--- title: "Package MKmisc" author: "Matthias Kohl" date: "`r Sys.Date()`" output: rmarkdown::html_vignette: toc: true vignette: > %\VignetteIndexEntry{MKmisc} %\VignetteEngine{knitr::rmarkdown} %\VignetteEncoding{utf8} --- ## Introduction Package MKmisc includes a collection of functions that I found useful in my daily work. It contains several functions for statistical data analysis; e.g. for sample size and power calculations, computation of confidence intervals, and generation of similarity matrices. We first load the package. ```{r} library(MKmisc) ``` ## Descriptive Statistics ### IQR I implemented function IQrange before the standard function IQR gained the type argument. Since 2010 (r53643, r53644) the function is identical to function IQR. ```{r} x <- rnorm(100) IQrange(x) IQR(x) ``` It is also possible to compute a standardized version of the IQR leading to a normal-consistent estimate of the standard deviation. ```{r} sIQR(x) sd(x) ``` ### Mean Absolute Deviation The mean absolute deviation under the assumption of symmetry is a robust alternative to the sample standard deviation. ```{r} meanAD(x) ``` ### Five Number Summary There is a function that computes a so-called five number summary which in contrast to function fivenum uses the first and third quartile instead of the lower and upper hinge. ```{r} fiveNS(x) ``` ### Coefficient of Variation (CV) There are functions to compute the (classical) coefficient of variation as well as two robust variants. In case of the robust variants, the mean is replaced by the median and the SD is replaced by the (standardized) MAD and the (standardized) IQR, respectively. ```{r} ## 5% outliers out <- rbinom(100, prob = 0.05, size = 1) sum(out) x <- (1-out)*rnorm(100, mean = 10, sd = 2) + out*25 CV(x) medCV(x) iqrCV(x) ``` ### Signal to Noise Ratio (SNR) There are functions to compute the (classical) signal to noise ratio as well as two robust variants. In case of the robust variants, the mean is replaced by the median and the SD is replaced by the (standardized) MAD and the (standardized) IQR, respectively. ```{r} SNR(x) medSNR(x) iqrSNR(x) ``` ### Box- and Whisker-Plot In contrast to the standard function boxplot which uses the lower and upper hinge for defining the box and the whiskers, the function qboxplot uses the first and third quartile. ```{r, fig.width=7, fig.height=7} x <- rt(10, df = 3) par(mfrow = c(1,2)) qboxplot(x, main = "1st and 3rd quartile") boxplot(x, main = "Lower and upper hinge") ``` The difference between the two versions often is hardly visible. ### OR, RR and Other Risk Measures Given the incidence of the outcome of interest in the nonexposed (p0) and exposed (p1) group, several risk measures can be computed. ```{r} ## Example from Wikipedia risks(p0 = 0.4, p1 = 0.1) risks(p0 = 0.4, p1 = 0.5) ``` Given p0 or p1 and OR, we can compute the respective RR. ```{r} or2rr(or = 1.5, p0 = 0.4) or2rr(or = 1/6, p1 = 0.1) ``` ### Generalized Logarithm The generalized logarithm may be useful as a variance stabilizing transformation when also negative values are present. ```{r} curve(log, from = -3, to = 5) curve(glog, from = -3, to = 5, add = TRUE, col = "orange") legend("topleft", fill = c("black", "orange"), legend = c("log", "glog")) ``` As in case of function log there is also glog10 and glog2. ```{r} curve(log10(x), from = -3, to = 5) curve(glog10(x), from = -3, to = 5, add = TRUE, col = "orange") legend("topleft", fill = c("black", "orange"), legend = c("log10", "glog10")) ``` There are also functions that compute the inverse of the generalized logarithm. ```{r} inv.glog(glog(10)) inv.glog(glog(10, base = 3), base = 3) inv.glog10(glog10(10)) inv.glog2(glog2(10)) ``` ### Simulate Correlated Variables To demonstrate