[BioC] Affy 1.5.8 functions not working on Mac OS X 10.3

Chad Wayne cwayne at bcm.tmc.edu
Tue Jan 25 22:43:51 CET 2005


My entire session follows. The previous .RData session was performed on  
another computer while I was learning the basics of R and affy. Prior  
to loading "timecourse.Rdata", I started X11 and changed the working  
directory to the proper working directory with my data in it.

 > load("/xxx/xxx/ArrayAnalysis/timecourse.RData")
 > ls()
  [1] "PMA.Calls"     "Pearson.cor"   "adjp"          "coc.phenodata"
  [5] "deg"           "eset"          "mynova.full"   "procs"
  [9] "raw"           "raw.pvalue"    "res"           "timecourse"
[13] "x"
 > library(affy)
Loading required package: Biobase
Loading required package: tools
Loading required package: reposTools

## Note here that everything loaded just fine, as far as I can tell.  
Knowing that these next two functions haven't worked for me, I used  
them first.

 > pData(eset)
Error: couldn't find function "pData"
 > ReadAffy(widget=T)
Loading required package: tkWidgets
Loading required package: widgetTools
Loading required package: tcltk
Loading required package: DynDoc
Error in AllButCelsForReadAffy(..., filenames = filenames, widget =  
widget,  :
	couldn't find function "read.phenoData"

## When I performed ReadAffy(widget=T), the widget did function  
normally. I selected the .cel files I wanted use (the ones that made up  
the eset above) and then after selecting them, the error message popped  
up.

 > packageDescription("affy", field="Version")
[1] "1.5.8"
 > version
          _
platform powerpc-apple-darwin6.8
arch     powerpc
os       darwin6.8
system   powerpc, darwin6.8
status
major    2
minor    0.1
year     2004
month    11
day      15
language R

## This search demonstrates the packages that are active at this point  
in the session.

 > search()
  [1] ".GlobalEnv"          "package:tkWidgets"   "package:DynDoc"
  [4] "package:widgetTools" "package:tcltk"       "package:affy"
  [7] "package:reposTools"  "package:Biobase"     "package:tools"
[10] "package:methods"     "package:stats"       "package:graphics"
[13] "package:grDevices"   "package:utils"       "package:datasets"
[16] "Autoloads"           "package:base"

## I wanted to demonstrate the functions and files associated with affy  
in my library. There are 116 of them.

 > ls("package:affy")
   [1] "$.AffyBatch"
   [2] "AffyRNAdeg"
   [3] "AllButCelsForReadAffy"
   [4] "MAplot"
   [5] "Mbox"
   [6] "ReadAffy"
   [7] "affy.scalevalue.exprSet"
   [8] "avdiff"
   [9] "barplot"
  [10] "barplot.ProbeSet"
  [11] "bg.adjust"
  [12] "bg.correct"
  [13] "bg.correct.mas"
  [14] "bg.correct.none"
  [15] "bg.correct.rma"
  [16] "bg.correct.rma2"
  [17] "bg.parameters"
  [18] "bgcorrect.methods"
  [19] "boxplot"
  [20] "cdfFromBioC"
  [21] "cdfFromData"
  [22] "cdfFromEnvironment"
  [23] "cdfFromLibPath"
  [24] "cdfName"
  [25] "cleancdfname"
  [26] "close"
  [27] "colnames"
  [28] "computeExprSet"
  [29] "debug.affy123"
  [30] "express.summary.stat"
  [31] "express.summary.stat.methods"
  [32] "expresso"
  [33] "expressoWidget"
  [34] "fit.li.wong"
  [35] "generateExprSet.methods"
  [36] "generateExprVal.method.avgdiff"
  [37] "generateExprVal.method.liwong"
  [38] "generateExprVal.method.mas"
  [39] "generateExprVal.method.medianpolish"
  [40] "generateExprVal.method.playerout"
  [41] "getCdfInfo"
  [42] "hist"
  [43] "hlog"
  [44] "image"
  [45] "indexProbes"
  [46] "indices2xy"
  [47] "intensity"
  [48] "intensity<-"
  [49] "just.rma"
  [50] "justRMA"
  [51] "li.wong"
  [52] "list.celfiles"
  [53] "loess.normalize"
  [54] "ma.plot"
  [55] "maffy.normalize"
  [56] "maffy.subset"
  [57] "mas5"
  [58] "mas5.detection"
  [59] "mas5calls"
  [60] "mas5calls.AffyBatch"
  [61] "mas5calls.ProbeSet"
  [62] "medianpolish"
  [63] "merge.AffyBatch"
  [64] "mm"
  [65] "mm<-"
  [66] "mmindex"
  [67] "multiloess"
  [68] "mva.pairs"
  [69] "ncol"
  [70] "normalize"
  [71] "normalize.AffyBatch.constant"
  [72] "normalize.AffyBatch.contrasts"
  [73] "normalize.AffyBatch.invariantset"
  [74] "normalize.AffyBatch.loess"
  [75] "normalize.AffyBatch.methods"
  [76] "normalize.AffyBatch.qspline"
  [77] "normalize.AffyBatch.quantiles"
  [78] "normalize.AffyBatch.quantiles.robust"
  [79] "normalize.constant"
  [80] "normalize.invariantset"
  [81] "normalize.loess"
  [82] "normalize.methods"
  [83] "normalize.qspline"
  [84] "normalize.quantiles"
  [85] "normalize.quantiles.robust"
  [86] "nrow"
  [87] "open"
  [88] "pairs.AffyBatch"
  [89] "playerout.costfunction"
  [90] "plot.ProbeSet"
  [91] "plotAffyRNAdeg"
  [92] "plotDensity"
  [93] "plotDensity.AffyBatch"
  [94] "plotLocation"
  [95] "pm"
  [96] "pm<-"
  [97] "pmcorrect.mas"
  [98] "pmcorrect.methods"
  [99] "pmcorrect.pmonly"
[100] "pmcorrect.subtractmm"
[101] "pmindex"
[102] "ppset.ttest"
[103] "ppsetApply"
[104] "probeNames"
[105] "probes"
[106] "probeset"
[107] "read.affybatch"
[108] "read.probematrix"
[109] "rma"
[110] "simplemultiLoess"
[111] "summaryAffyRNAdeg"
[112] "tukey.biweight"
[113] "tukeybiweight"
[114] "update"
[115] "whatcdf"
[116] "xy2indices"

