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Funcotator data source not correct

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    Genevieve Brandt (she/her)

    Hi Jamie Xie, there was an update to the Funcotator data sources in GATK 4.1.9.0 and 4.2.0.0, you can find those release notes here: https://github.com/broadinstitute/gatk/releases.

    Could you try updating to 4.2.0.0 and see if this issue is resolved?

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    Jamie Xie

    Hi Genevieve,

    Thanks for the response! I can't seem to find version 4.2.0.0 of the funcotator data sources or the workflow on Terra. The workflow listed here is still 4.1.7.0 and https://app.terra.bio/#workspaces/help-gatk/Variant-Functional-Annotation-With-Funcotator/workflows and this bucket gs://broad-public-datasets/funcotator only contains up to 4.1.7.0 as well.

    Where can I find the latest versions of these?

    Thanks,

    Jamie

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    Genevieve Brandt (she/her)

    Jamie,

    The version 4.2.0.0 is the GATK version, not the Funcotator data sources version, sorry about that!

    Genevieve

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    Jamie Xie

    Hi Genevieve,

    I tried re-running Funcotator workflow (version 4.1.7.0) with gatk docker version 4.2.0.0 but unfortunately I got the same warning. Is there something else I might be doing incorrectly?

    Thanks,

    Jamie

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    Genevieve Brandt (she/her)

    Hi Jamie,

    Thank you for checking that. Let's see if there is a different issue going on. I noticed that Funcotator did not process any variants:

    19:59:54.622 INFO  ProgressMeter - Traversal complete. Processed 0 total variants in 0.4 minutes.

    Do you know why this might be the case? How many variants are in your input file?

    Best,

    Genevieve

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    Jamie Xie

    Hi Genevieve,

    Yeah I don't know why it said it processed 0 variants. When running this in conjunction with Mutect2 (using the workflow I posted above), FilterMutectCalls processed around 4 million variants. The vcf files from Mutect2 are not empty.

    13:36:31.507 INFO  ProgressMeter - Traversal complete. Processed 4532912 total variants in 12.6 minutes.

    So even though there are clearly a lot of variants, the MAF output from Funcotator is empty even though the run was successful.

    Thanks,

    Jamie

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    Genevieve Brandt (she/her)

    Hi Jamie,

    Funcotator is usually run after FilterMutectCalls, could you check the output from FilterMutectCalls and verify there are variants in that file?

    Genevieve

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    Jamie Xie

    Hi Genevieve,

    Yes, in the Mutect2 workflow I'm referring to (https://dockstore.org/workflows/github.com/broadinstitute/gatk/mutect2:4.1.7.0) Funcotator runs after Mutect2 and FilterMutectCalls finishes. The filtered output from FilterMutectCalls does contain variants (it's a large file). I took a screenshot of the file size and the FilterMutectCalls log file.

