#### Created by Jean-Philippe Fortin
#### March 28, 2014
#' @importFrom RColorBrewer brewer.pal
#' @importFrom grDevices palette
#' @importFrom utils write.csv
#' @importFrom stats complete.cases lm
#' @importFrom graphics par
#' @importFrom shiny renderPlot renderPrint downloadHandler
#' @importFrom shiny reactive runApp headerPanel
server.shinyMethyl <- function(shinyMethylSet1,
shinyMethylSet2=NULL
){
function(input, output, session) {
betaQuantiles <- getBeta(shinyMethylSet1)
mQuantiles <- getM(shinyMethylSet1)
methQuantiles <- getMeth(shinyMethylSet1)
unmethQuantiles <- getUnmeth(shinyMethylSet1)
cnQuantiles <- getCN(shinyMethylSet1)
greenControls <- getGreenControls(shinyMethylSet1)
redControls <- getRedControls(shinyMethylSet1)
covariates <<- pData(shinyMethylSet1)
pca <- getPCA(shinyMethylSet1)$scores
sampleNames <- sampleNames(shinyMethylSet1)
slideNames <- substr(sampleNames,1,10)
arrayNames <- substr(sampleNames,12,17)
plateNames <- substr(sampleNames,1,6)
controlNames <- names(greenControls)
method <- shinyMethylSet1@originObject
## In the case covariates is empty:
if (ncol(covariates)==0){
covariates <- data.frame(slide=slideNames,
plate=plateNames)
rownames(covariates) <- sampleNames
covariates <<- covariates
}
## Global variables:
## that will be accessed by the knitr report
mouse.click.indices <- c()
mouse.click.indices <<- c()
colorSet <- "Set1" # Default color set
colorSet <<- "Set1" # Default color set
sampleColors <- as.numeric(as.factor(plateNames)) ## Default sample colors
sampleColors <<- as.numeric(as.factor(plateNames)) ## Default sample colors
genderCutoff <- -0.4 # Default cutoff for gender prediction
genderCutoff <<- -0.4 # Default cutoff for gender prediction
current.control.type <- "BISULFITE CONVERSION I"
current.control.type <<- "BISULFITE CONVERSION I"
current.probe.type <- "I Green"
current.probe.type <<- "I Green"
current.density.type <- "M-value"
current.density.type <<- "M-value"
#########################################################
########### Colors
#########################################################
# To change the global color scheme:
setColor <- reactive({
colorSet <<- input$colorChoice
})
set.palette <- function(n, name){
## The name of the colors are part of the RColorBrewer package
if (name != "Pault" & name != "Rainbow"){
palette(brewer.pal(n = n, name = name ))
## Custom palette
} else if (name == "Pault"){
colors <- c("#332288","#88CCEE","#44AA99","#117733","#999933",
"#DDCC77","#661100","#CC6677","#882255","#AA4499")
colors <- c("#4477AA","#CC6677","#DDCC77","#117733","#88CCEE",
"#AA4499","#44AA99","#999933","#882255","#661100",
"#6699CC", "#AA4466")
palette(colors)
} else if (name == "Rainbow"){
colors <- c("#781C81", "#3F56A7","#4B91C0","#5FAA9F","#91BD61",
"#D8AF3D","#E77C30","#D92120")
palette(colors)
}
}
## To choose the colors according to the phenotype:
sampleColors <- reactive({
return(as.numeric(as.factor(covariates[,match(input$phenotype,
colnames(covariates))])))
})
#########################################################
########### Computation of the densities
#########################################################
## Return x and y matrices of the densities for raw data
returnDensityMatrix <- reactive({
index <- match(input$probeType,c("I Green","I Red","II","X","Y"))
if (input$mOrBeta=="Beta-value"){
bw <- input$bandwidth
quantiles <- betaQuantiles[[index]]
} else {
bw <- input$bandwidth2
quantiles <- mQuantiles[[index]]
}
matrix <- apply(quantiles, 2, function(x){
