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Images were analyzed after background correction using the rolling ball radius tool (50 pixels).
The expression ratios were normalized without background correction using the print tip loess method [31] of the limma package [32] from Bioconductor.
LIMMA was used to download datasets from LIMMA, for background correction using the normexp model, and for within-array normalization using the print-old tip loess method.
Expression ratios (Cy5/Cy3) were calculated following background correction using the R programming language [ 17] and the BioConductor [ 18] package limma [ 19] to account for dye bias.
Following background correction using the median foreground minus the median background signal intensity for each channel, the log 2 ratios were calculated for each spot.
Urinary cadmium was measured by graphite furnace atomic absorption with Zeeman background correction, using the CDC modification of the method of Pruszkowska and colleagues [ 23].
Similar(51)
Depending on type of background correction used, the 5-year MFSR varied between 0.67 and 0.82 for the low-risk group, and between 0.47 and 0.53 for the high-risk group.
We tested the robustness of the results by repeating the procedure with and without background-correction, using the 15% top varying CpGs, resulting in highly similar epitypes.
To avoid the possibility that codon usage may be artefactually inflated for G and C ending codons, we masked repetitive elements in non-coding regions which are served for the background correction, using RepeatMasker.
Fluorescence intensities of cells encircled by ROIs were calculated by averaging corresponding pixel intensities in the original image, excluding pixels within encircled nuclei, after background correction using a blank (no cell) region of the image.
After background correction using a Normal plus Exponential convolution model, which adjusts the foreground to the background, we used an offset to dampen the variation of the log-ratios for intensities close to zero.
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