## I next performed the capabilities function. The only thing not  
available to me was GNOME

 > capabilities()
     jpeg      png    tcltk      X11    GNOME     libz http/ftp  sockets
     TRUE     TRUE     TRUE     TRUE    FALSE     TRUE     TRUE     TRUE
   libxml     fifo   cledit  IEEE754    bzip2     PCRE
     TRUE     TRUE     TRUE     TRUE     TRUE     TRUE

## I finally checked the library path for the R.framework. This is what  
I expected. I also physically checked the file heirarchy to make sure  
that in "library" there were files for R to use. There are.

 > .libPaths()
[1] "/Library/Frameworks/R.framework/Resources/library"


## If I type in eset, rather than getting an expression set matrix (?),  
I get the full list of the gene IDs + log values of the samples (45K+  
rows)

 > eset
list()
attr(,"exprs")
                                0h-1.cel  0h-2.cel 16h-1.cel 16h-2.cel
1415670_at                    10.783245 10.696067 11.372607 11.203421
1415671_at                    10.655126 10.972694 11.253215 11.287455
... etc.

## Also wanted to look at the attributes of the phenodata from the  
previous session to see whether the session was corrupted.

 > coc.phenodata
list()
attr(,"pData")
           time.after.hCG
0h-1.cel               0
0h-2.cel               0
16h-1.cel             16
16h-2.cel             16
8h-1.cel               8
8h-2.cel               8
attr(,"varLabels")
attr(,"varLabels")$"time after hCG"
[1] ""

attr(,"class")
[1] "phenoData"
attr(,"class")attr(,"package")
[1] "Biobase"

## Here we see another function error such that affy (BioC?) can't find  
the proper cdf file to compare the data against.

 > raw
Error in cdfFromBioC(cdfname, cur$where, verbose) :
	couldn't find function "testBioCConnection"
AffyBatch object
size of arrays=1002x1002 features (47068 kb)
cdf=Mouse430_2 (??? affyids)
Error: couldn't find function "exprs"
In addition: Warning message:
missing cdf environment ! in: show(list())
Error in ncol(exprs(x)) : Unable to find the argument "x" in selecting  
a method for function "ncol"

## End of session.

I suspect that there must be something wrong with either the install or  
that the calls used by affy became corrupt either at the source or  
during the install. I should offer one caveat here. I have experienced  
this problem with affy ever since I first installed it two weeks ago. I  
have attempted to do re-installs of the affy package and clean installs  
of R. Nothing has remedied this issue. And when I say clean install, I  
mean I have crawled through the system and deleted everything from the  
libraries to the install .pkg that OsX likes to hide. I also deleted  
preferences. I also check system level and user level libraries to  
ensure I got everything that was installed prior to the clean install.

