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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 169 (1999), S. 29-38 
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. (i) We have used a combined patch-clamp and fura-2 fluorescence technique to characterize a nonselective cation channel (NSC) in Ea.hy926 (EA) cells, an endothelial cell line derived from human umbilical vein. (ii) Stimulation with ATP, histamine and bradykinin activated slowly and with a long delay after application of the agonist, a nonselective cation current (I NSC) which is time- and voltage-independent. The permeability sequence for cations was P Na 〉 P Cs 〉〉 P NMDG , P Ca . In the absence of external Ca2+ and at rather high concentrations, La3+ and Gd3+ blocked I NSC . (iii) Single channel analysis revealed that ATP activates in the cell-attached configuration a nonselective cation channel with a conductance of approximately 24 pS and a permeation sequence identical to that of the macroscopic current. The channel activity disappeared after membrane excision. (iv) Activation of NSC required physiological intracellular Ca2+ levels (100 nm or higher). All agonists failed to activate NSC if cytosolic Ca2+ ([Ca2+] i ) was lowered by 10 mm BAPTA. Clamping internal Ca2+ at 1 μm sometimes (8 out of 17 cells) spontaneously activated I NSC in the absence of any additional stimulus. (v) Application of 2,5-di-tert-butylhydroquinone and internal perfusion of inositol 1,4,5-trisphosphate also activated I NSC . The phospholipase C inhibitor, U-73122 inhibited I NSC and the sustained Ca2+ plateau during agonist stimulation whereas the inactive analogue, U-73343 had no effect. (vi) These results indicate NSC may act as a Ca2+ entry pathway in endothelium. [Ca2+] i and inositol 1,4,5-trisphosphate play a role in the activation cascade of NSC, and possibly also store depletion.
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  • 2
    ISSN: 1432-1424
    Keywords: Key words: Endothelial cells — Volume-activated Cl− currents —ICln— Nucleosides — Gossypol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. We have measured the kinetic and pharmacological properties of volume-activated Cl− currents (I Cl,vol) in endothelial cells, and tried to correlate them with those of the already described volume-activated current I Cln. Both conductances show a similar permeability sequence for monovalent anions, and they are blocked by extracellular ATP. In the present report, we demonstrate by Western blot and RT-PCR that cultured endothelial cells from bovine pulmonary artery (CPAE) contain pI Cln. The expression of this protein has been shown to be closely associated with the I Cln current. I Cl,vol showed however, in contrast with I Cln, no striking inactivation at positive potentials. This property is also at variance with that of the volume-activated current related to MDR-1. Activation of I Cl,vol at potentials more negative than −80 mV was not time dependent, which excludes a major contribution of a ClC-2 related current. The antiviral nucleoside analogue AZT (3′-azido-3′-deoxythymidine) inhibited I Cl,vol by 21 ± 2.7% (n = 10), at a concentration of 100 μm. Another antiviral drug, acyclovir (ACV, 9-[(2-hydroxyethoxy)methyl]guanine) blocked I Cl,vol by 27 ± 6.2% at 100 μm (n = 11). Both blocking effects are much smaller than those reported for I Cln. The phenol derivative gossypol, which blocks I Cln-related currents, efficiently inhibited I Cl,vol in CPAE cells (67 ± 2.1% at 1 μm, n = 7, K I = 0.4 μm). The presence of pI Cln in CPAE cells and the similar qualitative pharmacological profile of I Cl,vol and I Cln support the hypothesis that pI Cln is a good molecular candidate for I Cl,vol in endothelial cells. The discrepant kinetic properties may indicate that these time-dependent currents at high positive or negative potentials are not intrinsic properties of the channels, but are caused by time-dependent depletion/accumulation phenomena due to the large amplitudes of these currents.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 163 (1998), S. 97-110 
    ISSN: 1432-1424
    Keywords: Key words: Patch clamp — Ca2+- and voltage-dependence — Regulatory Volume Decrease — Tamoxifen — Niflumic acid — Mg2+
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. A Ca2+-activated (I Cl,Ca) and a swelling-activated anion current (I Cl,vol) were investigated in Ehrlich ascites tumor cells using the whole cell patch clamp technique. Large, outwardly rectifying currents were activated by an increase in the free intracellular calcium concentration ([Ca2+] i ), or by hypotonic exposure of the cells, respectively. The reversal potential of both currents was dependent on the extracellular Cl− concentration. I Cl,Ca current density increased with increasing [Ca2+] i , and this current was abolished by lowering [Ca2+] i to 〈1 nm using 1,2-bis-(o-aminophenoxy)ethane-N,N,N′,N′-tetra-acetic acid (BAPTA). In contrast, activation of I Cl,vol did not require an increase in [Ca2+] i . The kinetics of I Cl,Ca and I Cl,vol were different: at depolarized potentials, I Cl,Ca as activated in a [Ca2+] i - and voltage-dependent manner, while at hyperpolarized potentials, the current was deactivated. In contrast, I Cl,vol exhibited time- and voltage-dependent deactivation at depolarized potentials and reactivation at hyperpolarized potentials. The deactivation of I Cl,vol was dependent on the extracellular Mg2+ concentration. The anion permeability sequence for both currents was I − 〉 Cl− 〉 gluconate. I Cl,Ca was inhibited by niflumic acid (100 μm), 5-Nitro-2-(3-phenylpropylamino)benzoic acid (NPPB, 100 μm) and 4,4′-diisothiocyano-2,2′-stilbenedisulfonic acid (DIDS, 100 μm), niflumic acid being the most potent inhibitor. In contrast, I Cl,vol was unaffected by niflumic acid (100 μm), but abolished by tamoxifen (10 μm). Thus, in Ehrlich cells, separate chloride currents, I Cl,Ca and I Cl,vol, are activated by an increase in [Ca2+] i and by cell swelling, respectively.
