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@ -0,0 +1,435 @@ |
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library(shiny) |
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library(openCyto) |
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library(flowCore) |
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library(flowWorkspace) |
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library(CytoML) |
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library(ggcyto) |
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# library(reshape2) |
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# library(CitFuns) |
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library(openxlsx) |
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library(tidyverse) |
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# Define UI for application that draws a histogram |
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ui <- fluidPage( |
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# Application title |
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titlePanel("Análisis Citometría ImmunoPreserve"), |
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# Sidebar with a slider input for number of bins |
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sidebarLayout( |
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sidebarPanel( |
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selectInput("phenotype", "Panel", selected="Panel1", choices=c("Panel1", "Panel2","Panel3","panel4")), |
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), |
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mainPanel( |
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textInput("cytopath", label="Directorio fenotipo", value=""), |
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actionButton("goButtonDir","Selecciona directorio fenotipo"), |
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textOutput("session"), |
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hr(), |
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actionButton("fcsconvert", "Convertir a fcs"), |
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hr(), |
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actionButton("pngexport", "Exportar informes"), |
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hr(), |
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textInput("dbpath", label="Ruta Base de Dades", value=""), |
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actionButton("goButtondbpath","Selecciona Ruta Base de Dades"), |
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textOutput("sessiondbpath"), |
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actionButton("popexport", "Actualizar BBDD") |
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) |
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) |
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) |
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# Define server logic required to draw a histogram |
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server <- function(input, output) { |
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observe({ |
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if(input$goButtonDir > 0){ |
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if (input$cytopath == ""){ |
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cito_dir<<-choose.dir() %>% gsub("\\","/",. ,fixed=T) %>% paste0("/") %>% stringi::stri_enc_tonative() |
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}else{ |
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cito_dir<<-input$cytopath %>% gsub("\\","/",. ,fixed=T) %>% gsub("/$", "", .) %>% paste0("/") %>% stringi::stri_enc_tonative() |
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} |
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output$session <- renderText( |
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cito_dir |
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) |
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} |
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}) |
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observe({ |
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if(input$goButtondbpath > 0){ |
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if (input$dbpath == ""){ |
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db_path<<-choose.dir() %>% gsub("\\","/",. ,fixed=T) %>% paste0("/") %>% stringi::stri_enc_tonative() |
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}else{ |
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db_path<<-input$dbpath %>% gsub("\\","/",. ,fixed=T) %>% gsub("/$", "", .) %>% paste0("/") %>% stringi::stri_enc_tonative() |
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} |
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output$sessiondbpath <- renderText( |
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db_path |
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) |
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} |
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}) |
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observeEvent(input$fcsconvert,{ |
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route<-cito_dir |
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files<-list.files(route, ".LMD") |
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for (lmd in files){ |
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fcs<-read.FCS(paste0(route,lmd), dataset = 2) |
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keyword(fcs)['$FIL']<-paste0(gsub(".LMD","",lmd), ".fcs") |
