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Engineering nanostructured probes for sensitive intracellular gene detection. 用于细胞内基因敏感检测的工程纳米结构探针。
Pub Date : 2004-03-01 DOI: 10.3970/MCB.2004.001.023
G. Bao, A. Tsourkas, P. Santangelo
The ability to detect, localize, quantify and monitor the expression of specific genes in living cells in real-time will offer unprecedented opportunities for advancement in molecular biology, disease pathophysiology, drug discovery, and medical diagnostics. However, current methods for quantifying gene expression employ either selective amplification (as in PCR) or saturation binding followed by removal of the excess probes (as in microarrays and in situ hybridization) to achieve specificity. Neither approach is applicable when detecting gene transcripts within living cells. Here we review the recent development in engineering nanostructured molecular probes for gene detection in vivo, describe probe design approaches and its structure-function relations, and discuss the critical issues and challenges in performing living cell gene detection with high specificity, sensitivity and signal-to-background ratio. The underlying biological and biochemical aspects are illustrated.
实时检测、定位、量化和监测活细胞中特定基因表达的能力,将为分子生物学、疾病病理生理学、药物发现和医学诊断领域的进步提供前所未有的机会。然而,目前定量基因表达的方法要么采用选择性扩增(如PCR),要么采用饱和结合,然后去除多余的探针(如微阵列和原位杂交),以达到特异性。这两种方法都不适用于检测活细胞内的基因转录物。本文综述了用于体内基因检测的工程纳米结构分子探针的最新进展,描述了探针的设计方法及其结构-功能关系,并讨论了实现高特异性、高灵敏度和高信本比的活细胞基因检测的关键问题和挑战。潜在的生物学和生物化学方面的说明。
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引用次数: 6
Y.C. "Bert" Fung: the father of modern biomechanics. Y.C.冯伯特:现代生物力学之父。
Ghassan S Kassab
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引用次数: 0
Flaw tolerant bulk and surface nanostructures of biological systems. 生物系统的容错体和表面纳米结构。
Huajian Gao, Baohua Ji, Markus J Buehler, Haimin Yao

Bone-like biological materials have achieved superior mechanical properties through hierarchical composite structures of mineral and protein. Gecko and many insects have evolved hierarchical surface structures to achieve extraordinary adhesion capabilities. We show that the nanometer scale plays a key role in allowing these biological systems to achieve their superior properties. We suggest that the principle of flaw tolerance may have had an overarching influence on the evolution of the bulk nanostructure of bone-like materials and the surface nanostructure of gecko-like animal species. We demonstrate that the nanoscale sizes allow the mineral nanoparticles in bone to achieve optimum fracture strength and the spatula nanoprotrusions in Gecko to achieve optimum adhesion strength. In both systems, strength optimization is achieved by restricting the characteristic dimension of the basic structure components to nanometer scale so that crack-like flaws do not propagate to break the desired structural link. Continuum modeling and atomistic simulations have been conducted to verify the concept of flaw tolerance at nanoscale. A simple tension-shear chain model has been developed to model the stiffness and fracture energy of biocomposites. It is found that, while the problem of low toughness of mineral crystals is alleviated by restricting the crystal size to nanoscale, the problem of low modulus of protein has been solved by adopting a large aspect ratio for the mineral platelets. The fracture energy of biocomposites is found to be proportional to the effective shear strain and the effective shear stress in protein along its path of deformation to fracture. The bioengineered mineral-protein composites are ideally suited for fracture energy dissipation as the winding paths of protein domain unfolding and slipping along protein-mineral interfaces lead to very large effective strain before fracture. The usual entropic elasticity of biopolymers may involve relatively small effective stress and may not be able to ensure simultaneous domain unfolding and interface slipping. Cross-linking mechanisms such as Ca++ induced sacrificial bonds in bone can increase the shear stress in protein and along the protein-mineral interface, effectively converting the behavior of entropic elasticity to one that resembles metal plasticity. The sacrificial bond mechanism not only builds up a large effective stress in protein but also allows protein deformation and interface slipping to occur simultaneously under similar stress levels, making it possible to engineer a very long range of deformation under significant stress in order to maximize energy absorption. Optimization of mineral platelets near theoretical strength is found to be crucial for allowing a large effective stress to be built up in protein via cross-linking mechanisms such as Ca++ induced sacrificial bonds. Similarly, for gecko adhesion, the strength optimization of individual spatulas is found to play a