d <- density(x, bw = bw, n =512)
c(d$x,d$y)
})
matrix.x <- matrix[1:512,]
matrix.y <- matrix[513:(2*512),]
return(list(matrix.x = matrix.x, matrix.y = matrix.y))
})
## Return x and y matrices of the densities for normalized data
returnDensityMatrixNorm <- reactive({
if (is.null(shinyMethylSet2)){
return(NULL)
} else {
index <- match(input$probeType,
c("I Green","I Red","II","X","Y"))
}
if (input$mOrBeta=="Beta-value"){
bw <- input$bandwidth
quantiles <- shinyMethylSet2@betaQuantiles[[index]]
} else {
bw <- input$bandwidth2
quantiles <- shinyMethylSet2@mQuantiles[[index]]
}
matrix <- apply(quantiles, 2, function(x){
d <- density(x, bw = bw, n =512)
c(d$x,d$y)
})
matrix.x <- matrix[1:512,]
matrix.y <- matrix[513:(2*512),]
return(list(matrix.x = matrix.x, matrix.y = matrix.y))
})
#########################################################
########### Mouse clicks section
#########################################################
## Check if the selected sample is already in the list or not:
check.mouse.clicks <- function(clickedIndex,
mouse.click.indices){
if (length(mouse.click.indices)!=0){
if (clickedIndex %in% mouse.click.indices){
mouse.click.indices <- mouse.click.indices[mouse.click.indices!=clickedIndex]
} else {
mouse.click.indices <- c(mouse.click.indices, clickedIndex)
}
} else {
mouse.click.indices <- c(mouse.click.indices, clickedIndex)
}
return(mouse.click.indices)
}
## To update the list of mouse clicks from internal controls plot:
updateMouseClicks <- reactive({
mouse.x <- input$controlsHover$x
mouse.y <- input$controlsHover$y
if (!is.null(mouse.x) & !is.null(mouse.y)){
y <- reactiveControlStat()
n <- length(y)
xDiff <- ((1:n)-rep(mouse.x, n)) / n
yDiff <- (y - mouse.y)/(1.3*max(y))
clickedIndex <- which.min(xDiff^2 + yDiff^2)
names(clickedIndex) <- shinyMethylSet1@sampleNames[clickedIndex]
if (current.control.type == as.character(input$controlType)){
mouse.click.indices <<- check.mouse.clicks(clickedIndex, mouse.click.indices)
}
current.control.type <<- as.character(input$controlType)
}
mouse.click.indices
})
## To update the list of mouse clicks from quality control plot:
updateMouseClicksQC <- reactive({
mouse.x <- input$qualityHover$x
mouse.y <- input$qualityHover$y
if (!is.null(mouse.x) & !is.null(mouse.y)){
med <- as.integer(nrow(methQuantiles[[3]])/2)
mediansU <- unlist(unmethQuantiles[[3]][med,])
mediansM <- unlist(methQuantiles[[3]][med,])
x <- log2(mediansU)
y <- log2(mediansM)
n <- length(y)
range.x <- max(x) - min(x)
range.y <- max(y) - min(y)
xDiff <- (x - mouse.x) / (1.4*range.x)
yDiff <- (y - mouse.y)/ (1.4*range.y)
clickedIndex <- which.min(xDiff^2 + yDiff^2)
names(clickedIndex) <- shinyMethylSet1@sampleNames[clickedIndex]
mouse.click.indices <<- check.mouse.clicks(clickedIndex,
mouse.click.indices)
}
mouse.click.indices
})
## To update the list of mouse clicks from densities plot:
updateMouseClicksDensities <- reactive({
mouse.x <- input$densitiesHover$x
mouse.y <- input$densitiesHover$y
#print(mouse.x)
#print(mouse.y)
if (!is.null(mouse.x) & !is.null(mouse.y)){
# If Beta values are selected
if (input$mOrBeta=="Beta-value") {
if (input$probeType == "II"){
ylim <- c(0,6)
} else {
ylim <- c(0,10)
}
xlim = c(-0.2,1.2)
} else {
xlim = c(-8,8)
ylim = c(0,0.35)
}
range.x <- xlim[2]-xlim[1]
range.y <- ylim[2]-ylim[1]
if (!is.null(mouse.x)){
density.matrix <- returnDensityMatrix()
x.matrix <- density.matrix[[1]]
y.matrix <- density.matrix[[2]]
x.matrix <- ((x.matrix - mouse.x)/range.x)^2
y.matrix <- ((y.matrix - mouse.y)/range.y)^2
