Ns126:Calendar/NOTES/2015-4-21: Difference between revisions
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{| class="wikitable" style="text-align: right; color: red;" | {| class="wikitable" style="text-align: right; color: red;" | ||
| ||cpg||cancer||gene||chr||start||end | | ||cpg||cancer||GSI||gene||chr||start||end | ||
|- | |- | ||
| | | cg18565473||cg18565473||BRCA||0.932682778025148||ETS1||11||128392042||128392162 | ||
|- | |- | ||
| | | cg23884187||cg23884187||BRCA||0.915793344077998||C20orf95||20||37275009||37275129 | ||
|- | |- | ||
| | | cg14052221||cg14052221||BRCA||0.912760995997153||PSAT1||9||80911998||80912118 | ||
|- | |- | ||
| | | cg24797187||cg24797187||BRCA||0.912059797561713||AFF3||2||100175708||100175828 | ||
|- | |- | ||
| | | cg18943693||cg18943693||BRCA||0.911675629661679||||1||155043501||155043621 | ||
|- | |- | ||
| | | cg12587766||cg12587766||COAD||0.962866407567279||LIFR||5||38556375||38556495 | ||
|- | |- | ||
| | | cg10157975||cg10157975||COAD||0.959068233395622||ZNF304||19||57862382||57862502 | ||
|- | |- | ||
| | | cg23977631||cg23977631||COAD||0.954738746472514||LONRF2||2||100938739||100938859 | ||
|- | |- | ||
| | | cg04117229||cg04117229||COAD||0.952980963623186||SPG20||13||36920753||36920873 | ||
|- | |- | ||
| | | cg09854653||cg09854653||COAD||0.952746178623677||QKI||6||163834843||163834963 | ||
|- | |- | ||
| | | cg03988778||cg03988778||HNSC||0.94124876913729||SVIP||11||22850831||22850951 | ||
|- | |- | ||
| | | cg08211306||cg08211306||HNSC||0.936441823565694||ENPP4||6||46097724||46097844 | ||
|- | |- | ||
| | | cg26968387||cg26968387||HNSC||0.932552091587754||ZNF420||19||37569208||37569328 | ||
|- | |- | ||
| | | cg03280624||cg03280624||HNSC||0.925140832107714||ZNF583||19||56915595||56915715 | ||
|- | |- | ||
| | | cg00471966||cg00471966||HNSC||0.92378591584013||ZNF420||19||37569290||37569410 | ||
|- | |- | ||
| | | cg26228351||cg26228351||KIRC||0.866821443647375||KIF21B||1||200992596||200992716 | ||
|- | |- | ||
| | | cg00593900||cg00593900||KIRC||0.840429026779539||ANGPTL6||19||10206686||10206806 | ||
|- | |- | ||
| | | cg11697226||cg11697226||KIRC||0.83932716910874||TNFRSF11A||18||59992325||59992445 | ||
|- | |- | ||
| | | cg09865339||cg09865339||KIRC||0.836396509994196||GPC2;STAG3||7||99774875||99774995 | ||
|- | |- | ||
| | | cg02632185||cg02632185||KIRC||0.824262102898211||MAST4||5||66299726||66299846 | ||
|- | |- | ||
| | | cg15598442||cg15598442||KIRP||0.926520443898138||||1||25175003||25175123 | ||
|- | |- | ||
| | | cg26622232||cg26622232||KIRP||0.907406091283148||OXR1||8||107669727||107669847 | ||
|- | |- | ||
| | | cg16326979||cg16326979||KIRP||0.898630310618579||OXR1||8||107670101||107670221 | ||
|- | |- | ||
| | | cg17031478||cg17031478||KIRP||0.897081288137618||HOXC4;HOXC5||12||54427113||54427233 | ||
|- | |- | ||
| | | cg17136799||cg17136799||KIRP||0.888400341253348||OXR1||8||107669723||107669843 | ||
|- | |- | ||
| | | cg16579555||cg16579555||LIHC||0.968046292345837||RNF135||17||29298292||29298412 | ||
|- | |- | ||
| | | cg26240185||cg26240185||LIHC||0.966346469016627||FAR1||11||13690097||13690217 | ||