    2021/03/26 13:23:18 Starting container setup.
    2021/03/26 13:23:20 Done container setup.
    2021/03/26 13:23:24 Starting localization.
    2021/03/26 13:23:31 Localization script execution started...
    2021/03/26 13:23:31 Localizing input gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeVCFs/RESPOND_60100256_T1_Malignant-unfiltered.vcf -> /cromwell_root/fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeVCFs/RESPOND_60100256_T1_Malignant-unfiltered.vcf
    2021/03/26 13:23:36 Localizing input gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeVCFs/RESPOND_60100256_T1_Malignant-unfiltered.vcf.idx -> /cromwell_root/fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeVCFs/RESPOND_60100256_T1_Malignant-unfiltered.vcf.idx
    Copying gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeVCFs/RESPOND_60100256_T1_Malignant-unfiltered.vcf.idx...
    / [0/1 files][    0.0 B/  3.0 MiB]   0% Done                                    
    / [1/1 files][  3.0 MiB/  3.0 MiB] 100% Done                                    
    Operation completed over 1 objects/3.0 MiB.                                      
    2021/03/26 13:23:37 Localizing input gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/script -> /cromwell_root/script
    2021/03/26 13:23:39 Localizing input gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeStats/merged.stats -> /cromwell_root/fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeStats/merged.stats
    2021/03/26 13:23:41 Localization script execution complete.
    2021/03/26 13:23:46 Done localization.
    2021/03/26 13:23:47 Running user action: docker run -v /mnt/local-disk:/cromwell_root --entrypoint=/bin/bash broadinstitute/gatk@sha256:192fedf9b9d65809da4a2954030269e3f311d296e6b5e4c6c7deec12c7fe84b2 /cromwell_root/script
    Picked up _JAVA_OPTIONS: -Djava.io.tmpdir=/cromwell_root/tmp.98cb9fe5
    13:23:52.091 INFO  NativeLibraryLoader - Loading libgkl_compression.so from jar:file:/gatk/gatk-package-4.1.7.0-local.jar!/com/intel/gkl/native/libgkl_compression.so
    13:23:52.377 INFO  FilterMutectCalls - ------------------------------------------------------------
    13:23:52.378 INFO  FilterMutectCalls - The Genome Analysis Toolkit (GATK) v4.1.7.0
    13:23:52.379 INFO  FilterMutectCalls - For support and documentation go to https://software.broadinstitute.org/gatk/
    13:23:52.380 INFO  FilterMutectCalls - Executing as root@d0dd8d871fcb on Linux v5.4.89+ amd64
    13:23:52.380 INFO  FilterMutectCalls - Java runtime: OpenJDK 64-Bit Server VM v1.8.0_212-8u212-b03-0ubuntu1.16.04.1-b03
    13:23:52.381 INFO  FilterMutectCalls - Start Date/Time: March 26, 2021 1:23:52 PM UTC
    13:23:52.381 INFO  FilterMutectCalls - ------------------------------------------------------------
    13:23:52.381 INFO  FilterMutectCalls - ------------------------------------------------------------
    13:23:52.383 INFO  FilterMutectCalls - HTSJDK Version: 2.21.2
    13:23:52.383 INFO  FilterMutectCalls - Picard Version: 2.21.9
    13:23:52.384 INFO  FilterMutectCalls - HTSJDK Defaults.COMPRESSION_LEVEL : 2
    13:23:52.384 INFO  FilterMutectCalls - HTSJDK Defaults.USE_ASYNC_IO_READ_FOR_SAMTOOLS : false
    13:23:52.385 INFO  FilterMutectCalls - HTSJDK Defaults.USE_ASYNC_IO_WRITE_FOR_SAMTOOLS : true
    13:23:52.385 INFO  FilterMutectCalls - HTSJDK Defaults.USE_ASYNC_IO_WRITE_FOR_TRIBBLE : false
    13:23:52.386 INFO  FilterMutectCalls - Deflater: IntelDeflater
    13:23:52.386 INFO  FilterMutectCalls - Inflater: IntelInflater
    13:23:52.386 INFO  FilterMutectCalls - GCS max retries/reopens: 20
    13:23:52.386 INFO  FilterMutectCalls - Requester pays: disabled
    13:23:52.387 INFO  FilterMutectCalls - Initializing engine
    13:23:54.186 INFO  FeatureManager - Using codec VCFCodec to read file file:///cromwell_root/fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeVCFs/RESPOND_60100256_T1_Malignant-unfiltered.vcf
    13:23:54.318 INFO  FilterMutectCalls - Done initializing engine
    13:23:54.436 INFO  ProgressMeter - Starting traversal
    13:23:54.436 INFO  ProgressMeter -        Current Locus  Elapsed Minutes    Variants Processed  Variants/Minute
    13:23:54.438 INFO  FilterMutectCalls - Starting pass 0 through the variants
    13:24:04.449 INFO  ProgressMeter -        chr1:42986564              0.2                 34000         203775.8
    13:24:14.536 INFO  ProgressMeter -       chr1:245222266              0.3                101000         301507.5
    13:24:24.540 INFO  ProgressMeter -       chr2:197693615              0.5                167000         332857.2
    13:24:34.541 INFO  ProgressMeter -       chr3:136061521              0.7                230000         344105.3
    13:24:44.567 INFO  ProgressMeter -       chr4:129088897              0.8                288000         344703.8
    13:24:54.601 INFO  ProgressMeter -       chr5:176891354              1.0                356000         355029.6
    13:25:04.660 INFO  ProgressMeter -       chr6:169434913              1.2                418000         357153.0
    13:25:14.800 INFO  ProgressMeter -          chr8:244125              1.3                486000         362853.6
    13:25:24.833 INFO  ProgressMeter -        chr9:89334980              1.5                554000         367719.5
    13:25:34.835 INFO  ProgressMeter -       chr10:94027559              1.7                616000         368134.8
    13:25:44.924 INFO  ProgressMeter -      chr11:114909723              1.8                677000         367648.4
    13:25:54.959 INFO  ProgressMeter -      chr12:116395048              2.0                733000         364912.6
    13:26:05.101 INFO  ProgressMeter -       chr14:90836185              2.2                800000         367354.4
    13:26:15.194 INFO  ProgressMeter -       chr16:24786480              2.3                865000         368720.6
    13:26:25.339 INFO  ProgressMeter -       chr17:47754984              2.5                934000         371366.8
    13:26:35.460 INFO  ProgressMeter -       chr19:13815219              2.7               1003000         373735.4
    13:26:45.566 INFO  ProgressMeter -       chr20:46583790              2.9               1068000         374454.4
    13:26:56.955 INFO  ProgressMeter -       chrX:154352329              3.0               1128000         370812.7
    13:26:57.715 INFO  FilterMutectCalls - Finished pass 0 through the variants
    13:26:58.029 INFO  FilterMutectCalls - Starting pass 1 through the variants
    13:27:06.960 INFO  ProgressMeter -       chr1:112422914              3.2               1190000         370864.8
    13:27:16.962 INFO  ProgressMeter -        chr2:40865689              3.4               1251000         370620.9
    13:27:26.964 INFO  ProgressMeter -       chr2:233846383              3.5               1315000         371248.7