clickedIndex <- arrayInd(which.min(x.matrix+y.matrix),
c(512,ncol(x.matrix)))[2]
names(clickedIndex) <- shinyMethylSet1@sampleNames[clickedIndex]
if (current.probe.type == as.character(input$probeType) &
current.density.type == as.character(input$mOrBeta)){
mouse.click.indices <<- check.mouse.clicks(clickedIndex,
mouse.click.indices)
}
current.probe.type <<- as.character(input$probeType)
current.density.type <<- as.character(input$mOrBeta)
}
}
mouse.click.indices
})
## To update the list of mouse clicks from normalized densities plot:
updateMouseClicksDensitiesNorm <- reactive({
mouse.x <- input$normHover$x
mouse.y <- input$normHover$y
if (!is.null(mouse.x) & !is.null(mouse.y)){
## If Beta values are selected
if (input$mOrBeta=="Beta-value") {
if (input$probeType == "II"){
ylim <- c(0,6)
} else {
ylim <- c(0,10)
}
xlim = c(-0.2,1.2)
} else {
xlim = c(-8,8)
ylim = c(0,0.35)
}
range.x <- xlim[2]-xlim[1]
range.y <- ylim[2]-ylim[1]
if (!is.null(mouse.x)){
density.matrix <- returnDensityMatrixNorm()
x.matrix <- density.matrix[[1]]
y.matrix <- density.matrix[[2]]
x.matrix <- ((x.matrix - mouse.x)/range.x)^2
y.matrix <- ((y.matrix - mouse.y)/range.y)^2
clickedIndex <- arrayInd(which.min(x.matrix+y.matrix),
c(512,ncol(x.matrix)))[2]
names(clickedIndex) <- shinyMethylSet2@sampleNames[clickedIndex]
mouse.click.indices <<- check.mouse.clicks(clickedIndex,
mouse.click.indices)
}
}
mouse.click.indices
})
## To update the list from all plots:
reactive.mouse.click.indices <- reactive({
updateMouseClicks()
updateMouseClicksQC()
updateMouseClicksDensities()
updateMouseClicksDensitiesNorm()
return(mouse.click.indices)
})
## Print the list of selected samples:
output$cumulativeListPrint <- renderPrint({
names <- names(reactive.mouse.click.indices())
n <- length(names)
if (n >=1){
cat("Selected samples: \n \n")
for (ii in 1:n){
cat(paste0(names[ii], "\n"))
}
}
})
cumulativeList <- reactive({
names <- names(reactive.mouse.click.indices())
return(names)
})
## To write the selected samples into a csv file:
output$selectedSamples <- downloadHandler(
filename <- "selectedSamples.csv",
content <- function(con){
write.csv(cumulativeList(),con)
}
)
#########################################################
########### Densities plots
#########################################################
## Densities plot for raw data
output$rawDensities <- renderPlot({
set.palette(n=8, name=setColor())
colors <- sampleColors()
lwd <- as.numeric(input$lwd)
lty <- as.numeric(input$lty)
index = match(input$probeType,c("I Green","I Red","II","X","Y"))
## Plot specifications
if (input$mOrBeta=="Beta-value"){
xlim <- c(-0.2,1.2)
if (input$probeType == "II"){
ylim <- c(0,6)
} else {
ylim <- c(0,10)
}
from = -4; to = 4;
main = "BETA-VALUE DENSITIES"
xlab = "Beta-values"
bw <- input$bandwidth
quantiles <- betaQuantiles[[index]]
} else {
xlim <- c(-8,8)
ylim <- c(0, 0.35)
from = -10; to = 10;
main = "M-VALUE DENSITIES"
xlab = "M-values"
quantiles <- mQuantiles[[index]]
bw <- input$bandwidth2
}
densitiesPlot(matrix.x = returnDensityMatrix()[[1]],
matrix.y = returnDensityMatrix()[[2]],
quantiles = quantiles,
sampleNames = sampleNames,
main = main, xlab = xlab,
xlim = xlim, ylim = ylim, col = colors,
mean = input$mean,
lwd = lwd, lty = lty,
from = from, to = to, bw = bw)
if (!input$mean){
addHoverDensity(selectedSamples = reactive.mouse.click.indices(),
sampleNames = sampleNames,
matrix.x = returnDensityMatrix()[[1]],
matrix.y = returnDensityMatrix()[[2]],
col = colors)
}
## To draw the lines:
if (input$mOrBeta=="Beta-value"){
