO analysis and inversion in which the angle gathers formed during the prstack migration are used, the proposed algorithm construct a linear equations during the prestack migration by the separable expression of the reflection/transmission operator. The unknowns of the linear equations are related to the elasticity of the medium, so the resolutions of them provided the elastic information of the medium.The proposed method of inversion is the same as AVO inversion in , the difference between them is only the method processing the amplitude via the incident angle and computational domain.