From b2b768929dfc1317af694ad166b6153f48e3422c Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Wed, 8 Jan 2025 09:18:09 -0800 Subject: [PATCH 01/23] DM-229 Consolidating parsing logic for load_processed functions. Still need to apply to export functions, and need to check that performance is unimpacted. --- dimelo/load_processed.py | 189 ++++++++++++++++++++------------------- 1 file changed, 95 insertions(+), 94 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 1c1e1a8..3e850eb 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -76,6 +76,60 @@ def regions_to_list( return results +def process_pileup_row( + row: str, + parsed_motif: utils.ParsedMotif, + region_start: int, + region_end: int, + region_strand: str, + single_strand: bool, + regions_5to3prime: bool, +) -> tuple[bool, int, int, int]: + """ + Returns: keep_basemod, pileup_coord_relative, modified_in_row, valid_in_row + """ + tabix_fields = row.split("\t") + pileup_basemod = tabix_fields[3] + pileup_strand = tabix_fields[5] + keep_basemod = False + if single_strand and pileup_strand.strip() != region_strand: + # We are on the wrong strand, skip the rest of the steps for this row + return keep_basemod, None, None, None + elif len(pileup_basemod.split(",")) == 3: + pileup_modname, pileup_motif, pileup_mod_coord = pileup_basemod.split(",") + if ( + pileup_motif == parsed_motif.motif_seq + and int(pileup_mod_coord) == parsed_motif.modified_pos + and pileup_modname in parsed_motif.mod_codes + ): + keep_basemod = True + elif len(pileup_basemod.split(",")) == 1: + if pileup_basemod in parsed_motif.mod_codes: + keep_basemod = True + else: + raise ValueError( + f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." + ) + if keep_basemod: + pileup_info = tabix_fields[9].split(" ") + genomic_coord = int(tabix_fields[1]) + if regions_5to3prime and region_strand == "-": + # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction + # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal + pileup_coord_relative = region_end - genomic_coord - 1 + else: + # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case + pileup_coord_relative = genomic_coord - region_start + + valid_in_row = int(pileup_info[0]) + modified_in_row = int(pileup_info[2]) + + return keep_basemod, pileup_coord_relative, modified_in_row, valid_in_row + + else: + return keep_basemod, None, None, None + + def pileup_counts_from_bedmethyl( bedmethyl_file: str | Path, motif: str, @@ -129,64 +183,35 @@ def pileup_counts_from_bedmethyl( # TODO: change to try-except if chromosome in source_tabix.contigs: for row in source_tabix.fetch(chromosome, start_coord, end_coord): - # TODO Consider using csv module - # TODO: probably this whole block should share logic with vectors_from_bedmethyl AND from export module functions - tabix_fields = row.split("\t") - pileup_basemod = tabix_fields[3] - pileup_strand = tabix_fields[5] - keep_basemod = False - if single_strand and pileup_strand != strand: - # This entry is on the wrong strand - skip it - continue - elif len(pileup_basemod.split(",")) == 3: - pileup_modname, pileup_motif, pileup_mod_coord = ( - pileup_basemod.split(",") - ) - if ( - pileup_motif == parsed_motif.motif_seq - and int(pileup_mod_coord) == parsed_motif.modified_pos - and pileup_modname in parsed_motif.mod_codes - ): - keep_basemod = True - elif len(pileup_basemod.split(",")) == 1: - if pileup_basemod in parsed_motif.mod_codes: - keep_basemod = True - else: - raise ValueError( - f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." + keep_basemod, _, modified_in_row, valid_in_row = ( + process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=start_coord, + region_end=end_coord, + region_strand=strand, + single_strand=single_strand, + regions_5to3prime=False, ) - # TODO: consolidate the above into a function; just do adding outside + ) if keep_basemod: - pileup_info = tabix_fields[9].split(" ") - valid_base_count += int(pileup_info[0]) - modified_base_count += int(pileup_info[2]) + valid_base_count += valid_in_row + modified_base_count += modified_in_row else: # Get counts from the whole input file for row in source_tabix.fetch(): - tabix_fields = row.split("\t") - pileup_basemod = tabix_fields[3] - keep_basemod = False - if len(pileup_basemod.split(",")) == 3: - pileup_modname, pileup_motif, pileup_mod_coord = pileup_basemod.split( - "," - ) - if ( - pileup_motif == parsed_motif.motif_seq - and int(pileup_mod_coord) == parsed_motif.modified_pos - and pileup_modname in parsed_motif.mod_codes - ): - keep_basemod = True - elif len(pileup_basemod.split(",")) == 1: - if pileup_basemod in parsed_motif.mod_codes: - keep_basemod = True - else: - raise ValueError( - f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." - ) + keep_basemod, _, modified_in_row, valid_in_row = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=start_coord, + region_end=end_coord, + region_strand=strand, + single_strand=single_strand, + regions_5to3prime=False, + ) if keep_basemod: - pileup_info = tabix_fields[9].split(" ") - valid_base_count += int(pileup_info[0]) - modified_base_count += int(pileup_info[2]) + valid_base_count += valid_in_row + modified_base_count += modified_in_row return (modified_base_count, valid_base_count) @@ -267,55 +292,31 @@ def pileup_vectors_from_bedmethyl( # center_coord = (start_coord+end_coord)//2 if chromosome in source_tabix.contigs: for row in source_tabix.fetch(chromosome, start_coord, end_coord): - # TODO: can we consolidate this with pileup_counts_from_bedmethyl? - # Just the checks? - # TODO: probably this whole block should share logic with counts_from_bedmethyl AND from export functions - tabix_fields = row.split("\t") - pileup_basemod = tabix_fields[3] - pileup_strand = tabix_fields[5] - keep_basemod = False - if single_strand and pileup_strand.strip() != strand: - # We are on the wrong strand, skip the rest of the steps for this row - continue - elif len(pileup_basemod.split(",")) == 3: - pileup_modname, pileup_motif, pileup_mod_coord = ( - pileup_basemod.split(",") - ) - if ( - pileup_motif == parsed_motif.motif_seq - and int(pileup_mod_coord) == parsed_motif.modified_pos - and pileup_modname in parsed_motif.mod_codes - ): - keep_basemod = True - elif len(pileup_basemod.split(",")) == 1: - if pileup_basemod in parsed_motif.mod_codes: - keep_basemod = True - else: - raise ValueError( - f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." - ) + ( + keep_basemod, + pileup_coord_relative, + modified_in_row, + valid_in_row, + ) = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=start_coord, + region_end=end_coord, + region_strand=strand, + single_strand=single_strand, + regions_5to3prime=regions_5to3prime, + ) if keep_basemod: - pileup_info = tabix_fields[9].split(" ") - genomic_coord = int(tabix_fields[1]) - if regions_5to3prime and strand == "-": - # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction - # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal - pileup_coord_relative = end_coord - genomic_coord - 1 - else: - # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case - pileup_coord_relative = genomic_coord - start_coord if pileup_coord_relative > region_len: print( f"WARNING: You have specified a region {chromosome}:{start_coord}-{end_coord} that is longer than the first region; the end of the region will be skipped. To make a profile plot with differently-sized region, consider using the window_size parameter to make a profile across centered windows." ) - break else: - valid_base_counts[pileup_coord_relative] += int( - pileup_info[0] - ) - modified_base_counts[pileup_coord_relative] += int( - pileup_info[2] + valid_base_counts[pileup_coord_relative] += valid_in_row + modified_base_counts[pileup_coord_relative] += ( + modified_in_row ) + return modified_base_counts, valid_base_counts From 3f3ec8549c1b66c99cfb8f46f92c7f0e36d03f15 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Wed, 8 Jan 2025 09:30:39 -0800 Subject: [PATCH 02/23] DM-231 Removed regions=None case for pileup_counts_from_bedmethyl because it will complicate parallelization and is not clearly useful. --- dimelo/load_processed.py | 64 +++++++++++++++------------------------- 1 file changed, 23 insertions(+), 41 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 3e850eb..95ee18d 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -133,7 +133,7 @@ def process_pileup_row( def pileup_counts_from_bedmethyl( bedmethyl_file: str | Path, motif: str, - regions: str | Path | list[str | Path] | None = None, + regions: str | Path | list[str | Path], window_size: int | None = None, single_strand: bool = False, cores: int | None = None, # currently unused @@ -172,46 +172,28 @@ def pileup_counts_from_bedmethyl( parsed_motif = utils.ParsedMotif(motif) - if regions is not None: - # Get counts from the specified regions - regions_dict = utils.regions_dict_from_input( - regions, - window_size, - ) - for chromosome, region_list in regions_dict.items(): - for start_coord, end_coord, strand in region_list: - # TODO: change to try-except - if chromosome in source_tabix.contigs: - for row in source_tabix.fetch(chromosome, start_coord, end_coord): - keep_basemod, _, modified_in_row, valid_in_row = ( - process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=start_coord, - region_end=end_coord, - region_strand=strand, - single_strand=single_strand, - regions_5to3prime=False, - ) - ) - if keep_basemod: - valid_base_count += valid_in_row - modified_base_count += modified_in_row - else: - # Get counts from the whole input file - for row in source_tabix.fetch(): - keep_basemod, _, modified_in_row, valid_in_row = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=start_coord, - region_end=end_coord, - region_strand=strand, - single_strand=single_strand, - regions_5to3prime=False, - ) - if keep_basemod: - valid_base_count += valid_in_row - modified_base_count += modified_in_row + # Get counts from the specified regions + regions_dict = utils.regions_dict_from_input( + regions, + window_size, + ) + for chromosome, region_list in regions_dict.items(): + for start_coord, end_coord, strand in region_list: + # TODO: change to try-except + if chromosome in source_tabix.contigs: + for row in source_tabix.fetch(chromosome, start_coord, end_coord): + keep_basemod, _, modified_in_row, valid_in_row = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=start_coord, + region_end=end_coord, + region_strand=strand, + single_strand=single_strand, + regions_5to3prime=False, + ) + if keep_basemod: + valid_base_count += valid_in_row + modified_base_count += modified_in_row return (modified_base_count, valid_base_count) From 91fb5e4abc317debb1a03716ec47b7f4c483773a Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Wed, 8 Jan 2025 09:40:23 -0800 Subject: [PATCH 03/23] DM-229 Adjusted export.pileup_to_bigwig to use new load_processed.process_pileup_row and renamed/configured a few variables to support this. --- dimelo/export.py | 88 +++++++++++++++++++----------------------------- 1 file changed, 34 insertions(+), 54 deletions(-) diff --git a/dimelo/export.py b/dimelo/export.py index c55010b..d3dabab 100644 --- a/dimelo/export.py +++ b/dimelo/export.py @@ -6,7 +6,7 @@ import pysam from tqdm.auto import tqdm -from . import utils +from . import load_processed, utils """ This module