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Furthermore, siRNA-transfected cells, grown under chondrogenic conditions did not display biologically significant changes in the expression of aggrecan, collagen type II or type X, or histology when grown in micropellets or monolayer cultures.
A 3-fold cut-off was chosen as a conservative estimate of biologically significant changes in gene expression.
For DNA methylation data, a second condition requiring a minimum difference of 1.5 between the two sample means was implemented in order to maximize the capture of biologically significant changes in methylation (Figure 2A).
In the case of the methylation data, a second criterion of a difference>1.5 between the mean of the two populations was added in order to increase the likelihood of detection of biologically significant changes in methylation levels.
By using an algorithm that detects metabolic modules based on biologically significant changes in gene expression [33], the 'reporter metabolites', i.e. the metabolites around which significant coordinated gene expression occurs, were identified.
We hypothesize that biologically significant changes in expression can be missed by expression arrays due to technical limitations, but might be captured by epigenomic studies by identifying genes at which promoter cytosine methylation or H3K9 acetylation differ and testing them with highly-quantitative techniques.
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Sample size could be as small as 400 g with a negligible change in sample variability, and without a biologically significant change in nutritive value concentrations.
However, in vivo, no biologically significant changes were observed in cytochrome P450 content or enzyme activity after once daily, oral dosing to rats for 28 days (unpublished data).
Notably, genes that don't seem differentially expressed at a statistically significant level may reveal biologically significant changes when studied in ontology groups [7].
An ongoing issue with complex proteomic experiments is the low statistical power of an experiment resulting in an inability to detect biologically significant changes.
To account for low and zero values in our data while still identifying biologically significant changes, differences were calculated relative to an RPKM of 0.1 when calculating fold changes from RPKM values of less than 0.1.
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