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David (http://david.abcc.ncifcrf.gov/) and KeggArray software (http://www.genome.jp/kegg/) was used for pathway analysis and to obtain P values of significantly regulated pathways for each comparison of 602 and NM strains against Coxiella burnetii RSA 493 (organism code: cbu) KEGG pathway database.
More than 1000 DEGs and over 200 regulated pathways for each stressor were identified.
Common gene expression levels of neighboring nodes in a metabolic network were calculated by averaging over all neighbors of a gene, and revealed several interesting regulated pathways for the human immune system [ 19].
We found consistent overlap between the human RH, mouse RH and TCGA data in terms of regulated pathways for genes with negative correlation between CNVs and expression data and attenuated response of X-linked genes in response to copy number increase.
Tests were run to identify significantly up- and down- regulated pathways for each tissue and their pairwise combinations against all other samples using Generally Applicable Gene-set Enrichment (GAGE) analyses [ 48], implemented as R/Bioconductor package [ 49], which is robust to small datasets with different sample sizes.
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For example, the most significantly regulated pathway for the regulated mRNA gene list was acute phase response signaling.
Most of the predicted regulators belonged to the protein families whose members were known to regulate pathways for carbohydrate metabolism, such as the AraC, GntR, LacI, and ROK families [ 46- 49].
In addition, maintenance/re-adjustment of cellular homeostasis and of the interactions between dehydration and other stress/hormone regulated pathways are considered critical for the survival [ 6, 31, 32].
Other promising methods that search for differentially regulated pathways rely on Petri Nets (Cabusora et al., 2005; Kueffner et al., 2000; Zien et al., 2000).
Another concern was a possibility of a bias due to decreased inherent variation within consistently reporting gene sets, as could be expected for tightly regulated pathways.
Expected differentially regulated pathways in cancer could be confirmed, e.g. the aggressive tumors showed significantly up-regulated pathways for purine and pyrimidine synthesis to maintain elevated proliferation, as well as the up-regulation of glycolysis and pyruvate metabolism to support energy supply for the tumor.
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