210 research outputs found
A Potts Neuron Approach to Communication Routing
A feedback neural network approach to communication routing problems is
developed with emphasis on Multiple Shortest Path problems, with several
requests for transmissions between distinct start- and endnodes. The basic
ingredients are a set of Potts neurons for each request, with interactions
designed to minimize path lengths and to prevent overloading of network arcs.
The topological nature of the problem is conveniently handled using a
propagator matrix approach. Although the constraints are global, the
algorithmic steps are based entirely on local information, facilitating
distributed implementations. In the polynomially solvable single-request case
the approach reduces to a fuzzy version of the Bellman-Ford algorithm. The
approach is evaluated for synthetic problems of varying sizes and load levels,
by comparing with exact solutions from a branch-and-bound method. With very few
exceptions, the Potts approach gives legal solutions of very high quality. The
computational demand scales merely as the product of the numbers of requests,
nodes, and arcs.Comment: 10 pages LaTe
High-Speed Quantum Dot Tracking in Plasma Membranes Reveals Short-Lived Small-Size Corralled Diffusion
Bulk and micropatterned conjugation of extracellular matrix proteins to characterized polyacrylamide substrates for cell mechanotransduction assays
Increasing numbers of cell mechanotransduction studies are currently utilizing elastic substrates fabricated from polyacrylamide in the form of thin gels. Their versatility depends on the ability to ensure the appropriate gel stiffness and control the uniformity and geometry of extracellular matrix protein coating of the gel. Beginning with a brief quantitative emphasis on the elastic properties of polyacrylamide gels, we present an inexpensive and highly reproducible method for uniform coating with a wide variety of extracellular matrix proteins. We used a reducing agent, hydrazine hydrate, to modify nonreactive amide groups in polyacrylamide to highly reactive hydrazide groups that can form covalent bonds with aldehyde or ketone groups in oxidized proteins. This simple conjugation method overcomes the limitations of previously used photoactivatable cross-linkers: nonuniform coating due to nonuniformity of irradiation and technically challenging procedures for micropatterning. As demonstrated in our study of cell polarity during constrained migration, this conjugation method is especially effective in gel micropatterning by manual microcontact printing of protein patterns as small as 5 microm and enables numerous studies of constrained cell attachment and migration that were previously unfeasible due to high cost or difficulty in controlling the protein coating
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
Journal Staff
Cells move along surfaces both as single cells and multi-cellular units. Recent research points toward pivotal roles for water flux through aquaporins (AQPs) in single cell migration. Their expression is known to facilitate this process by promoting rapid shape changes. However, little is known about the impact on migrating epithelial sheets during wound healing and epithelial renewal. Here, we investigate and compare the effects of AQP9 on single cell and epithelial sheet migration. To achieve this, MDCK-1 cells stably expressing AQP9 were subjected to migration assessment. We found that AQP9 facilitated cell locomotion at both the single and multi-cellular level. Furthermore, we identified major differences in the monolayer integrity and cell size upon expression of AQP9 during epithelial sheet migration, indicating a rapid volume-regulatory mechanism. We suggest a novel mechanism for epithelial wound healing based on AQP-induced swelling and expansion of the monolayer.Funding Agencies|Swedish Research Council for Medicine and Health|2007-34832009-66492010-3045|</p
Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS
Towards Truly “Global” Near Infrared Calibrations for Protein and Neutral Detergent Fibre in Dried Ground Forages
Over the past five years, Foss and DeLaval have sponsored the activities of a group of forage analysts with the aim of developing global Near Infrared (NIR) calibrations for parameters that are important in ruminant nutrition. The approach adopted has been based on the amalgamation of historical databases from centres worldwide and calibrations for protein and neutral detergent fibre (NDF) in dried ground forages have been developed based on databases that currently comprise approximately 30,000 records. Protein and NDF, while not the most important parameters in ruminant nutrition, were chosen for the initial calibration development exercise because of the amount of data available and because the methodologies adopted by different laboratories worldwide were relatively uniform. The aim was to create calibrations that would work for any forage type in any area of the world. Over the past two years, several trials have been carried out worldwide comparing the performance of global calibrations with the performance of locally developed calibrations for indigenous forages and based on reference values from local laboratories
Anti-human CD73 monoclonal antibody inhibits metastasis formation in human breast cancer by inducing clustering and internalization of CD73 expressed on the surface of cancer cells
High photon count rates improve the quality of super-resolution fluorescence fluctuation spectroscopy
Probing the diffusion of molecules has become a routine measurement across the life sciences, chemistry and physics. It provides valuable insights into reaction dynamics, oligomerisation, molecular (re-)organisation or cellular heterogeneities. Fluorescence correlation spectroscopy (FCS) is one of the widely applied techniques to determine diffusion dynamics in two and three dimensions. This technique relies on the temporal autocorrelation of intensity fluctuations but recording these fluctuations has thus far been limited by the detection electronics, which could not efficiently and accurately time-tag photons at high count rates. This has until now restricted the range of measurable dye concentrations, as well as the data quality of the FCS recordings, especially in combination with super-resolution stimulated emission depletion (STED) nanoscopy. Here, we investigate the applicability and reliability of (STED-)FCS at high photon count rates (average intensities of more than 1 MHz) using novel detection equipment, namely hybrid detectors and real-time gigahertz sampling of the photon streams implemented on a commercial microscope. By measuring the diffusion of fluorophores in solution and cytoplasm of live cells, as well as in model and cellular membranes, we show that accurate diffusion and concentration measurements are possible in these previously inaccessible high photon count regimes. Specifically, it offers much greater flexibility of experiments with biological samples with highly variable intensity, e.g. due to a wide range of expression levels of fluorescent proteins. In this context, we highlight the independence of diffusion properties of cytosolic GFP in a concentration range of approx. 0.01-1 µm. We further show that higher photon count rates also allow for much shorter acquisition times, and improved data quality. Finally, this approach also pronouncedly increases the robustness of challenging live cell STED-FCS measurements of nanoscale diffusion dynamics, which we testify by confirming a free diffusion pattern for a fluorescent lipid analogue on the apical membrane of adherent cells. © The Author(s). Published by IOP Publishing Ltd
- …