rRNA and expressed as a ratio to the 18S rRNA signal. Statistical analysis Differential expression of the microarray data was evaluated using the limma package. A linear model for the four age6diet groups was fit for each probe set. Differences between groups were then extracted from the model as contrasts. An empirical Bayes shrinkagemethod was employed on the standard errors to improve power for small sample sizes. Last, multiple test correction of P-values was done using the false discovery rate method. qRT-PCR data were analyzed using the General Linear Models procedure of SAS. Differences were considered significant when P,0.05. Supporting Information qRT-PCR analysis Quanitative reverse transcriptase-polymerase chain reaction was used to validate a subset of genes expressed differentially based on microarray results. Gene-specific primerprobe pairs were designed for each gene using Primer Express 2.0 software. cDNA was prepared from tissue RNA samples using the High Capacity cDNA Archive Kit. cDNA samples then were evaluated using real-time two-step qRT-PCR using an Applied Acknowledgments The authors wish to thank Carole Wilson and Jenny Drnevich for their assistance with microarray and statistical analyses. AML in children is a clinically and genetically heterogeneous disease characterized by differentiation arrest and malignant proliferation of clonal myeloid precursors. It is the second most frequent hematologic malignancy, accounting for 15 to 20% of all childhood leukemia. The overall survival rate of pediatric AML patients has been increased from approximately 30 to 73%, however, nearly half of the pediatric patients relapse. Therefore, 25279926 risk-group classifications including prognostic markers as well as more targeted therapeutic approaches for treating pediatric AML are urgently needed. In adult AML patients, miRNAs can be used as biomarkers, and recently, first studies investigating the expression of selected miRNAs in 50 and 80 pediatric AML samples suggest the same for children. miRNAs are small, non-coding, regulatory and highly conserved molecules found in humans, animals, plants and some viruses. They regulate a variety of developmental and physiological processes like cell differentiation, apoptosis and immune responses and their role in hematopoiesis is beginning to be appreciated. Often, miRNAs are located in fragile sites or common breakpoint regions for chromosome aberrations that involve oncogenes or tumor suppressor 
genes in cancer cells. Although around 70% of miRNAs are located in regions of Dipraglurant price 25395428″ target=_blank”>25395428 leukemia-associated cytogenetic changes, only a subset of these miRNAs are expressed in a study surveying a panel of acute myeloid leukemia cell lines. Loss of miR-145 and miR-146a results in a long-term myeloid disease in mice, and reintroduction of both miRNAs into AML cells significantly induced cell death and prevented growth in vitro. Additionally, it was shown that MiRNA Expression and Function in Pediatric AML expression profiles of miRNAs cannot only be used for distinction of leukemia of different lineages, but also for differentiation of cytogenetic subtypes of adult AML. Three independent studies demonstrate that the cytogenetic subtypes t, t and inv offer unique miRNA expression profiles. It was shown that miR-126 is highly over expressed in t and inv and miR-224, miR-368 and miR-382 are exclusively over expressed in t in adult AML patients. Just recently, the expression of a single miRNA, miR-125b, was analyz
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