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write.FCS(fcs, paste0(route, gsub(".LMD","",lmd), ".fcs")) |
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} |
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print("Conversión completada") |
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}) |
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observeEvent(input$pngexport,{ |
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# if (input$phenotype == "Pop"){ |
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# route<-cito_dir |
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# |
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# ws<-open_flowjo_xml(paste0(route,"Populations.wsp")) |
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# gs<-flowjo_to_gatingset(ws, name="All Samples") |
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# |
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# sampleNames(gs)<-sapply(sampleNames(gs), function(x) strsplit(x, "Pop ")[[1]][2]) %>% |
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# gsub("[[:space:]][0-9]*.fcs_.[0-9]*","", . , perl = T) |
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# |
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# for (samp in sampleNames(gs)){ |
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# print(samp) |
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# p<-autoplot(gs[[samp]], bins=64) |
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# ggsave(paste0(route, samp,".pop.png"),p,width = 10, height = 10) |
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# } |
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# } |
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if (input$phenotype == "Panel1"){ |
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route<-cito_dir |
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ws<-open_flowjo_xml(paste0(route,"Panel1.wsp")) |
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gs<-flowjo_to_gatingset(ws, name="All Samples") |
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sampleNames(gs)<-sapply(sampleNames(gs), function(x) strsplit(x, "Panel1 ")[[1]][2]) %>% |
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gsub("[[:space:]][0-9]*.fcs_.[0-9]*","", . , perl = T) |
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gs<-gs[sampleNames(gs)[!grepl("Iso|ISO|iso",sampleNames(gs))]] |
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bool.comb<-apply( |
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expand.grid(c("","!"), c("","!"), c("","!"), c("","!")), |
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1, |
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function(x) paste0(x[1],"CTLA4 & ",x[2],"LAG3 & ",x[3],"PD1 & ",x[4], "TIM3") |
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) |
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bool.name<-apply( |
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expand.grid(c("+","-"), c("+","-"), c("+","-"), c("+","-")), |
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1, |
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function(x) paste0("CTLA4",x[1]," LAG3",x[2]," PD1",x[3]," TIM3",x[4]) |
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) |
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print("Booleanos CD8") |
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for (i in 1:length(bool.comb)){ |
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call<-substitute(booleanFilter(v), list(v=as.symbol(bool.comb[i]))) |
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boolgate<-eval(call) |
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gs_pop_add(gs, boolgate, parent="_CD8", name = bool.name[i]) |
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} |
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print("Booleanos CD4") |
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for (i in 1:length(bool.comb)){ |
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call<-substitute(booleanFilter(v), list(v=as.symbol(bool.comb[i]))) |
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boolgate<-eval(call) |
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gs_pop_add(gs, boolgate, parent="_CD4", name = bool.name[i]) |
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} |
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recompute(gs) |
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names<-sampleNames(gs) # %>% gsub("ab|Ab|AB|iso|Iso|ISO| ","",.) %>% unique() |
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nodes<-gs_get_pop_paths(gs) |
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# nodes<-gsub("â\u0081»", "-", nodes) |
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# nodes<-gsub("â\u0081º", "+", nodes) |
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nodes<-nodes[grepl("CTLA4", nodes)] |
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nodes<-nodes[!grepl("_CD4$|_CD8$|CTLA4$|TIM3$|PD1$|LAG3$", nodes)] |
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pop<-gs_pop_get_stats(gs, nodes=nodes,type="percent") %>% as.data.frame %>% mutate(percent=percent*100) |
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pop$percent<-round(pop$percent, digits=2) |
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# pop$pop<-gsub("â\u0081»", "n", pop$pop) |
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# pop$pop<-gsub("â\u0081º", "p", pop$pop) |
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pop$pop<-gsub("-", "n", pop$pop, fixed=T) |
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pop$pop<-gsub("+", "p", pop$pop, fixed=T) |
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pop$pop<-gsub(" ", "_", pop$pop) |
