[BioC] "AveExp" column has dissappeared from limma topTable

Gordon K Smyth smyth at wehi.EDU.AU
Tue Apr 3 23:43:23 CEST 2007


Dear Amy,

See

  https://www.stat.math.ethz.ch/pipermail/bioconductor/2007-February/016132.html

Note that 'Amean' is the older name for 'AveExpr'.

Best wishes
Gordon

> Date: Mon, 2 Apr 2007 14:23:45 +0100
> From: "Mikhail, Amy" <a.mikhail at abdn.ac.uk>
> Subject: Re: [BioC] "AveExp" column has dissappeared from limma
> 	topTable
> To: "Marcus Davy" <mdavy at hortresearch.co.nz>
> Cc: bioconductor at stat.math.ethz.ch

>
> Hi Marcus,
>
> "Mosquito" is a subset of my original exprSet (with some probesets taken
> out).  So it is just a matrix and now that I look, there is no "Amean"
> element in my fit object.  I didn't know that Amean would not be
> derrived for matrices, thanks for pointing that out.
>
> Many thanks,
> Amy
>
> -----Original Message-----
> From: Marcus Davy [mailto:mdavy at hortresearch.co.nz]
> Sent: Monday, April 02, 2007 12:42 AM
> To: Mikhail, Amy; bioconductor at stat.math.ethz.ch
> Subject: Re: [BioC] "AveExp" column has dissappeared from limma topTable
>
>
> Have a look in your 'fit' object and the lmFit code for clues as to what
> is going wrong. When you create a fit object from lmFit, a list element
> called 'Amean' may not have been created. This is what is picked up by
> topTable.
>
> You didn't mention the structure of the argument object 'Mosquito'  in
> your lmFit call. Since you have an Affymetrix experiment, is it
> "PLMset", "exprSet" "ExpressionSet" or a matrix (unless you used the
> convert package to make an MAList)?
>
> If 'Mosquito' is a matrix then a list element for Amean is not created
> in your 'fit' object and hence not displayed in topTable
>
>>x <- matrix(1:20, nc=4)  #Any old numbers for illustration design <-
>>rep(1,4)  lmFit(x, design=rep(1,4))$Amean # Amean not created
> NULL
>> topTable(eBayes(lmFit(x, design)))
>   logFC        t     P.Value   adj.P.Val         B
> 5  12.5 3.220503 0.001279659 0.006398294 -2.286030
> 4  11.5 2.962863 0.003047925 0.007619813 -2.696287
> 3  10.5 2.705222 0.006825866 0.011376444 -3.072355
> 2   9.5 2.447582 0.014381831 0.017977289 -3.414236
> 1   8.5 2.189942 0.028528445 0.028528445 -3.721928
>
> Whereas an MAList (or "PLMset", "exprSet" "ExpressionSet") should give
> you the AveExpr column. Note: The Amean calculation is not adjusted for
> weights, whereas your coefficients are.
>
>>MA <- new("MAList")
>>MA$M <- x
>>MA$A  <- x
>>lmFit(MA, design)$Amean # Amean created
> [1]  8.5  9.5 10.5 11.5 12.5
>> topTable(eBayes(lmFit(MA, design)))
>   logFC AveExpr        t     P.Value   adj.P.Val         B
> 5  12.5    12.5 3.220503 0.001279659 0.006398294 -2.286030
> 4  11.5    11.5 2.962863 0.003047925 0.007619813 -2.696287
> 3  10.5    10.5 2.705222 0.006825866 0.011376444 -3.072355
> 2   9.5     9.5 2.447582 0.014381831 0.017977289 -3.414236
> 1   8.5     8.5 2.189942 0.028528445 0.028528445 -3.721928
>
> Hope that helps.
>
>
> Marcus
>
> On 31/3/07 9:57 AM, "Mikhail, Amy" <a.mikhail at abdn.ac.uk> wrote:
>
>> Dear Bioconductors,
>>
>> I have a problem with the "AveExp" column in topTable.  Usually it
>> appears in my toptable output, but during a limma session recently I
>> created one toptable (which did have the "AveExp" column in it) and
>> then another toptable (which was a subset).  In the second topTable,
>> the AveExp column was missing.  I have tried reloading R, and
>> reinstalling limma but the column seems now to be permanently missing
>> from my topTables.
>>
>> There was no error message or warning, so I really don't know what
>> I've done ;-(.  Sample code is below:
