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  • 1
    ISSN: 1432-1424
    Keywords: I sKpotassium channel ; Min K channel ; Disulfonic stilbene ; Inner ear ; Ion-selective vibrating probe ; Macropatch clamp technique
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Vestibular dark cell epithelium secretes K+ via I sKchannels in the apical membrane. The previous observation that disulfonic stilbenes increased the equivalent short circuit current (I sc) suggested that these agents might be useful investigative tools in this tissue. The present experiments were conducted to determine if the increase in I scwas associated with an increase in K+ flux and if the effect was directly on the I sKchannel or indirectly via a cytosolic intermediary. Measurements of transepithelial K+ flux with the K+-selective vibrating probe and of changes in net cellular solute flux by measurements of epithelial cell height showed that 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS) increased K+ flux by a factor of 1.96±0.71 and caused net solute efflux. The apical membrane was partitioned with a macropatch pipette and DIDS was applied either to the membrane outside the pipette, inside the pipette or to the entire apical membrane. DIDS inside the pipette increased the current across the patch, the membrane conductance, the slowly-inactivating (I sK) component of the membrane current and shifted the reversal voltage toward the equilibrium potential for K+. DIDS outside the patch decreased the patch current and conductance, consistent with shunting of current away from the membrane patch. These findings strongly support the notion that DIDS increases K+ secretion through I sKchannels in the apical membrane of vestibular dark cell epithelium by acting directly on the channels or on a tightly colocalized membrane component.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2277
    Keywords: Key words Donor antigen-specific reactivity ; Immunologic monitoring ; Mixed lymphocyte culture (MLC) ; Rejection ; Transplantation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Development of donor-specific proliferative hyporeactivity has been evaluated in many studies for its usefulness in identifying transplant recipients at low risk of immunological complications. These studies often result in controversial conclusions, however. The authors claim that the discrepancy in the predictive value of mixed lymphocyte culture- (MLC) reactivity might partly be due to differences in presentation and interpretation of results. The purpose of this study is to investigate the usefulness of a normalized evaluation of antigen-specific donor-reactivity in a small number of kidney transplant recipients. This could then serve as a basis for an extended clinical study. Ten cadaveric kidney recipients were tested for proliferative reactivity to donor- and third-party antigens up to 20 months posttransplantation. Expressing donor-specific reactivity as a relation between the percentage of pretransplant responses towards donor splenocytes and the percentage of pretransplant responses towards third-party donor cells should minimize influences of e. g. uremia, current immunosuppression or infections on the evaluation of specific reactivity and thus should allow an evaluation of the donor-specificity of T-cell alloresponses independently of fluctuations in global responsiveness. Four of ten recipients acquired a state of donor-specific hyporeactivity ( 〈 75 % relative specific reactivity) at 20 months posttransplantation (61 ± 12 %, mean ± SD). Six patients were classified non-hyporeactive (98 ± 10 % mean relative specific reactivity). Relative specific reactivity did not correlate with the levels of general reactivity. Three of the four hyporeactive and four of the six non-hyporeactive patients developed acute rejection. Stable graft function at 20 months posttransplantation (serum creatinine ≤ 2 mg/dl) was not closely related to the reactivity status, as five of eight patients with well-functioning grafts did not develop relative specific hyporeactivity. One recipient with chronic rejection was classified hyporeactive. One non-hyporeactive patient lost his graft due to non-immunological causes. Our data suggest that posttransplant relative specific reactivity does not predict acute rejection. Downregulation of donor-specific reactivity might not be a prerequisite for stable graft function but could help identifying recipients who require less immunosuppression. This, however, remains to be established in a prospective immunosuppression-weaning study.
    Type of Medium: Electronic Resource
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