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July 14, 2017

gnificant genes. adjusted p-values using Benjamini and Hochberg procedure, which control for false discovery rate, are used to decide the statistical significance of genes along with LFC. Disease-classification Genes The Meta Threshold Gradient MedChemExpress 80321-63-7 Directed Regularization method proposed by Ma and Huang was used to select genes which may distinguish LS and NL skin samples. MTGDR is an extension of the Threshold Gradient Directed Regularization , to the case where several studies are combined. For each study m, the independent variables Ym – defined as the binary indicators of group membership – is modeled through a logistic regression with the expression values for all genes as a covariates. MTGDR assumes that the regression coefficients of the logistic regression of the TGDR model may be different across studies but the sets of genes with nonzero coefficients are the same across studies. In microarray studies where more than thousands of genes are surveyed, only a small number of genes are actually associated with the outcome of interest. MTGDR tries to select such genes and estimates the corresponding coefficients simultaneously by maximum likelihood. The algorithm starts with initial values for the regression coefficients equal zero and in each iteration, updates only the coefficients associated with genes with large meta-gradients. Which and how many genes are updated in each iteration are determined by k and the tuning parameter t. A value of t = 1 indicates that only the gene with largest meta-gradient is updated whereas if t = 0, all genes will be updated. With a large t and a finite k, only a small number of genes will have nonzero coefficients. For the detailed descriptions on MTGDR, see Ma and Huang. Tuning parameters t and k were jointly determined by a 3-fold cross-validation. Samples from each study were randomly divided into 3 subsets and cross-validation was carried out by running MTGDR for a set of possible values for t and number of iterations up to 5000. Optimal parameters were selected as those that maximized the log-likelihood. The limit of 3fold cross-validation was due to the small size in one study. To evaluate the performance of the final classifier, we considered 5fold cross-validation. Samples in each study were randomly divided into 5 parts, 4 folds were used to run MTGDR, and PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22210479 then the resulting estimates were used to make prediction on the removed one fold. This procedure was repeated 5 times to produce class prediction on all samples, and the prediction error was computed. Prediction errors generated by 10 and 20-fold crossvalidation produced similar results. Meta Analysis The classic application of meta-analysis is to find a single outcome using published data where only the summary statistics are typically available. With microarray experiments, however, a more fortuitous situation of having the complete set of raw data available is commonly achievable. Thus, we took advantage of this feature and modeled the differences in expression values between LS and uninvolved skin pairs uniformly. The general model in a meta-analysis setting is as follows. Let Yij represent the measured effect for study j for a specific gene i. We have, Yij ~hij zeij, hij ~ mi zdij, eij N 0,s2 ij dij N 0,t2 i 1 where between-study variance ti2 represents the variability between studies, and it is usually estimated by the DerSimonian and Laird method. And s2 represents the within-study variance for the ith study. Both Yij and s2 are alre