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## Esto si no hay isotipo |
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pop_sp<-pop |
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pop_sp["Population"]<-str_extract(pop_sp$pop, "/_CD[4,8]{1}/") %>% gsub("/|_","",.) |
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pop_sp$pop<-sapply(strsplit(pop_sp$pop, "/"), tail, 1) |
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pop_sp<-pop_sp %>% spread(pop, percent) |
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## Esto si hay Isotipo |
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# pop["Type"]<-"ab" |
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# pop[grepl("iso|ISO|Iso",pop$sample),"Type"]<-"iso" |
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# pop$sample<-gsub("iso|ISO|Iso|ab|AB|Ab| ","",pop$sample) |
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# pop_sp<-pop %>% spread(Type, percent) |
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# pop_sp["Net"]<-pop_sp$ab |
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# pop_sp[!grepl("CTLA4n_LAG3n_PD1n_TIGITn_TIM3n",pop_sp$pop),"Net"]<-pop_sp[!grepl("CTLA4n_LAG3n_PD1n_TIGITn_TIM3n",pop_sp$pop),"ab"]-pop_sp[!grepl("CTLA4n_LAG3n_PD1n_TIGITn_TIM3n",pop_sp$pop),"iso"] |
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# pop_sp$Net[pop_sp$Net < 0]<-0 |
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# pop_sp["Population"]<-str_extract(pop_sp$pop, "/CD[4,8]{1}/") %>% gsub("/","",.) |
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# pop_sp$pop<-sapply(strsplit(pop_sp$pop, "/"), tail, 1) |
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# |
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# pop_sp<-pop_sp %>% select(-ab,-iso) %>% spread(pop,Net) |
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# pop_sp$CTLA4n_LAG3n_PD1n_TIGITn_TIM3n<- pop_sp %>% select(-CTLA4n_LAG3n_PD1n_TIGITn_TIM3n) %>% group_by(sample,Population) %>% |
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# gather(pop, value, -sample,-Population) %>% summarise(n=100-sum(value)) %>% pull(n) |
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# if (input$dbtype == "OV"){ |
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# pop_sp <- rename(pop_sp, "samples"="sample") |
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# } |
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# if (input$dbtype %in% c("UM", "CC")){ |
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# pop_sp <- rename(pop_sp, "CODIGO"="sample") |
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# |
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pop_sql<-read.xlsx(paste0(db_path,"Panel1.xlsx"), sheet = "IC") |
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pop_sp<-pop_sp %>% merge(pop_sql %>% slice(0), all=T) %>% select(colnames(pop_sql)) |
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for (id in names){ |
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print(id) |
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# iso<-sampleNames(gs)[grepl(id, sampleNames(gs)) & grepl("iso|Iso|ISO",sampleNames(gs))] |
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# ab<-sampleNames(gs)[grepl(id, sampleNames(gs)) & grepl("ab|Ab|AB",sampleNames(gs))] |
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data<-pop_sp %>% filter(sample == id) |
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data1<-data %>% gather(phen, value, -sample, -Population) |
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data1$phen<-gsub("p","+",data1$phen) |
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data1$phen<-gsub("n","-",data1$phen) |
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data1$phen<-gsub("_"," ",data1$phen) |
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data1$phen<-gsub("n","-",data1$phen, fixed = T) |
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data1$phen<-gsub("p","+",data1$phen, fixed = T) |
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data1$phen<-gsub("_"," ",data1$phen) |
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data1[data1$value < 1, "phen"]<-"Other" |
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data1$phen<-gsub("[A-Z]*-*[0-9T]- *", "", data1$phen) |
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data1$phen<-gsub("+ $", "", data1$phen) |
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data1$phen[data1$phen == ""]<-"All Negative" |
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data1["phen1"]<-"PD1" |
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data1[!grepl("PD1+", data1$phen),"phen1"]<-NA |
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data1["phen2"]<-"TIM3" |
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data1[!grepl("TIM3+", data1$phen),"phen2"]<-NA |
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data1["phen3"]<-"CTLA4" |
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data1[!grepl("CTLA4+", data1$phen),"phen3"]<-NA |
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data1["phen4"]<-"LAG3" |
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data1[!grepl("LAG3+", data1$phen),"phen4"]<-NA |
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data1<-data1 %>% arrange(desc(value)) |
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data2<-data1 %>% filter(!phen %in% c("All Negative","Other")) |
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data1<-rbind(data2, data1 %>% filter(phen %in% c("All Negative","Other")) %>% arrange(desc(phen))) |
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data_cd8<-data1 %>% filter(Population == "CD8") |
