---
title: "Smoking data"
author: "Rob Dunne"
date: "Saturday, August 8, 2020"
output:
rmarkdown::html_vignette:
toc: true
toc_depth: 2
vignette: >
%\VignetteIndexEntry{Smoking data}
%\VignetteEngine{knitr::rmarkdown}
%\VignetteEncoding{UTF-8}
bibliography: paper.bib
---
```{r, include = FALSE}
knitr::opts_chunk$set(
collapse = TRUE,
comment = "#>"
)
knitr::opts_chunk$set(fig.width=6, fig.height=6, dpi=300,echo = FALSE)
knitr::opts_chunk$set(fig.pos = "H", out.extra = "")
```
A small example using the smoking data set, containing normalized transcript measurements for 51 subjects (23 "never-smoked" and 34 "smokers")
and 22283 transcripts of lung tissue. See @Spira.et.al.2004
I have removed that data set temporarily until I solve a problem with upload the package to CRAN
# load the data
```{r, echo=TRUE, eval=FALSE}
library(RFlocalfdr.data)
data(smoking)
?smoking
y<-smoking$y
smoking_data<-smoking$rma
y.numeric <-ifelse((y=="never-smoked"),0,1)
```
```{r, echo=FALSE, eval=FALSE}
if (!require("BiocManager", quietly = TRUE))
install.packages("BiocManager")
BiocManager::install("GEOquery")
BiocManager::install("annotate")
install.packages("geneExpressionFromGEO")
library(geneExpressionFromGEO)
DF1 <- getGeneExpressionFromGEO("GSE994", FALSE, FALSE) #retrieveGeneSymbols, verbose = FALSE)
aa<-match(gsub("([A-Z0-9]*).*","\\1", rownames(smoking_data)), colnames(DF1))
all.equal(aa,1:57)
#[1] TRUE
temp <- t(DF1[1:57])
#but class temp is a data.frame not a ‘AffyBatch’ object. So how do we normalize it?
## library(affy)
## rma.data <- rma(data)
## or
## rma.data <- expresso(data,
## bgcorrect.method = "rma",
## normalize.method = "quantiles",
## pmcorrect.method = "pmonly",
## summary.method = "medianpolish")
```
# fit a ranger model
```{r, echo=TRUE, eval=FALSE}
library(ranger)
rf1 <-ranger(y=y.numeric ,x=smoking_data,importance="impurity",seed=123, num.trees = 10000,
classification=TRUE)
t2 <-count_variables(rf1)
imp<-log(rf1$variable.importance)
#png("./supp_figures/smoking_log_importances.png")
plot(density(imp),xlab="log importances",main="")
#dev.off()
```
```{r, echo=TRUE, eval=FALSE}
# starting from a randomForest model
library(RFlocalfdr)
library(randomForest)
sert.seed(123)
rf2 <-randomForest(y=factor(y.numeric) ,x=smoking_data,importance=TRUE, ntree = 10000)
imp.rf2 <- log(rf2$importance[,"MeanDecreaseGini"])
t2.rf2 <-varUsed(rf2)
plot(density(imp.rf2),xlab="log importances",main="")
cutoffs <- c(2,3,4,5)
res.con<- determine_cutoff(imp.rf2,t2.rf2 ,cutoff=cutoffs,plot=c(2,3,4,5))
plot(cutoffs,res.con[,3],pch=15,col="red",cex=1.5,ylab="max(abs(y - t1))")
cutoffs[which.min(res.con[,3])]
#2
```
<!-- ```{r log_importances, echo=FALSE, fig.cap="", fig.align="center", out.width = '50%'} -->
<!-- #knitr::include_graphics("./supp_figures/smoking_log_importances.png") -->
<!-- knitr::include_graphics("./supp_figures/smoking_data_determine_cutoff.png") -->
<!-- ``` -->
```{r simulation2, echo=FALSE, fig.cap="A small simulated data set. Each band contains blocks of size {1, 2, 4, 8, 16, 32, 64}, and each block consists of correlated (identical variables).", fig.align="center", out.width = '50%'}
knitr::include_graphics("./supp_figures/smoking_log_importances.png")
```
# Determine a cutoff to get a unimodal density.
See \@ref(fig:log_importances) for the log importances. They are clearly multimodal and we try to determine a cutoff so
that we are left with a unimodal distribution.