|- | |- | ||
| | | cg15375239||cg15375239||LIHC||0.961490202023512||SPINT2||19||38755227||38755347 | ||
|- | |- | ||
| | | cg15969216||cg15969216||LIHC||0.958031159935377||TSC22D1||13||45150202||45150322 | ||
|- | |- | ||
| | | cg03326059||cg03326059||LIHC||0.958023849811357||FAR1||11||13690100||13690220 | ||
|- | |- | ||
| | | cg13215643||cg13215643||LUAD||0.855326865879107||DACT1||14||59104765||59104885 | ||
|- | |- | ||
| | | cg07017994||cg07017994||LUAD||0.842561347568723||EPHB6||7||142552854||142552974 | ||
|- | |- | ||
| | | cg12487147||cg12487147||LUAD||0.839197022612119||HSD17B8||6||33172382||33172502 | ||
|- | |- | ||
| | | cg26615830||cg26615830||LUAD||0.837145673329791||MSX1||4||4861270||4861390 | ||
|- | |- | ||
| | | cg21929771||cg21929771||LUAD||0.820700381610812||PTPRU||1||29586520||29586640 | ||
|- | |- | ||
| | | cg07240673||cg07240673||LUSC||0.899039308699477||CLUAP1||16||3550848||3550968 | ||
|- | |- | ||
| | | cg18772127||cg18772127||LUSC||0.886204255597843||CMTM7||3||32443436||32443556 | ||
|- | |- | ||
| | | cg08562243||cg08562243||LUSC||0.874881316596866||CLUAP1||16||3551109||3551229 | ||
|- | |- | ||
| | | cg02566698||cg02566698||LUSC||0.872675000318173||CLUAP1||16||3550872||3550992 | ||
|- | |- | ||
| | | cg02379764||cg02379764||LUSC||0.869773727428307||CLUAP1||16||3550968||3551088 | ||
|- | |- | ||
| | | cg07635623||cg07635623||PRAD||0.955666568317178||SERPINB1||6||2841815||2841935 | ||
|- | |- | ||
| | | cg14283569||cg14283569||PRAD||0.947556926299886||||19||51416153||51416273 | ||
|- | |- | ||
| | | cg05098590||cg05098590||PRAD||0.94340784073766||ADD3||10||111767319||111767439 | ||
|- | |- | ||
| | | cg10938374||cg10938374||PRAD||0.942601773040438||IER3||6||30711998||30712118 | ||
|- | |- | ||
| | | cg16232979||cg16232979||PRAD||0.942431007327551||TPM4||19||16187571||16187691 | ||
|- | |- | ||
| | | cg27115721||cg27115721||THCA||0.881950689779005||FAM49A||2||16790278||16790398 | ||
|- | |- | ||
| | | cg04358131||cg04358131||THCA||0.877589155598355||MAFK||7||1572192||1572312 | ||
|- | |- | ||
| | | cg10540754||cg10540754||THCA||0.86094593963059||FAM49A||2||16790310||16790430 | ||
|- | |- | ||
| | | cg22749810||cg22749810||THCA||0.855202902841297||RNF213||17||78237329||78237449 | ||
|- | |- | ||
| | | cg12822074||cg12822074||THCA||0.849782546942327||RTN4RL2||11||57243805||57243925 | ||
|- | |- | ||
| | | cg03221247||cg03221247||UCEC||0.955580574481869||LYPLAL1||1||219347398||219347518 | ||
|- | |- | ||
| | | cg15494117||cg15494117||UCEC||0.951421840298937||TERC||3||169482835||169482955 | ||
|- | |- | ||
| | | cg15599946||cg15599946||UCEC||0.947489755922484||TERC||3||169482839||169482959 | ||
|- | |- | ||
| | | cg18985581||cg18985581||UCEC||0.946277345041878||||14||105512153||105512273 | ||
|- | |||
| cg02665570||cg02665570||UCEC||0.94585801514041||LYPLAL1||1||219347280||219347400 | |||
|- | |- | ||
|} | |} |
Revision as of 03:21, 22 April 2015
- Now, I do know how to insert figures, but I don't know how to insert tables into MediaWiki. Therefore, I write a small code to transfer routine table to wikitable.