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data_cd4<-data1 %>% filter(Population == "CD4") |
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data_cd8$ymax<-cumsum(data_cd8$value) |
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data_cd8$ymin<-c(0, head(data_cd8$ymax, n=-1)) |
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data_cd4$ymax<-cumsum(data_cd4$value) |
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data_cd4$ymin<-c(0, head(data_cd4$ymax, n=-1)) |
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data1<-rbind(data_cd8, data_cd4) |
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color<-c(c("CTLA4+ LAG3+ PD1+ TIM3+"="black","All Negative"="grey90","Other"="grey50", "PD1+"="#C07AFF", "CTLA4+"="#3EB3DE","TIM3+"="#5EF551","LAG3+"="#DEBB3E"), |
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c("CTLA4+ PD1+"="#6666FF","PD1+ TIM3+"="#849CA8", "LAG3+ PD1+"="#C47F9F", "CTLA4+ TIM3+"="#4ED498", "CTLA4+ LAG3+"="#8EB78E", "LAG3+ TIM3+"="#9ED848"), |
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c("CTLA4+ PD1+ TIM3+"="#B81515", "LAG3+ PD1+ TIM3+"="#0f5860")) |
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basic.color<-color[c("PD1+","TIM3+","CTLA4+","LAG3+")] |
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names(basic.color)<-c("PD1","TIM3","CTLA4","LAG3") |
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# Make the plot |
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g_coex<-ggplot(data1)+ |
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facet_grid(.~factor(Population, levels=c("CD8","CD4")))+ |
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geom_rect(aes(ymax=ymax, ymin=ymin, xmax=4.5, xmin=0), fill=color[data1$phen])+ |
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geom_rect(aes(ymax=ymax, ymin=ymin, xmax=5.4, xmin=5, fill=factor(phen1, levels=c("PD1","TIM3","CTLA4","LAG3"))))+ |
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geom_rect(aes(ymax=ymax, ymin=ymin, xmax=5.9, xmin=5.5, fill=factor(phen2, levels=c("PD1","TIM3","CTLA4","LAG3"))))+ |
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geom_rect(aes(ymax=ymax, ymin=ymin, xmax=6.4, xmin=6, fill=factor(phen3, levels=c("PD1","TIM3","CTLA4","LAG3"))))+ |
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geom_rect(aes(ymax=ymax, ymin=ymin, xmax=6.9, xmin=6.5, fill=factor(phen4, levels=c("PD1","TIM3","CTLA4","LAG3"))))+ |
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scale_fill_manual(values = basic.color, na.value="#FFFFFF00", drop=F, limits=c("PD1","TIM3","CTLA4","LAG3"), name="IC")+ |
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coord_polar(theta="y") + # Try to remove that to understand how the chart is built initially |
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xlim(c(0, 8)) +# Try to remove that to see how to make a pie chart |
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theme_classic()+ |
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theme(strip.background = element_blank(), |
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strip.text = element_text(size=12, face="bold"), |
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axis.line = element_blank(), |
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axis.ticks = element_blank(), |
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# plot.margin = margin(-200,0,0,0), |
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axis.text = element_blank()) |
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nodes<-gs_get_pop_paths(gs) |
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nodes_parent<-nodes[!grepl("CTLA4|LAG3|PD1|TIM3|root$", nodes)] |
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nodes_cd4<-nodes[grepl("CTLA4$|LAG3$|PD1$|TIM3$", nodes) & grepl("/_CD4/",nodes)] |
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nodes_cd8<-nodes[grepl("CTLA4$|LAG3$|PD1$|TIM3$", nodes) & grepl("/_CD8/",nodes)] |
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g1<-ggcyto_arrange(autoplot(gs[[id]], nodes_parent), nrow=2) |
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g2<-ggcyto_arrange(autoplot(gs[[id]], nodes_cd8), nrow=1) |
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g3<-ggcyto_arrange(autoplot(gs[[id]], nodes_cd4), nrow=1) |
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g_dots<-gridExtra::gtable_rbind(g1,g2,g3) |
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g_all<-ggpubr::ggarrange(g_dots, g_coex, ncol=1, heights=c(0.75,0.25)) |
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ggsave(paste0(route,id,".IC.png"), g_all, width = 10, height = 14) |
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ggsave(paste0(db_path, "Informes/",id,".IC.png"), g_all, width = 10, height = 14) |
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} |
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print("Informes finalizados!") |
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} |
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}) |
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observeEvent(input$popexport,{ |
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if (input$phenotype == "Pop"){ |
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route<-cito_dir |
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ws<-open_flowjo_xml(paste0(route,"Populations.wsp")) |
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gs<-flowjo_to_gatingset(ws, name="All Samples") |
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sampleNames(gs)<-sapply(sampleNames(gs), function(x) strsplit(x, "Pop ")[[1]][2]) %>% |
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gsub("[[:space:]][0-9]*.fcs_.[0-9]*","", . , perl = T) |