    13:27:36.994 INFO  ProgressMeter -       chr3:193608779              3.7               1381000         372309.1
    13:27:47.104 INFO  ProgressMeter -        chr5:31564663              3.9               1444000         372377.7
    13:27:57.104 INFO  ProgressMeter -        chr6:20171837              4.0               1501000         371125.9
    13:28:07.217 INFO  ProgressMeter -        chr7:42288088              4.2               1568000         372182.8
    13:28:17.358 INFO  ProgressMeter -        chr8:34388828              4.4               1631000         372203.1
    13:28:27.362 INFO  ProgressMeter -       chr9:114321725              4.5               1698000         373289.4
    13:28:37.472 INFO  ProgressMeter -      chr10:129746007              4.7               1765000         374160.0
    13:28:47.605 INFO  ProgressMeter -       chr12:26882636              4.9               1829000         374325.9
    13:28:57.695 INFO  ProgressMeter -       chr13:32368867              5.1               1886000         373147.6
    13:29:07.724 INFO  ProgressMeter -       chr15:43334929              5.2               1952000         373842.5
    13:29:17.750 INFO  ProgressMeter -       chr16:68616162              5.4               2015000         373941.0
    13:29:27.821 INFO  ProgressMeter -       chr17:69384288              5.6               2081000         374523.1
    13:29:37.833 INFO  ProgressMeter -       chr19:23087299              5.7               2148000         375310.1
    13:29:47.913 INFO  ProgressMeter -       chr21:10601785              5.9               2212000         375471.0
    13:29:57.982 INFO  ProgressMeter -        chrX:71902901              6.1               2257000         372498.6
    13:29:59.421 INFO  FilterMutectCalls - Finished pass 1 through the variants
    13:29:59.550 INFO  FilterMutectCalls - Starting pass 2 through the variants
    13:30:08.039 INFO  ProgressMeter -       chr1:111548156              6.2               2323000         373070.8
    13:30:18.058 INFO  ProgressMeter -        chr2:55726342              6.4               2391000         373962.8
    13:30:28.080 INFO  ProgressMeter -        chr3:11521735              6.6               2455000         374196.9
    13:30:38.112 INFO  ProgressMeter -         chr4:9721881              6.7               2523000         375005.6
    13:30:48.157 INFO  ProgressMeter -        chr5:76922639              6.9               2591000         375761.4
    13:30:58.224 INFO  ProgressMeter -        chr6:42852460              7.1               2647000         374763.7
    13:31:08.251 INFO  ProgressMeter -        chr7:71819459              7.2               2714000         375368.2
    13:31:18.361 INFO  ProgressMeter -        chr8:87223541              7.4               2781000         375875.1
    13:31:28.391 INFO  ProgressMeter -       chr9:135675692              7.6               2844000         375897.9
    13:31:38.472 INFO  ProgressMeter -       chr11:35850732              7.7               2912000         376523.3
    13:31:48.584 INFO  ProgressMeter -       chr12:65921536              7.9               2980000         377098.2
    13:31:58.668 INFO  ProgressMeter -       chr14:31090188              8.1               3044000         377175.4
    13:32:08.778 INFO  ProgressMeter -       chr15:86908424              8.2               3106000         376986.7
    13:32:18.798 INFO  ProgressMeter -       chr17:12787192              8.4               3175000         377705.7
    13:32:28.872 INFO  ProgressMeter -       chr18:53124794              8.6               3241000         378006.9
    13:32:38.961 INFO  ProgressMeter -       chr19:55293165              8.7               3311000         378744.1
    13:32:48.968 INFO  ProgressMeter -       chr22:41984997              8.9               3380000         379398.7
    13:32:51.849 INFO  FilterMutectCalls - Finished pass 2 through the variants
    13:32:51.891 INFO  FilterMutectCalls - Starting pass 3 through the variants
    13:32:59.173 INFO  ProgressMeter -        chr1:23328688              9.1               3418000         376475.9
    13:33:09.220 INFO  ProgressMeter -       chr1:150571067              9.2               3462000         374416.7
    13:33:19.311 INFO  ProgressMeter -        chr2:39023194              9.4               3517000         373570.0
    13:33:29.565 INFO  ProgressMeter -       chr2:206978301              9.6               3571000         372543.8
    13:33:39.714 INFO  ProgressMeter -        chr3:94994352              9.8               3617000         370798.8
    13:33:49.723 INFO  ProgressMeter -        chr4:75557406              9.9               3673000         370208.6
    13:33:59.754 INFO  ProgressMeter -        chr5:98918337             10.1               3730000         369723.6
    13:34:09.885 INFO  ProgressMeter -        chr6:36573890             10.3               3777000         368219.6
    13:34:20.036 INFO  ProgressMeter -        chr7:43800161             10.4               3835000         367807.5
    13:34:30.209 INFO  ProgressMeter -        chr8:23757197             10.6               3893000         367395.9
    13:34:40.270 INFO  ProgressMeter -        chr9:71408835             10.8               3947000         366689.2
    13:34:50.293 INFO  ProgressMeter -       chr10:62241897             10.9               4002000         366117.0
    13:35:00.295 INFO  ProgressMeter -       chr11:65020634             11.1               4057000         365573.4
    13:35:10.381 INFO  ProgressMeter -       chr12:48698210             11.3               4102000         364113.0
    13:35:20.475 INFO  ProgressMeter -       chr13:49721590             11.4               4159000         363740.8
    13:35:30.606 INFO  ProgressMeter -       chr15:39780250             11.6               4215000         363273.9
    13:35:40.738 INFO  ProgressMeter -       chr16:28479566             11.8               4267000         362480.0
    13:35:50.782 INFO  ProgressMeter -       chr17:36582310             11.9               4322000         362004.8
    13:36:00.808 INFO  ProgressMeter -       chr18:62347714             12.1               4377000         361550.8
    13:36:10.925 INFO  ProgressMeter -       chr19:36554365             12.3               4421000         360168.8
    13:36:21.035 INFO  ProgressMeter -       chr20:62852996             12.4               4476000         359712.1
    13:36:31.086 INFO  ProgressMeter -    KI270442.1:239384             12.6               4531000         359294.7
    13:36:31.433 INFO  FilterMutectCalls - Finished pass 3 through the variants
    13:36:31.506 INFO  FilterMutectCalls - No variants filtered by: AllowAllVariantsVariantFilter
    13:36:31.507 INFO  FilterMutectCalls - 0 read(s) filtered by: AllowAllReadsReadFilter 
    
