Measuring Receptor/Ligand Interaction at the Single-Bond Level: Experimental and Interpretative Issues

There is increased interest in measuring kinetic rates, lifetimes, and rupture forces of single receptor/ligand bonds. Valuable insights have been obtained from previous experiments attempting such measurements. However, it remains difficult to know with sufficient certainty that single bonds were indeed measured. Using exemplifying data, evidence supporting single-bond observation is examined and caveats in the experimental design and data interpretation are identified. Critical issues preventing definitive proof and disproof of single-bond observation include complex binding schemes, multimeric interactions, clustering, and heterogeneous surfaces. It is concluded that no single criterion is sufficient to ensure that single bonds are actually observed. However, a cumulative body of evidence may provide reasonable confidence. 0 2002 Biomedical Engineering Society.

Notch sensitivity of circular rings subjected to impact loading

'Notch-sensitive regions' have been observed during a series of experimental investigations into the dynamic plastic behaviour and failure of thin-walled metallic radially notched circular rings with are-shaped supports subjected to concentrated impact loads. The experimental results show that the exterior notches at some regions have no effect on the deformation of the rings, but do have effect at the remaining regions. The notch-sensitive region is theoretically determined by using the equivalent structures technique; fairly good agreement has been reached between the simple theory and the experimental results. Both dimensional and theoretical analyses prove that whether a plastic hinge formed or not at the notched section does not depend on the mean radius of the ring and the input kinetic energy. It depends on the weak coefficient of the notched section and the angle of the support. Generally speaking, there are mainly three failure modes for a notched circular ring with are-shaped support under impact loading: Mode I, large inelastic deformation when the notch is outside the sensitive region, in this case the ring deforms as a normal one; Mode II, large inelastic deformation only at some part of the ring and tearing occurred at the notched sections; Mode III, large inelastic deformation and total rupture occurred at the notched sections. It is believed that the present study could assist the understanding of the dynamic behaviour and failure of other kinds of nonstraight components with macroscopic imperfections under impulsive loading.

Singular behaviour near the tip of a sharp V-notch in a power law hardening material

This paper presents an asymptotic analysis of the near-tip stress and strain fields of a sharp V-notch in a power law hardening material. First, the asymptotic solutions of the HRR type are obtained for the plane stress problem under symmetric loading. It is found that the angular distribution function of the radial stress sigma(r) presents rapid variation with the polar angle if the notch angle beta is smaller than a critical notch angle; otherwise, there is no such phenomena. Secondly, the asymptotic solutions are developed for antisymmetric loading in the cases of plane strain and plane stress. The accurate calculation results and the detailed comparisons are given as well. All results show that the singular exponent s is changeable for various combinations of loading condition and plane problem.

Fatigue crack-growth from a circular notch under high-levels of biaxial stress

A series of experiments have been conducted on cruciform specimens to investigate fatigue crack growth from circular notches under high levels of biaxial stress. Two stress levels (Δσ₁= 380 and 560 MPa) and five stress biaxialities (λ=+1.0, +0.5, 0, −0.5 and −1.0; where λ=σ₂/σ₁ were adopted in the fatigue tests in type 316 stainless steel having a monotonic yield strength of 243 MPa. The results reveal that fatigue crack growth rates are markedly influenced by both the stress amplitude and the stress biaxiality. A modified model has been developed to describe fatigue crack growth under high levels of biaxial stress.

Complex stress functions and plastic zone sizes for notch cracks subjected to various loading conditions

In this paper, the conformal mapping method is used to solve the plane problem of an infinite plate containing a central lip-shaped notch subjected to biaxial loading at a remote boundary or a surface uniform pressure on the notch. The stress intensity factors KI and KII are obtained by the derived complex stress functions. The simple analytical expressions can be applied to the situation of cracks originating from a circular or an elliptical notch. The plastic zone sizes for such notch cracks are subsequently evaluated in light of the Dugdale strip yield concept. The results are consistent with available numerical data.

Static and fatigue failure behavior of carbon-fiber reinforced epoxy composite-materials with edge notch

A study of carbon fiber reinforced epoxy composite material with 0° ply or ±45°ply(unnotched or with edge notch) was carried out under static tensile and tension-tensioncyclic loading testing. Static and fatigue behaviour and damage failure modes in unnotched/notched specimens plied in different manners were analysed and compared with each other.A variety of techniques (acoustic emission, two types of strain extensometer, high speed pho-tography, optical microscopy, scanning electron microscope, etc.) were used to examine thedamage of the laminates. Experimental results show that when these carbon/epoxy laminateswith edge notch normal to the direction of the load are axially loaded in static or fatiguetension, the crack does not propagate along the length of notch but is in the interface (fiberdirection). The notch has no substantial effect on the stresses at the unnotched portion. Thedamage failure mechanism is discussed.

Soluble Angiopoietin Receptor Tie-2 In Patients With Acute Myocardial Infarction And Its Detection By Optical Protein-Chip

The Tie-2 receptor has been shown to play a role in angiogenesis in atherosclerosis. The conventional method assaying the level of soluble Tie-2 (sTie-2) was ELISA. However, this method has some disadvantages. The aims of this research are to establish a more simple detection method, the optical protein-chip based on imaging ellipsomtry (OPC-IE) applying to Tie-2 assay. The sTie-2 biosensor surface on silicon wafer was prepared first, and then serum levels of sTie-2 in 38 patients with AMI were measured on admission (day 1), day 2, day 3 and day 7 after onset of chest pain and 41 healthy controls by ELISA and OPC-IE in parallel. Median level of sTie-2 increased significantly in the AMI patients when compared with the controls. Statistics showed there was a significant correlation in sTie-2 results between the two methods (r=0.923, P0.01). The result of this study showed that the level of sTie-2 increased in AMI, and OPC-IE assay was a fast, reliable, and convenient technique to measure sTie-2 in serum.

Parametric Analysis for Monitoring 2D Kinetics of Receptor-Ligand binding

Thermal fluctuation approach is widely used to monitor association kinetics of surface-bound receptor-ligand interactions. Various protocols such as sliding standard deviation (SD) analysis (SSA) and Page's test analysis (PTA) have been used to estimate two-dimensional (2D) kinetic rates from the time course of displacement of molecular carrier. In the current work, we compared the estimations from both SSA and modified PTA using measured data from an optical trap assay and simulated data from a random number generator. Our results indicated that both SSA and PTA were reliable in estimating 2D kinetic rates. Parametric analysis also demonstrated that such the estimations were sensitive to parameters such as sampling rate, sliding window size, and threshold. These results furthered the understandings in quantifying the biophysics of receptor-ligand interactions.

Molecular evolution of CXCR1, a G protein-coupled receptor involved in signal transduction of neutrophils

Human neutrophils are a type of white blood cell, which forms an early line of defense against bacterial infections. Neutrophils are highly responsive to the chemokine, interleukin-8 (IL-8) due to the abundant distribution of CXCR1, one of the IL-8 receptors on the neutrophil cell surface. As a member of the GPCR family, CXCR1 plays a crucial role in the IL-8 signal transduction pathway in neutrophils. We sequenced the complete coding region of the CXCR1 gene in worldwide human populations and five representative nonhuman primate species. Our results indicate accelerated protein evolution in the human lineage, which was likely caused by Darwinian positive selection. The sliding window analysis and the codon-based neutrality test identified signatures of positive selection at the N-terminal ligand/receptor recognition domain of human CXCR1.