old Java version on run_test.sh micro ValueError: Chain results file TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort

[LinuxPersonEC]

Not really a bug but using an older version of Java results in the below. Perhaps a more graceful Java version detection and/or warning/error message? Newer Java of course works.

#### STEP 4: Classify chains using gradient boosting model

Classifying chains
classify_chains: loaded dataframe of size 0
classify_chains: total number of transcripts: 0
classify_chains: 0 rows with spanning chains
classify_chains: filtered dataset contains 0 records
classify_chains: omputing additional features...
classify_chains: WARNING! The final df for classification is empty
classify_chains: df for single-exon model contains 0 records
classify_chains: df for multi-exon model contains 0 records
classify_chains: loading models at /path/to/me/TOGA/./models/se_model.dat (SE) and /path/to/me/TOGA/./models/me_model.dat (ME)
classify_chains: applying models to SE and ME datasets...
classify_chains: applying -1.0 score to the spanning chains
classify_chains: applying -2.0 score to the processed pseudogene alignments
classify_chains: number of processed pseudogene alignments: 0
classify_chains: arranging the final output
classify_chains: classification result stats:
* orthologs: 0
* paralogs: 0
* spanning chains: 0
* processed pseudogenes: 0
classify_chains: using 0.5 as a threshold to separate orthologs from paralogs
classify_chains: combining results for 0 individual transcripts
classify_chains: saving the classification to /path/to/me/TOGA/micro_test_out/temp/trans_to_chain_classes.tsv
classify_chains: found no classifiable chains for 0 transcripts
classify_chains: saving these transcripts to: /path/to/me/TOGA/micro_test_out/temp/rejected/classify_chains_rejected.txt
Chain results file /path/to/me/TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort.
Traceback (most recent call last):
  File "/path/to/me/TOGA/./toga.py", line 1742, in <module>
    main()
  File "/path/to/me/TOGA/./toga.py", line 1738, in main
    toga_manager.run()
  File "/path/to/me/TOGA/./toga.py", line 621, in run
    self.__classify_chains()
  File "/path/to/me/TOGA/./toga.py", line 847, in __classify_chains
    check_chains_classified(self.chain_results_df)
  File "/path/to/me/TOGA/modules/sanity_check_functions.py", line 169, in check_chains_classified
    raise ValueError(msg)
ValueError: Chain results file /path/to/me/TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort

Edit: I do get this error message from the 2nd test on the page:

./toga.py test_input/hg38.mm10.chr11.chain test_input/hg38.genCode27.chr11.bed test_input/hg38.2bit test_input/mm10.2bit --kt --pn test -i supply/hg38.wgEncodeGencodeCompV34.isoforms.txt --nc /path/to/me/TOGA/nextflow_config_files --cb 3,5 --cjn 500 --u12 supply/hg38.U12sites.tsv --ms

#### STEP 4: Classify chains using gradient boosting model

Classifying chains
classify_chains: loaded dataframe of size 0
classify_chains: total number of transcripts: 0
classify_chains: 0 rows with spanning chains
classify_chains: filtered dataset contains 0 records
classify_chains: omputing additional features...
classify_chains: WARNING! The final df for classification is empty
classify_chains: df for single-exon model contains 0 records
classify_chains: df for multi-exon model contains 0 records
classify_chains: loading models at /path/to/me/TOGA/./models/se_model.dat (SE) and /path/to/me/TOGA/./models/me_model.dat (ME)
classify_chains: applying models to SE and ME datasets...
classify_chains: applying -1.0 score to the spanning chains
classify_chains: applying -2.0 score to the processed pseudogene alignments
classify_chains: number of processed pseudogene alignments: 0
classify_chains: arranging the final output
classify_chains: classification result stats:
* orthologs: 0
* paralogs: 0
* spanning chains: 0
* processed pseudogenes: 0
classify_chains: using 0.5 as a threshold to separate orthologs from paralogs
classify_chains: combining results for 0 individual transcripts
classify_chains: saving the classification to /path/to/me/TOGA/test/temp/trans_to_chain_classes.tsv
classify_chains: found no classifiable chains for 0 transcripts
classify_chains: saving these transcripts to: /path/to/me/TOGA/test/temp/rejected/classify_chains_rejected.txt
Chain results file /path/to/me/TOGA/test/temp/chain_results_df.tsv is empty! Abort.
Traceback (most recent call last):
  File "/path/to/me/TOGA/./toga.py", line 1742, in <module>
    main()
  File "/path/to/me/TOGA/./toga.py", line 1738, in main
    toga_manager.run()
  File "/path/to/me/TOGA/./toga.py", line 621, in run
    self.__classify_chains()
  File "/path/to/me/TOGA/./toga.py", line 847, in __classify_chains
    check_chains_classified(self.chain_results_df)
  File "/path/to/me/TOGA/modules/sanity_check_functions.py", line 169, in check_chains_classified
    raise ValueError(msg)
ValueError: Chain results file /path/to/me/TOGA/test/temp/chain_results_df.tsv is empty! Abort.