- I must say: mediawiki is really a excellent tool to manage the lab and monitor the progress or task to help us complete the project quickly.
perl table2wikitable.pl input.table.txt
#!/usr/bin/perl #table2wikitable.pl use strict; use Cwd; chdir getcwd; my $input=@ARGV[0]; open F,$input; print "{| class=\"wikitable\" style=\"text-align: right; color: red;\"\n"; while(<F>){ chomp; my @line=split /\t/; my $tmp=join("||",@line); print "| $tmp\n|-\n"; } print"|}\n";
- corresponding table is as the following. Pan-Cancer Methylation 450K dataset were collected from TCGA Project to identify optional padloc regions.
Symbol | Case | Control | Cancer | 肿瘤名称 |
KIRC | 160 | 160 | Kidney renal clear cell carcinoma | 肾透明细胞癌 |
BRCA | 92 | 92 | Breast invasive carcinoma | 浸润性乳腺癌 |
THCA | 56 | 56 | Thyroid carcinoma | 甲状腺癌 |
HNSC | 50 | 50 | Head and Neck squamous cell carcinoma | 头颈部鳞状细胞癌 |
PRAD | 49 | 49 | Prostate adenocarcinoma | 前列腺癌 |
LIHC | 49 | 49 | Liver hepatocellular carcinoma | 肝癌 |
KIRP | 45 | 45 | Kidney renal papillary cell carcinoma | 肾乳头状细胞癌 |
LUSC | 41 | 41 | Lung squamous cell carcinoma | 肺腺癌 |
COAD | 39 | 39 | Colon adenocarcinoma | 结肠癌 |
UCEC | 30 | 30 | Uterine Corpus Endometrial carcinoma | 子宫内膜癌 |
LUAD | 26 | 26 | Lung adenocarcinoma | 肺鳞癌 |
- In order to publish MONOD paper to a good paper quickly. 1) fast 2) large data
- collect methylation 450 data of cancer tissues and normals from TCGA Project
- sample size
- clinical information: gender, age
- collect methylation 450 data of cancer tissues and normals from TCGA Project
tar xvf 08177614-f305-4fbf-84ca-fd2fbfe26755.tar # tar xvf 548dd1cf-84a1-4d96-86f0-b47d6300daa6.tar # tar xvf 9cd95b1c-782c-478d-9ec9-de7c9c441cc4.tar # tar xvf c5da6cd3-0266-4ebb-bd11-713ffe9b5ef9.tar #
- focus on 3 cancers which we have preliminary data (lung, colon, pancreatic)
library("stringr") for(cancer in c("COAD","LUAD","LUSC","PAAD")){ dir<-paste("/home/sguo/monod/data/",tolower(cancer),"/DNA_Methylation/JHU_USC__HumanMethylation450/Level_3",sep="") setwd(dir) pattern=paste("jhu-usc.edu_",cancer,".*",sep="") print (pattern) file=list.files(pattern=pattern) idv<-unique(as.array(str_extract(file,"TCGA-[0-9|a-z|A-Z]*-[0-9|a-z|A-Z]*"))) pairidv<-c() for (i in 1:length(idv)){ t1<-paste(idv[i],"-01",sep="") t2<-paste(idv[i],"-11",sep="") if(all(any(grepl(t1,file)),any(grepl(t2,file)))){ pairidv<-c(pairidv,t1,t2) } } l1<-length(pairidv) l2<-length(file) id1<-lapply(lapply(strsplit(file,"[.]"),function(x) x[6]),function(x) substr(x,1,15)) id2<-lapply(lapply(strsplit(file,"[.]"),function(x) x[6]),function(x) substr(x,14,15)) sam<-lapply(lapply(strsplit(file,"[.]"),function(x) x[6]),function(x) substr(x,1,15)) tab<-table(unlist(lapply(lapply(strsplit(file,"[.]"),function(x) x[6]),function(x) substr(x,14,15)))) c1<-tab[which(names(tab)=="01")] c2<-tab[which(names(tab)=="11")] c3<-length(pairidv) print(c((c1),(c2),Pair=c3)) }
- The sample size for the genome-wide DNA methyaltion dataset for 3 cancers in TCGA Project are as the following:
Cancer | Normal | Paired | Total | |
COAD | 312 | 38 | 76 | 350 |
LUAD | 473 | 32 | 58 | 505 |
LUSC | 370 | 42 | 80 | 412 |
PAAD | 184 | 10 | 20 | 194 |
Sum | 1339 | 122 | 234 | 1461 |
- totally, 1339 cancer tissues and 122 normal tissues were used to refine the padloc regions.