abline(v=0,lty=3,lwd=3)
abline(v=1,lty=3,lwd=3)
} else {
abline(v=0,lty=3,lwd=3)
}
})
## Density plot for normalized data:
output$normDensities <- renderPlot({
if (is.null(shinyMethylSet2)){
return(NULL)
} else {
set.palette(n=8, name=setColor())
colors <- sampleColors()
lwd <- as.numeric(input$lwd)
lty <- as.numeric(input$lty)
index <- match(input$probeType,
c("I Green","I Red","II","X","Y"))
## Plot specifications
if (input$mOrBeta=="Beta-value"){
xlim <- c(-0.2,1.2)
if (input$probeType == "II"){
ylim <- c(0,6)
} else {
ylim <- c(0,10)
}
from = -4; to = 4;
main = "Normalized data (Beta values)"
xlab = "Beta-values"
bw <- input$bandwidth
quantiles = shinyMethylSet2@betaQuantiles[[index]]
} else {
xlim <- c(-8,8)
ylim <- c(0, 0.35)
from = -10; to = 10;
main = "Normalized data (M values)"
xlab = "M-values"
quantiles = shinyMethylSet2@mQuantiles[[index]]
bw <- input$bandwidth2
}
densitiesPlot(matrix.x = returnDensityMatrixNorm()[[1]],
matrix.y = returnDensityMatrixNorm()[[2]],
quantiles = quantiles,
sampleNames = sampleNames,
main = main, xlab = xlab,
xlim = xlim, ylim = ylim, col = colors,
mean = input$mean,
lwd = lwd, lty = lty,
from = from, to = to, bw=bw)
if (!input$mean){
addHoverDensity(selectedSamples = reactive.mouse.click.indices(),
sampleNames = sampleNames,
matrix.x = returnDensityMatrixNorm()[[1]],
matrix.y = returnDensityMatrixNorm()[[2]],
col = colors)
}
## To draw the lines:
if (input$mOrBeta=="Beta-value"){
abline(v=0,lty=3,lwd=3)
abline(v=1,lty=3,lwd=3)
} else {
abline(v=0,lty=3,lwd=3)
}
}
})
#########################################################
########### Control probes and QC plot
#########################################################
## Return a summary measure of the selected control probes:
reactiveControlStat <- reactive({
controlStat <- returnControlStat(input$controlType,
greenControls = greenControls,
redControls = redControls,
controlNames = controlNames
)
return(controlStat)
})
## Internal controls plots
output$internalControls <- renderPlot({
colors <- sampleColors()
set.palette(n=8, name=setColor())
controlStat <- reactiveControlStat()
plotInternalControls(y = controlStat,
col = colors,
main = input$controlType,
sampleNames = sampleNames)
addHoverPoints(y = controlStat,
selectedSamples = reactive.mouse.click.indices(),
sampleNames = sampleNames)
})
## Quality control plot
output$medianChannels <- renderPlot({
set.palette(n=8, name=setColor())
colors <- sampleColors()
plotQC(unmethQuantiles = unmethQuantiles,
methQuantiles = methQuantiles,
sampleNames = sampleNames,
col = colors)
controlStat <- reactiveControlStat()
addHoverQC(y = controlStat,
selectedSamples = reactive.mouse.click.indices(),
unmethQuantiles = unmethQuantiles,
methQuantiles = methQuantiles)
})
#########################################################
########### Sex plot
#########################################################
## To change the gender cutoff for prediction:
setGenderCutoff <- reactive({
if (!is.null(input$genderCutoff)){
genderCutoff <<- input$genderCutoff$x
}
})
output$diffPrint <- renderPrint({
if (ncol(data())!=3){
cat("No gender was provided in the phenotype data")
} else {
diff.genders <- diffGenders()
diff.genders <- diff.genders[!is.na(diff.genders)]
if (!is.null(diffGenders())){
n <- length(diffGenders())
for (i in 1:n){
cat(paste0(diffGenders()[i]),"\n")
}
} else {
cat("The provided gender agrees with the predicted gender for all samples")
}
}
})
## Sex plot
output$genderClustering <- renderPlot({
setGenderCutoff()