TOGA/test/temp/chain_results_df.tsv 
gene	gene_overs	chain	synt	gl_score	gl_exo	chain_len	exon_qlen	loc_exo	exon_cover	intr_cover	gene_len	ex_num	ex_fract	intr_fract	flank_cov

[lynnyummy]

Hi, I have a similar error of the chain_results_df.tsv

####` Initiating TOGA class ####
Version 1.1.6
Commit: affa067aaf3beeb8c06471fcddf1bb24deff5bc0
Branch: master

Calling cmd:
./modules/chain_score_filter test_input/align_micro_sample.chain 15000 > /nfs/users//TOGA/micro_test_out/temp/genome_alignment.chain

Command finished with exit code 0.
Continue without isoforms file: not provided
Found 1 sequences in /nfs/users//TOGA/test_input/hg38.micro_sample.2bit
Found 1 sequences in /nfs/users//TOGA/test_input/hg38.micro_sample.2bit
Found 2 sequences in /nfs/users//TOGA/test_input/q2bit_micro_sample.2bit
Saving output to /nfs/users//TOGA/micro_test_out
Arguments stored in /nfs/users//TOGA/micro_test_out/project_args.json


#### STEP 0: making chain and bed file indexes

Started chain indexing...
chain_bst_index: indexing 3 chains
chain_bst_index: Saved chain /nfs/users//TOGA/micro_test_out/temp/genome_alignment.chain index to /nfs/users//TOGA/micro_test_out/temp/genome_alignment.bst
Started bed file indexing...
bed_hdf5_index: indexed 3 transcripts


#### STEP 1: Generate extract chain features jobs

Calling cmd:
./split_chain_jobs.py /nfs/users//TOGA/micro_test_out/temp/genome_alignment.chain /nfs/users//TOGA/micro_test_out/temp/toga_filt_ref_annot.bed /nfs/users//TOGA/micro_test_out/temp/toga_filt_ref_annot.hdf5 --log_file /nfs/users//TOGA/micro_test_out/toga_2023_12_21_at_11_46.log --parallel_logs_dir /nfs/users//TOGA/micro_test_out/temp_logs --jobs_num 1 --jobs /nfs/users//TOGA/micro_test_out/temp/chain_classification_jobs --jobs_file /nfs/users//TOGA/micro_test_out/temp/chain_class_jobs_combined --results_dir /nfs/users//TOGA/micro_test_out/temp/chain_classification_results --rejected /nfs/users//TOGA/micro_test_out/temp/rejected/SPLIT_CHAIN_REJ.txt

split_chain_jobs: Use bed file /nfs/users//TOGA/micro_test_out/temp/toga_filt_ref_annot.bed and chain file /nfs/users//TOGA/micro_test_out/temp/genome_alignment.chain
split_chain jobs: the run data overview is:

* vv: False
* jobs: /nfs/users//TOGA/micro_test_out/temp/chain_classification_jobs
* results_dir: /nfs/users//TOGA/micro_test_out/temp/chain_classification_results
* errors_dir: None
* chain_file: /nfs/users//TOGA/micro_test_out/temp/genome_alignment.chain
* bed_file: /nfs/users//TOGA/micro_test_out/temp/toga_filt_ref_annot.bed
* index_file: /nfs/users//TOGA/micro_test_out/temp/genome_alignment.chain_ID_position
* job_size: None
* jobs_num: 1
* bed_index: /nfs/users//TOGA/micro_test_out/temp/toga_filt_ref_annot.hdf5
* jobs_file: /nfs/users//TOGA/micro_test_out/temp/chain_class_jobs_combined
* ref: hg38
* on_cluster: True
split_chain_jobs: searching for intersections between reference transcripts and chains
split_chain_jobs: chains-to-transcripts dict contains 2 records
split_chain_jobs: skipped 0 transcripts that do not intersect any chain
split_chain_jobs: preparing 2 commands
split_chain_jobs: command size of 3 for each cluster job
split_chain_jobs: results in 1 cluster jobs
split_chain_jobs: estimated time: 0:00:00.013618
Command finished with exit code 0.