If I try to do a reinstall over the current package I get the following  
session:

trying URL `http://www.bioconductor.org/src/contrib/PACKAGES'
Content type `text/plain' length 46409 bytes
opened URL
==================================================
downloaded 45Kb

trying URL `http://www.bioconductor.org/src/contrib/PACKAGES'
Content type `text/plain' length 46409 bytes
opened URL
==================================================
downloaded 45Kb

trying URL `http://www.bioconductor.org/src/contrib/affy_1.5.8.tar.gz'
Content type `application/x-tar' length 1076622 bytes
opened URL
==================================================
downloaded 1051Kb

* Installing *source* package 'affy' ...
creating cache ./config.cache
checking how to run the C preprocessor... /lib/cpp
checking for main in -lz... no
updating cache ./config.cache
creating ./config.status
creating src/Makevars
** libs
/Library/Frameworks/R.framework/Resources/bin/SHLIB: line 1: make:  
command not found
** Removing  
'/Library/Frameworks/R.framework/Versions/2.0.1/Resources/library/affy'
** Restoring previous  
'/Library/Frameworks/R.framework/Versions/2.0.1/Resources/library/affy'
ERROR: compilation failed for package 'affy'

 >
Delete downloaded files (y/N)? y             ##I deleted the file this  
time, but have and haven't at other times.

 >

It is at this point, I toss up my hands and beg for help. Sorry for the  
length of this I wanted to be as thorough as possible could. I really  
do appreciate the help.

Chad.

On Jan 25, 2005, at 1:57 PM, Robert Gentleman wrote:

