Quantifying the Effects of Contact Duration, Loading Rate, and Approach Velocity on P-Selectin-PSGL-1 Interactions Using AFM

Kinetics and its regulation by extrinsic physical factors govern selectin-ligand interactions that mediate tethering and rolling of circulating cells on the vessel wall under hemodynamic forces. While the force regulation of off-rate for dissociation of selectin-ligand bonds has been extensively studied, much less is known about how transport impacts the on-rate for association of these bonds and their stability. We used atomic force microscopy (AFM) to quantify how the contact duration, loading rate, and approach velocity affected kinetic rates and strength of bonds of P-selectin interacting with P-selectin glycoprotein ligand I (PSGL-1). We found a saturable relationship between the contact time and the rupture force, a biphasic relationship between the adhesion probability and the retraction velocity, a piece-wise linear relationship between the rupture force and the logarithm of the loading rate, and a threshold relationship between the approach velocity and the rupture force. These results provide new insights into how physical factors regulate receptor-ligand interactions.

Quantifying Cell Binding Kinetics Mediated By Surface-Bound Blood Type B Antigen To Immobilized Antibodies

Cell adhesion is crucial to many biological processes, such as inflammatory responses, tumor metastasis and thrombosis formation. Recently a commercial surface plasmon resonance (SPR)-based BIAcore biosensor has been extended to determine cell binding mediated by surface-bound biomolecular interactions. How such cell binding is quantitatively governed by kinetic rates and regulating factors, however, has been poorly understood. Here we developed a novel assay to determine the binding kinetics of surface-bound biomolecular interactions using a commercial BIAcore 3000 biosensor. Human red blood cells (RBCs) presenting blood group B antigen and CM5 chip bearing immobilized anti-B monoclonal antibody (mAb) were used to obtain the time courses of response unit, or sensorgrams, when flowing RBCs over the chip surface. A cellular kinetic model was proposed to correlate the sensorgrams with kinetic rates. Impacts of regulating factors, such as cell concentration, flow duration and rate, antibody-presenting level, as well as pH value and osmotic pressure of suspending medium were tested systematically, which imparted the confidence that the approach can be applied to kinetic measurements of cell adhesion mediated by surface-bound biomolecular interactions. These results provided a new insight into quantifying cell binding using a commercial SPR-based BIAcore biosensor.

Quantifying the effectiveness of SiO2/Au light trapping nanoshells for thin film poly-Si solar cells

In order to enhance light absorption of thin film poly-crystalline silicon (TF poly-Si) solar cells over a broad spectral range, and quantify the effectiveness of nanoshell light trapping structure over the full solar spectrum in theory, the effective photon trapping flux (EPTF) and effective photon trapping efficiency (EPTE) were firstly proposed by considering both the external quantum efficiency of TF poly-Si solar cell and scattering properties of light trapping structures. The EPTF, EPTE and scattering spectrum exhibit different behaviors depending on the geometric size and density of nanoshells that form the light trapping layer. With an optimum size and density of SiO2/Au nanoshell light trapping layer, the EPTE could reach up to 40% due to the enhancement of light trapping over a broad spectral range, especially from 500 to 800 nm.

中国砂鼠利什曼原虫（Leishmania gerbilli）的细微结构及动质体碱性昼白的细胞化学特性的初步研究

本文以中国砂鼠利会曼原虫（Leishmanina grbilli）为材料，对动质体的细微结构及动质体碱性蛋白的细胞化学特性作了研究。已知真细菌类染色质体中有碱性的HU蛋白与DNA结合着，动质体中是否也有碱性蛋白与DNA相结合，一直是有争议的问题。de Souza等（1978-83）证明锥虫的动质体中确有碱性蛋白存在，但迄今止，国际上还没有关于利会曼原虫动质体碱性蛋白的报导。用热三氯醋酸-偶氮洋红G法、热三氯醋酸-固绿法、碱性比布列希猩法、碳酸氨银反应（AS反应）、经亚硝酸处理后作AS反应和经碱性蛋白不同成份抽提后作AS反应等光学显微细胞化学方法以及酒精钨酸染色（E-PTA）、用乙醇-醋酸固定、DNA酶处理后作E-PTA染色和碳酸氨银反应等电镜水平的细胞化学方法进行了研究。得到了如下的结果：（1）L.gerbilli的动质体中确实含有大量的酸溶性碱性蛋白。（2）对用E—PTA染色后，动质体只有周边着色的现象作了探讨，认为是假期象。动质体中碱性蛋白与DNA的分布是一致的。（3）动质体碱性蛋白对热三氯醋酸（TCA）处理的耐受性比HU蛋白的强比组慢白的弱；动质体碱性蛋白对亚硝酸处理的耐受性和与AS液的亲和力也比组蛋白弱。（4）可以抽提富含精氨酸的组蛋白的80%Alc-0.18NHCl，和可以抽抻富含赖氨酸的组蛋白的10%Alc-0.18NHCl，都不能把动质体碱性昼白全部抽提掉。（5）与真细菌类的HU蛋白相比较。L.gerbilli的动质体的碱性蛋白看来含有较多的精氨酸成份。作超薄切片，用电镜观察了L.gerbilli的质膜及质膜下微管、鞭毛、细胞核、动质体、线粒体、复片层结构等，发现：①与其它利会曼原虫和锥虫的鞭毛基部的结构不同，L.gerbilli的鞭毛基部没有基板存在。②L.gerbilli的复片层结构特别发达，是由膜组成的同心圆或同心椭圆结构。③L.gerbilli的动质体在通常情况下作“棒”状，但我们发现它实际上是一个环状结构。这在细胞膨胀时表现得非常突出。

Quantifying intrinsic specificity: A potential complement to affinity in drug screening

We report here the investigation of a novel description of specificity in protein-ligand binding based on energy landscape theory. We define a new term, intrinsic specificity ratio (ISR), which describes the level of discrimination in binding free energies of the native basin for a protein-ligand complex from the weaker binding states of the same ligand. We discuss the relationship between the intrinsic specificity we defined here and the conventional definition of specificity. In a docking study of molecules with the enzyme COX-2, we demonstrate a statistical correspondence between ISR value and geometrical shapes of the small molecules binding to COX-2. We further observe that the known selective (nonselective) inhibitors of COX-2 have higher (lower) ISR values. We suggest that intrinsic specificity ratio may be a useful new criterion and a complement to affinity in drug screening and in searching for potential drug lead compounds.