#### STEP 2: Extract chain features: parallel step

Extracting chain features, project name: chain_feats__micro_test_out_at_1703159205
Project path: ./nextflow_logs/chain_feats__micro_test_out_at_1703159205
Selected parallelization strategy: nextflow
Parallel manager: pushing job nextflow /nfs/users//TOGA/execute_joblist.nf --joblist /nfs/users//TOGA/micro_test_out/temp/chain_class_jobs_combined
Logs from individual chain runner jobs are show below


#### STEP 3: Merge step 2 output

Reading /nfs/users//TOGA/micro_test_out/temp/toga_filt_ref_annot.bed
merge_chains_output: got data for 3 transcripts
merge_chains_output: Loading the results...
merge_chains_output: There are 0 result files to combine
merge_chains_output: got 0 keys in chain_genes_data
merge_chains_output: got 0 keys in chain_raw_data
merge_chains_output: There were 0 transcript lines and 0 chain lines
merge_chains_output: chain_genes_data dict reverted, there are 0 keys now
merge_chains_output: Combining the data...
merge_chains_output: got combined dict with 0 keys
merge_chains_output: Writing output to /nfs/users//TOGA/micro_test_out/temp/chain_results_df.tsv
merge_chains_output: total runtime: 0:00:03.469617


#### STEP 4: Classify chains using gradient boosting model

Classifying chains
classify_chains: loaded dataframe of size 0
classify_chains: total number of transcripts: 0
classify_chains: 0 rows with spanning chains
classify_chains: filtered dataset contains 0 records
classify_chains: omputing additional features...
classify_chains: WARNING! The final df for classification is empty
classify_chains: df for single-exon model contains 0 records
classify_chains: df for multi-exon model contains 0 records
classify_chains: loading models at ./models/se_model.dat (SE) and ./models/me_model.dat (ME)
classify_chains: applying models to SE and ME datasets...
classify_chains: applying -1.0 score to the spanning chains
classify_chains: applying -2.0 score to the processed pseudogene alignments
classify_chains: number of processed pseudogene alignments: 0
classify_chains: arranging the final output
classify_chains: classification result stats:
* orthologs: 0
* paralogs: 0
* spanning chains: 0
* processed pseudogenes: 0
classify_chains: using 0.5 as a threshold to separate orthologs from paralogs
classify_chains: combining results for 0 individual transcripts
classify_chains: saving the classification to /nfs/users//TOGA/micro_test_out/temp/trans_to_chain_classes.tsv
classify_chains: found no classifiable chains for 0 transcripts
classify_chains: saving these transcripts to: /nfs/users//TOGA/micro_test_out/temp/rejected/classify_chains_rejected.txt
Chain results file /nfs/users//TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort.

I don't know if you figure out this issue?
Thanks!

[guo-cheng]

I also have this problem. Do you know how to solve this?

[lynnyummy]

I also have this problem. Do you know how to solve this?

not yet....
have you sorted it out?

[MichaelHiller]

@kirilenkobm Can you pls look into this?
What I don't understand is what Java has to do with any of this, as TOGA doesn't use Java??

[kirilenkobm]

@MichaelHiller @lynnyummy @SomePersonSomeWhereInTheWorld @guo-cheng
The Chain results file /nfs/users//TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort. message is misleading -> in fact, nextflow does not even start the jobs. However, it does not return any non-0 error code, so TOGA crashes due to a bit general sanity check.
Nextflow indeed requires Java, and, of course, some pairs of versions are incompatible :(.
Please try downgrading nextflow. I'd recommend trying 23.10.0

[LinuxPersonEC]

@MichaelHiller @lynnyummy @SomePersonSomeWhereInTheWorld @guo-cheng The Chain results file /nfs/users//TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort. message is misleading -> in fact, nextflow does not even start the jobs. However, it does not return any non-0 error code, so TOGA crashes due to a bit general sanity check. Nextflow indeed requires Java, and, of course, some pairs of versions are incompatible :(. Please try downgrading nextflow. I'd recommend trying 23.10.0

Already using that:

nextflow -v
nextflow version 23.10.0.5889

Same results:

classify_chains: using 0.5 as a threshold to separate orthologs from paralogs
classify_chains: combining results for 0 individual transcripts
classify_chains: saving the classification to /path/to/me/TOGA/test/temp/trans_to_chain_classes.tsv
classify_chains: found no classifiable chains for 0 transcripts
classify_chains: saving these transcripts to: /path/to/me/TOGA/test/temp/rejected/classify_chains_rejected.txt
Chain results file /path/to/me/TOGA/test/temp/chain_results_df.tsv is empty! Abort.
Traceback (most recent call last):
  File "/path/to/me/TOGA/toga.py", line 1742, in <module>
    main()
  File "/path/to/me/TOGA/toga.py", line 1738, in main
    toga_manager.run()
  File "/path/to/me/TOGA/toga.py", line 621, in run
    self.__classify_chains()
  File "/path/to/me/TOGA/toga.py", line 847, in __classify_chains
    check_chains_classified(self.chain_results_df)
  File "/path/to/me/TOGA/modules/sanity_check_functions.py", line 169, in check_chains_classified
    raise ValueError(msg)
ValueError: Chain results file /path/to/me/TOGA/test/temp/chain_results_df.tsv is empty! Abort.
rk3199@g029:~/TOGA$ ls  /path/to/me/TOGA/test/temp/chain_results_df.tsv
/path/to/me/TOGA/test/temp/chain_results_df.tsv