>
> On Jan 25, 2005, at 9:35 AM, Chad Wayne wrote:
>
>> I appreciate you taking the time to answer my questions.
>>
>>>
>>> On Jan 24, 2005, at 12:55 PM, Chad Wayne wrote:
>>>
>>>> It was recommended that I post this problem to the mailing list. I  
>>>> am unable to get affy to work, e.g. the library loads into R, but  
>>>> the functions don't seem to work. I was hoping someone may have  
>>>> seen this problem before and may have a solution. I am working in  
>>>> the correct working directory.
>>>>
>>>> I am using version v2.0.1 of R with the default Bioconductor  
>>>> package installed. Current package of Affy is 1.5.8. I am using Mac  
>>>> OS X 10.3.7 and running X11 (X11 1.0 - XFree86 4.3.0). This is a  
>>>> shared environment and all packages were installed in the top  
>>>> library, not in the User library.
>>>>
>>>> On prompt:
>>>> > load("/nnn/nnn/nnn/timecourse.RData")
>>>
>>>> > library(affy)
>>>> Loading required package: Biobase
>>>> Loading required package: tools
>>>> Loading required package: reposTools
>>>
>>> So, if you go
>>>   search()
>>> at this point you should get something like:
>>> > search()
>>>  [1] ".GlobalEnv"         "package:affy"       "package:reposTools"
>>>  [4] "package:Biobase"    "package:tools"      "package:methods"
>>>  [7] "package:stats"      "package:graphics"   "package:grDevices"
>>> [10] "package:utils"      "package:datasets"   "Autoloads"
>>> [13] "package:base"
>>>
>>> The important things are for affy and Biobase to be there (at least  
>>> given what you are saying below)
>>>
>>
>> I loaded affy and did the search as you suggested:
>>
>> > library(affy)
>> Loading required package: Biobase
>> Loading required package: tools
>> Loading required package: reposTools
>> > search()
>>  [1] ".GlobalEnv"         "package:affy"       "package:reposTools"
>>  [4] "package:Biobase"    "package:tools"      "package:methods"
>>  [7] "package:stats"      "package:graphics"   "package:grDevices"
>> [10] "package:utils"      "package:datasets"   "Autoloads"
>> [13] "package:base"
>>
>> Appears to be there along with biobase.
>>
>>
>>>> > ReadAffy(widget=T)
>>>> Error: couldn't find function "ReadAffy"
>>>> > ReadAffy()
>>>> Error: couldn't find function "ReadAffy"
>>>> > pData(eset)
>>>> Error: couldn't find function "pData"
>>>>
>>>
>>>
>>> You can also try something like
>>> ls("package:affy")
>>>
>>> which should show about 116 things (mostly functions) and one should  
>>> be ReadAffy.
>>>
>>
>> I also performed the ls("package:affy") and retrieved 116 items. I  
>> attempted again to try the ReadAffy function and it failed.
>>
>
>   Hmm, this seems really odd.  So a whole bunch of questions in the  
> hopes that one of them sheds some light on what your problem is.  
> Basically, if the library is attached and the function is listed, it  
> is there (or ought to be). Please show the entire transcript of you  
> session (in answering the questions below, not just the part you want  
> to) [possibly skipping to the end, as I now suspect that you do not  
> have enough system software installed - we should think about how to  
> find that out more automatically]
>
>  If you just type ReadAffy, what happens?
> what if you type the name of one of the other functions (in any of the  
> packages)?
> what is .libPaths() reporting?
>
>
>>> If not, then the installation is not correct. Are you sure they have  
>>> global read permission etc on the R library directory and all  
>>> contents? Have you checked to see if there are any files
>>> in those directories?
>>>
>>
>> Do I need to go to terminal to change the permissions or is there a  
>> nice simple way to check permissions and change them accordingly if  
>> that is the problem. Sorry that this appears so dense, but I haven't  
>> quite got the hang of the unix side of OsX yet.
>>
>
> I doubt I was right about that; but there is nothing special going on  
> on OS X, you do need a lot of stuff installed though - and that might  
> be your problem.
>
> try this:
> > capabilities()
>     jpeg      png    tcltk      X11 http/ftp  sockets   libxml     fifo
>     TRUE     TRUE     TRUE     TRUE     TRUE     TRUE     TRUE     TRUE
>   cledit  IEEE754    iconv
>     TRUE     TRUE     TRUE
>
> And in particular you are going to need X11 and tcltk (and you want  
> most of the others). The R FAQ for OS X provides explicit details on  
> how to do this.
>
>  Robert
>
>
>> Thanks again,
>>
>> Chad.
>>
>>>  Robert
>>>
>>>> I've also attempted to "start from scratch" from the original .cel  
>>>> files and get the same type of error message that the function  
>>>> couldn't be found.
>>>>
>>>> I have checked to ensure that affy is installed and loaded, and  
>>>> based on the Package Manager, it is .
>>>>
>>>>
>>>> I've also tried using limma (v1.8.10), and had no problems, other  
>>>> than I can't read in the eset.
>>>>
>>>> > packageDescription("limma", field="Version")
>>>> [1] "1.8.10"
>>>> > packageDescription("affy", field="Version")
>>>> [1] "1.5.8"
>>>> > version
>>>>          _
>>>> platform powerpc-apple-darwin6.8
>>>> arch     powerpc
>>>> os       darwin6.8
>>>> system   powerpc, darwin6.8
>>>> status
>>>> major    2
>>>> minor    0.1
>>>> year     2004
>>>> month    11
>>>> day      15
>>>> language R
>>>>
>>>>
>>>> Thanks for the assistance.
>>>>
>>>> Chad Wayne
>>>>
>>>> Chad M. Wayne, Ph. D.
>>>> Baylor College of Medicine
>>>>
>>>> _______________________________________________
>>>> Bioconductor mailing list
>>>> Bioconductor at stat.math.ethz.ch
>>>> https://stat.ethz.ch/mailman/listinfo/bioconductor
>>>>
>>>>
>>> +-------------------------------------------------------------------- 
>>> -------------------+
>>> | Robert Gentleman              phone: (206) 667-7700                 
>>>             |
>>> | Head, Program in Computational Biology   fax:  (206) 667-1319   |
>>> | Division of Public Health Sciences       office: M2-B865            
>>>          |
>>> | Fred Hutchinson Cancer Research Center                              
>>>             |
>>> | email: rgentlem at fhcrc.org                                           
>>>                             |
>>> +-------------------------------------------------------------------- 
>>> -------------------+
>>>
>>>
>>
>>
> +---------------------------------------------------------------------- 
> -----------------+
> | Robert Gentleman              phone: (206) 667-7700                   
>           |
> | Head, Program in Computational Biology   fax:  (206) 667-1319   |
> | Division of Public Health Sciences       office: M2-B865              
>        |
> | Fred Hutchinson Cancer Research Center                                
>           |
> | email: rgentlem at fhcrc.org                                             
>                           |
> +---------------------------------------------------------------------- 
> -----------------+
>
>



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