- firstly, we should check the methylation status of the CpG sites/regions in the normal tissues.
1) hypermethylation in cancer/patient plasma 2) hypomethylation in normal/health plasma 3) large methylation difference (delta beta) 4) high group specificity index (GSI) 5) high ratio release to plasma
- s(j): average methylation in individual group j
- S(max): average methylation in the group with highest methylation level
# group specificity index,GSI setwd("../pan") load("PanPairMethData.RData") pheno=data$pheno xmean <- rowsum(data[,2:ncol(data)], pheno)/table(pheno) gsi<-apply(xmean,2,function(x) (length(x)-sum(x)/max(x))/(length(x)-1)) rlt<-data.frame(pheno=names(table(pheno)),xmean[,match(names(sort(gsi,decreasing=T)[1:5]),colnames(xmean))]) write.table(rlt,file="pancancer.high.gsi.site.txt",sep="\t",quote=F,col.names=NA,row.names=T)
pdf("gsi.distribution.pdf") hist(gsi,main="Histogram of Group Specific Index") dev.off()
num90<-sum(gsi>0.85) xmean90<-xmean[,match(names(sort(gsi,decreasing=T)[1:num90]),colnames(xmean))] xx1<-table(data$pheno)[as.numeric(names(table(apply(xmean90,2,function(x) which.max(x)))))] xx2<-table(apply(xmean90,2,function(x) which.max(x))) rlt2<-data.frame(sample_size=xx1,xx2) write.table(rlt2,file="target.status.specifi.txt",sep="\t",quote=F,col.names=NA,row.names=T)
- run the code and you can find the strongest group specific index CpGs with TCGA Pancancer dataset.
Sample_size | cg16579555 | cg26240185 | cg12587766 | cg15375239 | cg10157975 |
BRCA-01 | 0.01 | 0.014 | 0.021 | 0.012 | 0.011 |
BRCA-11 | 0.01 | 0.011 | 0.02 | 0.011 | 0.011 |
COAD-01 | 0.011 | 0.012 | 0.605 | 0.011 | 0.353 |
COAD-11 | 0.011 | 0.015 | 0.033 | 0.012 | 0.022 |
HNSC-01 | 0.011 | 0.012 | 0.021 | 0.013 | 0.031 |
HNSC-11 | 0.011 | 0.011 | 0.022 | 0.012 | 0.012 |
KIRC-01 | 0.009 | 0.01 | 0.016 | 0.019 | 0.011 |
KIRC-11 | 0.009 | 0.009 | 0.016 | 0.01 | 0.009 |
KIRP-01 | 0.01 | 0.026 | 0.021 | 0.016 | 0.01 |
KIRP-11 | 0.01 | 0.011 | 0.022 | 0.012 | 0.011 |
LIHC-01 | 0.396 | 0.498 | 0.042 | 0.468 | 0.028 |
LIHC-11 | 0.057 | 0.099 | 0.024 | 0.133 | 0.015 |
LUAD-01 | 0.01 | 0.014 | 0.022 | 0.012 | 0.012 |
LUAD-11 | 0.011 | 0.013 | 0.024 | 0.013 | 0.013 |
LUSC-01 | 0.009 | 0.01 | 0.017 | 0.01 | 0.009 |
LUSC-11 | 0.008 | 0.01 | 0.018 | 0.011 | 0.01 |
PRAD-01 | 0.011 | 0.011 | 0.02 | 0.012 | 0.011 |
PRAD-11 | 0.01 | 0.012 | 0.019 | 0.012 | 0.011 |
THCA-01 | 0.011 | 0.013 | 0.028 | 0.013 | 0.013 |
THCA-11 | 0.011 | 0.013 | 0.024 | 0.012 | 0.013 |
UCEC-01 | 0.013 | 0.014 | 0.02 | 0.012 | 0.026 |
UCEC-11 | 0.011 | 0.015 | 0.022 | 0.012 | 0.014 |
- clearly, we can find cancer specific hypermethylated genes with previous method. Then, how about the distribution of the GSI.
- 745 CpGs or regions whose GSI>0.9. these CpGs/regions showed high tissue/status specific.
- 3181 CpGs or regions whose GSI>0.85. these CpGs/regions showed high tissue/status specific.
