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101.
Ca2+ oscillations induced by hormonal stimulation of individual fura-2-loaded hepatocytes 总被引:10,自引:0,他引:10
T Kawanishi L M Blank A T Harootunian M T Smith R Y Tsien 《The Journal of biological chemistry》1989,264(22):12859-12866
Isolated rat hepatocytes were loaded with the Ca2+ indicator fura-2 to measure cytosolic free Ca2+ concentrations ([Ca2+]i) in individual cells by digital ratio imaging microscopy. Stimulation with 0.1 nM vasopressin, 0.5 microM phenylephrine, or 0.5 microM ATP caused repetitive spikes of high [Ca2+]i in a high percentage of cells, in agreement with Woods et al. (Woods, N. M., Cuthbertson, K. S. R., and Cobbold, P. H. (1986) Nature 319, 600-602), but unlike the results of Monck et al. (Monck, J. R., Reynolds, E. E., Thomas, A. P., and Williamson, J. R. (1988) J. Biol. Chem. 263, 4569-4575). Reduction in extracellular [Ca2+] decreased the frequency but not the amplitude of the spikes, suggesting that the spikes result from dumping of intracellular stores and that the entry of extracellular Ca2+ affects only the rate of replenishment of those stores. Membrane depolarization failed to elevate [Ca2+]i and had an effect similar to removal of extracellular Ca2+ in decreasing the frequency of agonist-evoked [Ca2+]i oscillations or inhibiting them altogether, arguing against any significant role for voltage-operated Ca2+ channels. 相似文献
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Response of fishes and aquatic habitats to sand-bed stream restoration using large woody debris 总被引:4,自引:1,他引:3
Effects of habitat rehabilitation of Little Topashaw Creek, a sinuous, sand-bed stream draining 37 km2 in northwest Mississippi are described. The rehabilitation project consisted of placing 72 large woody debris structures along eroding concave banks and planting 4000 willow cuttings in sandbars. Response was measured by monitoring flow, channel geometry, physical aquatic habitat, and fish populations. Initially, debris structures reduced high flow velocities at concave bank toes, preventing further erosion and inducing deposition. Physical response during the first year following construction included creation of sand berms along eroding banks and slight increases in base flow water width and depth. Fish collections showed assemblages typical of incising streams within the region, but minor initial responses to debris addition were evident. Progressive failure of the structures and renewed erosion were observed during the second year after construction. 相似文献
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On the mechanism of action of griseofulvin in dermatophytosis 总被引:1,自引:0,他引:1
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V C Blank A Sterin-Prync L Retegui A Vidal M Criscuolo L P Roguin 《European journal of biochemistry》1999,265(1):11-19
Four monoclonal antibodies (mAbs) directed against the recombinant human interferon-alpha2b (IFN-alpha2b) were used as probes to study the interaction of the IFN molecule to its receptors. The [125I]IFN-alpha2b binding to immobilized mAbs was completely inhibited by IFN-alpha2b and IFN-alpha2a but neither IFNbeta nor IFNgamma showed any effect. Gel-filtration HPLC of the immune complexes formed by incubating [125I]IFN-alpha2b with paired mAbs revealed the lack of simultaneous binding of two different antibodies to the tracer, suggesting that all mAbs recognize the same IFN antigenic domain. Furthermore, the mAbs were also able to neutralize the IFN-alpha2b anti-viral and anti-proliferative activities as well as [125I]IFN-alpha2b binding to WISH cell-membranes. As [125I]mAbs did not recognize IFN exposed epitopes in the IFN:receptor complexes, mAb induction of a conformational change in the IFN binding domain impairing its binding to receptors was considered unlikely. In order to identify the IFN region recognized by mAbs, IFN-alpha2b was digested with different proteolytic enzymes. Immunoreactivity of the resulting peptides was examined by Western blot and their sequences were established by Edman degradation after blotting to poly(vinylidene difluoride) membranes. Data obtained indicated that the smallest immunoreactive region recognized by mAbs consisted of residues 107-132 or 107-146. As this zone includes the sequence 123-140, which has been involved in the binding to receptors, and our mAbs did not show an allosteric behaviour, it is concluded that they are directed to overlapping epitopes located close to or even included in the IFN binding domain. 相似文献