plotPredictedGender(cutoff = genderCutoff,
cnQuantiles = cnQuantiles)
plotDiscrepancyGenders(cutoff = genderCutoff,
cnQuantiles = cnQuantiles,
covariates = covariates)
})
data <- reactive({
setGenderCutoff()
predictedGender <- returnPredictedGender(cutoff = genderCutoff,
cnQuantiles = cnQuantiles)
givenGender <- returnGivenGender(covariates)
dataToReturn <- data.frame(predictedGender = predictedGender)
if (!is.null(givenGender)){
non.matching.samples <- predictedGender != givenGender
na.samples <- is.na(givenGender)
n <- length(predictedGender)
agree <- rep("YES",n)
agree[non.matching.samples] <- "NO"
agree[na.samples] <- NA
dataToReturn$givenGender <- givenGender
dataToReturn$agree <- agree
}
rownames(dataToReturn) <- sampleNames
return(dataToReturn)
})
output$downloadClusters <- downloadHandler(
filename <- "predictedGender.csv",
content <- function(con){
write.csv(data(),con)
})
diffGenders <- reactive({
non.matching.samples <- c()
diffs <- data()
if (ncol(diffs)==3){
non.matching.samples <- shinyMethylSet1@sampleNames[diffs$agree=="NO"]
non.matching.samples <- non.matching.samples[complete.cases(non.matching.samples)]
}
return(non.matching.samples)
})
output$diffPrint <- renderPrint({
if (ncol(data())!=3){
cat("No gender was provided in the phenotype data")
} else {
diff.genders <- diffGenders()
diff.genders <- diff.genders[!is.na(diff.genders)]
if (!is.null(diffGenders())){
n <- length(diffGenders())
for (i in 1:n){
cat(paste0(diffGenders()[i]),"\n")
}
} else {
cat("The provided gender agrees with the predicted gender for all samples")
}
}
})
## Densities plot for gender X
output$densitiesGenderX <- renderPlot({
setGenderCutoff()
lwd <- as.numeric(input$lwd)
lty <- as.numeric(input$lty)
bw <- input$bandwidth
index <- match("X",c("I Green","I Red","II","X","Y"))
matrix <- apply(betaQuantiles[[index]], 2, function(x){
d <- density(x, bw = bw, n =512)
c(d$x,d$y)
})
matrix.x <- matrix[1:512,]
matrix.y <- matrix[513:(2*512),]
predictedGender <- returnPredictedGender(cutoff = genderCutoff,
cnQuantiles = cnQuantiles)
colors <- rep("lightskyblue", length(predictedGender))
colors[predictedGender=="F"] <- "orange"
densitiesPlot(matrix.x = matrix.x,
matrix.y = matrix.y,
quantiles = betaQuantiles[[index]],
sampleNames = sampleNames,
main = "X Chromosome",
xlab = "Beta-values",
xlim = c(-0.2,1.2),
ylim = c(0,7),
bw = input$bandwidth,
lwd = lwd,
lty = lty,
mean = FALSE,
col = colors,
from=-4,
to =4)
abline(v=0,lty=3,lwd=3)
abline(v=1,lty=3,lwd=3)
})
#########################################################
########### PCA plot
#########################################################
output$pcaPlot <- renderPlot({
set.palette(n=8, name=setColor())
##palette(brewer.pal(8,setColor()))
if (method!="Merging"){
plotPCA(pca = pca,
pc1 = input$pc1,
pc2 = input$pc2,
col = sampleColors(),
covariates = covariates,
selectedCov = input$phenotype
)
}
})
## Print the summary of the PCA regression:
output$modelPrint <- renderPrint({
if (method!="Merging"){
y <- shinyMethylSet1@pca$scores[,as.numeric(input$pcToExplore)]
cov <- covariates[match(rownames(shinyMethylSet1@pca$scores),
rownames(covariates)),]
x <- (as.factor(cov[,match(input$covToRegress,colnames(cov))]))
model <- lm(y~ x)
return(summary(model))
}
else {
return("Merging was used to create the shinyMethylSet1; no PCA was performed. ")
}
})
#########################################################
########### Design plot
#########################################################
output$arrayDesign <- renderPlot({