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nodes<-sapply(strsplit(gs_get_pop_paths(gs), "/"), tail, 1) |
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nodes<-nodes[grepl("_",nodes)] |
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pop<-gs_pop_get_stats(gs, nodes=nodes,type="percent") %>% as.data.frame %>% mutate(percent=percent*100) |
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pop[,"pop"]<-gsub("_","",pop$pop) |
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pop$pop<-gsub(" ","_",pop$pop) |
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pop$pop<-gsub("+","pos",pop$pop, fixed=T) |
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pop$pop<-gsub("-","neg",pop$pop, fixed=T) |
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pop<-rename(pop, "samples"="sample") |
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pop$percent<-round(pop$percent, digits=2) |
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pop_sp<-pop %>% spread(pop, percent) |
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pop_sql<-sqlFetch(dta, "POPULATIONS") %>% slice(0) |
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pop_sp<-pop_sp %>% merge(pop_sql, all=T) %>% select(colnames(pop_sql)) |
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vartypes<-rep("Number", pop_sp %>% select(-samples) %>% colnames %>% length) |
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names(vartypes)<-pop_sp %>% select(-samples) %>% colnames |
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sqlSave(dta, pop_sp, tablename="POPULATIONS", append = T, varTypes = vartypes, rownames = F) |
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print("Tabla POPULATIONS sincronizada.") |
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} |
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if (input$phenotype == "Panel1"){ |
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route<-cito_dir |
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ws<-open_flowjo_xml(paste0(route,"Panel1.wsp")) |
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gs<-flowjo_to_gatingset(ws, name="All Samples") |
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sampleNames(gs)<-sapply(sampleNames(gs), function(x) strsplit(x, "Panel1 ")[[1]][2]) %>% |
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gsub("[[:space:]][0-9]*.fcs_.[0-9]*","", . , perl = T) |
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gs<-gs[sampleNames(gs)[!grepl("Iso|ISO|iso",sampleNames(gs))]] |
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bool.comb<-apply( |
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expand.grid(c("","!"), c("","!"), c("","!"), c("","!")), |
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1, |
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function(x) paste0(x[1],"CTLA4 & ",x[2],"LAG3 & ",x[3],"PD1 & ",x[4], "TIM3") |
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) |
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bool.name<-apply( |
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expand.grid(c("+","-"), c("+","-"), c("+","-"), c("+","-")), |
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1, |
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function(x) paste0("CTLA4",x[1]," LAG3",x[2]," PD1",x[3]," TIM3",x[4]) |
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) |
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print("Booleanos CD8") |
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for (i in 1:length(bool.comb)){ |
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call<-substitute(booleanFilter(v), list(v=as.symbol(bool.comb[i]))) |
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boolgate<-eval(call) |
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gs_pop_add(gs, boolgate, parent="_CD8", name = bool.name[i]) |
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} |
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print("Booleanos CD4") |
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for (i in 1:length(bool.comb)){ |
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call<-substitute(booleanFilter(v), list(v=as.symbol(bool.comb[i]))) |
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boolgate<-eval(call) |
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gs_pop_add(gs, boolgate, parent="_CD4", name = bool.name[i]) |
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} |
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recompute(gs) |
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names<-sampleNames(gs) # %>% gsub("ab|Ab|AB|iso|Iso|ISO| ","",.) %>% unique() |
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nodes<-gs_get_pop_paths(gs) |
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# nodes<-gsub("â\u0081»", "-", nodes) |
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# nodes<-gsub("â\u0081º", "+", nodes) |
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nodes<-nodes[grepl("CTLA4", nodes)] |
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nodes<-nodes[!grepl("_CD4$|_CD8$|CTLA4$|TIM3$|PD1$|LAG3$", nodes)] |
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pop<-gs_pop_get_stats(gs, nodes=nodes,type="percent") %>% as.data.frame %>% mutate(percent=percent*100) |
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pop$percent<-round(pop$percent, digits=2) |
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# pop$pop<-gsub("â\u0081»", "n", pop$pop) |
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# pop$pop<-gsub("â\u0081º", "p", pop$pop) |
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pop$pop<-gsub("-", "n", pop$pop, fixed=T) |
