7th Drug hypersensitivity meeting: part two

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ATP release via anion channels

ATP serves not only as an energy source for all cell types but as an ‘extracellular messenger-for autocrine and paracrine signalling. It is released from the cell via several different purinergic signal efflux pathways. ATP and its Mg2+ and/or H+ salts exist in anionic forms at physiological pH and may exit cells via some anion channel if the pore physically permits this. In this review we survey experimental data providing evidence for and against the release of ATP through anion channels. CFTR has long been considered a probable pathway for ATP release in airway epithelium and other types of cells expressing this protein, although non-CFTR ATP currents have also been observed. Volume-sensitive outwardly rectifying (VSOR) chloride channels are found in virtually all cell types and can physically accommodate or even permeate ATP4- in certain experimental conditions. However, pharmacological studies are controversial and argue against the actual involvement of the VSOR channel in significant release of ATP. A large-conductance anion channel whose open probability exhibits a bell-shaped voltage dependence is also ubiquitously expressed and represents a putative pathway for ATP release. This channel, called a maxi-anion channel, has a wide nanoscopic pore suitable for nucleotide transport and possesses an ATP-binding site in the middle of the pore lumen to facilitate the passage of the nucleotide. The maxi-anion channel conducts ATP and displays a pharmacological profile similar to that of ATP release in response to osmotic, ischemic, hypoxic and salt stresses. The relation of some other channels and transporters to the regulated release of ATP is also discussed

Voltage-dependent anion-selective channel (VDAC) interacts with the dynein light chain Tctex 1 and the heat-shock protein PBP74

The voltage-dependent anion-selective channel 1 (VDAC1), i.e. eukaryotic porin, functions as a channel in membranous structures as described for the outer mitochondrial membrane. the cell membrane, endosomes, caveolae, the sarcoplasmatic reticulum, synaptosomes. and post-synaptic density fraction. The identification of VDAC1 interacting proteins may be a promising approach for better understanding the biological context and function of the channel protein. In this study human VDAC1 was used as a bait protein in a two-hybrid screening, which is based on the Sos recruitment system (SRS). hVDAC1 interacts with the dynein light chain Tctex-1 and the heat-shock protein peptide-binding protein 74 (PBP74)/mitochondrial heat-shock protein 70 (mtHSP70)/glucose- regulated protein 75 (GRP75)/mortalin in vivo. Both interactions were confirmed by overlay-assays using recombinant partner proteins and purified hVDAC1. Indirect immunofluorescence on HeLa cells indicates a co-localisation of hVDAC1 with the dynein light chain and the PBP74. In addition, HeLa cells were transfected transiently with enhanced green fluorescent protein (EGFP)-hVDAC1 fusion proteins, which also clearly co-localise with both proteins. The functional relevance of the identified protein interactions was analysed in planar lipid bilayer (PLB) experiments. In these experiments both recombinant binding partners altered the electrophysiological properties of hVDAC1, While rTctex-1 increases the voltage-dependence of hVDAC1 slightly, the rPBP74 drastically minimises the voltage-dependence, indicating a modulation of channel properties in each case. Since the identified proteins are known to be involved in the transport or processing of proteins, the results of this study represent additional evidence of membrane-associated trafficking of the voltage-dependent anion-selective channel 1. (C) 2002 Elsevier Science Ltd. All rights reserved

Endosomes: another extra-mitochondrial location of type-1 porin/voltage-dependent anion-selective channels (VDAC)

Endocytotic vesicles (EV) isolated from rat renal cortex were subjected to SDS-polyacrylamide gel electrophoresis and Western blotting. A monoclonal antibody against human type-1 porin (31 kDa) detected a strong band of 31 kDa. The same antibody has been used as the primary antibody in indirect immunocytochemistry. Light microscopy of cryostat sections of rat renal cortex showed a heavy staining of EV underneath the brush-border membrane. Electron microscopy was performed by ”preembedding immunogold staining” of rat renal cortex, the sections of which showed an extensive labelling of EV with gold particles. These results demonstrate that the expression of type-1 porin is not restricted to outer mitochondrial membranes. The biological function of endosomal type-1 porin has as yet to be ascertained

Fast variational alignment of non-flat 1D displacements for applications in neuroimaging

Background:In the context of signal analysis and pattern matching, alignment of 1D signals for the comparison of signal morphologies is an important problem. For image processing and computer vision, 2D optical flow (OF) methods find wide application for motion analysis and image registration and variational OF methods have been continuously improved over the past decades.New method:We propose a variational method for the alignment and displacement estimation of 1D signals. We pose the estimation of non-flat displacements as an optimization problem with a similarity and smoothness term similar to variational OF estimation. To this end, we can make use of efficient optimization strategies that allow real-time applications on consumer grade hardware.Results:We apply our method to two applications from functional neuroimaging: The alignment of 2-photon imaging line scan recordings and the denoising of evoked and event-related potentials in single trial matrices. We can report state of the art results in terms of alignment quality and computing speeds.Existing methods:Existing methods for 1D alignment target mostly constant displacements, do not allow native subsample precision or precise control over regularization or are slower than the proposed method.Conclusions:Our method is implemented as a MATLAB toolbox and is online available. It is suitable for 1D alignment problems, where high accuracy and high speed is needed and non-constant displacements occur

Intention concepts and brain-machine interfacing

Intentions, including their temporal properties and semantic content, are receiving increased attention, and neuroscientific studies in humans vary with respect to the topography of intention-related neural responses. This may reflect the fact that the kind of intentions investigated in one study may not be exactly the same kind investigated in the other. Fine-grained intention taxonomies developed in the philosophy of mind may be useful to identify the neural correlates of well-defined types of intentions, as well as to disentangle them from other related mental states, such as mere urges to perform an action. Intention-related neural signals may be exploited by brain-machine interfaces (BMIs) that are currently being developed to restore speech and motor control in paralyzed subjects. Such BMI devices record the brain activity of the agent, interpret (‘decode’) the agent’s intended action, and send the corresponding execution command to an artificial effector system, e.g., a computer cursor or a robotic arm. In the present paper, we evaluate the potential of intention concepts from philosophy of mind to improve the performance and safety of BMIs based on higher-order, intention-related control signals. To this end, we address the distinction between future-, present-directed, and motor intentions, as well as the organization of intentions in time, specifically to what extent it is sequential or hierarchical. This has consequences as to whether these different types of intentions can be expected to occur simultaneously or not. We further illustrate how it may be useful or even necessary to distinguish types of intentions exposited in philosophy, including yes- vs. no-intentions and oblique vs. direct intentions, to accurately decode the agent’s intentions from neural signals in practical BMI applications