the .tsv file does have:

chain_results_df.tsv
gene	gene_overs	chain	synt	gl_score	gl_exo	chain_len	exon_qlen	loc_exo	exon_cover	intr_cover	gene_len	ex_num	ex_fract	intr_fract	flank_cov

java --version
java 20.0.1 2023-04-18

[lynnyummy]

@MichaelHiller @lynnyummy @SomePersonSomeWhereInTheWorld @guo-cheng The Chain results file /nfs/users//TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort. message is misleading -> in fact, nextflow does not even start the jobs. However, it does not return any non-0 error code, so TOGA crashes due to a bit general sanity check. Nextflow indeed requires Java, and, of course, some pairs of versions are incompatible :(. Please try downgrading nextflow. I'd recommend trying 23.10.0

Hi, I've checked my Nextflow is 23.10.0 and the Java is openjdk 21-internal 2023-09-19.

I checked those error files again, I think probably it is not enough memory for Java,
N E X T F L O W ~ version 23.10.0 There is insufficient memory for the Java Runtime Environment to continue. Native memory allocation (malloc) failed to allocate 1424 bytes for AllocateHeap

Can we modify the run_test.sh and specify Java in the script? any suggestion?

thanks!

[kirilenkobm]

@lynnyummy

The error message is quite misleading - it appears that the problem might be related to Java's heap memory settings rather than a lack of physical memory (it failed to allocate just 1424 bytes, it's nothing).
Pls try to increase Java heap size by doing
export NXF_OPTS='-Xms1g -Xmx4g'

[lynnyummy]

export NXF_OPTS='-Xms1g -Xmx4g

Hi, I tried to put this at the end of the command, but still failed and reported the same error.
TOGA/micro_test_out/temp/chain_results_df.tsv is empty! Abort.

[kirilenkobm]

Seems like I should give this a push #104
so that nextflow logs are carefully arranged and we can analyse what exactly is going on

[lynnyummy]

@lynnyummy

The error message is quite misleading - it appears that the problem might be related to Java's heap memory settings rather than a lack of physical memory (it failed to allocate just 1424 bytes, it's nothing). Pls try to increase Java heap size by doing export NXF_OPTS='-Xms1g -Xmx4g'

I sorted out this issue, just specify the memory in my own script.

[lynnyummy]

Hi I got the same error for the second test with the human and mouse datasets. I checked the log file, and indicated that
`Mar-05 15:41:35.927 [Task submitter] ERROR nextflow.processor.TaskProcessor - Error executing process > 'execute_jobs (2)'

Caused by:
java.io.IOException: Cannot run program "sbatch" (in directory "~/TOGA/nextflow_logs/work/15/f2b3a7ba56330223cecf3946904a73"): error=2, No such file or directory

Command executed:

sbatch .command.run

Command exit status:

Command output:
(empty)

Work dir:
~/TOGA/nextflow_logs/work/15/f2b3a7ba56330223cecf3946904a73

Tip: when you have fixed the problem you can continue the execution adding the option -resume to the run command line
Mar-05 15:41:35.928 [Task submitter] DEBUG nextflow.Session - Session aborted -- Cause: Error submitting process 'execute_jobs (2)' for execution
Mar-05 15:41:35.944 [main] DEBUG nextflow.Session - Session await > all processes finished
Mar-05 15:41:35.944 [main] DEBUG nextflow.Session - Session await > all barriers passed
Mar-05 15:41:35.945 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: slurm) - terminating tasks monitor poll loop
Mar-05 15:41:35.949 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=0; failedCount=217; ignoredCount=0; cachedCount=0; pendingCount=317; submittedCount=0; runningCount=-217; retriesCount=217; abortedCount=0; succeedDuration=0ms; failedDuration=0ms; cachedDuration=0ms;loadCpus=-217; loadMemory=0; peakRunning=0; peakCpus=0; peakMemory=0; ]
Mar-05 15:41:36.158 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done
Mar-05 15:41:36.174 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye`

Seems like I should give this a push #104 so that nextflow logs are carefully arranged and we can analyse what exactly is going on

It can still generate the file genome_alignment.chain, but not chain_results_df.tsv. Can you help to check the issue?