类骨生物材料通过矿物质和蛋白质的层次化复合结构获得了优异的力学性能。壁虎和许多昆虫进化出了分层的表面结构,以获得非凡的粘附能力。我们表明,纳米尺度在允许这些生物系统实现其优越性能方面起着关键作用。我们认为,缺陷容忍原理可能对类骨材料的体纳米结构和类壁虎动物物种的表面纳米结构的演变产生了总体影响。我们证明了纳米尺度的尺寸可以使骨中的矿物纳米颗粒达到最佳的断裂强度,而壁虎中的刮刀纳米突出物可以达到最佳的粘附强度。在这两种体系中,强度优化都是通过将基本结构部件的特征尺寸限制在纳米尺度来实现的,从而使类裂纹缺陷不会传播而破坏所需的结构环节。通过连续体模型和原子模拟验证了纳米尺度上的缺陷容限概念。建立了一个简单的拉伸-剪切链模型来模拟生物复合材料的刚度和断裂能。研究发现,通过将晶体尺寸限制在纳米尺度,可以缓解矿物晶体韧性低的问题,而采用大长宽比的矿物血小板则可以解决蛋白质模量低的问题。研究发现,生物复合材料的断裂能与蛋白质在变形至断裂过程中的有效剪切应变和有效剪切应力成正比。生物工程的矿物-蛋白质复合材料非常适合断裂能量耗散,因为蛋白质结构域沿蛋白质-矿物界面展开和滑动的缠绕路径导致断裂前的有效应变非常大。生物聚合物通常的熵弹性可能涉及相对较小的有效应力,并且可能无法同时保证域展开和界面滑动。骨中的Ca++诱导的牺牲键等交联机制可以增加蛋白质和蛋白质-矿物界面的剪切应力,有效地将熵弹性行为转化为类似金属塑性的行为。牺牲键机制不仅在蛋白质中建立了一个大的有效应力,而且允许蛋白质变形和界面滑动在相似的应力水平下同时发生,使得在显著应力下设计一个很长的变形范围成为可能,以最大限度地吸收能量。矿物血小板接近理论强度的优化被发现是允许通过交联机制(如Ca++诱导的牺牲键)在蛋白质中建立一个大的有效应力的关键。同样,对于壁虎的粘附,单个刮刀的强度优化在更高层次的粘附能量提升中起着至关重要的作用。
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引用次数: 0
Yuan-Cheng Fung: A scientific giant and a kind man. 冯元成:科学巨人,善良的人。
Pin Tong
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引用次数: 0
Cantilever arrays for multiplexed mechanical analysis of biomolecular reactions. 用于生物分子反应多重力学分析的悬臂阵列。
Pub Date : 2004-01-01 DOI: 10.3970/MCB.2004.001.211
Min Yue, J. Stachowiak, A. Majumdar
Microchips containing arrays of cantilever beams have been used to mechanically detect and quantitatively analyze multiple reactions of DNA hybridization and antigen-antibody binding simultaneously. The reaction-induced deflection of a cantilever beam reflects the interplay between strain energy increase of the beam and the free energy reduction of a reaction, providing an ideal tool for investigating the connection between mechanics and chemistry of biomolecular reactions. Since free energy reduction is common for all reactions, the cantilever array forms a universal platform for label-free detection of various specific biomolecular reactions. A few such reactions and their implications in biology and biotechnology are discussed.
包含悬臂梁阵列的微芯片已被用于同时机械检测和定量分析DNA杂交和抗原抗体结合的多种反应。悬臂梁的反应引起的挠曲反映了梁的应变能增加与反应的自由能减少之间的相互作用,为研究生物分子反应的力学和化学之间的联系提供了理想的工具。由于自由能还原对所有反应都是常见的,因此悬臂阵列形成了各种特定生物分子反应的无标记检测的通用平台。本文讨论了一些这样的反应及其在生物学和生物技术中的意义。
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引用次数: 12
A Stochastic Analysis of a Brownian Ratchet Model for Actin-Based Motility and Integrate-and-Firing Neurons 基于肌动蛋白的运动和整合-放电神经元的布朗棘轮模型的随机分析
Pub Date : 2001-06-20 DOI: 10.3970/mcb.2004.001.267
H. Qian
In recent single-particle tracking (SPT) measurements on {it Listeria monocytogenes} motility {em in vitro}, the actin-based stochastic dynamics of the bacterium movement is analyzed statistically (Kuo and McGrath, 2000). The mean-square displacement (MSD) of the detrended trajectory exhibit a linear behavior; it has been suggested that a corresponding analysis for the Brownian ratchet model (Peskin, Odell, & Oster, 1993) leads to a non-monotonic MSD. A simplified version of the Brownian ratchet, when its motion is limited by the bacterium movement, is proposed and analyzed stochastically. Analytical results for the simple model are obtained and statistical data analysis is investigated. The MSD of the stochastic bacterium movement is a quadratic function while the MSD for the detrended trajectory is shown to be linear. The mean velocity and effective diffusion constant of the propelled bacterium in the long-time limit, and the short-time relaxation are obtained from the MSD analysis. The MSD of the gap between actin and the bacterium exhibits an oscillatory behavior when there is a large resistant force from the bacterium. The stochastic model for actin-based motility is also mathematically equivalent to a model for integrate-and-firing neurons. Hence our mathematical results have applications in other biological problems. For comparison, a continuous formalism of the BR model with great analytical simplicity is also studied.
在最近对体外单核增生李斯特菌运动的单粒子跟踪(SPT)测量中,对细菌运动的基于肌动蛋白的随机动力学进行了统计分析(Kuo和McGrath, 2000)。非趋势轨迹的均方位移(MSD)表现为线性行为;有人提出,对布朗棘轮模型(Peskin, Odell, & Oster, 1993)的相应分析导致非单调MSD。当布朗棘轮的运动受到细菌运动的限制时,提出并随机分析了布朗棘轮的简化版本。给出了简单模型的分析结果,并对统计数据进行了分析。随机细菌运动的MSD是一个二次函数,而非趋势轨迹的MSD是线性的。通过MSD分析,得到了被推进细菌在长时间极限下的平均速度和有效扩散常数,以及短时间弛豫。当有较大的细菌阻力时,肌动蛋白与细菌之间间隙的MSD表现出振荡行为。基于肌动蛋白的运动的随机模型在数学上也等同于整合-发射神经元的模型。因此,我们的数学结果在其他生物学问题上也有应用。为了比较,本文还研究了一种分析简单的连续型BR模型。
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引用次数: 6
期刊
Mechanics & chemistry of biosystems : MCB
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