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  • 1
    Electronic Resource
    Electronic Resource
    Eine einfache, empfindliche Mikromethode zur Bestimmung der Plasma-Renin-Konzentration beim Menschen (1972)
    Springer
    Journal of molecular medicine 50 (1972), S. 833-840 
    Details
    ISSN: 1432-1440
    Keywords: Plasma renin concentration in humans ; micromethod for plasma renin ; sheep angiotensinogen ; plasma renin concentration in babies ; Plasma-Renin-Konzentration beim Menschen ; Mikromethode zur Plasma-Renin-Bestimmung ; Schaf-Angiotensinogen ; Plasma-Renin-Konzentration bei Säuglingen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Es wird ein einfaches, empfindliches Verfahren zur Bestimmung der Plasma-Renin-Konzentration beim Menschen beshrieben, das auf der von Boucheret al. [Canad. J. Physiol. Pharmacol.45, 881 (1967)] angegebenen Methode beruht. 1,0 ml Plasma, 1.0 ml Dowex 50WX2-(NH4)+ und Schaf-Angiotensinogen im Überschuß — gelöst in 2,0 ml EDTA- und NaN2-haltigem Trisphosphatpuffer — werden 15 Std bei 37°C inkubiert. Die Umsetzung des Human-Renins mit dem Schafsubstrat folgte einer Michaelis-Menten-Kinetik und ergab eine 1,5–2fach höhere Angiotensinausbeute als bei der entsprechenden Reaktion mit homologem Substrat. Die Präzision bei Doppelbestimmungen betrug 7,0±6,2% des kleineren Wertes. Die Angiotensin II-Wiederfindungsrate war in dem untersuchten Bereich von 25–200 ng linear und betrug im Mittel 84,4±10%. Bei gesunden Erwachsenen unter freier Kochsalzzufuhr wurde folgender Normbereich ermittelt: 0,9±0,6 ng Angiotensin/ml/h in Ruhe bzw. 2,1±1,0 ng/ml/h nach aktiver Orthostase. Bei gesunden Säuglingen betrug die Plasma-Renin-Konzentration unter ambulanten Bedingungen 3,4±1,6 ng/ml/h und war damit signifikant höher als die Normalwerte der Erwachsenen in Ruhe und nach Orthostase (P〈0,001).
    Notes: Summary A simple, sensitive method for determination of human plasma renin concentration is described. The procedure is based on the method of Boucheret al. [Canad. J. Physiol. Pharmacol.45, 881 (1967)]. One ml plasma and 1.0 ml Dowex 50WX2-(NH4)+ are incubated for 15 hours at 37°C with 150 mg sheep angiotensinogen in 2.0 ml trisphosphate buffer containing EDTA and NaN3. The reaction of the human renin with the sheep angiotensinogen followed the Michaelis-Menten kinetics and yielded a 1.5 to 2 fold amount of angiotensin compared to the reaction with homologous substrate; the recovery of angiotensin was 84.4±10% being linear in the range studied (25–200 ng angiotensin II). The normal values of 22 healthy adults during a free diet were 0.9±0.6 ng angiotensin/ml/h in recumbent and 2.1±1.0 ng/ml/h in upright posture. During ambulatory conditions the plasma renin concentration in healthy babies was 3.4±1.6 ng/ml/h. These values were significantly higher than those in adults in recumbent and upright posture (p〈0.001).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01496341
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  • 2
    Electronic Resource
    Electronic Resource
    Clusters of Cl− channels in CFTR-expressing Sf9 cells switch spontaneously between slow and fast gating modes (1996)
    Springer
    Pflügers Archiv 432 (1996), S. 528-537 
    Details
    ISSN: 1432-2013
    Keywords: Key words Cl ; channels ; CFTR ; cAMP ; Forskolin ; Insect chloride channels ; Sf9 cells ; Patch clamp
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The Sf9 insect Spodoptora frugiperda cell line was used for heterologous expression of the cloned human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA, or the cloned β-galactosidase gene, using the baculovirus Autographa califonica as the infection vector. Using application of the patch-clamp technique, evidence for functional expression of CFTR was obtained according to the following three criteria. Firstly, whole-cell currents recorded 2 days after infection with CFTR revealed a statistically significant increase of membrane conductance, ≈25 times above that of mock-infected control cells, with the reversal potential of the major current component being governed by the chloride equilibrium potential (E Cl). Secondly, in contrast to uninfected cells and cells infected with β-galactosidase, the membrane conductance to chloride of CFTR-injected cells was stimulated by cytosolic adenosine 3´,5´-cyclic monophosphate (cAMP), which was raised by exposing the cells to 10 μM forskolin. Thirdly, recordings of currents through single channels in excised outside-out membrane patches of CFTR-infected cells revealed channels which were clearly different from the native insect chloride channel. Excised outside-out patches of CFTR-infected and forskolin-stimulated cells exhibited wave-like gating kinetics of well-resolved current transitions. All-point Gaussian distributions revealed contributions from several (five to nine) identical channels. Such channels, in excised outside-out patches, studied with a pipette [Cl−] = 40 mM and a bath [Cl−] = 150 mM, rectified the current in agreement with simple electrodiffusion and with a single-channel Goldman-Hodgkin-Katz permeability, P Cl = 1.34⋅10−14 ± 0.23⋅ 10−14 cm3/s (n = 5), corresponding to a physiological single-channel conductance of 2.8 ± 0.5 pS (V M = E Cl) and a limiting conductance, γ150/150, = 7.7 ± 1.3 pS ([Cl−]Bath = [Cl−]Cell = 150 mM). Currents recorded from multichannel excised outside-out patches could shift from the above mode of resolvable unitary conductance transitions to one which was too fast to reveal the dwell-times of closed and open states. During periods characterized by noisy currents, the variance (σ2) of current fluctuations about their stationary mean value depicted a U-shaped function of membrane potential, with a minimum value at a pipette potential where the chloride current was shown to be zero. Thus, it can be concluded that the current fluctuations are caused by fast gating of channels specific for chloride ions. Switching back and forth between the two gating modes of clusters of chloride channels occurred from moment to moment in excised patches when the membrane potential was held at a constant value indicating cooperative gating as a result of interaction between neighbouring chloride channels.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004240050166
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    Paper (German National Licenses)
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