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  • 1
    Electronic Resource
    Electronic Resource
    Pentosan Polysulfate Regulates Scavenger Receptor-Mediated, But Not Fluid-Phase, Endocytosis in Immortalized Cerebral Endothelial Cells (2000)
    Springer
    Cellular and molecular neurobiology 20 (2000), S. 731-745 
    Details
    ISSN: 1573-6830
    Keywords: blood–brain barrier ; dextran sulfate@keyword = fluid-phase endocytosis ; fucoidan ; heparin ; pentosan polysulfate ; receptor-mediated endocytosis ; scavenger receptors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract SUMMARY 1. Effects of pentosan polysulfate (PPS) and the structurally related sulfated polyanions dextran sulfate, fucoidan, and heparin on the scavenger receptor-mediated and fluid-phase endocytosis in GP8 immortalized rat brain endothelial cells were investigated. 2. Using 1,1′-dioctadecyl-3,3,3,3′-tetramethylindocarboxyamine perchlorate-labeled acetylated low-density lipoprotein (DiI-AcLDL), we found a binding site with high affinity and low binding capacity, and another one with low affinity and high binding capacity. Increasing ligand concentrations could not saturate DiI-AcLDL uptake. DiI-AcLDL uptake, but not binding, was sensitive to pretreatment with filipin, an inhibitor of caveola formation. 3. PPS (20–200 μg/ml) significantly reduced the binding of DiI-AcLDL after coincubation for 3 hr, though this effect was less expressed after 18 hr. Among other polyanions, only fucoidan decreased the DiI-AcLDL binding after 3 hr, whereas dextran sulfate significantly increased it after 18 hr. PPS treatment induced an increase in DiI-AcLDL uptake, whereas other polysulfated compounds caused a significant reduction. 4. Fluid-phase endocytosis determined by the accumulation of Lucifer yellow was concentration and time dependent in GP8 cells. Coincubation with PPS or other sulfated polyanions could not significantly alter the rate of Lucifer yellow uptake. 5. In conclusion, PPS decreased the binding and increased the uptake of DiI-AcLDL in cerebral endothelial cells, an effect not mimicked by the other polyanions investigated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007007026500
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  • 2
    Electronic Resource
    Electronic Resource
    Early Appearance but Lagged Accumulation of Detergent-Insoluble Prion Protein in the Brains of Mice Inoculated with a Mouse-Adapted Creutzfeldt–Jakob Disease Agent (2000)
    Springer
    Cellular and molecular neurobiology 20 (2000), S. 717-730 
    Details
    ISSN: 1573-6830
    Keywords: prion diseases ; prion protein@keyword = conformational change ; immunoblotting ; infectivity ; replication ; spongiform neurodegeneration ; astrogliosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract SUMMARY 1. To elucidate mechanisms for the generation of the detergent-insoluble, proteinase K-resistant prion protein (PrPSc) from the detergent-soluble, proteinase K-sensitive PrP (PrPC) and the replication of the infectious agent in prion diseases, we followed the kinetics of detergent-insoluble PrP and PrPSc levels, infectious titers, and associated pathological changes in the brains of mice inoculated with a mouse-adapted Creutzfeldt–Jakob disease agent. 2. PrPSc in brain homogenate and detergent-insoluble PrP enriched by two-cycle ultracentrifugation were detected by immunoblotting and their relative amounts were estimated according to a standard curve plotted between the amount of PrP and signal intensity on immunoblotting. The titer of infectivity was determined by the incubation periods of mice inoculated with the unfractionated homogenate on the basis of a standard curve plotted between the titer and incubation period. 3. Detergent-insoluble PrP became detectable 4 weeks postinoculation (p.i.) well before the detection of PrPSc. The low level of detergent-insoluble PrP continued until dramatic accumulation occurred at 14 weeks p.i., correlating well with the accumulation of PrPSc and development of pathological changes. The infectious titer was undetectable at 4 weeks p.i. and its logarithmic increase occurred 10 weeks p.i. preceding the logarithmic accumulation of PrPs. 4. The lag time of detergent-insoluble PrP accumulation and the discrepancy between infectious titers and PrPs observed during the early period after inoculation suggest a slow and rate-limiting step for the detergent-insoluble PrP to become the infectious agent-associated PrPSc.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007054909662
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