contains code to export indexed and compressed parse output files to other formats that may be helpful for downstream analysis. @@ -57,8 +57,8 @@ def pileup_to_bigwig( # Because we need to set up the bigwig header for we start writing data to it, we need to pre-index the length of each contig tabix = pysam.TabixFile(str(bedmethyl_file)) - contig_lengths_tuples = [] - lines_by_contig = {} + contig_lengths = {} + contig_lines = {} parsed_motif = utils.ParsedMotif(motif) @@ -84,11 +84,11 @@ def pileup_to_bigwig( )[0] fields = last_row.split("\t") max_coord = int(fields[2]) - contig_lengths_tuples.append((contig, max_coord)) - lines_by_contig[contig] = rows_count + contig_lengths[contig] = max_coord + contig_lines[contig] = rows_count with pyBigWig.open(str(output_file_path), "w") as bw: - bw.addHeader(contig_lengths_tuples) + bw.addHeader([(contig, length) for contig, length in contig_lengths.items()]) for contig in tqdm( tabix.contigs, desc=f"Step 2: Writing {Path(bedmethyl_file).name} contents to {Path(output_file_path).name}", @@ -100,57 +100,37 @@ def pileup_to_bigwig( for row in tqdm( tabix.fetch(contig), desc=f"Writing {contig}.", - total=lines_by_contig[contig], + total=contig_lines[contig], leave=False, ): - # TODO: This code is copied from load_processed.pileup_counts_from_bedmethyl and should probably be consolidated at some point - tabix_fields = row.split("\t") - pileup_basemod = tabix_fields[3] - pileup_strand = tabix_fields[5] - keep_basemod = False - if (strand != ".") and (pileup_strand != strand): - # This entry is on the wrong strand - skip it - continue - elif len(pileup_basemod.split(",")) == 3: - pileup_modname, pileup_motif, pileup_mod_coord = ( - pileup_basemod.split(",") + keep_basemod, genomic_coord, modified_in_row, valid_in_row = ( + load_processed.process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=0, + region_end=contig_lengths[contig], + region_strand=strand, + single_strand=(strand != "."), + regions_5to3prime=False, ) - if ( - pileup_motif == parsed_motif.motif_seq - and int(pileup_mod_coord) == parsed_motif.modified_pos - and pileup_modname in parsed_motif.mod_codes - ): - keep_basemod = True - elif len(pileup_basemod.split(",")) == 1: - if pileup_basemod in parsed_motif.mod_codes: - keep_basemod = True - else: - raise ValueError( - f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." - ) - # TODO: consolidate the above into a function; just do adding outside - if keep_basemod: - pileup_info = tabix_fields[9].split(" ") - valid_base_counts = int(pileup_info[0]) - modified_base_counts = int(pileup_info[2]) - if valid_base_counts > 0: - genomic_coord = int(tabix_fields[1]) - contig_list.append(contig) - start_list.append(genomic_coord) - end_list.append(genomic_coord + 1) - values_list.append(modified_base_counts / valid_base_counts) - - if len(values_list) > chunk_size: - bw.addEntries( - contig_list, # Contig names - start_list, # Start positions - ends=end_list, # End positions - values=values_list, # Corresponding values - ) - contig_list = [] - start_list = [] - end_list = [] - values_list = [] + ) + if keep_basemod and valid_in_row > 0: + contig_list.append(contig) + start_list.append(genomic_coord) + end_list.append(genomic_coord + 1) + values_list.append(modified_in_row / valid_in_row) + + if len(values_list) > chunk_size: + bw.addEntries( + contig_list, # Contig names + start_list, # Start positions + ends=end_list, # End positions + values=values_list, # Corresponding values + ) + contig_list = [] + start_list = [] + end_list = [] + values_list = [] bw.addEntries( contig_list, # Contig names start_list, # Start positions From 53d039bcebc71e0bbcfb7dee46f3d23b2cdf43d9 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 9 Jan 2025 10:01:12 -0800 Subject: [PATCH 04/23] DM-192 Implemented parallelization core including shared memory and chunk generation, currently only for load_processed.pileup_vectors_from_bedmethyl. pileup_counts_from_bedmethyl is a simpler case. Passes pre-existing tests including when chunk_size 1: - with ProcessPoolExecutor(max_workers=cores_to_run) as executor: + with concurrent.futures.ProcessPoolExecutor( + max_workers=cores_to_run + ) as executor: # Use functools.partial to pre-fill arguments process_partial = partial( process_region, function_handle=function_handle, cores=1, **kwargs @@ -82,19 +86,25 @@ def process_pileup_row( region_start: int, region_end: int, region_strand: str, - single_strand: bool, - regions_5to3prime: bool, -) -> tuple[bool, int, int, int]: + subregion_start: int | None = None, + subregion_end: int | None = None, + single_strand: bool = False, + regions_5to3prime: bool = False, +) -> tuple[bool, int, int, int, int]: """ Returns: keep_basemod, pileup_coord_relative, modified_in_row, valid_in_row """ + if subregion_start is None: + subregion_start = region_start + if subregion_end is None: + subregion_end = region_end tabix_fields = row.split("\t") pileup_basemod = tabix_fields[3] pileup_strand = tabix_fields[5] keep_basemod = False if single_strand and pileup_strand.strip() != region_strand: # We are on the wrong strand, skip the rest of the steps for this row - return keep_basemod, None, None, None + return keep_basemod, 0, 0, 0, 0 elif len(pileup_basemod.split(",")) == 3: pileup_modname, pileup_motif, pileup_mod_coord = pileup_basemod.split(",") if ( @@ -110,24 +120,28 @@ def process_pileup_row( raise ValueError( f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." ) - if keep_basemod: - pileup_info = tabix_fields[9].split(" ") - genomic_coord = int(tabix_fields[1]) - if regions_5to3prime and region_strand == "-": - # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction - # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal - pileup_coord_relative = region_end - genomic_coord - 1 - else: - # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case - pileup_coord_relative = genomic_coord - region_start - - valid_in_row = int(pileup_info[0]) - modified_in_row = int(pileup_info[2]) - - return keep_basemod, pileup_coord_relative, modified_in_row, valid_in_row + pileup_info = tabix_fields[9].split(" ") + genomic_coord = int(tabix_fields[1]) + if regions_5to3prime and region_strand == "-": + # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction + # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal + pileup_coord_relative = subregion_end - genomic_coord - 1 + subregion_offset = region_end - subregion_end else: - return keep_basemod, None, None, None + # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case + pileup_coord_relative = genomic_coord - subregion_start + subregion_offset = subregion_start - region_start + valid_in_row = int(pileup_info[0]) + modified_in_row = int(pileup_info[2]) + + return ( + keep_basemod, + pileup_coord_relative, + modified_in_row, + valid_in_row, + subregion_offset, + ) def pileup_counts_from_bedmethyl( @@ -177,19 +191,25 @@ def pileup_counts_from_bedmethyl( regions, window_size, ) + + cores_to_run = utils.cores_to_run(cores) + _ = cores_to_run + for chromosome, region_list in regions_dict.items(): for start_coord, end_coord, strand in region_list: # TODO: change to try-except if chromosome in source_tabix.contigs: for row in source_tabix.fetch(chromosome, start_coord, end_coord): - keep_basemod, _, modified_in_row, valid_in_row = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=start_coord, - region_end=end_coord, - region_strand=strand, - single_strand=single_strand, - regions_5to3prime=False, + keep_basemod, _, modified_in_row, valid_in_row, _ = ( + process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=start_coord, + region_end=end_coord, + region_strand=strand, + single_strand=single_strand, + regions_5to3prime=False, + ) ) if keep_basemod: valid_base_count += valid_in_row @@ -209,6 +229,76 @@ def counts_from_fake(*args, **kwargs) -> tuple[int, int]: return test_data.fake_peak_enrichment(halfsize=window_halfsize, peak_height=0.15) +def pileup_vectors_process_chunk( + bedmethyl_file, + parsed_motif, + chunk, + region_len, + shm_name_modified, + shm_name_valid, + lock, + single_strand, + regions_5to3prime, +) -> None: + source_tabix = pysam.TabixFile(str(bedmethyl_file)) + existing_valid = shared_memory.SharedMemory(name=shm_name_valid) + existing_modified = shared_memory.SharedMemory(name=shm_name_modified) + valid_base_counts = np.ndarray( + (region_len,), dtype=np.int32, buffer=existing_valid.buf + ) + modified_base_counts = np.ndarray( + (region_len,), dtype=np.int32, buffer=existing_modified.buf + ) + + chromosome = chunk["chromosome"] + region_start = chunk["region_start"] + region_end = chunk["region_end"] + subregion_start = chunk["subregion_start"] + subregion_end = chunk["subregion_end"] + strand = chunk["strand"] + + subregion_offset = 0 + + valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) + modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) + + for row in source_tabix.fetch(chunk["chromosome"], subregion_start, subregion_end): + ( + keep_basemod, + pileup_coord_relative, + modified_in_row, + valid_in_row, + subregion_offset, + ) = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=region_start, + region_end=region_end, + subregion_start=subregion_start, + subregion_end=subregion_end, + region_strand=strand, + single_strand=single_strand, + regions_5to3prime=regions_5to3prime, + ) + if keep_basemod: + if pileup_coord_relative > region_len: + print( + f"WARNING: You have specified a region starting at {chromosome}:{subregion_start - subregion_offset} that is longer than the first region; the end of the region will be skipped. To make a profile plot with differently-sized region, consider using the window_size parameter to make a profile across centered windows." + ) + else: + valid_base_subregion[pileup_coord_relative] += valid_in_row + modified_base_subregion[pileup_coord_relative] += modified_in_row + + with lock: + print(subregion_offset) + valid_base_counts[ + subregion_offset : subregion_offset + abs(subregion_end - subregion_start) + ] += valid_base_subregion + modified_base_counts[ + subregion_offset : subregion_offset + abs(subregion_end - subregion_start) + ] += modified_base_subregion + + def pileup_vectors_from_bedmethyl( bedmethyl_file: str | Path, motif: str, @@ -254,52 +344,65 @@ def pileup_vectors_from_bedmethyl( tuple containing (modified_base_counts, valid_base_counts) """ - source_tabix = pysam.TabixFile(str(bedmethyl_file)) + print(f"function call: {bedmethyl_file}, {regions}, {motif}") parsed_motif = utils.ParsedMotif(motif) regions_dict = utils.regions_dict_from_input(regions, window_size) + chunks_list = utils.process_chunks_from_regions_dict( + regions_dict, chunk_size=10_000 + ) + + print(regions_dict) + print(chunks_list) + + cores_to_run = utils.cores_to_run(cores) # Peek at a region to figure out what size the vectors should be first_key = next(iter(regions_dict)) first_tuple = regions_dict[first_key][0] region_len = first_tuple[1] - first_tuple[0] - valid_base_counts = np.zeros(region_len, dtype=int) - modified_base_counts = np.zeros(region_len, dtype=int) + shm_valid = shared_memory.SharedMemory( + create=True, size=(region_len) * np.dtype(np.int32).itemsize + ) + shm_modified = shared_memory.SharedMemory( + create=True, size=(region_len) * np.dtype(np.int32).itemsize + ) - for chromosome, region_list in regions_dict.items(): - for start_coord, end_coord, strand in region_list: - # TODO: This is not used anywhere; disabling for now - # center_coord = (start_coord+end_coord)//2 - if chromosome in source_tabix.contigs: - for row in source_tabix.fetch(chromosome, start_coord, end_coord): - ( - keep_basemod, - pileup_coord_relative, - modified_in_row, - valid_in_row, - ) = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=start_coord, - region_end=end_coord, - region_strand=strand, - single_strand=single_strand, - regions_5to3prime=regions_5to3prime, - ) - if keep_basemod: - if pileup_coord_relative > region_len: - print( - f"WARNING: You have specified a region {chromosome}:{start_coord}-{end_coord} that is longer than the first region; the end of the region will be skipped. To make a profile plot with differently-sized region, consider using the window_size parameter to make a profile across centered windows." - ) - else: - valid_base_counts[pileup_coord_relative] += valid_in_row - modified_base_counts[pileup_coord_relative] += ( - modified_in_row - ) - - return modified_base_counts, valid_base_counts + manager = multiprocessing.Manager() + lock = manager.Lock() + + with concurrent.futures.ProcessPoolExecutor(max_workers=cores_to_run) as executor: + futures = [ + executor.submit( + pileup_vectors_process_chunk, + bedmethyl_file, + parsed_motif, + chunk, + region_len, + shm_modified.name, + shm_valid.name, + lock, + single_strand, + regions_5to3prime, + ) + for chunk in chunks_list + ] + for future in tqdm( + concurrent.futures.as_completed(futures), total=len(futures) + ): + try: + future.result() + except Exception as err: + raise RuntimeError("Subprocess failed.") from err + + modified_base_counts = np.ndarray( + (region_len,), dtype=np.int32, buffer=shm_modified.buf + ) + valid_base_counts = np.ndarray((region_len,), dtype=np.int32, buffer=shm_valid.buf) + + return np.copy(modified_base_counts), np.copy(valid_base_counts) def vector_from_fake(window_size: int, *args, **kwargs) -> np.ndarray: diff --git a/dimelo/test/dimelo_test.py b/dimelo/test/dimelo_test.py index 787aa83..87ff71e 100644 --- a/dimelo/test/dimelo_test.py +++ b/dimelo/test/dimelo_test.py @@ -317,7 +317,7 @@ class TestLoadProcessed: outputs section of dimelo/test/generate_test_targets.ipynb. """ - def test_unit__regions_list_list( + def test_unit__regions_to_list( self, test_case, kwargs, diff --git a/dimelo/utils.py b/dimelo/utils.py index aa85e55..83e9ce1 100644 --- a/dimelo/utils.py +++ b/dimelo/utils.py @@ -98,6 +98,28 @@ def adjust_threshold( return thresh +def process_chunks_from_regions_dict( + regions_dict: dict, + chunk_size: int = 10_000, +): + chunk_list = [] + for chromosome, region_list in regions_dict.items(): + for start_coord, end_coord, strand in region_list: + for subregion_start in range(start_coord, end_coord, chunk_size): + subregion_end = min(end_coord, subregion_start + chunk_size) + chunk_list.append( + { + "chromosome": chromosome, + "region_start": start_coord, + "region_end": end_coord, + "subregion_start": subregion_start, + "subregion_end": subregion_end, + "strand": strand, + } + ) + return chunk_list + + def regions_dict_from_input( regions: str | Path | list[str | Path] | None = None, window_size: int | None = None, From caf9dcdb988cce84333eef49acef03b5393e24b9 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 9 Jan 2025 10:41:09 -0800 Subject: [PATCH 05/23] DM-232 Added strand information to regions .bed files. Re-generated read_vectors_from_hdf5 test targets to pass tests; other test targets are the same for now. --- dimelo/test/data/ctcf_demo_not_peak.bed | 400 +++++++++--------- dimelo/test/data/ctcf_demo_peak.bed | 200 ++++----- .../test/data/test_targets/test_matrix.pickle | Bin 3144815 -> 3144853 bytes 3 files changed, 300 insertions(+), 300 deletions(-) diff --git a/dimelo/test/data/ctcf_demo_not_peak.bed b/dimelo/test/data/ctcf_demo_not_peak.bed index 920b114..edfe622 100644 --- a/dimelo/test/data/ctcf_demo_not_peak.bed +++ b/dimelo/test/data/ctcf_demo_not_peak.bed @@ -1,200 +1,200 @@ -chr14 44123158 44123308 + 465.63275 . -chr14 44127026 44127176 + 465.63275 . -chr14 44123159 44123308 - 465.63275 . -chr14 44127026 44127175 - 465.63275 . -chr1 114356587 114356736 - 441.00102 . -chr1 114360454 114360603 - 441.00102 . -chr1 114356586 114356736 + 441.00102 . -chr1 114360454 114360604 + 441.00102 . -chrX 9700334 9700484 - 418.31102 . -chrX 9704202 9704352 - 418.31102 . -chr15 54632158 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Adjusted generate_targets.py to properly handle copying over new kwargs from cases.py when re-running some but not all targets: if all targets are re-run, then the old test_matrix.pkl is discarded. If only some are re-run, we need the old test_matrix.pkl, but still need to copy over all the kwargs for all cases that won't be re-run and still need to copy over old results, which will be overridden for re-run targets. --- dimelo/test/cases.py | 10 ++++++++++ .../test/data/test_targets/test_matrix.pickle | Bin 3144853 -> 3144762 bytes dimelo/test/generate_targets.py | 18 ++++++++++++++++-- 3 files changed, 26 insertions(+), 2 deletions(-) diff --git a/dimelo/test/cases.py b/dimelo/test/cases.py index 2a717ec..85366d9 100644 --- a/dimelo/test/cases.py +++ b/dimelo/test/cases.py @@ -31,6 +31,8 @@ "sort_by": ["read_start", "read_name", "motif"], "smooth_window": 1, "title": "megalodon_peaks_190", + "single_strand": False, + "regions_5to3prime": False, }, # outputs dict function:values {}, # populated in subsequent cells @@ -48,6 +50,8 @@ "sort_by": ["read_start", "read_name", "motif"], "smooth_window": 10, "title": "megalodon_single_190", + "single_strand": False, + "regions_5to3prime": False, }, # outputs dict function:values {}, # populated in subsequent cells @@ -65,6 +69,8 @@ "sort_by": ["read_start", "read_name", "motif"], "smooth_window": 100, "title": "megalodon_single_and_peaks_190", + "single_strand": True, + "regions_5to3prime": True, }, # outputs dict function:values {}, # populated in subsequent cells @@ -82,6 +88,8 @@ "sort_by": ["read_start", "read_name", "motif"], "smooth_window": 100, "title": "megalodon_peaks_nothresh", + "single_strand": True, + "regions_5to3prime": False, }, # outputs dict function:values {}, # populated in subsequent cells @@ -99,6 +107,8 @@ "sort_by": ["read_start", "read_name", "motif"], "smooth_window": 1, "title": "megalodon_single_nothresh", + "single_strand": False, + "regions_5to3prime": True, }, # outputs dict function:values {}, # populated in subsequent cells diff --git a/dimelo/test/data/test_targets/test_matrix.pickle b/dimelo/test/data/test_targets/test_matrix.pickle index 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open(RelativePath(args.initial_target_pickle), "rb") as file: old_test_matrix = pickle.load(file) - for key, value in old_test_matrix.items(): + # loop through the old test matrix + for key, (old_kwargs, old_results) in old_test_matrix.items(): + # if the old test case is in the new test matrix, bring over the results + # either the test targets will be regenerated, and the results replaced, or they won't be regenerated, and we'll need the old results if key in test_matrix: - test_matrix[key] = value + new_kwargs = test_matrix[key][0] + # if case will be covered, bring in new kwargs + if args.case_subset is None or key in args.case_subset: + test_matrix[key] = (new_kwargs, old_results) + # if case will not be covered, keep old kwargs + else: + test_matrix[key] = (old_kwargs, old_results) print( f"Running {'all cases' if args.case_subset is None else args.case_subset} through {args.target_subset} to supplement test targets from {args.initial_target_pickle}. Any new tests in cases.py will be added to the test matrix." ) @@ -171,6 +180,11 @@ def main(test_matrix): f"Cannot run subset {args.target_subset} without a pre-existing complete set of generated targets, {args.initial_target_pickle} does not exist. Either specify an --initial-target-pickle that does exist or run without subsetting." ) + print("Generating targets for the following test matrix kwargs") + for test_name, (kwargs, _) in test_matrix.items(): + print(test_name) + print(kwargs) + for subset in args.target_subset: function_name = f"generate_{subset}" globals()[function_name](test_matrix, args.case_subset) From caa3e59a9863e9214f5d15333027dbd0b7a2e37f Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 9 Jan 2025 12:36:21 -0800 Subject: [PATCH 07/23] DM-192 Fixed logic to properly handle single_strand case within process_pileup_row. Passing all tests. --- dimelo/export.py | 2 +- dimelo/load_processed.py | 21 +++++++++------------ 2 files changed, 10 insertions(+), 13 deletions(-) diff --git a/dimelo/export.py b/dimelo/export.py index d3dabab..4256954 100644 --- a/dimelo/export.py +++ b/dimelo/export.py @@ -103,7 +103,7 @@ def pileup_to_bigwig( total=contig_lines[contig], leave=False, ): - keep_basemod, genomic_coord, modified_in_row, valid_in_row = ( + keep_basemod, genomic_coord, modified_in_row, valid_in_row, _ = ( load_processed.process_pileup_row( row=row, parsed_motif=parsed_motif, diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 537345e..e951d10 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -101,10 +101,10 @@ def process_pileup_row( tabix_fields = row.split("\t") pileup_basemod = tabix_fields[3] pileup_strand = tabix_fields[5] - keep_basemod = False + if single_strand and pileup_strand.strip() != region_strand: - # We are on the wrong strand, skip the rest of the steps for this row - return keep_basemod, 0, 0, 0, 0 + # We are on the wrong strand, can't keep this position + keep_basemod = False elif len(pileup_basemod.split(",")) == 3: pileup_modname, pileup_motif, pileup_mod_coord = pileup_basemod.split(",") if ( @@ -113,9 +113,10 @@ def process_pileup_row( and pileup_modname in parsed_motif.mod_codes ): keep_basemod = True + else: + keep_basemod = False elif len(pileup_basemod.split(",")) == 1: - if pileup_basemod in parsed_motif.mod_codes: - keep_basemod = True + keep_basemod = pileup_basemod in parsed_motif.mod_codes else: raise ValueError( f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." @@ -123,6 +124,8 @@ def process_pileup_row( pileup_info = tabix_fields[9].split(" ") genomic_coord = int(tabix_fields[1]) + # TODO: consider moving this logic back outside of process_pileup_row, to avoid unecessary operations for both pileup_counts + # and for long regions that only need the offset value once if regions_5to3prime and region_strand == "-": # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal @@ -290,7 +293,6 @@ def pileup_vectors_process_chunk( modified_base_subregion[pileup_coord_relative] += modified_in_row with lock: - print(subregion_offset) valid_base_counts[ subregion_offset : subregion_offset + abs(subregion_end - subregion_start) ] += valid_base_subregion @@ -349,12 +351,7 @@ def pileup_vectors_from_bedmethyl( parsed_motif = utils.ParsedMotif(motif) regions_dict = utils.regions_dict_from_input(regions, window_size) - chunks_list = utils.process_chunks_from_regions_dict( - regions_dict, chunk_size=10_000 - ) - - print(regions_dict) - print(chunks_list) + chunks_list = utils.process_chunks_from_regions_dict(regions_dict, chunk_size=1_000) cores_to_run = utils.cores_to_run(cores) From ac3657e7aaf4abbe049931b9dbf67255dd8d9bb3 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 9 Jan 2025 13:04:23 -0800 Subject: [PATCH 08/23] DM-192 Further refactor to simplify process_pileup_row and keep regions_5to3prime logic within the pileup_vectors_from_bedmethyl function, which is the only one that actually needs it. Runs faster, still passes all tests. --- dimelo/export.py | 3 +- dimelo/load_processed.py | 86 ++++++++++++++++++---------------------- 2 files changed, 39 insertions(+), 50 deletions(-) diff --git a/dimelo/export.py b/dimelo/export.py index 4256954..c3fd6c1 100644 --- a/dimelo/export.py +++ b/dimelo/export.py @@ -103,7 +103,7 @@ def pileup_to_bigwig( total=contig_lines[contig], leave=False, ): - keep_basemod, genomic_coord, modified_in_row, valid_in_row, _ = ( + keep_basemod, genomic_coord, modified_in_row, valid_in_row = ( load_processed.process_pileup_row( row=row, parsed_motif=parsed_motif, @@ -111,7 +111,6 @@ def pileup_to_bigwig( region_end=contig_lengths[contig], region_strand=strand, single_strand=(strand != "."), - regions_5to3prime=False, ) ) if keep_basemod and valid_in_row > 0: diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index e951d10..e9441c7 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -86,18 +86,11 @@ def process_pileup_row( region_start: int, region_end: int, region_strand: str, - subregion_start: int | None = None, - subregion_end: int | None = None, single_strand: bool = False, - regions_5to3prime: bool = False, -) -> tuple[bool, int, int, int, int]: +) -> tuple[bool, int, int, int]: """ - Returns: keep_basemod, pileup_coord_relative, modified_in_row, valid_in_row + Returns: keep_basemod, genomic_coord, modified_in_row, valid_in_row """ - if subregion_start is None: - subregion_start = region_start - if subregion_end is None: - subregion_end = region_end tabix_fields = row.split("\t") pileup_basemod = tabix_fields[3] pileup_strand = tabix_fields[5] @@ -124,26 +117,14 @@ def process_pileup_row( pileup_info = tabix_fields[9].split(" ") genomic_coord = int(tabix_fields[1]) - # TODO: consider moving this logic back outside of process_pileup_row, to avoid unecessary operations for both pileup_counts - # and for long regions that only need the offset value once - if regions_5to3prime and region_strand == "-": - # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction - # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal - pileup_coord_relative = subregion_end - genomic_coord - 1 - subregion_offset = region_end - subregion_end - else: - # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case - pileup_coord_relative = genomic_coord - subregion_start - subregion_offset = subregion_start - region_start valid_in_row = int(pileup_info[0]) modified_in_row = int(pileup_info[2]) return ( keep_basemod, - pileup_coord_relative, + genomic_coord, modified_in_row, valid_in_row, - subregion_offset, ) @@ -203,16 +184,13 @@ def pileup_counts_from_bedmethyl( # TODO: change to try-except if chromosome in source_tabix.contigs: for row in source_tabix.fetch(chromosome, start_coord, end_coord): - keep_basemod, _, modified_in_row, valid_in_row, _ = ( - process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=start_coord, - region_end=end_coord, - region_strand=strand, - single_strand=single_strand, - regions_5to3prime=False, - ) + keep_basemod, _, modified_in_row, valid_in_row = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=start_coord, + region_end=end_coord, + region_strand=strand, + single_strand=single_strand, ) if keep_basemod: valid_base_count += valid_in_row @@ -260,7 +238,17 @@ def pileup_vectors_process_chunk( subregion_end = chunk["subregion_end"] strand = chunk["strand"] - subregion_offset = 0 + flip_coords = regions_5to3prime and strand == "-" + + if flip_coords: + subregion_offset = region_end - subregion_end + else: + subregion_offset = subregion_start - region_start + + if region_end - region_start > region_len: + print( + f"WARNING: You have specified a region at {chromosome}:{region_start}-{region_end} that is longer than the first region; the end of the region will be skipped. To make a profile plot with differently-sized region, consider using the window_size parameter to make a profile across centered windows." + ) valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) @@ -268,29 +256,28 @@ def pileup_vectors_process_chunk( for row in source_tabix.fetch(chunk["chromosome"], subregion_start, subregion_end): ( keep_basemod, - pileup_coord_relative, + genomic_coord, modified_in_row, valid_in_row, - subregion_offset, ) = process_pileup_row( row=row, parsed_motif=parsed_motif, - region_start=region_start, - region_end=region_end, - subregion_start=subregion_start, - subregion_end=subregion_end, + region_start=subregion_start, + region_end=subregion_end, region_strand=strand, single_strand=single_strand, - regions_5to3prime=regions_5to3prime, ) if keep_basemod: - if pileup_coord_relative > region_len: - print( - f"WARNING: You have specified a region starting at {chromosome}:{subregion_start - subregion_offset} that is longer than the first region; the end of the region will be skipped. To make a profile plot with differently-sized region, consider using the window_size parameter to make a profile across centered windows." - ) + if flip_coords: + # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction + # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal + pileup_coord_in_subregion = subregion_end - genomic_coord - 1 else: - valid_base_subregion[pileup_coord_relative] += valid_in_row - modified_base_subregion[pileup_coord_relative] += modified_in_row + # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case + pileup_coord_in_subregion = genomic_coord - subregion_start + if pileup_coord_in_subregion < (subregion_end - subregion_start): + valid_base_subregion[pileup_coord_in_subregion] += valid_in_row + modified_base_subregion[pileup_coord_in_subregion] += modified_in_row with lock: valid_base_counts[ @@ -308,7 +295,8 @@ def pileup_vectors_from_bedmethyl( window_size: int | None = None, single_strand: bool = False, regions_5to3prime: bool = False, - cores: int | None = None, # currently unused + cores: int | None = None, + chunk_size: int = 100, ) -> tuple[np.ndarray, np.ndarray]: """ Extract per-position pileup counts at valid motifs across one or more superimposed regions. @@ -351,7 +339,9 @@ def pileup_vectors_from_bedmethyl( parsed_motif = utils.ParsedMotif(motif) regions_dict = utils.regions_dict_from_input(regions, window_size) - chunks_list = utils.process_chunks_from_regions_dict(regions_dict, chunk_size=1_000) + chunks_list = utils.process_chunks_from_regions_dict( + regions_dict, chunk_size=chunk_size + ) cores_to_run = utils.cores_to_run(cores) From a9648dfa8b1e88154db8c2a87b56c62f4e853460 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 9 Jan 2025 13:08:56 -0800 Subject: [PATCH 09/23] DM-192 Added chunk_size for load_processes parallelization to cases.py, re-ran targets for pileup_counts_from_bedmethyl and pileup_vectors_from_bedmethyl. --- dimelo/test/cases.py | 5 +++++ .../test/data/test_targets/test_matrix.pickle | Bin 3144762 -> 3144835 bytes 2 files changed, 5 insertions(+) diff --git a/dimelo/test/cases.py b/dimelo/test/cases.py index 85366d9..001238c 100644 --- a/dimelo/test/cases.py +++ b/dimelo/test/cases.py @@ -33,6 +33,7 @@ "title": "megalodon_peaks_190", "single_strand": False, "regions_5to3prime": False, + "chunk_size": 1_000_000, }, # outputs dict function:values {}, # populated in subsequent cells @@ -52,6 +53,7 @@ "title": "megalodon_single_190", "single_strand": False, "regions_5to3prime": False, + "chunk_size": 100, }, # outputs dict function:values {}, # populated in subsequent cells @@ -71,6 +73,7 @@ "title": "megalodon_single_and_peaks_190", "single_strand": True, "regions_5to3prime": True, + "chunk_size": 10_000, }, # outputs dict function:values {}, # populated in subsequent cells @@ -90,6 +93,7 @@ "title": "megalodon_peaks_nothresh", "single_strand": True, "regions_5to3prime": False, + "chunk_size": 1000, }, # outputs dict function:values {}, # populated in subsequent cells @@ -109,6 +113,7 @@ "title": "megalodon_single_nothresh", "single_strand": False, "regions_5to3prime": True, + "chunk_size": 100, }, # outputs dict function:values {}, # populated in subsequent cells diff --git a/dimelo/test/data/test_targets/test_matrix.pickle b/dimelo/test/data/test_targets/test_matrix.pickle index f2cd6f3550a117c60cd672ee1a92dfb19c61ffec..b40735983ebb5f6b9f70d35e6d15f6ae2a655c08 100644 GIT binary patch delta 19922 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Added progress bars and quiet toggles for pileup loaders. Set default chunk size to 1 MB after whole-chromosome speed testing. Added option to parallelize within rather than between regions with regions_to_list. --- dimelo/load_processed.py | 200 +++++++++++++++++++++++++---------- dimelo/plot_depth_profile.py | 1 + 2 files changed, 144 insertions(+), 57 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index e9441c7..4da5f64 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -25,7 +25,9 @@ def regions_to_list( function_handle, regions, window_size: int | None = None, + quiet: bool = True, cores: int | None = None, + parallelize_within_regions: bool = False, **kwargs, ): """ @@ -53,29 +55,26 @@ def regions_to_list( cores_to_run = utils.cores_to_run(cores) - if cores_to_run > 1: - with concurrent.futures.ProcessPoolExecutor( - max_workers=cores_to_run - ) as executor: - # Use functools.partial to pre-fill arguments - process_partial = partial( - process_region, function_handle=function_handle, cores=1, **kwargs - ) + with concurrent.futures.ProcessPoolExecutor( + max_workers=1 if parallelize_within_regions else cores_to_run + ) as executor: + # Use functools.partial to pre-fill arguments + process_partial = partial( + process_region, + function_handle=function_handle, + quiet=quiet or not parallelize_within_regions, + cores=cores_to_run if parallelize_within_regions else 1, + **kwargs, + ) - # Use executor.map without lambda - results = list( - tqdm( - executor.map(process_partial, region_tuples), - total=len(region_tuples), - desc=f"Processing regions in parallel across {cores_to_run}", - ) + results = list( + tqdm( + executor.map(process_partial, region_tuples), + total=len(region_tuples), + desc=f"Processing regions in parallel across {cores_to_run}", + disable=quiet or parallelize_within_regions, ) - else: - # Single-threaded fallback - results = [ - process_region(region, function_handle, cores=1, **kwargs) - for region in tqdm(region_tuples, desc="Processing regions") - ] + ) return results @@ -128,13 +127,63 @@ def process_pileup_row( ) +def pileup_counts_process_chunk( + bedmethyl_file, + parsed_motif, + chunk, + shm_name_modified, + shm_name_valid, + lock, + single_strand, +) -> None: + source_tabix = pysam.TabixFile(str(bedmethyl_file)) + existing_valid = shared_memory.SharedMemory(name=shm_name_valid) + existing_modified = shared_memory.SharedMemory(name=shm_name_modified) + valid_base_counts = np.ndarray((1,), dtype=np.int32, buffer=existing_valid.buf) + modified_base_counts = np.ndarray( + (1,), dtype=np.int32, buffer=existing_modified.buf + ) + + chromosome = chunk["chromosome"] + subregion_start = chunk["subregion_start"] + subregion_end = chunk["subregion_end"] + strand = chunk["strand"] + + valid_base_subregion_counts = 0 + modified_base_subregion_counts = 0 + + for row in source_tabix.fetch(chromosome, subregion_start, subregion_end): + ( + keep_basemod, + _, + modified_in_row, + valid_in_row, + ) = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_start=subregion_start, + region_end=subregion_end, + region_strand=strand, + single_strand=single_strand, + ) + if keep_basemod: + valid_base_subregion_counts += valid_in_row + modified_base_subregion_counts += modified_in_row + + with lock: + valid_base_counts[0] += valid_base_subregion_counts + modified_base_counts[0] += modified_base_subregion_counts + + def pileup_counts_from_bedmethyl( bedmethyl_file: str | Path, motif: str, regions: str | Path | list[str | Path], window_size: int | None = None, single_strand: bool = False, - cores: int | None = None, # currently unused + quiet: bool = True, + cores: int | None = None, + chunk_size: int = 1_000_000, ) -> tuple[int, int]: """ Extract number of modified bases and total number of bases from the given bedmethyl file. @@ -157,46 +206,74 @@ def pileup_counts_from_bedmethyl( window_size: (currently disabled) window around center of region, +-window_size single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions - cores: cores across which to parallelize processes (currently unused) + quiet: disables progress bars + cores: cores across which to parallelize processes + chunk_size: size of genomic subregions to assign out to each process Returns: tuple containing counts of (modified_bases, total_bases) """ - source_tabix = pysam.TabixFile(str(bedmethyl_file)) - # Don't need vectors, just need counts; also not guaranteed that windows are the same length - valid_base_count = 0 - modified_base_count = 0 - parsed_motif = utils.ParsedMotif(motif) - # Get counts from the specified regions - regions_dict = utils.regions_dict_from_input( - regions, - window_size, + regions_dict = utils.regions_dict_from_input(regions, window_size) + chunks_list = utils.process_chunks_from_regions_dict( + regions_dict, chunk_size=chunk_size ) cores_to_run = utils.cores_to_run(cores) - _ = cores_to_run - - for chromosome, region_list in regions_dict.items(): - for start_coord, end_coord, strand in region_list: - # TODO: change to try-except - if chromosome in source_tabix.contigs: - for row in source_tabix.fetch(chromosome, start_coord, end_coord): - keep_basemod, _, modified_in_row, valid_in_row = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=start_coord, - region_end=end_coord, - region_strand=strand, - single_strand=single_strand, - ) - if keep_basemod: - valid_base_count += valid_in_row - modified_base_count += modified_in_row - return (modified_base_count, valid_base_count) + shm_valid = shared_memory.SharedMemory( + create=True, size=np.dtype(np.int32).itemsize + ) + shm_modified = shared_memory.SharedMemory( + create=True, size=np.dtype(np.int32).itemsize + ) + + manager = multiprocessing.Manager() + lock = manager.Lock() + + with concurrent.futures.ProcessPoolExecutor(max_workers=cores_to_run) as executor: + futures = [ + executor.submit( + pileup_counts_process_chunk, + bedmethyl_file, + parsed_motif, + chunk, + shm_modified.name, + shm_valid.name, + lock, + single_strand, + ) + for chunk in chunks_list + ] + for future in tqdm( + concurrent.futures.as_completed(futures), + total=len(futures), + disable=quiet, + desc=f"Loading genomic chunks, approx. {chunk_size/1000}kb per chunk", + ): + try: + future.result() + except Exception as err: + raise RuntimeError("Subprocess failed.") from err + + # Directly convert shared memory buffers to integers + modified_base_count = int.from_bytes( + shm_modified.buf[:4], byteorder="little", signed=True + ) + valid_base_count = int.from_bytes( + shm_valid.buf[:4], byteorder="little", signed=True + ) + + return modified_base_count, valid_base_count + + # We need to convert these shared memory buffers to ints that are no longer tied to the buffer, + # so they can be passed up without any seg fault type errors + modified_base_counts = np.ndarray((1,), dtype=np.int32, buffer=shm_modified.buf) + valid_base_counts = np.ndarray((1,), dtype=np.int32, buffer=shm_valid.buf) + + return int(np.copy(modified_base_counts[0])), int(np.copy(valid_base_counts[0])) def counts_from_fake(*args, **kwargs) -> tuple[int, int]: @@ -253,7 +330,7 @@ def pileup_vectors_process_chunk( valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) - for row in source_tabix.fetch(chunk["chromosome"], subregion_start, subregion_end): + for row in source_tabix.fetch(chromosome, subregion_start, subregion_end): ( keep_basemod, genomic_coord, @@ -295,8 +372,9 @@ def pileup_vectors_from_bedmethyl( window_size: int | None = None, single_strand: bool = False, regions_5to3prime: bool = False, + quiet: bool = True, cores: int | None = None, - chunk_size: int = 100, + chunk_size: int = 1_000_000, ) -> tuple[np.ndarray, np.ndarray]: """ Extract per-position pileup counts at valid motifs across one or more superimposed regions. @@ -328,14 +406,14 @@ def pileup_vectors_from_bedmethyl( single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping - cores: cores across which to parallelize processes (currently unused) + quiet: disables progress bars + cores: cores across which to parallelize processes + chunk_size: size of genomic subregions to assign out to each process Returns: tuple containing (modified_base_counts, valid_base_counts) """ - print(f"function call: {bedmethyl_file}, {regions}, {motif}") - parsed_motif = utils.ParsedMotif(motif) regions_dict = utils.regions_dict_from_input(regions, window_size) @@ -377,13 +455,19 @@ def pileup_vectors_from_bedmethyl( for chunk in chunks_list ] for future in tqdm( - concurrent.futures.as_completed(futures), total=len(futures) + concurrent.futures.as_completed(futures), + total=len(futures), + disable=quiet, + desc=f"Loading genomic chunks, approx. {chunk_size/1000}kb per chunk", ): try: future.result() except Exception as err: raise RuntimeError("Subprocess failed.") from err + # We need to convert these shared memory buffers to numpy arrays which + # we then copy, so that they no longer reference the shared memory which + # will soon be de-allocated modified_base_counts = np.ndarray( (region_len,), dtype=np.int32, buffer=shm_modified.buf ) @@ -416,6 +500,7 @@ def read_vectors_from_hdf5( sort_by: str | list[str] = ["chromosome", "region_start", "read_start"], calculate_mod_fractions: bool = True, cores: int | None = None, # currently unused + quiet: bool = True, # currently unused ) -> tuple[list[tuple], list[str], dict | None]: """ Pulls a list of read data out of an .h5 file containing processed read vectors, formatted @@ -458,6 +543,7 @@ def read_vectors_from_hdf5( include chromosome, region_start, region_end, read_start, read_end, and motif. More to be added in future. cores: cores across which to parallelize processes (currently unused) + quiet: silences progress bars (currently unused) Returns: a list of tuples, each tuple containing all datasets corresponding to an individual read that diff --git a/dimelo/plot_depth_profile.py b/dimelo/plot_depth_profile.py index d2d9ef3..e4b3327 100644 --- a/dimelo/plot_depth_profile.py +++ b/dimelo/plot_depth_profile.py @@ -198,6 +198,7 @@ def get_depth_profiles( window_size=window_size, single_strand=single_strand, regions_5to3prime=regions_5to3prime, + quiet=False, ) trace = valid_base_counts.astype(float) trace[trace == 0] = np.nan From 56e6333d8c51b09496e9c13112af488a9e242039 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 9 Jan 2025 20:25:59 -0800 Subject: [PATCH 11/23] DM-192,DM-236 Properly close and delink shared memory when appropriate to avoid intermittent crashes. Adjust tabix fetch to avoid negative start values. Tweak wording on progress bars. --- dimelo/load_processed.py | 41 +++++++++++++++++++++++++--------------- 1 file changed, 26 insertions(+), 15 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 4da5f64..cb0b018 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -152,7 +152,7 @@ def pileup_counts_process_chunk( valid_base_subregion_counts = 0 modified_base_subregion_counts = 0 - for row in source_tabix.fetch(chromosome, subregion_start, subregion_end): + for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): ( keep_basemod, _, @@ -173,6 +173,9 @@ def pileup_counts_process_chunk( with lock: valid_base_counts[0] += valid_base_subregion_counts modified_base_counts[0] += modified_base_subregion_counts + # Close the file descriptor/handle to the shared memory + existing_valid.close() + existing_modified.close() def pileup_counts_from_bedmethyl( @@ -251,7 +254,7 @@ def pileup_counts_from_bedmethyl( concurrent.futures.as_completed(futures), total=len(futures), disable=quiet, - desc=f"Loading genomic chunks, approx. {chunk_size/1000}kb per chunk", + desc=f"Loading genomic chunks, up to {chunk_size/1000}kb per chunk", ): try: future.result() @@ -265,16 +268,14 @@ def pileup_counts_from_bedmethyl( valid_base_count = int.from_bytes( shm_valid.buf[:4], byteorder="little", signed=True ) + # Close and unlink shared memory - not fully handled by garbage collection otherwise + shm_modified.close() + shm_modified.unlink() + shm_valid.close() + shm_valid.unlink() return modified_base_count, valid_base_count - # We need to convert these shared memory buffers to ints that are no longer tied to the buffer, - # so they can be passed up without any seg fault type errors - modified_base_counts = np.ndarray((1,), dtype=np.int32, buffer=shm_modified.buf) - valid_base_counts = np.ndarray((1,), dtype=np.int32, buffer=shm_valid.buf) - - return int(np.copy(modified_base_counts[0])), int(np.copy(valid_base_counts[0])) - def counts_from_fake(*args, **kwargs) -> tuple[int, int]: """ @@ -330,7 +331,7 @@ def pileup_vectors_process_chunk( valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) - for row in source_tabix.fetch(chromosome, subregion_start, subregion_end): + for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): ( keep_basemod, genomic_coord, @@ -363,6 +364,9 @@ def pileup_vectors_process_chunk( modified_base_counts[ subregion_offset : subregion_offset + abs(subregion_end - subregion_start) ] += modified_base_subregion + # Close the file descriptor/handle to the shared memory + existing_modified.close() + existing_valid.close() def pileup_vectors_from_bedmethyl( @@ -458,7 +462,7 @@ def pileup_vectors_from_bedmethyl( concurrent.futures.as_completed(futures), total=len(futures), disable=quiet, - desc=f"Loading genomic chunks, approx. {chunk_size/1000}kb per chunk", + desc=f"Loading genomic chunks, up to {chunk_size/1000}kb per chunk", ): try: future.result() @@ -468,12 +472,19 @@ def pileup_vectors_from_bedmethyl( # We need to convert these shared memory buffers to numpy arrays which # we then copy, so that they no longer reference the shared memory which # will soon be de-allocated - modified_base_counts = np.ndarray( - (region_len,), dtype=np.int32, buffer=shm_modified.buf + modified_base_counts = np.copy( + np.ndarray((region_len,), dtype=np.int32, buffer=shm_modified.buf) + ) + valid_base_counts = np.copy( + np.ndarray((region_len,), dtype=np.int32, buffer=shm_valid.buf) ) - valid_base_counts = np.ndarray((region_len,), dtype=np.int32, buffer=shm_valid.buf) + # Close and unlink shared memory - not fully handled by garbage collection otherwise + shm_modified.close() + shm_modified.unlink() + shm_valid.close() + shm_valid.unlink() - return np.copy(modified_base_counts), np.copy(valid_base_counts) + return modified_base_counts, valid_base_counts def vector_from_fake(window_size: int, *args, **kwargs) -> np.ndarray: From 97efec1bd44e5e0baac70e63f83191814f5c4be3 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 9 Jan 2025 21:08:40 -0800 Subject: [PATCH 12/23] DM-191 Re-arrange contents of load_processed for better clarity. Added some comments. --- dimelo/export.py | 2 - dimelo/load_processed.py | 483 +++++++++++++++++++++++---------------- 2 files changed, 291 insertions(+), 194 deletions(-) diff --git a/dimelo/export.py b/dimelo/export.py index c3fd6c1..ce64027 100644 --- a/dimelo/export.py +++ b/dimelo/export.py @@ -107,8 +107,6 @@ def pileup_to_bigwig( load_processed.process_pileup_row( row=row, parsed_motif=parsed_motif, - region_start=0, - region_end=contig_lengths[contig], region_strand=strand, single_strand=(strand != "."), ) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index cb0b018..509cc4b 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -14,11 +14,9 @@ from . import test_data, utils - -def process_region(region_tuple, function_handle, **kwargs): - chromosome, start_coord, end_coord, strand = region_tuple - single_region_str = f"{chromosome}:{start_coord}-{end_coord},{strand}" - return function_handle(regions=single_region_str, **kwargs) +################################################################################################################ +#### Loader wrappers #### +################################################################################################################ def regions_to_list( @@ -31,6 +29,8 @@ def regions_to_list( **kwargs, ): """ + User-facing function. + Run any standard load_processed pileup or extract loader loading each region from the region specifier into a new element of a list. @@ -40,6 +40,9 @@ def regions_to_list( window_size: window around centers of regions, defaults to None cores: process count across which to parallelize. Each individual region will only ever get one core. **kwargs: all necessary keyword arguments to pass down to the loader + + Returns: + list([function_handle returns for region in regions]) """ regions_dict = utils.regions_dict_from_input( regions, @@ -79,103 +82,26 @@ def regions_to_list( return results -def process_pileup_row( - row: str, - parsed_motif: utils.ParsedMotif, - region_start: int, - region_end: int, - region_strand: str, - single_strand: bool = False, -) -> tuple[bool, int, int, int]: - """ - Returns: keep_basemod, genomic_coord, modified_in_row, valid_in_row +def process_region(region_tuple, function_handle, **kwargs): """ - tabix_fields = row.split("\t") - pileup_basemod = tabix_fields[3] - pileup_strand = tabix_fields[5] - - if single_strand and pileup_strand.strip() != region_strand: - # We are on the wrong strand, can't keep this position - keep_basemod = False - elif len(pileup_basemod.split(",")) == 3: - pileup_modname, pileup_motif, pileup_mod_coord = pileup_basemod.split(",") - if ( - pileup_motif == parsed_motif.motif_seq - and int(pileup_mod_coord) == parsed_motif.modified_pos - and pileup_modname in parsed_motif.mod_codes - ): - keep_basemod = True - else: - keep_basemod = False - elif len(pileup_basemod.split(",")) == 1: - keep_basemod = pileup_basemod in parsed_motif.mod_codes - else: - raise ValueError( - f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." - ) - - pileup_info = tabix_fields[9].split(" ") - genomic_coord = int(tabix_fields[1]) - valid_in_row = int(pileup_info[0]) - modified_in_row = int(pileup_info[2]) - - return ( - keep_basemod, - genomic_coord, - modified_in_row, - valid_in_row, - ) + Helper function for regions_to_list. Takes in the region tuple, creates a string, and runs the loader function. + Args: + region_tuple: chromosome, start, end, strand + function_handle: a function that takes a regions specifier and loads processed data + **kwargs: the remaining kwargs necessary to run the function in question -def pileup_counts_process_chunk( - bedmethyl_file, - parsed_motif, - chunk, - shm_name_modified, - shm_name_valid, - lock, - single_strand, -) -> None: - source_tabix = pysam.TabixFile(str(bedmethyl_file)) - existing_valid = shared_memory.SharedMemory(name=shm_name_valid) - existing_modified = shared_memory.SharedMemory(name=shm_name_modified) - valid_base_counts = np.ndarray((1,), dtype=np.int32, buffer=existing_valid.buf) - modified_base_counts = np.ndarray( - (1,), dtype=np.int32, buffer=existing_modified.buf - ) - - chromosome = chunk["chromosome"] - subregion_start = chunk["subregion_start"] - subregion_end = chunk["subregion_end"] - strand = chunk["strand"] - - valid_base_subregion_counts = 0 - modified_base_subregion_counts = 0 + Returns: + Whatever function_handle returns + """ + chromosome, start_coord, end_coord, strand = region_tuple + single_region_str = f"{chromosome}:{start_coord}-{end_coord},{strand}" + return function_handle(regions=single_region_str, **kwargs) - for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): - ( - keep_basemod, - _, - modified_in_row, - valid_in_row, - ) = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=subregion_start, - region_end=subregion_end, - region_strand=strand, - single_strand=single_strand, - ) - if keep_basemod: - valid_base_subregion_counts += valid_in_row - modified_base_subregion_counts += modified_in_row - with lock: - valid_base_counts[0] += valid_base_subregion_counts - modified_base_counts[0] += modified_base_subregion_counts - # Close the file descriptor/handle to the shared memory - existing_valid.close() - existing_modified.close() +################################################################################################################ +#### Pileup loaders #### +################################################################################################################ def pileup_counts_from_bedmethyl( @@ -189,6 +115,8 @@ def pileup_counts_from_bedmethyl( chunk_size: int = 1_000_000, ) -> tuple[int, int]: """ + User-facing function. + Extract number of modified bases and total number of bases from the given bedmethyl file. Called by plotters or by the user. @@ -259,7 +187,7 @@ def pileup_counts_from_bedmethyl( try: future.result() except Exception as err: - raise RuntimeError("Subprocess failed.") from err + raise RuntimeError("pileup_counts_process_chunk failed.") from err # Directly convert shared memory buffers to integers modified_base_count = int.from_bytes( @@ -277,98 +205,6 @@ def pileup_counts_from_bedmethyl( return modified_base_count, valid_base_count -def counts_from_fake(*args, **kwargs) -> tuple[int, int]: - """ - Generates a fake set of enrichment counts. Ignores all arguments. - - Returns: - tuple containing counts of (modified_bases, total_bases) - """ - window_halfsize = 500 - return test_data.fake_peak_enrichment(halfsize=window_halfsize, peak_height=0.15) - - -def pileup_vectors_process_chunk( - bedmethyl_file, - parsed_motif, - chunk, - region_len, - shm_name_modified, - shm_name_valid, - lock, - single_strand, - regions_5to3prime, -) -> None: - source_tabix = pysam.TabixFile(str(bedmethyl_file)) - existing_valid = shared_memory.SharedMemory(name=shm_name_valid) - existing_modified = shared_memory.SharedMemory(name=shm_name_modified) - valid_base_counts = np.ndarray( - (region_len,), dtype=np.int32, buffer=existing_valid.buf - ) - modified_base_counts = np.ndarray( - (region_len,), dtype=np.int32, buffer=existing_modified.buf - ) - - chromosome = chunk["chromosome"] - region_start = chunk["region_start"] - region_end = chunk["region_end"] - subregion_start = chunk["subregion_start"] - subregion_end = chunk["subregion_end"] - strand = chunk["strand"] - - flip_coords = regions_5to3prime and strand == "-" - - if flip_coords: - subregion_offset = region_end - subregion_end - else: - subregion_offset = subregion_start - region_start - - if region_end - region_start > region_len: - print( - f"WARNING: You have specified a region at {chromosome}:{region_start}-{region_end} that is longer than the first region; the end of the region will be skipped. To make a profile plot with differently-sized region, consider using the window_size parameter to make a profile across centered windows." - ) - - valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) - modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) - - for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): - ( - keep_basemod, - genomic_coord, - modified_in_row, - valid_in_row, - ) = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_start=subregion_start, - region_end=subregion_end, - region_strand=strand, - single_strand=single_strand, - ) - if keep_basemod: - if flip_coords: - # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction - # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal - pileup_coord_in_subregion = subregion_end - genomic_coord - 1 - else: - # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case - pileup_coord_in_subregion = genomic_coord - subregion_start - if pileup_coord_in_subregion < (subregion_end - subregion_start): - valid_base_subregion[pileup_coord_in_subregion] += valid_in_row - modified_base_subregion[pileup_coord_in_subregion] += modified_in_row - - with lock: - valid_base_counts[ - subregion_offset : subregion_offset + abs(subregion_end - subregion_start) - ] += valid_base_subregion - modified_base_counts[ - subregion_offset : subregion_offset + abs(subregion_end - subregion_start) - ] += modified_base_subregion - # Close the file descriptor/handle to the shared memory - existing_modified.close() - existing_valid.close() - - def pileup_vectors_from_bedmethyl( bedmethyl_file: str | Path, motif: str, @@ -381,6 +217,8 @@ def pileup_vectors_from_bedmethyl( chunk_size: int = 1_000_000, ) -> tuple[np.ndarray, np.ndarray]: """ + User-facing function. + Extract per-position pileup counts at valid motifs across one or more superimposed regions. Called by profile plotters, can also be used by a user directly. @@ -467,7 +305,7 @@ def pileup_vectors_from_bedmethyl( try: future.result() except Exception as err: - raise RuntimeError("Subprocess failed.") from err + raise RuntimeError("pileup_vectors_process_chunk failed.") from err # We need to convert these shared memory buffers to numpy arrays which # we then copy, so that they no longer reference the shared memory which @@ -487,8 +325,23 @@ def pileup_vectors_from_bedmethyl( return modified_base_counts, valid_base_counts +def counts_from_fake(*args, **kwargs) -> tuple[int, int]: + """ + Test helper function. + + Generates a fake set of enrichment counts. Ignores all arguments. + + Returns: + tuple containing counts of (modified_bases, total_bases) + """ + window_halfsize = 500 + return test_data.fake_peak_enrichment(halfsize=window_halfsize, peak_height=0.15) + + def vector_from_fake(window_size: int, *args, **kwargs) -> np.ndarray: """ + Tes helper function. + Generates a fake peak trace. Ignores all arguments except window_size. Args: @@ -502,6 +355,233 @@ def vector_from_fake(window_size: int, *args, **kwargs) -> np.ndarray: ) +def pileup_vectors_process_chunk( + bedmethyl_file, + parsed_motif, + chunk, + region_len, + shm_name_modified, + shm_name_valid, + lock, + single_strand, + regions_5to3prime, +) -> None: + """ + Helper function to allow pileup_vectors_from_bedmethyl to operate in a parallized fashion. + + Sum up modified and valid counts for a subregion chunk in a bedmethyl file. + + Args: + bedmethyl_file: Path to bedmethyl file + parsed_motif: ParsedMotif object + chunk: a dict containing subregion chunk information + shm_name_modified: the name string for the shared memory location containing the modified counts array + shm_name_valid: the name string for the shared memory location containing the valid counts array + lock: a manager.Lock object to allow synchronization in accessing shared memory + single_strand: True if only single-strand mods are desired + + Returns: + None. Counts are added in-place to shared memory. + """ + source_tabix = pysam.TabixFile(str(bedmethyl_file)) + existing_valid = shared_memory.SharedMemory(name=shm_name_valid) + existing_modified = shared_memory.SharedMemory(name=shm_name_modified) + valid_base_counts = np.ndarray( + (region_len,), dtype=np.int32, buffer=existing_valid.buf + ) + modified_base_counts = np.ndarray( + (region_len,), dtype=np.int32, buffer=existing_modified.buf + ) + + chromosome = chunk["chromosome"] + region_start = chunk["region_start"] + region_end = chunk["region_end"] + subregion_start = chunk["subregion_start"] + subregion_end = chunk["subregion_end"] + strand = chunk["strand"] + + flip_coords = regions_5to3prime and strand == "-" + + if flip_coords: + subregion_offset = region_end - subregion_end + else: + subregion_offset = subregion_start - region_start + + if region_end - region_start > region_len: + print( + f"WARNING: You have specified a region at {chromosome}:{region_start}-{region_end} that is longer than the first region; the end of the region will be skipped. To make a profile plot with differently-sized region, consider using the window_size parameter to make a profile across centered windows." + ) + + valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) + modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) + + for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): + ( + keep_basemod, + genomic_coord, + modified_in_row, + valid_in_row, + ) = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_strand=strand, + single_strand=single_strand, + ) + if keep_basemod: + if flip_coords: + # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction + # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal + pileup_coord_in_subregion = subregion_end - genomic_coord - 1 + else: + # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case + pileup_coord_in_subregion = genomic_coord - subregion_start + if pileup_coord_in_subregion < (subregion_end - subregion_start): + valid_base_subregion[pileup_coord_in_subregion] += valid_in_row + modified_base_subregion[pileup_coord_in_subregion] += modified_in_row + + with lock: + valid_base_counts[ + subregion_offset : subregion_offset + abs(subregion_end - subregion_start) + ] += valid_base_subregion + modified_base_counts[ + subregion_offset : subregion_offset + abs(subregion_end - subregion_start) + ] += modified_base_subregion + # Close the file descriptor/handle to the shared memory + existing_modified.close() + existing_valid.close() + + +def pileup_counts_process_chunk( + bedmethyl_file, + parsed_motif, + chunk, + shm_name_modified, + shm_name_valid, + lock, + single_strand, +) -> None: + """ + Helper function to allow pileup_counts_from_bedmethyl to operate in a parallized fashion. + + Sum up modified and valid counts for a subregion chunk in a bedmethyl file. + + Args: + bedmethyl_file: Path to bedmethyl file + parsed_motif: ParsedMotif object + chunk: a dict containing subregion chunk information + shm_name_modified: the name string for the shared memory location containing the modified counts sum + shm_name_valid: the name string for the shared memory location containing the valid counts sum + lock: a manager.Lock object to allow synchronization in accessing shared memory + single_strand: True if only single-strand mods are desired + + Returns: + None. Counts are added in-place to shared memory. + """ + source_tabix = pysam.TabixFile(str(bedmethyl_file)) + existing_valid = shared_memory.SharedMemory(name=shm_name_valid) + existing_modified = shared_memory.SharedMemory(name=shm_name_modified) + valid_base_counts = np.ndarray((1,), dtype=np.int32, buffer=existing_valid.buf) + modified_base_counts = np.ndarray( + (1,), dtype=np.int32, buffer=existing_modified.buf + ) + + chromosome = chunk["chromosome"] + subregion_start = chunk["subregion_start"] + subregion_end = chunk["subregion_end"] + strand = chunk["strand"] + + valid_base_subregion_counts = 0 + modified_base_subregion_counts = 0 + + for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): + ( + keep_basemod, + _, + modified_in_row, + valid_in_row, + ) = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_strand=strand, + single_strand=single_strand, + ) + if keep_basemod: + valid_base_subregion_counts += valid_in_row + modified_base_subregion_counts += modified_in_row + + with lock: + valid_base_counts[0] += valid_base_subregion_counts + modified_base_counts[0] += modified_base_subregion_counts + + # Close the file descriptor/handle to the shared memory + existing_valid.close() + existing_modified.close() + + +def process_pileup_row( + row: str, + parsed_motif: utils.ParsedMotif, + region_strand: str, + single_strand: bool = False, +) -> tuple[bool, int, int, int]: + """ + Helper function designed for pileup_counts_from_bedmethyl via pileup_counts_process_chunk, pileup_vectors_from_bedmethyl + via pileup_vectors_process_chunk, and export.pileup_to_bigwig; changed to logic here may impact some or all of + these. + + Process a row from a pileup, determining whether the basemod is relevant and passing back its coordinate, + modification count, and valid read count. + + Args: + row: a string row from a bedmethyl file + parsed_motif: a ParsedMotif object + region_strand: the strand from the query region + single_strand: True if only mods on the region_strand are to be kept + + Returns: keep_basemod, genomic_coord, modified_in_row, valid_in_row + """ + tabix_fields = row.split("\t") + pileup_basemod = tabix_fields[3] + pileup_strand = tabix_fields[5] + + if single_strand and pileup_strand.strip() != region_strand: + # We are on the wrong strand, can't keep this position + keep_basemod = False + elif len(pileup_basemod.split(",")) == 3: + pileup_modname, pileup_motif, pileup_mod_coord = pileup_basemod.split(",") + if ( + pileup_motif == parsed_motif.motif_seq + and int(pileup_mod_coord) == parsed_motif.modified_pos + and pileup_modname in parsed_motif.mod_codes + ): + keep_basemod = True + else: + keep_basemod = False + elif len(pileup_basemod.split(",")) == 1: + keep_basemod = pileup_basemod in parsed_motif.mod_codes + else: + raise ValueError( + f"Unexpected format in bedmethyl file: {row} contains {pileup_basemod} which cannot be parsed." + ) + + pileup_info = tabix_fields[9].split(" ") + genomic_coord = int(tabix_fields[1]) + valid_in_row = int(pileup_info[0]) + modified_in_row = int(pileup_info[2]) + + return ( + keep_basemod, + genomic_coord, + modified_in_row, + valid_in_row, + ) + + +################################################################################################################ +#### Single read loaders #### +################################################################################################################ + + def read_vectors_from_hdf5( file: str | Path, motifs: list[str], @@ -514,6 +594,8 @@ def read_vectors_from_hdf5( quiet: bool = True, # currently unused ) -> tuple[list[tuple], list[str], dict | None]: """ + User-facing function. + Pulls a list of read data out of an .h5 file containing processed read vectors, formatted for read-by-read vector processing downstream use cases. @@ -747,6 +829,8 @@ def readwise_binary_modification_arrays( cores: int | None = None, # currently unused ) -> tuple[list[np.ndarray], np.ndarray[int], np.ndarray[str], dict | None]: """ + Primarily designed as a helper function for single-read plotting, but can be used by a user. + Pulls a list of read data out of a file containing processed read vectors, formatted with seaborn plotting in mind. Currently we only support .h5 files. @@ -900,6 +984,8 @@ def reads_from_fake( file: Path, regions: Path, motifs: list[str] ) -> tuple[list[np.ndarray], np.ndarray[int], np.ndarray[str], dict]: """ + Helper function to support testing. + TODO: What does the bed file represent in this method? This one is breaking my brain a bit. TODO: Variable names in this method stink. TODO: Currently assumes mod calling (thresholding probabilities) was already performed elsewhere @@ -952,17 +1038,27 @@ def reads_from_fake( def convert_bytes_to_strings(tup): - """Convert all bytes elements in a tuple to strings.""" + """ + Helper function for single read loading. + Convert all bytes elements in a tuple to strings. + """ return tuple(item.decode() if isinstance(item, bytes) else item for item in tup) # tuple(convert_bytes(item) for item in tup) def adjust_mod_probs_in_arrays(mod_array, val_array): + """ + Helper function to correct for an idiosyncracy in modkit single-read parsing wherein 0-255 + "mod quality" values are parsed as floating-point values from 1/512 to 511/512. + """ mod_array[np.flatnonzero(val_array)] += 1 / 512 return mod_array def adjust_mod_probs_in_tuples(tup, mod_idx, val_idx): + """ + Helper function to apply mod prob adjustments + """ return tuple( item if index != mod_idx else adjust_mod_probs_in_arrays(item, tup[val_idx]) for index, item in enumerate(tup) @@ -970,6 +1066,9 @@ def adjust_mod_probs_in_tuples(tup, mod_idx, val_idx): def binary_to_np_array(compressed_bytes, dtype, decompressor, binarized, int8tofloat): + """ + Helper function to decompress binary data to boolean or floating point arrays + """ if binarized: return np.frombuffer(decompressor(compressed_bytes), dtype=dtype).astype(bool) elif int8tofloat: From a8ed1ae5b598dc4b6dad87d92549375109151204 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Fri, 17 Jan 2025 12:09:33 -0800 Subject: [PATCH 13/23] DM-238 Add quiet and cores parameters and corresponding documentation to all pileup plotters. A few other small tweaks. --- dimelo/load_processed.py | 44 ++++++++++++++++++++++--------- dimelo/plot_depth_histogram.py | 12 +++++++-- dimelo/plot_depth_profile.py | 13 ++++++++- dimelo/plot_enrichment.py | 10 +++++++ dimelo/plot_enrichment_profile.py | 10 +++++++ 5 files changed, 73 insertions(+), 16 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 509cc4b..43ebef7 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -38,7 +38,11 @@ def regions_to_list( function_handle: the loader function you want to run. regions: the region specifier window_size: window around centers of regions, defaults to None - cores: process count across which to parallelize. Each individual region will only ever get one core. + quiet: disables progress bars + cores: CPU cores across