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#26887861   2016/03/09 Save this To Up

Exposure to a 50-Hz magnetic field induced ceramide generation in cultured cells.

Purpose To investigate the effects of a 50-Hz magnetic field (MF) exposure on ceramide metabolism, as well as the cascade downstream signaling pathways in human amniotic (FL) cells. Materials and methods FL cells were exposed to MF at 0.4 mT for different durations (from 5-60 min). The ceramides levels were analyzed with high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The activity of cathepsin D was assayed using a fluorometric assay kit, and the activity of protein phosphatase 2A (PP2A) was examined by Western blotting. After exposing to MF at 0.4 mT for 60 min with sequential culture for different durations (0, 3, 6, 12 or 36 h), the rate of cell apoptosis was assessed by flow cytometry. Results Exposing cells to MF at 0.4 mT for different durations caused a significant increase in ceramide production via de novo synthesis and hydrolysis of sphingomyelin (SM), and the effect was different according to the exposure time. However, no significant change in cell apoptosis was detected after MF exposure for 60 min with sequentially culturing for up to 36 h. In addition, increase in ceramide did not activate its downstream signal molecules, cathepsin D and PP2A, which are usually closely related to apoptosis of cells. Conclusions Exposure to a 50-Hz MF could raise ceramide levels but had no significant effect on apoptosis in cultured cells.

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#1394123   1992/10/29 Save this To Up

Cathepsin D as a prognostic indicator for node-negative breast cancer patients using both immunoassays and enzymatic assays.

This is a retrospective study on 162 node-negative patients, with both biochemical and clinical factors being measured for determination of prognostic markers. Steroid receptors were measured on all tumors, while tumor size, histological grade, ploidy status, and cell cycle kinetics indicators could not be found or measured on 25 or less of the patient group. The primary focus of this study was the measurement of cathepsin D, analyzed by two different procedures, and 161 of the 162 patients had at least one value. The antigenic assay was performed using the US-CIS kit, and it was sensitive and reproducible. A biochemical assay using the enzymatic activity of cathepsin D was developed, and it gave proportional values, compared to the antigenic assay values (r2 = 0.79). Our results indicated that the mean antigenic levels were 20% higher than the biochemical assay levels (P = 0.001). High levels of cathepsin D by the antigenic assay predicted poor relapse-free (P = 0.0001) and overall (P = 0.0004) survival. High levels of cathepsin D by the biochemical assay also predicted poor relapse-free (P = 0.031) and overall (P = 0.0013) survival. The cathepsin D values were still useful as predictors of outcome after multivariate analysis. Several other factors, such as grade and S phase, were useful as additional prognostic indicators. In conclusion, cathepsin D is the most useful marker in node-negative patients, and the analysis can be performed by both a biochemical and an antigenic assay.

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