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  • 4
    ISSN: 1432-1424
    Keywords: Volume regulation ; Multidrug resistance ; Cancer cells ; Chloride channels ; MDR-1 gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The characteristics of volume-activated chloride currents, drug transport function and levels of P-glycoprotein (PgP) expression were compared between two human chronic erythroleukemia cell lines: a parental (K562) cell line and a derivative obtained by vinblastine selection (K562 VBL400). Parental K562 cells showed no detectable P-glycoprotein expression, measured at the protein level (immunofluorescence labeling with monoclonal antibodies), and had very low levels of MDR-1 mRNA expression (RT-PCR analysis), when compared with levels measured in K562 VBL400. Differences in Pgp-mediated transport were estimated by comparing the rates of Fluo3 accumulation. The higher drug-transport function of K562 VBL400 cells (e.g., lower Fluo3 accumulation) correlated with their elevated levels of MDR-1. The rate of dye transport was sensitive to verapamil but was not affected by the tonicity of the extracellular medium. In contrast to the clear differences in transport function, the characteristics of chloride currents induced by cell swelling were indistinguishable between the two cell lines. Currents measured in the whole-cell configuration were outwardly rectifying, had a higher permeability to iodide than to chloride (SCN− 〉 I− 〉 Cl− 〉 gluconate), were potently blocked by NPPB and were unresponsive to verapamil. The percentage of responding cells and the mean current density were nearly identical in both cell lines. In addition, activation of the volume-sensitive current was not prevented during whole-cell recordings obtained with pipettes containing high concentration of cytotoxic drugs (vincristine or vinblastine). These results do not lend support to the previously reported association between Pgp expression and volume-sensitive chloride channels, and suggest that a different protein is responsible for this type of chloride channel in K562 cells.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 177 (2000), S. 13-22 
    ISSN: 1432-1424
    Keywords: Key words: Endothelium — Volume-regulated anion channels — Open pore properties — Extracellular and intracellular pH — Volume regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. We have investigated the effects of extracellular and intracellular pH on single channel and macroscopic (macropatches) currents through volume-regulated anion channels (VRAC) in endothelial cells. Protonation of extracellular binding sites with an apparent pK of 4.6 increased voltage independent of the single-channel amplitude. Cytosolic acidification had a dual effect on VRAC currents: on the one hand, it increased single channel conductance by ∼20% due to protonation of a group with an apparent pK of 6.5 and a Hill coefficient of 2. On the other hand, it reduced channel activity due to protonation of a group with an apparent pK of 6.3 and a Hill coefficient of 2.1. This dual effect enhances the macroscopic current at a slightly acidic pH but inhibits it at more acidic pH. Cytosolic alkalization also reduced channel activity with a pK of 8.4 and a Hill coefficient of 1.9, but apparently did not affect single-channel conductance. These data show that VRAC channels are maintained in an active state in a narrow pH range around the normal physiological pH and shut down outside this range. They also show that HEPES-buffered pipette solutions do not effectively buffer pH in the vicinity of the VRAC channels.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Applied physics 66 (1998), S. S519 
    ISSN: 1432-0630
    Keywords: PACS: 61.16, 68.35, 68.90
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Physics
    Notes: 2 on Ru(0001) has been studied in the temperature range between 100 K and 300 K by means of low-temperature scanning tunnelling microscopy (STM). At temperatures below 130 K the existence of a molecular precursor state leads to the formation of dense, disordered islands (partly with a (1×1) configuration). Between 160 K and 170 K the onset of diffusion of the chemisorbed O atoms allows a local ordering process in the adsorbate layer. Above 210 K the system reaches a well-ordered equilibrium state, formed by (2×2) and (2×1) structures. From the analysis of the island density versus adsorption temperature data the diffusion energy for the physisorbed precursor state is estimated.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    European biophysics journal 19 (1990), S. 19-23 
    ISSN: 1432-1017