```{r ,echo=TRUE, eval=FALSE}
cutoffs <- c(2,3,4,5)
#png("./supp_figures/smoking_data_determine_cutoff.png")
res.con<- determine_cutoff(imp,t2,cutoff=cutoffs,plot=c(2,3,4,5))
#dev.off()
#png("./supp_figures/smoking_data_determine_cutoffs_2.png")
plot(cutoffs,res.con[,3],pch=15,col="red",cex=1.5,ylab="max(abs(y - t1))")
#dev.off()
cutoffs[which.min(res.con[,3])]
```
<!-- ```{r smoking_data_determine_cutoff, echo=FALSE, fig.align="center", fig.show='hold', fig.cap="", out.width = '40%', out.height = '40%'} -->
<!-- knitr::include_graphics("./supp_figures/smoking_data_determine_cutoff.png") -->
<!-- knitr::include_graphics("./supp_figures/smoking_data_determine_cutoffs_2.png") -->
<!-- ``` -->
<!-- why cant I get them side by side -->
<!-- https://stackoverflow.com/questions/25415365/insert-side-by-side-png-images-using-knitr -->
# fit RFlocalfdr
We select a cutoff of 3 and fit the RFlocalfdr model
```{r , echo=TRUE,eval=FALSE}
temp<-imp[t2 > 3]
temp <- temp - min(temp) + .Machine$double.eps
qq <- plotQ(temp,debug.flag = 1)
ppp<-run.it.importances(qq,temp,debug.flag = 0)
#png("./supp_figures/smoking_significant_genes.png")
#aa<-significant.genes(ppp,temp,cutoff=0.05,do.plot=1)
aa<-significant.genes(ppp,temp,cutoff=0.05,debug.flag=0,do.plot=TRUE,use_95_q=TRUE)
#dev.off()
length(aa$probabilities) # 17
aa<-significant.genes(ppp,temp,cutoff=0.05,debug.flag=0,do.plot=TRUE,use_95_q=FALSE)
length(aa$probabilities) # 19
```
The option **do.plot=1** returns a plot containing the
- histogram of the importances in magenta
- fdr in black -- axis scale on right
- alpha value for significance -- axis scale on right
The option **do.plot=2** returns the same plot with the addition of
- fitted curve using estimates_C_0.95 in red
- 0.95 quantile of the fitted distribution, also in red
- cutoff for significant genes (in orange)
- abline(v = object$C_0.95, lwd = 2, col = "blue") what are these
- abline(v = object$cc, lwd = 2, col = "purple")
<!-- ```{r smoking_significant_genes, echo=FALSE, fig.cap="", fig.align="center", out.width = '50%'} -->
<!-- knitr::include_graphics("./supp_figures/smoking_significant_genes.png") -->
<!-- ``` -->
```{r , echo=TRUE,eval=FALSE}
sessionInfo()
```
```{r , echo=TRUE,eval=FALSE}
devtools::install_github("parsifal9/RFlocalfdr", build_vignettes = TRUE, force = TRUE)
library(RFlocalfdr)
data(smoking)
?smoking
y<-smoking$y
smoking_data<-smoking$rma
y.numeric <-ifelse((y=="never-smoked"),0,1)
library(ranger)
rf1 <-ranger(y=y.numeric ,x=smoking_data,importance="impurity",seed=123, num.trees = 10000,
classification=TRUE)
t2 <-count_variables(rf1)
imp<-log(rf1$variable.importance)
#png("./supp_figures/smoking_log_importances.png")
plot(density(imp),xlab="log importances",main="")
#dev.off()
cutoffs <- c(2,3,4,5)
#png("./supp_figures/smoking_data_determine_cutoff.png")
res.con<- determine_cutoff(imp,t2,cutoff=cutoffs,plot=c(2,3,4,5))
#dev.off()
#png("./supp_figures/smoking_data_determine_cutoffs_2.png")
plot(cutoffs,res.con[,3],pch=15,col="red",cex=1.5,ylab="max(abs(y - t1))")
#dev.off()
cutoffs[which.min(res.con[,3])]
temp<-imp[t2 > 3]
temp <- temp - min(temp) + .Machine$double.eps
qq <- plotQ(temp,debug.flag = 1)
ppp<-run.it.importances(qq,temp,debug.flag = 0)
#png("./supp_figures/smoking_significant_genes.png")
#aa<-significant.genes(ppp,temp,cutoff=0.05,do.plot=1)
aa<-significant.genes(ppp,temp,cutoff=0.05,debug.flag=0,do.plot=TRUE,use_95_q=TRUE)
#dev.off()
length(aa$probabilities) # 17 -- Roc gets 30
aa<-significant.genes(ppp,temp,cutoff=0.05,debug.flag=0,do.plot=TRUE,use_95_q=FALSE)
length(aa$probabilities) # 19-- Roc gets 30
```