Pearson correlation in my lectures, I have written this simple function to simulate correlated variables and to generate a scatter plot of the data. ```{r, fig.width=7, fig.height=7} res <- simCorVars(n = 500, r = 0.8) cor(res$Var1, res$Var2) ``` ### Plot TSH, fT3 and fT4 Values The thyroid function is usually investigated by determining the values of TSH, fT3 and fT4. The function thyroid can be used to visualize the measured values as relative values with respect to the provided reference ranges. ```{r, fig.width=7, fig.height=7} thyroid(TSH = 1.5, fT3 = 2.5, fT4 = 14, TSHref = c(0.2, 3.0), fT3ref = c(1.7, 4.2), fT4ref = c(7.6, 15.0)) ``` ### Generalized and Negative Logarithm as Transformations We can use the generalized logarithm for transforming the axes in ggplot2 plots. ```{r} library(ggplot2) data(mpg) p1 <- ggplot(mpg, aes(displ, hwy)) + geom_point() p1 p1 + scale_x_log10() p1 + scale_x_glog10() p1 + scale_y_log10() p1 + scale_y_glog10() ``` The negative logrithm is for instance useful for displaying p values. The interesting values are on the top. This is for instance used in a so-called volcano plot. ```{r} x <- matrix(rnorm(1000, mean = 10), nrow = 10) g1 <- rep("control", 10) y1 <- matrix(rnorm(500, mean = 11.25), nrow = 10) y2 <- matrix(rnorm(500, mean = 9.75), nrow = 10) g2 <- rep("treatment", 10) group <- factor(c(g1, g2)) Data <- rbind(x, cbind(y1, y2)) pvals <- apply(Data, 2, function(x, group) t.test(x ~ group)$p.value, group = group) ## compute log-fold change logfc <- function(x, group){ res <- tapply(x, group, mean) log2(res[1]/res[2]) } lfcs <- apply(Data, 2, logfc, group = group) ps <- data.frame(pvals = pvals, logfc = lfcs) ggplot(ps, aes(x = logfc, y = pvals)) + geom_point() + geom_hline(yintercept = 0.05) + scale_y_neglog10() + geom_vline(xintercept = c(-0.1, 0.1)) + xlab("log-fold change") + ylab("-log10(p value)") + ggtitle("A Volcano Plot") ``` ### Change Data from Wide to Long Often it's better to have the data in a long format than in a wide format; e.g., when plotting with package ggplot2. The necessary transformation can be done with function melt.long. ```{r, fig.width=7, fig.height=7} library(ggplot2) ## some random data test <- data.frame(x = rnorm(10), y = rnorm(10), z = rnorm(10)) test.long <- melt.long(test) test.long ggplot(test.long, aes(x = variable, y = value)) + geom_boxplot(aes(fill = variable)) ## introducing an additional grouping variable group <- factor(rep(c("a","b"), each = 5)) test.long.gr <- melt.long(test, select = 1:2, group = group) test.long.gr ggplot(test.long.gr, aes(x = variable, y = value, fill = group)) + geom_boxplot() ``` ## Confidence Intervals ### Binomial Proportion There are several functions for computing confidence intervals. We can compute 10 different confidence intervals for binomial proportions; e.g. ```{r} ## default: "wilson" binomCI(x = 12, n = 50) ## Clopper-Pearson interval binomCI(x = 12, n = 50, method = "clopper-pearson") ## identical to binom.test(x = 12, n = 50)$conf.int ``` For all intervals implemented see the help page of function binomCI. ### Mean and SD We can compute confidence intervals for mean and SD of a normal distribution. ```{r} x <- rnorm(50, mean = 2, sd = 3) ## mean and SD unknown normCI(x) ## SD known normCI(x, sd = 3) ## mean known normCI(x, mean = 2) ``` ### Difference in Means We can compute confidence interval for the difference of means assuming normal distributions. ```{r} x <- rnorm(20) y <- rnorm(20, sd = 2) ## paired normDiffCI(x, y, paired = TRUE) ## compare normCI(x-y) ## unpaired y <- rnorm(10, mean = 1, sd = 2) ## classical normDiffCI(x, y, method = "classical") ## Welch (default as in case of function t.test) normDiffCI(x, y, method = "welch") ## Hsu normDiffCI(x, y, method = "hsu") ``` In case of unequal variances and unequal sample sizes per group the classical confidence interval may have a bad coverage (too long or too short), as is indicated by the small Monte-Carlo simulation study below. ```{r} M <- 100 CIhsu <- CIwelch <- CIclass <- matrix(NA, nrow = M, ncol = 2) for(i in 1:M){ x <- rnorm(10) y <- rnorm(30, sd = 0.1) CIclass[i,] <- normDiffCI(x, y, method = "classical")$conf.int CIwelch[i,] <- normDiffCI(x, y, method = "welch")$conf.int CIhsu[i,] <- normDiffCI(x, y, method = "hsu")$conf.int } ## coverage probabilies ## classical sum(CIclass[,1] < 0 & 0 < CIclass[,2])/M ## Welch sum(CIwelch[,1] < 0 & 0 < CIwelch[,2])/M ## Hsu sum(CIhsu[,1] < 0 & 0 < CIhsu[,2])/M ``` ### Coefficient of Variation We provide 11 different confidence intervals for the (classical) coefficient of variation; e.g. ```{r} x <- rnorm(100, mean = 10, sd = 2) # CV = 0.2 ## default: "miller" cvCI(x) ## Gulhar et al. (2012) cvCI(x, method = "gulhar") ``` For all intervals implemented see the help page of function cvCI. ### Quantiles, Median and MAD We start with the computation of confidence intervals for quantiles. ```{r} x <- rexp(100, rate = 0.5) ## exact quantileCI(x = x, prob = 0.95) ## asymptotic quantileCI(x = x, prob = 0.95, method = "asymptotic") ``` Next, we consider the median. ```{r} ## exact medianCI(x = x) ## asymptotic medianCI(x = x, method = "asymptotic") ``` It often happens that quantile confidence intervals are not unique. Here the minimum length interval might be of interest. ```{r} medianCI(x = x, minLength = TRUE) ``` Finally, we take a look at MAD (median absolute deviation) where by default the standardized MAD is used (see function mad). ```{r} ## exact madCI(x = x) ## aysymptotic madCI(x = x, method = "asymptotic") ## unstandardized madCI(x = x, constant = 1) ``` ### Relative Risk There is also a function for computing an approximate confidence interval for the relative risk (RR). ```{r} ## Example from Wikipedia rrCI(a = 15, b = 135, c = 100, d = 150) rrCI(a = 75, b = 75, c = 100, d = 150) ``` ## Sample Size ### Welch Two-Sample t-Test For computing the sample size of the Welch t-test, we only consider the situation of equal group size (balanced design). ```{r} ## identical results as power.t.test, since sd = sd1 = sd2 = 1 power.welch.t.test(n = 20, delta = 1) power.welch.t.test(power = .90, delta = 1) power.welch.t.test(power = .90, delta = 1, alternative = "one.sided") ## sd1 = 0.5, sd2 = 1 power.welch.t.test(delta = 1, sd1 = 0.5, sd2 = 1, power = 0.9) ``` ### Hsu Two-Sample t-Test For computing the sample size of the Hsu t-test, we only consider the situation of equal group size (balanced design). ```{r} ## slightly more conservative than Welch t-test power.hsu.t.test(n = 20, delta = 1) power.hsu.t.test(power = .90, delta = 1) power.hsu.t.test(power = .90, delta = 1, alternative = "one.sided") ## sd1 = 0.5, sd2 = 1 power.welch.t.test(delta = 0.5, sd1 = 0.5, sd2 = 1, power = 0.9) power.hsu.t.test(delta = 0.5, sd1 = 0.5, sd2 = 1, power = 0.9) ``` ### Two Negative Binomial Rates When we consider two negative binomial rates, we can compute sample size or power applying function power.nb.test. ```{r} ## examples from Table III in Zhu and Lakkis (2014) power.nb.test(mu0 = 5.0, RR = 2.0, theta = 1/0.5, duration = 1, power = 0.8, approach = 1) power.nb.test(mu0 = 5.0, RR = 2.0, theta = 1/0.5, duration = 1, power = 0.8, approach = 2) power.nb.test(mu0 = 5.0, RR = 2.0, theta = 1/0.5, duration = 1, power = 0.8, approach = 3) ``` ## Moderated Tests Based on Package limma ### Moderated t-Test The function to compute the moderated t-test was motivated by the fact that my students have problems to understand and correctly adapt the code of the