- The question is that whether they can be found in patients plasma? (Dr.Zhang might have some information)
- 3181 tissue and status specific hypermethylation CpG sites were showed as the following table.
Sample_size | Var1 | Freq | |
BRCA-01 | 92 | 1 | 33 |
BRCA-11 | 92 | 2 | 1 |
COAD-01 | 39 | 3 | 1695 |
COAD-11 | 39 | 4 | 16 |
HNSC-01 | 50 | 5 | 143 |
KIRC-01 | 160 | 7 | 1 |
KIRC-11 | 160 | 8 | 1 |
KIRP-01 | 45 | 9 | 20 |
KIRP-11 | 45 | 10 | 1 |
LIHC-01 | 49 | 11 | 768 |
LIHC-11 | 49 | 12 | 40 |
LUAD-01 | 26 | 13 | 1 |
LUSC-01 | 41 | 15 | 8 |
PRAD-01 | 49 | 17 | 271 |
PRAD-11 | 49 | 18 | 1 |
THCA-01 | 56 | 19 | 4 |
THCA-11 | 56 | 20 | 7 |
UCEC-01 | 30 | 21 | 168 |
UCEC-11 | 30 | 22 | 2 |
- Here, I choose top 5 regions to represent such specific tissue and it's specific status (cancer or normal), as the following:
cpg | cancer | GSI | gene | chr | start | end | |
cg18565473 | cg18565473 | BRCA | 0.932682778025148 | ETS1 | 11 | 128392042 | 128392162 |
cg23884187 | cg23884187 | BRCA | 0.915793344077998 | C20orf95 | 20 | 37275009 | 37275129 |
cg14052221 | cg14052221 | BRCA | 0.912760995997153 | PSAT1 | 9 | 80911998 | 80912118 |
cg24797187 | cg24797187 | BRCA | 0.912059797561713 | AFF3 | 2 | 100175708 | 100175828 |
cg18943693 | cg18943693 | BRCA | 0.911675629661679 | 1 | 155043501 | 155043621 | |
cg12587766 | cg12587766 | COAD | 0.962866407567279 | LIFR | 5 | 38556375 | 38556495 |
cg10157975 | cg10157975 | COAD | 0.959068233395622 | ZNF304 | 19 | 57862382 | 57862502 |
cg23977631 | cg23977631 | COAD | 0.954738746472514 | LONRF2 | 2 | 100938739 | 100938859 |
cg04117229 | cg04117229 | COAD | 0.952980963623186 | SPG20 | 13 | 36920753 | 36920873 |
cg09854653 | cg09854653 | COAD | 0.952746178623677 | QKI | 6 | 163834843 | 163834963 |
cg03988778 | cg03988778 | HNSC | 0.94124876913729 | SVIP | 11 | 22850831 | 22850951 |
cg08211306 | cg08211306 | HNSC | 0.936441823565694 | ENPP4 | 6 | 46097724 | 46097844 |
cg26968387 | cg26968387 | HNSC | 0.932552091587754 | ZNF420 | 19 | 37569208 | 37569328 |
cg03280624 | cg03280624 | HNSC | 0.925140832107714 | ZNF583 | 19 | 56915595 | 56915715 |
cg00471966 | cg00471966 | HNSC | 0.92378591584013 | ZNF420 | 19 | 37569290 | 37569410 |
cg26228351 | cg26228351 | KIRC | 0.866821443647375 | KIF21B | 1 | 200992596 | 200992716 |
cg00593900 | cg00593900 | KIRC | 0.840429026779539 | ANGPTL6 | 19 | 10206686 | 10206806 |
cg11697226 | cg11697226 | KIRC | 0.83932716910874 | TNFRSF11A | 18 | 59992325 | 59992445 |
cg09865339 | cg09865339 | KIRC | 0.836396509994196 | GPC2;STAG3 | 7 | 99774875 | 99774995 |
cg02632185 | cg02632185 | KIRC | 0.824262102898211 | MAST4 | 5 | 66299726 | 66299846 |
cg15598442 | cg15598442 | KIRP | 0.926520443898138 | 1 | 25175003 | 25175123 | |
cg26622232 | cg26622232 | KIRP | 0.907406091283148 | OXR1 | 8 | 107669727 | 107669847 |
