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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 639 (1991), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 588 (1990), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 779 (1996), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: There have been only a few studies of how allergens cross the airway epithelium to cause allergic sensitization. House dust mite fecal pellets (HDMFP) contain several proteolytic enzymes. Group 1 allergens are cysteine peptidases, whilst those of groups 3, 6 and 9 have catalytic sites indicative of enzymes that mechanistically behave as serine peptidases. We have previously shown that the group 1 allergen Der p 1 leads to cleavage of tight junctions (TJs), allowing allergen delivery to antigen presenting cells.In this study we determined whether HDMFP serine peptidases similarly compromise the airway epithelium by attacking TJs, desmosomes and adherens junctions.Experiments were performed in monolayers of MDCK, Calu-3 or 16HBE14o-epithelial cells. Cell junction morphology was examined by 2-photon molecular excitation microscopy and digital image analysis. Barrier function was measured as mannitol permeability. Cleavage of cell adhesion proteins was studied by immunoblotting and mass spectrometry.HDMFP serine peptidases led to a progressive cleavage of TJs and increased epithelial permeability. Desmosomal puncta became more concentrated. Cleavage of TJs involved proteolysis of the TJ proteins, occludin and ZO-1. This was associated with activation of intracellular proteolysis of ZO-1. In contrast to occludin, E-cadherin of adherens junctions was cleaved less extensively. Although Calu-3 and 16HBE14o-cells expressed tethered ligand receptors for serine peptidases, these were not responsible for transducing the changes in TJs.HDMFP serine peptidases cause cleavage of TJs. This study identifies a second general class of HDM peptidase capable of increasing epithelial permeability and thereby creating conditions that would favour transepithelial delivery of allergens.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 398 (1983), S. 165-168 
    ISSN: 1432-2013
    Keywords: Photoprotein ; Light ; Aequorin ; Photomultiplier tubes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Bioluminescent indicators have found many uses in, for example, the detection of ATP and free calcium levels. However, such probes often emit only very low levels of light and it is important to optimise the efficiency of the system used to detect this light. We describe some of the problems encountered in using photomultiplier tubes for detecting low levels of light, and some ways of overcoming these problems. We have developed a versatile photomultiplier light detection system which is both efficient and physically small. This system is described and details of its fabrication are given.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2013
    Keywords: Chamber ; Perfusion ; Superfusion ; Bath ; Voltage-clamp ; Single cell ; Patch-clamp ; Heart
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We describe a simple and inexpensive experimental chamber that is designed to overcome several problems encountered when doing electrical and optical studies on single cells or on isolated membrane patches. The bath is small enough to fit on the stage of a standard inverted microscope. It includes a novel solution level-detector, the output of which is used to actively control the level of solution in the experimental chamber. A Peltier-effect device is located adjacent to the flow-chamber and heats or cools the inflowing solution. Solutions can be rapidly switched using two electrically actuated microvalves. The attraction of this system is that, with appropriately quiet power supplies, not only is the bath solution-level held at a fixed height, but the temperature of the bathing solution can also be set over a wide temperature range (minimum range is 15 to 45°C), and solutions can be rapidly changed. All of the construction details are supplied as are appropriate electrical circuits. Without modification, the chamber can be used for applications as diverse as fluorescence microscopy of living cells, time-lapse photomicroscopy and single-cell motion detection as well as single cell voltage-clamp and isolated membrane patch-clamp. With simple modification the system can be adapted for use in experiments on multicellular preparations.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 432 (1996), S. 555-561 
    ISSN: 1432-2013
    Keywords: Key words Microscopy ; Confocal microscope ; Laser ; Two-photon ; Fluorescence ; Imaging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The construction of a two-photon/confocal microscope system is described in detail. For two-photon illumination, a Ti:sapphire modelocked laser generating 62-fs pulses at 715 nm was used. The effect of the optical train on illumination pulse width was examined and the observed increase in pulse duration was almost completely removed by the addition/adjustment of a prism compressor system. The imaging capabilities of the two-photon microscope are demonstrated and it is shown that the imaging performance of the two-photon microscope is similar to that of a conventional confocal microscope. With two-photon illumination, the resolution (full width at half-maximum intensity) was 0.42 μm (x–y) and 0.81 μm axially, while with single-photon illumination (at 488 nm in the same instrument with a confocal pinhole detector) the resolution was 0.3 μm (x–y) and 0.75 μm axially. The results are discussed with regard to the general problem of femtosecond pulse distortion in an optical system and a simple procedure for optimal pulse restoration is described.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 428 (1994), S. 415-417 
    ISSN: 1432-2013
    Keywords: Heart ; Cardiac muscle ; Contraction ; E-C coupling ; Calcium channels ; Ryanodine receptors ; Intracellular calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This paper examines the [Ca2+]i transient in isolated rat heart cells using a laser scanning confocal microscope and the calcium indicator fluo-3. We find that the depolarization-evoked [Ca2+]i transient is activated synchronously near the surface and in the middle of the heart cell with similar kinetics of activation. The time of rise of the transient did not depend on whether the sarcoplasmic reticulum (SR) Ca-release was abolished (by thapsigargin and ryanodine). The synchrony of activation and the similarity of levels of [Ca2+]i at the peripheral and deeper myoplasm (regardless of the availability of SR Ca-release) shows that sarcolemmal Ca channels and SR Ca-release channels are distributed throughout the rat heart cell and that the propagation of the action potential into the interior of the cell is rapid. In addition, the activation of calcium release from the SR by CICR is rapid (≪2 ms) when compared to the time-course of calcium influx via the sarcolemmal Ca channel.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2013
    Keywords: Key words Na-Ca exchange ; Na-K pump ; Calcium ; Sodium ; Cardiac myocytes ; Heart
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Measurement of Na-Ca exchange activity was used to examine subsarcolemmal sodium levels ([Na+]s) in single, voltage-clamped guinea-pig cardiac myocytes while Na-K pump activity was modulated pharmacologically. Changes in Nas were evaluated from phase-plane analysis of the changes in intracellular calcium, measured using the fluorescent indicators Fura-red and Fluo-3. Activation of β-adrenoceptors with 1 μM isoprenaline resulted in activation of the cAMP-dependent chloride current, but had no effect on the calcium transient mediated via the Na-Ca exchanger, regardless of whether the Na-K pump was active or inhibited (with strophanthidin). The ability of Na-Ca exchange activity to report [Na+]s was demonstrated by the effect of changing the extracellular rubidium concentration from 1 to 5.4 mM to modulate Na-K pump activity. We suggest that β-adrenergic stimulation does not directly affect either the Na-K pump or the Na-Ca exchanger and that the Na-Ca exchanger can be used as a sensitive indicator of changes in [Na+]s and Na-K pump activity.
    Type of Medium: Electronic Resource
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