Self-consistent theory of reversible ligand binding to a spherical cell

In this article, we study the kinetics of reversible ligand binding to receptors on a spherical cell surface using a self-consistent stochastic theory. Binding, dissociation, diffusion and rebinding of ligands are incorporated into the theory in a systematic manner. We derive explicitly the time evolution of the ligand-bound receptor fraction p(t) in various regimes . Contrary to the commonly accepted view, we find that the well-known Berg-Purcell scaling for the association rate is modified as a function of time. Specifically, the effective on-rate changes non-monotonically as a function of time and equals the intrinsic rate at very early as well as late times, while being approximately equal to the Berg-Purcell value at intermediate times. The effective dissociation rate, as it appears in the binding curve or measured in a dissociation experiment, is strongly modified by rebinding events and assumes the Berg-Purcell value except at very late times, where the decay is algebraic and not exponential. In equilibrium, the ligand concentration everywhere in the solution is the same and equals its spatial mean, thus ensuring that there is no depletion in the vicinity of the cell. Implications of our results for binding experiments and numerical simulations of ligand-receptor systems are also discussed.Comment: 23 pages with 4 figure

Endothelial Cell Capture of Heparin-Binding Growth Factors under Flow

Circulation is an important delivery method for both natural and synthetic molecules, but microenvironment interactions, regulated by endothelial cells and critical to the molecule's fate, are difficult to interpret using traditional approaches. In this work, we analyzed and predicted growth factor capture under flow using computer modeling and a three-dimensional experimental approach that includes pertinent circulation characteristics such as pulsatile flow, competing binding interactions, and limited bioavailability. An understanding of the controlling features of this process was desired. The experimental module consisted of a bioreactor with synthetic endothelial-lined hollow fibers under flow. The physical design of the system was incorporated into the model parameters. The heparin-binding growth factor fibroblast growth factor-2 (FGF-2) was used for both the experiments and simulations. Our computational model was composed of three parts: (1) media flow equations, (2) mass transport equations and (3) cell surface reaction equations. The model is based on the flow and reactions within a single hollow fiber and was scaled linearly by the total number of fibers for comparison with experimental results. Our model predicted, and experiments confirmed, that removal of heparan sulfate (HS) from the system would result in a dramatic loss of binding by heparin-binding proteins, but not by proteins that do not bind heparin. The model further predicted a significant loss of bound protein at flow rates only slightly higher than average capillary flow rates, corroborated experimentally, suggesting that the probability of capture in a single pass at high flow rates is extremely low. Several other key parameters were investigated with the coupling between receptors and proteoglycans shown to have a critical impact on successful capture. The combined system offers opportunities to examine circulation capture in a straightforward quantitative manner that should prove advantageous for biologicals or drug delivery investigations

Effects of receptor clustering on ligand dissociation: Theory and simulations

Receptor-ligand binding is a critical first step in signal transduction and the duration of the interaction can impact signal generation. In mammalian cells, clustering of receptors may be facilitated by heterogeneous zones of lipids, known as lipid rafts. In vitro experiments show that disruption of rafts significantly alters the dissociation of fibroblast growth factor-2 (FGF-2) from heparan sulfate proteoglycans, co-receptors for FGF-2. In this paper, we develop a continuum stochastic formalism in order to (i) study how rebinding affects the dissociation of ligands from a planar substrate, and (ii) address the question of how receptor clustering influences ligand rebinding. We find that clusters reduce the effective dissociation rate dramatically when the clusters are dense and the overall surface density of receptors is low. The effect is much less pronounced in the case of high receptor density and shows non-monotonic behavior with time. These predictions are verified via lattice Monte Carlo simulations. Comparison with experimental results suggests that the theory does not capture the complete biological system. We speculate that additional co-operative mechanisms might be present in order to increase ligand retention, and present one possible ``internal diffusion'' model.Comment: Expanded text and added figures, revised version to appear in Biophys.

Electron probe microanalysis of ion exchange of selected elements between dentine and adhesive restorative materials

The document attached has been archived with permission from the Australian Dental Association. An external link to the publisher’s copy is included.Background: There have been numerous attempts to demonstrate the phenomenon of ion exchange between auto cure glass ionomer cements (GICs) and dentine. The purpose of this study was to employ an electron probe microanalysis (EPMA) technique to examine the interchange of elements between non-demineralized dentine and two types of restorative material, auto cure GICs and a resin composite. Methods: Restorations of auto cure GICs (Riva Fast, Fuji IX Fast, Ketac Molar Quick and Fuji VII) and a bonded composite resin were placed in each of 10 recently extracted human third molar teeth. After two weeks the restorations were sectioned and prepared for EPMA. Percentage weights of calcium, phosphorus aluminum, strontium and fluoride were calculated in the restorations 200μm from the restorative interface and 200μm into the dentine at 5μm intervals. Results: There was evidence of calcium and phosphorus in all five auto cure GICs to a depth of 50μm. Aluminum and strontium ions were also present in dentine except subjacent to Ketac Molar restorations. There was evidence of element transfer into composite resin and resin-bonded dentine. Conclusions: The findings of this paper support the concept of ion exchange as a bonding mechanism between auto cure GIC and dentine. Element penetration into tooth structure and GIC exceeded beyond the “ion exchange layer” observed in scanning electron microscopy studies. Penetration of calcium and phosphorus into composite resin from dentine likely occurred as a result of the self-etching process dissolving calcium and phosphorus and incorporating these elements into the hybrid layer. The presence of Al and Sr ions in dentine were likely to be associated with resin tags extending into the dentine.GM Knight, JM McIntyre, GG Craig and Mulyan