cg16326979 | cg16326979 | KIRP | 0.898630310618579 | OXR1 | 8 | 107670101 | 107670221 |
cg17031478 | cg17031478 | KIRP | 0.897081288137618 | HOXC4;HOXC5 | 12 | 54427113 | 54427233 |
cg17136799 | cg17136799 | KIRP | 0.888400341253348 | OXR1 | 8 | 107669723 | 107669843 |
cg16579555 | cg16579555 | LIHC | 0.968046292345837 | RNF135 | 17 | 29298292 | 29298412 |
cg26240185 | cg26240185 | LIHC | 0.966346469016627 | FAR1 | 11 | 13690097 | 13690217 |
cg15375239 | cg15375239 | LIHC | 0.961490202023512 | SPINT2 | 19 | 38755227 | 38755347 |
cg15969216 | cg15969216 | LIHC | 0.958031159935377 | TSC22D1 | 13 | 45150202 | 45150322 |
cg03326059 | cg03326059 | LIHC | 0.958023849811357 | FAR1 | 11 | 13690100 | 13690220 |
cg13215643 | cg13215643 | LUAD | 0.855326865879107 | DACT1 | 14 | 59104765 | 59104885 |
cg07017994 | cg07017994 | LUAD | 0.842561347568723 | EPHB6 | 7 | 142552854 | 142552974 |
cg12487147 | cg12487147 | LUAD | 0.839197022612119 | HSD17B8 | 6 | 33172382 | 33172502 |
cg26615830 | cg26615830 | LUAD | 0.837145673329791 | MSX1 | 4 | 4861270 | 4861390 |
cg21929771 | cg21929771 | LUAD | 0.820700381610812 | PTPRU | 1 | 29586520 | 29586640 |
cg07240673 | cg07240673 | LUSC | 0.899039308699477 | CLUAP1 | 16 | 3550848 | 3550968 |
cg18772127 | cg18772127 | LUSC | 0.886204255597843 | CMTM7 | 3 | 32443436 | 32443556 |
cg08562243 | cg08562243 | LUSC | 0.874881316596866 | CLUAP1 | 16 | 3551109 | 3551229 |
cg02566698 | cg02566698 | LUSC | 0.872675000318173 | CLUAP1 | 16 | 3550872 | 3550992 |
cg02379764 | cg02379764 | LUSC | 0.869773727428307 | CLUAP1 | 16 | 3550968 | 3551088 |
cg07635623 | cg07635623 | PRAD | 0.955666568317178 | SERPINB1 | 6 | 2841815 | 2841935 |
cg14283569 | cg14283569 | PRAD | 0.947556926299886 | 19 | 51416153 | 51416273 | |
cg05098590 | cg05098590 | PRAD | 0.94340784073766 | ADD3 | 10 | 111767319 | 111767439 |
cg10938374 | cg10938374 | PRAD | 0.942601773040438 | IER3 | 6 | 30711998 | 30712118 |
cg16232979 | cg16232979 | PRAD | 0.942431007327551 | TPM4 | 19 | 16187571 | 16187691 |
cg27115721 | cg27115721 | THCA | 0.881950689779005 | FAM49A | 2 | 16790278 | 16790398 |
cg04358131 | cg04358131 | THCA | 0.877589155598355 | MAFK | 7 | 1572192 | 1572312 |
cg10540754 | cg10540754 | THCA | 0.86094593963059 | FAM49A | 2 | 16790310 | 16790430 |
cg22749810 | cg22749810 | THCA | 0.855202902841297 | RNF213 | 17 | 78237329 | 78237449 |
cg12822074 | cg12822074 | THCA | 0.849782546942327 | RTN4RL2 | 11 | 57243805 | 57243925 |
cg03221247 | cg03221247 | UCEC | 0.955580574481869 | LYPLAL1 | 1 | 219347398 | 219347518 |
cg15494117 | cg15494117 | UCEC | 0.951421840298937 | TERC | 3 | 169482835 | 169482955 |
cg15599946 | cg15599946 | UCEC | 0.947489755922484 | TERC | 3 | 169482839 | 169482959 |
cg18985581 | cg18985581 | UCEC | 0.946277345041878 | 14 | 105512153 | 105512273 | |
cg02665570 | cg02665570 | UCEC | 0.94585801514041 | LYPLAL1 | 1 | 219347280 | 219347400 |