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  • 1
    ISSN: 1432-2013
    Keywords: Ca2+ influx ; Nystatin perforated patchclamp technique ; Fura-2 ; HT29 ; ATP ; Thapsigargin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Indirect evidence has accumulated indicating a voltage dependence of the agonist-stimulated Ca2+ influx into epithelial cells. Manoeuvres expected to depolarise the membrane voltage during agonist stimulation resulted in: (1) a decrease of the sustained phase of the adenosine triphosphate (ATP, 10−5 mol/l)-induced intracellular Ca2+ transient, (2) a reduced fura-2 Mn2+-quenching rate, and (3) prevention of the refilling of the agonist-sensitive store. To quantify the change in intracellular Ca2+ as a function of membrane voltage, we measured simultaneously the intracellular Ca2+ activity ([Ca2+]i) with fura-2 and the electrical properties using the nystatin perforated patch-clamp technique in single HT29 cells. Ca2+ influx was either stimulated by ATP (10−5 mol/l) or thapsigargin (TG, 10−8 mol/l). After [Ca2+]i reached the sustained plateau phase we clamped the membrane voltage in steps of 10 mV in either direction. A stepwise depolarisation resulted in a stepwise reduction of [Ca2+]i. Similarly a stepwise hyperpolarisation resulted in a stepwise increase of [Ca2+]i (ATP: 27.5±10 nmol/l per 10 mV, n=6; TG: 19 ±7.9 nmol/l per 10 mV, n=12). The summarised data show a linear relationship between the Δ fluorescence ratio 340/380 nm change and the applied holding voltage. In unstimulated cells the same voltage-clamp protocol did not change [Ca2+]i (n=9). Under extracellular Ca2+-free conditions [Ca2+]i remained unaltered when changing the membrane voltage. These data provide direct evidence that the Ca2+ influx in epithelial cells is membrane voltage dependent. Our data indicate that small changes in membrane voltage lead to substantial changes in [Ca2+]i. This may be due either to a change of driving force for Ca2+ into the cell, or may reflect voltage-dependent regulation of the respective Ca2+ entry mechanism.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0932
    Keywords: Biomécanique ; Rachis cervical ; Fracture de l'odontoïde ; Stabilité ; Ostéosynthèse de l'odontoïde par vis ; Biomechanics ; Cervical spine ; Odontoid fracture ; Stability ; Odontoid screw fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Fresh odontoid fractures of types II and III and in some cases nonunions of the odontoid can be repaired by direct anterior screw fixation preserving C1–C2 motion. The goal of this experimental study was to investigate the biomechanical stability of the fragment achieved by this direct odontoid osteosynthesis according to Böhler. Sixteen human C1–C2 segments with fractures of type II or type III were biomechanically tested in vitro under standardized conditions. Flexion and extension moments, anterior and posterior shear forces were applied, and the motion of the refixed odontoid fragment relative to C2 was determined. The results show that direct screw fixation of the odontoid under the experimental conditions provides sufficient stability for the dens fragment.
    Notes: Résumé Les fractures fraîches de l'odontoïde de type II et III et certains cas de pseudarthrose de l'odontoïde peuvent être réparés par une ostéosynthèse antérieure directe préservant la mobilité C1–C2. Le but de cette étude expérimentale était d'explorer la stabilité biomécanique permise par cette ostéosynthèse directe de l'odontoïde selon Böhler. 16 segments C1–C2 présentant des fractures de type II et III ont été testés in vitro dans des conditions bien définies. On a appliqué des moments de flexion et d'extension et des forces de cisaillement antérieur et postérieur et l'on a déterminé la mobilité du fragment odontoïdien refixé par rapport à C2. Les résultats ont montré que la fixation directe par vissage de l'odontoïde assure dans les conditions expérimentales une stabilité suffisante au fragment odontoïdien.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0932
    Keywords: Biomécanique ; Rachis cervical ; Fracture de l'odontoïde ; Instabilité atlanto-axoïdienne ; Arthrodèse atlanto-axoïdienne postérieure ; Biomechanics ; Cervical spine ; Odontoid fracture ; Atlantoaxial instability ; Posterior atlantoaxial fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Unstable C1-C2 segments are generally treated surgically. Depending on the indication a direct screw fixation of the odontoid or a C1-C2 arthrodesis is a possible technique. In this experimental in vitro study the three different atlantoaxial fusion techniques by Gallie, Brooks, and Magerl were compared biomechanically. Sixteen human C1-C2 segments with odontoid fractures of type II and III were investigated under standardized conditions. Flexion and extension moments, anterior, and posterior shear forces, left and right torsional moments were applied, and the motion of C1 relative to C2 was determined. The results of this investigation show clearly that the segments treated with the technique by Magerl with transarticular screws achieved the highest stiffness, compared to the wiring methods of Brooks and Gallie. These differences were most evident for posterior shear forces and for torsional moments. For these load conditions the ratio of stiffness Magerl: Brooks: Gallie was about 10:2:1. Significant differences for the plastic deformation of the differently fixed C1-C2 segments were found within the first few load/unload cycles, which give information about the relationship between primary and long-term stability.