limma package. ```{r} ## One-sample test X <- matrix(rnorm(10*20, mean = 1), nrow = 10, ncol = 20) mod.t.test(X) ## Two-sample test set.seed(123) X <- rbind(matrix(rnorm(5*20), nrow = 5, ncol = 20), matrix(rnorm(5*20, mean = 1), nrow = 5, ncol = 20)) g2 <- factor(c(rep("group 1", 10), rep("group 2", 10))) mod.t.test(X, group = g2) ## Paired two-sample test mod.t.test(X, group = g2, paired = TRUE) ``` ### Moderated 1-Way ANOVA The function to compute a moderated 1-way ANOVA was motivated by the fact that my students have problems to understand and correctly adapt the code of the limma package. ```{r} set.seed(123) X <- rbind(matrix(rnorm(5*20), nrow = 5, ncol = 20), matrix(rnorm(5*20, mean = 1), nrow = 5, ncol = 20)) gr <- factor(c(rep("A1", 5), rep("B2", 5), rep("C3", 5), rep("D4", 5))) mod.oneway.test(X, gr) ``` ### Pairwise moderated t-tests As a optional post-hoc analysis after mod.oneway.test one can use pairwise moderated t-tests. One should carefully think about the adjustment of p values in this context. ```{r} pairwise.mod.t.test(X, gr) ``` ## Hsu Two-Sample t-Test The Hsu two-sample t-test is an alternative to the Welch two-sample t-test using a different formula for computing the degrees of freedom of the respective t-distribution. The following code is taken and adapted from the help page of the t.test function. ```{r} t.test(1:10, y = c(7:20)) # P = .00001855 t.test(1:10, y = c(7:20, 200)) # P = .1245 -- NOT significant anymore hsu.t.test(1:10, y = c(7:20)) hsu.t.test(1:10, y = c(7:20, 200)) ## Traditional interface with(sleep, t.test(extra[group == 1], extra[group == 2])) with(sleep, hsu.t.test(extra[group == 1], extra[group == 2])) ## Formula interface t.test(extra ~ group, data = sleep) hsu.t.test(extra ~ group, data = sleep) ``` ## Multiple Imputation t-Test Function mi.t.test can be used to compute a multiple imputation t-test by applying the approch of Rubin (1987) in combination with the adjustment of Barnard and Rubin (1999). ```{r} ## Generate some data set.seed(123) x <- rnorm(25, mean = 1) x[sample(1:25, 5)] <- NA y <- rnorm(20, mean = -1) y[sample(1:20, 4)] <- NA pair <- c(rnorm(25, mean = 1), rnorm(20, mean = -1)) g <- factor(c(rep("yes", 25), rep("no", 20))) D <- data.frame(ID = 1:45, variable = c(x, y), pair = pair, group = g) ## Use Amelia to impute missing values library(Amelia) res <- amelia(D, m = 10, p2s = 0, idvars = "ID", noms = "group") ## Per protocol analysis (Welch two-sample t-test) t.test(variable ~ group, data = D) ## Intention to treat analysis (Multiple Imputation Welch two-sample t-test) mi.t.test(res$imputations, x = "variable", y = "group") ## Per protocol analysis (Two-sample t-test) t.test(variable ~ group, data = D, var.equal = TRUE) ## Intention to treat analysis (Multiple Imputation two-sample t-test) mi.t.test(res$imputations, x = "variable", y = "group", var.equal = TRUE) ## Specifying alternatives mi.t.test(res$imputations, x = "variable", y = "group", alternative = "less") mi.t.test(res$imputations, x = "variable", y = "group", alternative = "greater") ## One sample test t.test(D$variable[D$group == "yes"]) mi.t.test(res$imputations, x = "variable", subset = D$group == "yes") mi.t.test(res$imputations, x = "variable", mu = -1, subset = D$group == "yes", alternative = "less") mi.t.test(res$imputations, x = "variable", mu = -1, subset = D$group == "yes", alternative = "greater") ## paired test t.test(D$variable, D$pair, paired = TRUE) mi.t.test(res$imputations, x = "variable", y = "pair", paired = TRUE) ``` ## Imputation of Standard Deviations for Changes from Baseline The function imputeSD can be used to impute standard deviations for changes from baseline adopting the approach of Section 16.1.3.2 of the Cochrane handbook (2011). ```{r} SD1 <- c(0.149, 0.022, 0.036, 0.085, 0.125, NA, 0.139, 0.124, 0.038) SD2 <- c(NA, 0.039, 0.038, 0.087, 0.125, NA, 0.135, 0.126, 0.038) SDchange <- c(NA, NA, NA, 0.026, 0.058, NA, NA, NA, NA) imputeSD(SD1, SD2, SDchange) ``` ## AUC ### Estimation There are two functions that can be used to calculate and test AUC values. First function AUC, which computes the area under the receiver operating characteristic curve (AUC under ROC curve) using the connection of AUC to the Wilcoxon rank sum test. We use some random data and groups to demonstrate the use of this function. ```{r} x <- c(runif(50, max = 0.6), runif(50, min = 0.4)) g <- c(rep(0, 50), rep(1, 50)) AUC(x, group = g) ``` Sometimes the labels of the group should be switched to avoid an AUC smaller than 0.5, which represents a result worse than a pure random choice. ```{r} g <- c(rep(1, 50), rep(0, 50)) AUC(x, group = g) ## no switching AUC(x, group = g, switchAUC = FALSE) ``` ### Testing We can also perform statistical tests for AUC. First, the one-sample test which corresponds to the Wilcoxon signed rank test. ```{r} g <- c(rep(0, 50), rep(1, 50)) AUC.test(pred1 = x, lab1 = g) ``` We can also compare two AUC using the test of Hanley and McNeil (1982). ```{r} x2 <- c(runif(50, max = 0.7), runif(50, min = 0.3)) g2 <- c(rep(0, 50), rep(1, 50)) AUC.test(pred1 = x, lab1 = g, pred2 = x2, lab2 = g2) ``` ### Pairwise There is also a function for pairwise comparison if there are more than two groups. ```{r} x3 <- c(x, x2) g3 <- c(g, c(rep(2, 50), rep(3, 50))) pairwise.auc(x = x3, g = g3) ``` In addition to the pairwise.auc there are further functions for pairwise comparisons. ## Pairwise Comparisons Often we are in a situation that we want to compare more than two groups pairwise. ### FC and logFC In the analysis of omics data, the FC or logFC are important measures and are often used in combination with (adjusted) p values. ```{r} x <- rnorm(100) ## assumed as log-data g <- factor(sample(1:4, 100, replace = TRUE)) levels(g) <- c("a", "b", "c", "d") ## modified FC pairwise.fc(x, g) ## "true" FC pairwise.fc(x, g, mod.fc = FALSE) ## without any transformation pairwise.logfc(x, g) ``` The function returns a modified FC. That is, if the FC is smaller than 1 it is transformed to -1/FC. One can also use other functions than the mean for the aggregation of the data. ```{r} pairwise.logfc(x, g, ave = median) ``` ### Arbitrary Criteria Furthermore, function pairwise.fun enables the application of arbitrary functions for pairwise comparisons. ```{r} pairwise.wilcox.test(airquality$Ozone, airquality$Month, p.adjust.method = "none") ## To avoid the warnings library(exactRankTests) pairwise.fun(airquality$Ozone, airquality$Month, fun = function(x, y) wilcox.exact(x, y)$p.value) ``` ## Binary Classification ### PPV and NPV In case of medical diagnostic tests, usually sensitivity and specificity of the tests are known and there is also at least a rough estimate of the prevalence of the tested disease. In the practival application, the positive predictive value (PPV) and the negative predictive value are of crucial importance. ```{r} ## Example: HIV test ## 1. ELISA screening test (4th generation) predValues(sens = 0.999, spec = 0.998, prev = 0.001) ## 2. Western-Plot confirmation test predValues(sens = 0.998, spec = 0.999996, prev = 1/3) ``` ### Performance Measures and Scores In the development of diagnostic tests and more general in binary classification a variety of performance measures and scores can be found in literature. Functions perfMeasures and prefScores compute several of them. ```{r} ## example from dataset infert fit <- glm(case ~ spontaneous+induced, data = infert, family = binomial()) pred <- predict(fit, type = "response") ## with group numbers perfMeasures(pred, truth = infert$case, namePos = 1) perfScores(pred, truth = infert$case, namePos = 1) ## with group names my.case <- factor(infert$case, labels = c("control", "case")) perfMeasures(pred, truth = my.case, namePos = "case") perfScores(pred, truth = my.case, namePos = "case") ## using weights perfMeasures(pred, truth = infert$case, namePos = 1, weight = 0.3) perfScores(pred, truth = infert$case, namePos = 1, weight = 0.3) ``` ### Optimal Cutoff The function optCutoff computes the optimal cutoff for various performance weasures for binary classification. More precisely, all performance measures that are implemented in function perfMeasures. ```{r} ## example from dataset infert fit <- glm(case ~ spontaneous+induced, data = infert, family = binomial()) pred <- predict(fit, type = "response") optCutoff(pred, truth = infert$case, namePos = 1) ``` The computation of an optimal cut-off doesn't make any sense for continuous scoring rules as their computation does not involve any cut-off (discretization/dichotomization). ### Hosmer-Lemeshow and le Cessie-van Houwelingen-Copas-Hosmer These tests are used to investigate the goodness of fit in logistic regression. ```{r} ## Hosmer-Lemeshow goodness of fit tests for C and H statistic HLgof.test(fit = pred, obs = infert$case) ## e Cessie-van Houwelingen-Copas-Hosmer global goodness of fit test HLgof.test(fit = pred, obs = infert$case, X = model.matrix(case ~ spontaneous+induced, data = infert)) ``` ### Sample Size Calculation Given an expected sensitivity and specificity we can compute sample size, power, delta or significance level of diagnostic test. ```{r} ## see n2 on page 1202 of Chu and Cole (2007) power.diagnostic.test(sens = 0.99, delta = 0.14, power = 0.95) # 40 power.diagnostic.test(sens = 0.99, delta = 0.13, power = 0.95) # 43 power.diagnostic.test(sens = 0.99, delta = 0.12, power = 0.95) # 47 ``` The sample size planning for developing binary classifiers in case of high dimensional data, we can apply function ssize.pcc, which is based on the probability of correct classification (PCC). ```{r} ## see Table 2 of Dobbin et al. (2008) g <- 0.1 fc <- 1.6 ssize.pcc(gamma = g, stdFC = fc, nrFeatures = 22000) ``` ## Omics Data ### Aggregating Technical Replicates In case of omics experiments it is often the case that technical replicates are determined and hence it is part of the preprocessing of the raw data to aggregate these technical replicates. This is the purpose of function repMeans. ```{r} M <- matrix(rnorm(100), ncol = 5) FL <- matrix(rpois(100, lambda = 10), ncol = 5) # only for this example repMeans(x = M, flags = FL, use.flags = "max", ndups = 5, spacing = 4) ``` ### 1- and 2-Way ANOVA Functions oneWayAnova and twoWayAnova return a function that can be used to perform a 1- or 2-way ANOVA, respectively. ```{r} af <- oneWayAnova(c(rep(1,5),rep(2,5))) ## p value af(rnorm(10)) x <- matrix(rnorm(12*10), nrow = 10) ## 2-way ANOVA with interaction af1 <- twoWayAnova(c(rep(1,6),rep(2,6)), rep(c(rep(1,3), rep(2,3)), 2)) ## p values apply(x, 1, af1) ## 2-way ANOVA without interaction af2 <- twoWayAnova(c(rep(1,6),rep(2,6)), rep(c(rep(1,3), rep(2,3)), 2), interaction = FALSE) ## p values apply(x, 1, af2) ``` ### Correlation Distance Matrix In the analysis of omics data correlation and absolute correlation distance matrices are often used during quality control. Function corDist can compute the Pearson, Spearman, Kendall or Cosine sample correlation and absolute correlation as well as the minimum covariance determinant or the orthogonalized Gnanadesikan-Kettenring correlation and absolute correlation. ```{r} M <- matrix(rcauchy(1000), nrow = 5) ## Pearson corDist(M) ## Spearman corDist(M, method = "spearman") ## Minimum Covariance Determinant corDist(M, method = "mcd") ``` ### MAD Matrix In case of outliers the MAD may be useful as dispersion measure. ```{r} madMatrix(t(M)) ``` ### Similarity Matrices First, we can plot a similarity matrix based on correlation. ```{r, fig.width=8, fig.height=7} M <- matrix(rnorm(1000), ncol = 20) colnames(M) <- paste("Sample", 1:20) M.cor <- cor(M) corPlot(M.cor, minCor = min(M.cor), labels = colnames(M)) ``` Next, we can use the MAD. ```{r, fig.width=8, fig.height=7} ## random data x <- matrix(rnorm(1000), ncol = 10) ## outliers x[1:20,5] <- x[1:20,5] + 10 madPlot(x, new = TRUE, maxMAD = 2.5, labels = TRUE, title = "MAD: Outlier visible") ## in contrast corPlot(x, new = TRUE, minCor = -0.5, labels = TRUE, title = "Correlation: Outlier masked") ``` ### Colors for Heatmaps Nowadays there are better solutions e.g. provided by Bioconductor package complexHeatmaps. This was my solution to get a better coloring of heatmaps. ```{r, fig.width=7, fig.height=9} ## generate some random data data.plot <- matrix(rnorm(100*50, sd = 1), ncol = 50) colnames(data.plot) <- paste("patient", 1:50) rownames(data.plot) <- paste("gene", 1:100) data.plot[1:70, 1:30] <- data.plot[1:70, 1:30] + 3 data.plot[71:100, 31:50] <- data.plot[71:100, 31:50] - 1.4 data.plot[1:70, 31:50] <- rnorm(1400, sd = 1.2) data.plot[71:100, 1:30] <- rnorm(900, sd = 1.2) nrcol <- 128 ## Load required packages library(gplots) library(RColorBrewer) myCol <- rev(colorRampPalette(brewer.pal(10, "RdBu"))(nrcol)) heatmap.2(data.plot, col = myCol, trace = "none", tracecol = "black", main = "standard colors") farbe <- heatmapCol(data = data.plot, col = myCol, lim = min(abs(range(data.plot)))-1) heatmap.2(data.plot, col = farbe, trace = "none", tracecol = "black", main = "heatmapCol colors") ``` ## String Alignment ### String Distances In Bioinformatics the (pairwise and multiple) alignment of strings is an important topic. For this one can use the distance or similarity between strings. The Hamming and the the Levenshtein (edit) distance are implemented in function stringDist. ```{r} x <- "GACGGATTATG" y <- "GATCGGAATAG" ## Hamming distance stringDist(x, y) ## Levenshtein distance d <- stringDist(x, y) d ``` In case of the Levenshtein (edit) distance, the respective scoring and traceback matrices are attached as attributes to the result. ```{r} attr(d, "ScoringMatrix") attr(d, "TraceBackMatrix") ``` The characters in the trace-back matrix reflect insertion of a gap in string y (d: deletion), match (m), mismatch (mm), and insertion of a gap in string x (i). ### String Similarities The function stringSim computes the optimal alignment scores for global (Needleman-Wunsch) and local (Smith-Waterman) alignments with constant gap penalties. Scoring and trace-back matrix are again attached as attributes to the results. ```{r} ## optimal global alignment score d <- stringSim(x, y) d attr(d, "ScoringMatrix") attr(d, "TraceBackMatrix") ## optimal local alignment score d <- stringSim(x, y, global = FALSE) d attr(d, "ScoringMatrix") attr(d, "TraceBackMatrix") ``` The entry stop indicates that the minimum similarity score has been reached. ### Optimal Alignment Finally, the function traceBack computes an optimal global or local alignment based on a trace back matrix as provided by function stringDist or stringSim. ```{r} x <- "GACGGATTATG" y <- "GATCGGAATAG" ## Levenshtein distance d <- stringDist(x, y) ## optimal global alignment traceBack(d) ## Optimal global alignment score d <- stringSim(x, y) ## optimal global alignment traceBack(d) ## Optimal local alignment score d <- stringSim(x, y, global = FALSE) ## optimal local alignment traceBack(d, global = FALSE) ``` ## sessionInfo ```{r} sessionInfo() ```