Fluoride content and recharge ability of five glassionomer dental materials

<p>Abstract</p> <p>Background</p> <p>The relationship between fluoride content and fluoride release for glass-ionomer cements is not well understood. The aim of this laboratory study was: to determine the fluoride concentrations at the surfaces of glass-ionomer materials with respect to different storage media and different pH environments; to examine the recharge ability of the materials after NaF immersion; and to assess the morphological changes at the material surfaces using scanning electron microscope and energy dispersive spectroscopic techniques (SEM/EDS).</p> <p>Methods</p> <p>Five glass-ionomer materials, Fuji Triage (FT), Fuji II LC (FII), Fuji VIII (FVIII), Fuji IX GP (FIX), and Ketac N100 (KN), were analyzed in this study. Resin-based fluoride releasing material Helioseal F (HSF) was used as a comparison material. The sample consisted of 120 cured cement disks (n = 20 disks of each tested material, 10 × 1.5 mm). Five disks of each material were stored in 4 different storage media (I- saline, II- acidic solution ph = 2.5, III- acid solution ph = 5.5, IV- NaF solution (c = 500/106). After 7 days, two disks of each material were transferred from media I, II and III to the NaF solution for 3 min. EDS analysis was conducted in 3 randomly selected spots of each experimental disk. SEM was used to determine morphological characteristics of the material surface. Differences between the experimental groups have been analyzed using Student's t-test with the level of significance set at p < 0.001.</p> <p>Results</p> <p>FT showed the highest fluoride content at the surface of the material. The lowest amounts of fluoride ions were detected at the surfaces of the FT disks stored at low pH environments, and this difference was statistically significant (p < 0.001). Glass-ionomers showed significantly higher fluoride concentrations when compared to the HSF (p < 0.001). After immersion in the NaF solution, fluoride concentrations at the surfaces of the disks increased when compared with previous storage media (FT>FVIII>KN>FII>FIX). SEM analysis of the surface morphology revealed numerous voids, cracks and microporosities in all experimental groups, except for KN and HSF. More homogenous material structure with more discrete cracks was observed in samples stored at neutral pH environment, compared to disks stored in acidic solutions.</p> <p>Conclusion</p> <p>The tested materials could be considered as promising dental materials with potential prophylactic characteristics due to their relatively high fluoride content, but also the ability to extensively reabsorb fluoride ions, especially in acidic environments.</p

Cheek Tooth Morphology and Ancient Mitochondrial DNA of Late Pleistocene Horses from the Western Interior of North America: Implications for the Taxonomy of North American Late Pleistocene Equus

Horses were a dominant component of North American Pleistocene land mammal communities and their remains are well represented in the fossil record. Despite the abundant material available for study, there is still considerable disagreement over the number of species of Equus that inhabited the different regions of the continent and on their taxonomic nomenclature. In this study, we investigated cheek tooth morphology and ancient mtDNA of late Pleistocene Equus specimens from the Western Interior of North America, with the objective of clarifying the species that lived in this region prior to the end-Pleistocene extinction. Based on the morphological and molecular data analyzed, a caballine (Equus ferus) and a non-caballine (E. conversidens) species were identified from different localities across most of the Western Interior. A second non-caballine species (E. cedralensis) was recognized from southern localities based exclusively on the morphological analyses of the cheek teeth. Notably the separation into caballine and non-caballine species was observed in the Bayesian phylogenetic analysis of ancient mtDNA as well as in the geometric morphometric analyses of the upper and lower premolars. Teeth morphologically identified as E. conversidens that yielded ancient mtDNA fall within the New World stilt-legged clade recognized in previous studies and this is the name we apply to this group. Geographic variation in morphology in the caballine species is indicated by statistically different occlusal enamel patterns in the specimens from Bluefish Caves, Yukon Territory, relative to the specimens from the other geographic regions. Whether this represents ecomorphological variation and/or a certain degree of geographic and genetic isolation of these Arctic populations requires further study

Chondroitin sulfates and their binding molecules in the central nervous system

Chondroitin sulfate (CS) is the most abundant glycosaminoglycan (GAG) in the central nervous system (CNS) matrix. Its sulfation and epimerization patterns give rise to different forms of CS, which enables it to interact specifically and with a significant affinity with various signalling molecules in the matrix including growth factors, receptors and guidance molecules. These interactions control numerous biological and pathological processes, during development and in adulthood. In this review, we describe the specific interactions of different families of proteins involved in various physiological and cognitive mechanisms with CSs in CNS matrix. A better understanding of these interactions could promote a development of inhibitors to treat neurodegenerative diseases