    Notes: Résumé L'instabilité C1-C2 est le plus souvent traitée chirurgicalement. Selon l'indication, peuvent être proposées soit une ostéosynthèse par vissage direct de l'odontoïde, soit une arthrodèse C1-C2. Dans cette étude expérimentale in vitro, on a comparé au plan biomécanique, les trois différentes techniques d'arthrodèse atlanto-axoïdiennes, selon Gallie, Brooks et Magerl. 16 segments C1-C2 présentant des fractures de type II et III ont été testés in vitro dans des conditions bien définies. On a appliqué des moments de flexion et d'extension, des forces de cisaillement antérieur et postérieur et des moments de torsion vers la droite et vers la gauche, et l'on a analysé la mobilité de C1-C2. Les résultats de cette investigation ont clairement démontré que les segments traités par la technique de Magerl avec des vis transarticulaires, présentaient la plus grande rigidité, en comparaison des méthodes de cerclage de Brooks et de Gallie. Ces différences étaient plus évidentes dans le cas du cisaillement postérieur et de la torsion. Dans ces conditions de contrainte, le rapport de rigidité Magerl/Brooks/Gallie était de 10/2/1. Des différences significatives de déformation plastique ont été retrouvées entre les différentes fixations C1-C2 au cours des premiers cycles de mise en charge et décharge, qui renseignent sur la relation entre la stabilité primaire et à long terme.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2013
    Keywords: Key words CO2/HCO3 ; NH3/NH4+ ; pHi ; [Ca2+]i ; Fura-2 ; BCECF ; Ca2+ store ; Ca2+ influx ; Inositol 1 ; 4 ; 5-trisphosphate ; Epithelia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The influence of intracellular pH (pHi) on intracellular Ca2+ activity ([Ca2+]i) in HT29 cells was examined microspectrofluorometrically. pHi was changed by replacing phosphate buffer by the diffusible buffers CO2/HCO3 –or NH3/NH4 + (pH 7.4). CO2/HCO3 –buffers at 2,5 or 10% acidified pHi by 0.1, 0.32 and 0.38 pH units, respectively, and increased [Ca2+]i by 8–15 nmol/l. This effect was independent of the extracellular Ca2+ activity and the filling state of thapsigargin-sensitive Ca2+ stores. Removing the CO2/HCO3 –buffer alkalinized pHi by 0.14 (2%), 0.27 (5%), and 0.38 (10%) units and enhanced [Ca2+]i to a peak value of 20, 65, and 143 nmol/l, respectively. Experiments carried out with Ca2+-free solution and with thapsigargin showed that the [Ca2+]i transient was due to release from intracellular pools and stimulated Ca2+ entry. NH3/NH4 + (20 mmol/l) induced a transient intracellular alkalinization by 0.6 pHunits and increased [Ca2+]i to a peak (Δ [Ca2+]i = 164 nmol/l). The peak [Ca2+]i increase was not influenced by removal of external Ca2+, but the decline to basal [Ca2+]i was faster. Neither the phospholipase C inhibitor U73122 nor the inositol 1,4,5-trisphosphate (InsP 3) antagonist theophylline had any influence on the NH3/NH4 +-stimulated [Ca2+]i increase, whereas carbachol-induced [Ca2+]i transients were reduced by more than 80% and 30%, respectively. InsP 3 measurements showed no change of InsP 3 during exposure to NH3/NH4 +, whereas carbachol enhanced the InsP 3 concentration, and this effect was abolished by U73122. The pHi influence on ”capacitative” Ca2+ influx was also examined. An acid pHi attenuated, and an alkaline pHi enhanced, carbachol- and thapsigargin-induced [Ca2+]i influx. We conclude that: (1) an alkaline pHi releases Ca2+ from InsP 3-dependent intracellular stores; (2) the store release is InsP 3 independent and occurs via an as yet unknown mechanism; (3) the store release stimulates capacitative Ca2+ influx; (4) the capacitative Ca2+ influx activated by InsP 3 agonists is decreased by acidic and enhanced by alkaline pHi. The effects of pHi on [Ca2+]i should be of relevance under many physiological conditions.
    Type of Medium: Electronic Resource
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