set.palette(n=8, name=setColor())
color <- covariates[,match(input$phenotype,colnames(covariates))]
plotDesign450k(as.character(sampleNames), covariates = color ,
legendTitle = input$phenotype)
})
output$arrayDesignLegend <- renderPlot({
set.palette(n=8, name=setColor())
color <- covariates[,match(input$phenotype,colnames(covariates))]
plotLegendDesign450k(as.character(sampleNames), covariates =color ,
legendTitle = input$phenotype)
})
#########################################################
########### Probe Bias Plot
#########################################################
## Densities plot for raw data
output$probeBiasPlot <- renderPlot({
set.palette(n=8, name=setColor())
colors <- sampleColors()
lwd <- as.numeric(input$lwd)
lty <- as.numeric(input$lty)
index = match(input$probeType,c("I Green","I Red","II","X","Y"))
selectedSample <- as.character(input$selectedSampleBias)
sampleIndex <- match(selectedSample, sampleNames)
indexIGreen <- 1
indexIRed <- 2
indexII <- 3
bw <- input$bandwidth
quantilesIGreen <- betaQuantiles[[indexIGreen]][,sampleIndex]
quantilesIRed <- betaQuantiles[[indexIRed]][,sampleIndex]
quantilesII <- betaQuantiles[[indexII]][,sampleIndex]
quantilesIRed <- as.vector(quantilesIRed)
quantilesIGreen <- as.vector(quantilesIGreen)
quantilesII <- as.vector(quantilesII)
## Plot specifications
xlim <- c(-0.2,1.2)
if (input$probeType == "II"){
ylim <- c(0,6)
} else {
ylim <- c(0,10)
}
from = -4; to = 4;
main = "Probe Type Differences - Raw Data"
xlab = "Beta-values"
bw <- input$bandwidth
plot(density(quantilesIGreen, bw=bw),
main = main,
ylab = "Density",
xlab = xlab,
col = "olivedrab",
xlim = xlim,
ylim = ylim, lty=lty, lwd=6)
lines(density(quantilesII, bw=bw), col="black", lty=lty, lwd=6)
lines(density(quantilesIRed, bw=bw), col="brown2", lty=lty, lwd=6)
## To draw the lines:
abline(v=0,lty=3,lwd=3)
abline(v=1,lty=3,lwd=3)
legend(x=0.4, y=8,
c("Type I Red","Type I Green","Type II"),
col=c("brown2","olivedrab","black"), lwd=6, lty=1, bty="n")
})
## Densities plot for raw data
output$probeBiasPlotNorm <- renderPlot({
if (is.null(shinyMethylSet2)){
return(NULL)
} else {
set.palette(n=8, name=setColor())
colors <- sampleColors()
lwd <- as.numeric(input$lwd)
lty <- as.numeric(input$lty)
index = match(input$probeType,c("I Green","I Red","II","X","Y"))
selectedSample <- as.character(input$selectedSampleBias)
sampleIndex <- match(selectedSample, sampleNames)
indexIGreen <- 1
indexIRed <- 2
indexII <- 3
bw = input$bandwidth
quantilesIGreen <- shinyMethylSet2@betaQuantiles[[indexIGreen]][,sampleIndex]
quantilesIRed <- shinyMethylSet2@betaQuantiles[[indexIRed]][,sampleIndex]
quantilesII <- shinyMethylSet2@betaQuantiles[[indexII]][,sampleIndex]
quantilesIRed <- as.vector(quantilesIRed)
quantilesIGreen <- as.vector(quantilesIGreen)
quantilesII <- as.vector(quantilesII)
## Plot specifications
xlim <- c(-0.2,1.2)
if (input$probeType == "II"){
ylim <- c(0,6)
} else {
ylim <- c(0,10)
}
from = -4; to = 4;
main = "Probe Type Differences - Normalized Data"
xlab = "Beta-values"
bw <- input$bandwidth
plot(density(quantilesIGreen, bw=bw),
main = main,
ylab = "Density",
xlab = xlab,
col = "olivedrab",
xlim = xlim,
ylim = ylim, lty=lty, lwd=6)
lines(density(quantilesII, bw=bw), col="black", lty=lty, lwd=6)
lines(density(quantilesIRed, bw=bw), col="brown2", lty=lty, lwd=6)
legend(x=0.4, y=8,
c("Type I Red","Type I Green","Type II"),
col=c("brown2","olivedrab","black"), lwd=6, lty=1, bty="n")
# To draw the lines:
abline(v=0,lty=3,lwd=3)
abline(v=1,lty=3,lwd=3)
}
})
## End
}
}