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pop$pop<-gsub("+", "p", pop$pop, fixed=T) |
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pop$pop<-gsub(" ", "_", pop$pop) |
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## Esto si no hay isotipo |
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pop_sp<-pop |
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pop_sp["Population"]<-str_extract(pop_sp$pop, "/_CD[4,8]{1}/") %>% gsub("/|_","",.) |
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pop_sp$pop<-sapply(strsplit(pop_sp$pop, "/"), tail, 1) |
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pop_sp<-pop_sp %>% spread(pop, percent) |
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## Esto si hay Isotipo |
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# pop["Type"]<-"ab" |
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# pop[grepl("iso|ISO|Iso",pop$sample),"Type"]<-"iso" |
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# pop$sample<-gsub("iso|ISO|Iso|ab|AB|Ab| ","",pop$sample) |
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# pop_sp<-pop %>% spread(Type, percent) |
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# pop_sp["Net"]<-pop_sp$ab |
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# pop_sp[!grepl("CTLA4n_LAG3n_PD1n_TIGITn_TIM3n",pop_sp$pop),"Net"]<-pop_sp[!grepl("CTLA4n_LAG3n_PD1n_TIGITn_TIM3n",pop_sp$pop),"ab"]-pop_sp[!grepl("CTLA4n_LAG3n_PD1n_TIGITn_TIM3n",pop_sp$pop),"iso"] |
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# pop_sp$Net[pop_sp$Net < 0]<-0 |
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# pop_sp["Population"]<-str_extract(pop_sp$pop, "/CD[4,8]{1}/") %>% gsub("/","",.) |
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# pop_sp$pop<-sapply(strsplit(pop_sp$pop, "/"), tail, 1) |
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# |
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# pop_sp<-pop_sp %>% select(-ab,-iso) %>% spread(pop,Net) |
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# pop_sp$CTLA4n_LAG3n_PD1n_TIGITn_TIM3n<- pop_sp %>% select(-CTLA4n_LAG3n_PD1n_TIGITn_TIM3n) %>% group_by(sample,Population) %>% |
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# gather(pop, value, -sample,-Population) %>% summarise(n=100-sum(value)) %>% pull(n) |
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# if (input$dbtype == "OV"){ |
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# pop_sp <- rename(pop_sp, "samples"="sample") |
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# } |
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# if (input$dbtype %in% c("UM", "CC")){ |
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# pop_sp <- rename(pop_sp, "CODIGO"="sample") |
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# |
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pop_sql<-read.xlsx(paste0(db_path,"Panel1.xlsx"), sheet = "IC") |
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pop_sp<-pop_sp %>% merge(pop_sql %>% slice(0), all=T) %>% select(colnames(pop_sql)) |
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ic_sp<-rbind(pop_sql, pop_sp) |
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nodes<-sapply(strsplit(gs_get_pop_paths(gs), "/"), tail, 1) |
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nodes<-gs_get_pop_paths(gs)[grepl("_",nodes)] |
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pop<-gs_pop_get_stats(gs, nodes=nodes,type="percent") %>% as.data.frame %>% mutate(percent=percent*100) |
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pop<-pop %>% mutate(Population=str_extract(pop, "/_CD[4,8]{1}/"), |
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Population=case_when(is.na(Population)~"", |
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|
Population == "/_CD4/"~"_CD4", |
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|
Population == "/_CD8/"~"_CD8", |
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|
TRUE~Population), |
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pop=gsub("_","",pop), |
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|
pop=paste0(pop,Population)) %>% select(-Population) |
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pop$pop<-sapply(strsplit(pop$pop, "/"), tail, 1) |
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pop$pop<-gsub(" ","_",pop$pop) |
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# pop$pop<-gsub("+","pos",pop$pop, fixed=T) |
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# pop$pop<-gsub("-","neg",pop$pop, fixed=T) |
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# pop<-rename(pop, "samples"="sample") |
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|
pop$percent<-round(pop$percent, digits=2) |
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|
pop_sp<-pop %>% spread(pop, percent) |
|
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|
pop_sql<-read.xlsx(paste0(db_path,"Panel1.xlsx"), sheet = "POPULATIONS") |
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|
pop_sp<-pop_sp %>% merge(pop_sql %>% slice(0), all=T) %>% select(colnames(pop_sql)) |
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|
pop_sp<-rbind(pop_sql,pop_sp) |
|
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|
write.xlsx(list("IC"=ic_sp, "POPULATIONS"=pop_sp), paste0(db_path, "Panel1.xlsx")) |
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|
|
print("Tabla Panel1 sincronizada.") |
|
|
|
} |
|
|
|
}) |
|
|
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|
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|
|
} |
|
|
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|
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|
|
# Run the application |
|
|
|
shinyApp(ui = ui, server = server) |