    13:36:31.507 INFO  ProgressMeter -   KI270728.1:1585634             12.6               4532912         359246.5
    13:36:31.507 INFO  ProgressMeter - Traversal complete. Processed 4532912 total variants in 12.6 minutes.
    13:36:31.724 INFO  FilterMutectCalls - Shutting down engine
    [March 26, 2021 1:36:31 PM UTC] org.broadinstitute.hellbender.tools.walkers.mutect.filtering.FilterMutectCalls done. Elapsed time: 12.66 minutes.
    Runtime.totalMemory()=3106406400
    Using GATK jar /root/gatk.jar defined in environment variable GATK_LOCAL_JAR
    Running:
        java -Dsamjdk.use_async_io_read_samtools=false -Dsamjdk.use_async_io_write_samtools=true -Dsamjdk.use_async_io_write_tribble=false -Dsamjdk.compression_level=2 -Xmx7500m -jar /root/gatk.jar FilterMutectCalls -V /cromwell_root/fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeVCFs/RESPOND_60100256_T1_Malignant-unfiltered.vcf -R gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/GRCh38.primary_assembly.genome.fa -O RESPOND_60100256_T1_Malignant-filtered.vcf -stats /cromwell_root/fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-MergeStats/merged.stats --filtering-stats filtering.stats --min-reads-per-strand 5 --min-allele-fraction 5.0
    2021/03/26 13:36:34 Starting delocalization.
    2021/03/26 13:36:36 Delocalization script execution started...
    2021/03/26 13:36:36 Delocalizing output /cromwell_root/memory_retry_rc -> gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/memory_retry_rc
    2021/03/26 13:36:36 Delocalizing output /cromwell_root/rc -> gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/rc
    2021/03/26 13:36:37 Delocalizing output /cromwell_root/stdout -> gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/stdout
    2021/03/26 13:36:38 Delocalizing output /cromwell_root/stderr -> gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/stderr
    2021/03/26 13:36:40 Delocalizing output /cromwell_root/filtering.stats -> gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/filtering.stats
    2021/03/26 13:36:41 Delocalizing output /cromwell_root/RESPOND_60100256_T1_Malignant-filtered.vcf -> gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/RESPOND_60100256_T1_Malignant-filtered.vcf
    2021/03/26 13:36:46 Delocalizing output /cromwell_root/RESPOND_60100256_T1_Malignant-filtered.vcf.idx -> gs://fc-9828bda3-33d6-40b9-ad27-544fb296a8e0/37a63de4-948f-45a9-bd5a-75b8ee8fb165/Mutect2/09c6ec7f-abc9-40b0-9435-6e9270471089/call-Filter/RESPOND_60100256_T1_Malignant-filtered.vcf.idx
    2021/03/26 13:36:48 Delocalization script execution complete.
    2021/03/26 13:36:49 Done delocalization.