>>
>>> targets <- readTargets("BaseAgeAffyTargets.txt")
>>> targets
>>   Name                                   FileName             Target
>> 1 R2s2  AM_041006_R2S2v1_Plasmodium_Anopheles.CEL Adults age 15 days
>> 2 R2s6    AM_041006_R2S2_Plasmodium_Anopheles.CEL Adults age 43 days
>> 3 R3s2    AM_041006_R3S2_Plasmodium_Anopheles.CEL Adults age 15 days
>> 4 R3s6    AM_041006_R3S6_Plasmodium_Anopheles.CEL Adults age 43 days
>> 5 R4s2 AM_041006_BAR4Sii_Plasmodium_Anopheles.CEL Adults age 15 days
>> 6 R4s6    AM_041006_R4S6_Plasmodium_Anopheles.CEL Adults age 43 days
>>>
>>> design <- cbind(M15d=c(1,0,1,0,1,0),M43d=c(0,1,0,1,0,1))
>>> fit <- lmFit(Mosquito, design)
>>> cont.matrix <- makeContrasts(M15dvsM43d=M43d-M15d, levels=design)
>>> fit2 <- contrasts.fit(fit, cont.matrix)
>>> fit2 <- eBayes(fit2)
>>> Lowvar.toptable <- topTable(fit2[ Lowvar.genes, ], number=100,
>> adjust="BH")
>>
>>> Lowvar.toptable[1:8,]
>>                       ID           logFC  t
>> P.Value  adj.P.Val B
>> 8    Ag.2L.2099.0_CDS_at 0.0525816225 0.613337707
>> 0.5608673 0.9967802 -5.844476
>> 57 Ag.UNKN.1771.0_CDS_at 0.0514293902 0.602622882
>> 0.5675464 0.9967802 -5.851185
>> 46   Ag.3R.2483.0_CDS_at 0.0460176987 0.540864707
>> 0.6069533 0.9967802 -5.887786
>> 24   Ag.2R.3361.0_CDS_at -0.0461903587 -0.540335772
>> 0.6072973 0.9967802 -5.888084
>> 60  Ag.UNKN.363.0_CDS_at -0.0394899270 -0.464386924 0.6577950
>> 0.9967802 -5.928101
>> 64   Ag.X.217.0_CDS_a_at -0.0393985675 -0.455020526
>> 0.6641680 0.9967802 -5.932647
>> 58 Ag.UNKN.1863.0_CDS_at 0.0359376810 0.415978011
>> 0.6910563 0.9967802 -5.950663
>> 30   Ag.3L.1321.0_CDS_at -0.0334439607 -0.387588909
>> 0.7109213 0.9967802 -5.962804
>>
>> I also note that "A" is also missing from my fit object;
>>
>>> fit2$Amean
>> NULL
>>
>> I would be most grateful for any suggestions as to what could be
>> happening here....
>> I realise that I am using R2.4.0 with a more recent version of limma,
>> but this has not caused me any problems up until now.
>>
>> Looking forward to any suggestions,
>> Best wishes,
>> Amy
>>
>>
>>> sessionInfo()
>> R version 2.4.0 (2006-10-03)
>> i386-pc-mingw32
>>
>> locale:
>> LC_COLLATE=English_United States.1252;LC_CTYPE=English_United
>> States.1252;LC_MONETARY=English_United
>> States.1252;LC_NUMERIC=C;LC_TIME=English_United States.1252
>>
>> attached base packages:
>> [1] "splines"   "tools"     "methods"   "stats"     "graphics"
>> "grDevices" "utils"     "datasets"  "base"
>>
>> other attached packages:
>>    mixtools     mvtnorm agahomology     affyPLM       gcrma
> matchprobes
>> affydata     annaffy        KEGG          GO  genefilter
>>     "0.1.0"     "0.7-5"    "1.14.2"    "1.10.0"     "2.6.0"
> "1.6.0"
>> "1.10.0"     "1.6.1"    "1.14.0"    "1.14.0"    "1.12.0"
>>    survival       limma geneplotter    annotate        affy
> affyio
>> Biobase
>>      "2.30"     "2.9.8"    "1.12.0"    "1.12.1"    "1.12.2"
> "1.2.0"
>> "1.12.2"
>>
>> **********************************************************************
>> **
>> *********************************************************
>>
>> Amy Mikhail
>> Research student
>> University of Aberdeen
>> Zoology Building
>> Tillydrone Avenue
>> Aberdeen
>> AB24 2TZ
>> Scotland
>> Email: a.mikhail at abdn.ac.uk
>> Phone: 00-44-1224-273256 (office)


----------------------------
Dr Gordon K Smyth, Research Fellow, Bioinformatics,
Walter and Eliza Hall Institute of Medical Research,
1G Royal Parade, Parkville, Vic 3050, Australia
Tel: (03) 9345 2326, Fax (03) 9347 0852,
Email: smyth at wehi.edu.au, www: http://www.statsci.org



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