    Keywords: Cardiac sodium channels ; Patch clamp ; Block ; Intracellular magnesium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Currents through single cardiac sodium channels have been measured in inside-out patches from guinea pig ventricular cells. To abolish the fast inactivation, Na channels were modified by DPI 201–106. In symmetrical Na solutions, a diminution of outward sodium currents can be observed that depends on the intracellular magnesium concentration and the membrane potential. Inward currents were not altered by the concentrations of magnesium used (between 0 and 22.5 mmol/1). In Mg free solutions a linear current-voltage relation can also be measured in the range of outward Na currents. At +60 mV (symmetrical Na solutions, single channel conductance 24 pS) a half maximal block of cardiac Na channels by intracellular magnesium was found at 2.1 mmol/l. From the analysis of single channel current-voltage relationships the concentration and voltage-dependent block by intracellular magnesium of cardiac sodium channels could be described as binding of Mg at one site with a K d value of 5.1 mmol/1 at 0 mV. The site is located at an electrical distance of 0.18 from the inside.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    European biophysics journal 15 (1987), S. 117-127 
    ISSN: 1432-1017
    Keywords: Myocardial mouse cells ; voltage clamp ; sodium channels ; inactivation kinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Inactivation of sodium currents is investigated in single myocardial mouse cells by the use of a one suction pipette voltage clamp technique. Semilogarithmic plots of the decay phase of sodium currents show two phases and analysis yields a fast (τ h1) and a slow (τ h2) time constant. At increased depolarization the contribution of τ h2 is decreased. Increasing the temperature from 22.0±0.5°C to 36.5±0.5°C influences τ h1 and τ h2 to the same extent. The Q 10 values were 2.28±0.32 for τ h1 and 2.46±0.35 (mean±SD) for τ h2. Between-50 and-20 mV the time course of current decay is substantially faster than inactivation induced by prepulses. Onset of inactivation by the prepulse protocol needs two time constants (τ c1τ c2) for a satisfactory description. Both decrease steeply with increasing depolarization. For small depolarizations, τ c2 values are in the range of several seconds. Recovery from inactivation by short prepulses (40 ms) could be described by two exponentials (τ r1τ r2). For longer prepulses (1.000 ms) a very slow component, τ r3, was observed indicating a history dependence of inactivation. Delay time constants for the onset of inactivation (τd2) are determined between-50 and-20 mV. The more depolarizing voltages generate smaller delay times (0.55±0.10 ms at-20 mV, mean±SD) but larger deviations from the exponential time course. Delay in recovery from inactivation (time constant τd2) has the value 2.0±0.7 ms (mean±SD) at-80 mV and decreases at more hyperpolarizing potentials. The remaining series resistance at maximum compensation was calculated as 80 kΩ. Its influence on the sensitive delay in the double pulse inactivation is discussed.
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  • 9
    Electronic Resource
    Electronic Resource
    s.l. ; Stafa-Zurich, Switzerland
    Solid state phenomena Vol. 103-104 (Apr. 2005), p. 381-384 
    ISSN: 1662-9779
    Source: Scientific.Net: Materials Science & Technology / Trans Tech Publications Archiv 1984-2008
    Topics: Physics
    Notes: A Post-Etch-Residue (PER) removal process for tank and spray tools has been developed using a new inorganic aqueous based chemistry. The performance of this new type of polymer remover, Inosolv 400 Fotopur®, on process wafers is compared with other inorganic chemistries such as DSP (Dilute Sulphuric acid hydrogen Peroxide) and DSP+, containing traces of HF. Inosolv 400 Fotopur®has improved polymer removal capabilities. Furthermore Inosolv 400 Fotopur® does not show any attack of the metal or dielectric layers and is inorganic based and thusenvironmentally friendly
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 22 (1984), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Defects have been found in a colorimetric assay developed by its inventors for determining rapidly and quantitatively whether strains of Escherichia coli are resistant to the lethality of mammalian serum. Results gained with the colorimetric assay on the serum resistance of E. coli strains bearing colicin V plasmids differed from results gained with an assay in which the rate of growth of the bacterial strains in a medium containing rabbit serum was estimated turbidimetrically. The colorimetric assay was intended by its developers to facilitate studies of the genetics of serum resistance in pathogenic bacteria. When used as originally described, the assay is likely to mislead in such studies.
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