    Best,

    Jamie

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    Genevieve Brandt (she/her)

    Jamie, 

    Okay great! Thank you for posting the details. Could make sure that there are variants that passed the filtering (marked as PASS in the FILTER column)? It's still so strange to me that there were no variants processed in the Funcotator progress meter. You can run SelectVariants with --exclude-filtered to get only variants that have passed the filtering steps. 

    Let me know the output that you get from that.

    Best,

    Genevieve

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    Jamie Xie

    Hi Genevieve,

    I think you found the problem! I looked through the vcf file and none of the variants were marked with PASS, and because I set this filter argument to true, none of the variants were annotated.

    As a follow up question, are we calling FilterMutectCalls correctly in the workflow? I couldn't find any documentation as to how this field should be formatted.

    Thanks for the help!

    Jamie

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    Genevieve Brandt (she/her)

    Hi Jamie,

    For the funco_filter_funcotations, I think it would be fine to leave that as true. It is best to only annotate the PASS variants because the filtered variants don't need to be analyzed with this step. You should be able to get variants that PASS your FilterMutectCalls command by removing your FilterMutectCalls options, they are too stringent.

    For the --min-reads-per-strand, here is a forum discussion related to that option. It is an advanced filter and we don't recommend it for most users.

    The --min-allele-fraction can be useful if you are looking for certain variants above a certain allele fraction (filters out variants with low allele fraction). Generally AF values are between 0 and 1 and so with 5.0 you would be filtering out all of your variants.

    FilterMutectCalls generally does quite well with the default filters, and then if you are seeing strange results you can go back and change the filtering options depending on what you find.

    This should fix your issue! 

    Genevieve

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    Jamie Xie

    Hi Genevieve,

    Thanks for the clarification, that makes a lot of sense now! We thought the AF was a percentage rather than a fraction.

    Regarding the default arguments for FilterMutectCalls, how would I input those? Or is FilterMutectCalls always called after Mutect2 with the default arguments?

    Best,

    Jamie

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    Genevieve Brandt (she/her)

    I believe if you just leave the FilterMutectCalls step of the Mutect2 WDL as-is, it should be using the recommended options for most use cases. You probably won't need to change the m2_extra_filtering_args parameter from what it was originally.

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