A Cellular Model To Monitor Proteasome Dysfunction by α-Synuclein

Impairment of the ubiquitin-proteasome degradation system has recently been suggested to be related to the onset of neurodegenerative disorders such as Alzheimer′s disease and Parkinson's disease. In this study, we investigated whether intracellular α-synuclein affects proteasome activity in SH-SY5Y cells. To monitor intracellular proteasome activity, we used a reporter consisting of a short peptide degron fused to the carboxyl-terminus of green fluorescent protein (GFP-CL1), which is known to be degraded by proteasome. The level of intact GFP-CL1 was dramatically increased by coexpression of GFP-CL1 and α-synuclein, as judged by confocal microscopic and immunoblot analyses. Expression of two pathogenic mutants of α-synuclein, A30P and A53T, and phosphomimetic S129D mutant increased the intensities of GFP more effectively than did wild-type α-synuclein. GFP fluorescence in cells transfected with Δ73-83 mutant or β-synuclein, which does not assemble into filaments <i>in vitro</i>, was not changed as compared with that in cells expressing GFP-CL1 alone. Thus, the ability of α-synuclein to inhibit proteasome activity is related to its propensity to assemble into filaments. Furthermore, we observed that some compounds inhibiting α-synuclein filament formation <i>in vitro</i> prevented the α-synuclein-mediated proteasome dysfunction in cells transfected with both GFP-CL1 and α-synuclein. The cellular model expressing both GFP-CL1 and α-synuclein may be a useful tool to screen compounds protecting neurons from α-synuclein-mediated proteasome dysfunction

Ciclesonide uptake and metabolism in human alveolar type II epithelial cells (A549)-2

<p><b>Copyright information:</b></p><p>Taken from "Ciclesonide uptake and metabolism in human alveolar type II epithelial cells (A549)"</p><p>http://www.biomedcentral.com/1471-2210/7/12</p><p>BMC Pharmacology 2007;7():12-12.</p><p>Published online 27 Sep 2007</p><p>PMCID:PMC2048954.</p><p></p>of ciclesonide, des-CIC, and fatty acid esters of des-CIC. Results represent the mean ± standard deviation from 5 dishes per time point. *** p < 0.001 for ciclesonide fluticasone propionate (Aspin-Welch t-test)

Ciclesonide uptake and metabolism in human alveolar type II epithelial cells (A549)-1

<p><b>Copyright information:</b></p><p>Taken from "Ciclesonide uptake and metabolism in human alveolar type II epithelial cells (A549)"</p><p>http://www.biomedcentral.com/1471-2210/7/12</p><p>BMC Pharmacology 2007;7():12-12.</p><p>Published online 27 Sep 2007</p><p>PMCID:PMC2048954.</p><p></p

Ciclesonide uptake and metabolism in human alveolar type II epithelial cells (A549)-0

<p><b>Copyright information:</b></p><p>Taken from "Ciclesonide uptake and metabolism in human alveolar type II epithelial cells (A549)"</p><p>http://www.biomedcentral.com/1471-2210/7/12</p><p>BMC Pharmacology 2007;7():12-12.</p><p>Published online 27 Sep 2007</p><p>PMCID:PMC2048954.</p><p></p>of ciclesonide, des-CIC, and fatty acid esters of des-CIC. Results represent the mean ± standard deviation from 5 dishes per time point. *** p < 0.001 for ciclesonide fluticasone propionate (Aspin-Welch t-test)

Univariate analysis of severe side effects during adjuvant chemotherapy.

<p>Univariate analysis of severe side effects during adjuvant chemotherapy.</p

Clinicopathological characteristics of patients stratified by the CAR.

<p>Clinicopathological characteristics of patients stratified by the CAR.</p

Kaplan-Meier curves of the effect of the CAR on DFS and OS.

<p>Disease-free survival is better in the L-group than in the H-group (p = 0.03). However, there is no significant difference in overall survival between the two groups (p = 0.25) (Fig 1B). CAR, CRP to albumin ratio; OS, overall survival; DFS, disease-free survival.</p

Surgical and scoring characteristics of patients stratified by the CAR.

<p>Surgical and scoring characteristics of patients stratified by the CAR.</p

Additional file 1: Table S1. of Effectiveness of adjuvant chemotherapy for elderly patients with lymph node-positive colorectal cancer

Reasons for withholding adjuvant chemotherapy. (DOC 46 kb

Immunoblotting analysis of abnormal tau in the sarkosyl-insoluble fraction.

<p>(A) Immunoblotting analysis was visualized using AT8 antibody for the sarkosyl-insoluble fraction. The numbers indicate individual mice: 1–15, MB 1 mg/kg/day group; 16–30, MB 0.3 mg/kg/day group; and 31–44, water only group. Molecular weight markers are shown on the right (kDa). P, positive control (P301L tau transgenic mouse, 20 month-old female). (B) A comparison of relative AT8 expression levels of the MB-treated groups and the water only group. The data were compared with the AT8 band intensity. (C) A comparison of relative phosphorylated tau (AT8)/total tau (HT7) levels of the MB-treated groups and the water only group. The data were compared with the AT8 band intensity, which was normalized with the total tau (HT7) band intensity. The central lines indicate medians and the vertical lines represent 25^th and 75^th percentiles. P<0.01 was considered to represent a statistically significant difference. a.u., arbitrary unit. N.S., no significant difference.</p