which to parallelize processing + parallelize_within_regions: if True, regions will be run sequentially in parallelized chunks. If False, + each individual region's chunks will be run sequentially but there will be parallelization across + regions, i.e. each core will be assigned one region at a time by the executor **kwargs: all necessary keyword arguments to pass down to the loader Returns: @@ -57,7 +61,12 @@ def regions_to_list( ] cores_to_run = utils.cores_to_run(cores) - + # quiet and cores logic below is driven by the following: + # If the parallelization is within regions: + # (1) progress bars should happen within regions if at all, because we assume regions are + # large if they make sense to parallelize + # (2) the cores_to_run will be allocated to within-region parallelization, and the top-level + # jobs sequence is run sequentially with concurrent.futures.ProcessPoolExecutor( max_workers=1 if parallelize_within_regions else cores_to_run ) as executor: @@ -65,8 +74,10 @@ def regions_to_list( process_partial = partial( process_region, function_handle=function_handle, - quiet=quiet or not parallelize_within_regions, - cores=cores_to_run if parallelize_within_regions else 1, + quiet=quiet or not parallelize_within_regions, # + cores=cores_to_run + if parallelize_within_regions + else 1, # if parallelization is within region **kwargs, ) @@ -110,7 +121,7 @@ def pileup_counts_from_bedmethyl( regions: str | Path | list[str | Path], window_size: int | None = None, single_strand: bool = False, - quiet: bool = True, + quiet: bool = False, cores: int | None = None, chunk_size: int = 1_000_000, ) -> tuple[int, int]: @@ -138,7 +149,7 @@ def pileup_counts_from_bedmethyl( single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions quiet: disables progress bars - cores: cores across which to parallelize processes + cores: CPU cores across which to parallelize processing chunk_size: size of genomic subregions to assign out to each process Returns: @@ -212,7 +223,7 @@ def pileup_vectors_from_bedmethyl( window_size: int | None = None, single_strand: bool = False, regions_5to3prime: bool = False, - quiet: bool = True, + quiet: bool = False, cores: int | None = None, chunk_size: int = 1_000_000, ) -> tuple[np.ndarray, np.ndarray]: @@ -249,7 +260,7 @@ def pileup_vectors_from_bedmethyl( the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping quiet: disables progress bars - cores: cores across which to parallelize processes + cores: CPU cores across which to parallelize processing chunk_size: size of genomic subregions to assign out to each process Returns: @@ -379,9 +390,10 @@ def pileup_vectors_process_chunk( shm_name_valid: the name string for the shared memory location containing the valid counts array lock: a manager.Lock object to allow synchronization in accessing shared memory single_strand: True if only single-strand mods are desired + regions_5to3prime: True means negative strand regions get flipped, False means no flipping Returns: - None. Counts are added in-place to shared memory. + None. Counts are added to arrays in-place to shared memory. """ source_tabix = pysam.TabixFile(str(bedmethyl_file)) existing_valid = shared_memory.SharedMemory(name=shm_name_valid) @@ -526,7 +538,7 @@ def process_pileup_row( ) -> tuple[bool, int, int, int]: """ Helper function designed for pileup_counts_from_bedmethyl via pileup_counts_process_chunk, pileup_vectors_from_bedmethyl - via pileup_vectors_process_chunk, and export.pileup_to_bigwig; changed to logic here may impact some or all of + via pileup_vectors_process_chunk, and export.pileup_to_bigwig; changes to logic here may impact some or all of these. Process a row from a pileup, determining whether the basemod is relevant and passing back its coordinate, @@ -538,7 +550,7 @@ def process_pileup_row( region_strand: the strand from the query region single_strand: True if only mods on the region_strand are to be kept - Returns: keep_basemod, genomic_coord, modified_in_row, valid_in_row + Returns: keep_basemod, genomic_coord, modified_in_row, valid_in_row. Values are provided even if keep_basemod is False. """ tabix_fields = row.split("\t") pileup_basemod = tabix_fields[3] @@ -590,8 +602,8 @@ def read_vectors_from_hdf5( single_strand: bool = False, sort_by: str | list[str] = ["chromosome", "region_start", "read_start"], calculate_mod_fractions: bool = True, + quiet: bool = True, # currently unused; change to default False when pbars are implemented cores: int | None = None, # currently unused - quiet: bool = True, # currently unused ) -> tuple[list[tuple], list[str], dict | None]: """ User-facing function. @@ -613,6 +625,8 @@ def read_vectors_from_hdf5( After this processing, we calculate modification fractions, sort, and return. + TODO: Implement progress bars and parallelization as with pileup loaders + Args: file: Path to an hdf5 (.h5) file containing modification data for single reads, stored in datasets read_name, chromosome, read_start, @@ -635,8 +649,8 @@ def read_vectors_from_hdf5( sort_by: Read properties by which to sort, either one string or a list of strings. Options include chromosome, region_start, region_end, read_start, read_end, and motif. More to be added in future. - cores: cores across which to parallelize processes (currently unused) quiet: silences progress bars (currently unused) + cores: cores across which to parallelize processes (currently unused) Returns: a list of tuples, each tuple containing all datasets corresponding to an individual read that @@ -826,6 +840,7 @@ def readwise_binary_modification_arrays( sort_by: str | list[str] = ["chromosome", "region_start", "read_start"], thresh: float | None = None, relative: bool = True, + quiet: bool = True, # currently unused; change to default False when pbars are implemented cores: int | None = None, # currently unused ) -> tuple[list[np.ndarray], np.ndarray[int], np.ndarray[str], dict | None]: """ @@ -842,6 +857,8 @@ def readwise_binary_modification_arrays( coordinates. If positions are relative, regions_5to3prime can be used to show all regions as upstream-to-downstream along their respective strands. + TODO: Implement progress bars and parallelization as with pileup loaders + Args: file: Path to an hdf5 (.h5) file containing modification data for single reads, stored in datasets read_name, chromosome, read_start, @@ -870,6 +887,7 @@ def readwise_binary_modification_arrays( in the genomes, centered at the center of the region. If False, absolute coordinates are provided. There is not currently a check for all reads being on the same chromosome if relative=False, but this could create unexpected behaviour for a the standard visualizations. + quiet: silences progress bars (currently unused) cores: cores across which to parallelize processes (currently unused) Returns: diff --git a/dimelo/plot_depth_histogram.py b/dimelo/plot_depth_histogram.py index 655e163..949b4ec 100644 --- a/dimelo/plot_depth_histogram.py +++ b/dimelo/plot_depth_histogram.py @@ -14,7 +14,8 @@ def plot_depth_histogram( window_size: int, single_strand: bool = False, average_within_region: bool = False, - cores=None, + quiet: bool = False, + cores: int | None = None, **kwargs, ) -> Axes: """ @@ -40,6 +41,8 @@ def plot_depth_histogram( single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions average_within_region: if True, each region will only report a single depth value, averaging across all non-zero depths + quiet: disables progress bars + cores: CPU cores across which to parallelize processing kwargs: other keyword parameters passed through to utils.line_plot Returns: @@ -55,6 +58,7 @@ def plot_depth_histogram( window_size=window_size, single_strand=single_strand, average_within_region=average_within_region, + quiet=quiet, cores=cores, ) @@ -159,7 +163,8 @@ def get_depth_counts( window_size: int, single_strand: bool = False, average_within_region: bool = False, - cores=1, + quiet: bool = False, + cores: int | None = None, ) -> list[np.ndarray]: """ Get the depth counts, ready for plotting. @@ -180,6 +185,8 @@ def get_depth_counts( the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping smooth_window: size of the moving window to use for smoothing. If set to None, no smoothing is performed + quiet: disables progress bars + cores: CPU cores across which to parallelize processing Returns: List of depth histogram traces @@ -201,6 +208,7 @@ def get_depth_counts( motif=motif, window_size=window_size, single_strand=single_strand, + quiet=quiet, cores=cores, ) # places where read depth is zero are assumed to not have the motif present - this may not always be true, diff --git a/dimelo/plot_depth_profile.py b/dimelo/plot_depth_profile.py index e4b3327..0e59db1 100644 --- a/dimelo/plot_depth_profile.py +++ b/dimelo/plot_depth_profile.py @@ -15,6 +15,8 @@ def plot_depth_profile( single_strand: bool = False, regions_5to3prime: bool = False, smooth_window: int | None = None, + quiet: bool = False, + cores: int | None = None, **kwargs, ) -> Axes: """ @@ -41,6 +43,8 @@ def plot_depth_profile( the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping smooth_window: size of the moving window to use for smoothing. If set to None, no smoothing is performed + quiet: disables progress bars + cores: CPU cores across which to parallelize processing kwargs: other keyword parameters passed through to utils.line_plot Returns: @@ -57,6 +61,8 @@ def plot_depth_profile( single_strand=single_strand, regions_5to3prime=regions_5to3prime, smooth_window=smooth_window, + quiet=quiet, + cores=cores, ) axes = make_depth_profile_plot( @@ -157,6 +163,8 @@ def get_depth_profiles( single_strand: bool = False, regions_5to3prime: bool = False, smooth_window: int | None = None, + quiet: bool = False, + cores: int | None = None, ) -> list[np.ndarray]: """ Get the depth profile traces, ready for plotting. @@ -177,6 +185,8 @@ def get_depth_profiles( the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping smooth_window: size of the moving window to use for smoothing. If set to None, no smoothing is performed + quiet: disables progress bars + cores: CPU cores across which to parallelize processing Returns: List of depth profile traces @@ -198,7 +208,8 @@ def get_depth_profiles( window_size=window_size, single_strand=single_strand, regions_5to3prime=regions_5to3prime, - quiet=False, + quiet=quiet, + cores=cores, ) trace = valid_base_counts.astype(float) trace[trace == 0] = np.nan diff --git a/dimelo/plot_enrichment.py b/dimelo/plot_enrichment.py index 74c80d6..b30e8e4 100644 --- a/dimelo/plot_enrichment.py +++ b/dimelo/plot_enrichment.py @@ -12,6 +12,8 @@ def plot_enrichment( sample_names: list[str], window_size: int | None = None, single_strand: bool = False, + quiet: bool = False, + cores: int | None = None, **kwargs, ) -> Axes: """ @@ -31,6 +33,8 @@ def plot_enrichment( window_size: (currently disabled) window around center of region, +-window_size//2 single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions + quiet: disables progress bars + cores: CPU cores across which to parallelize processing kwargs: other keyword parameters passed through to utils.bar_plot Returns: @@ -143,6 +147,8 @@ def get_enrichments( motifs: list[str], window_size: int | None = None, single_strand: bool = False, + quiet: bool = False, + cores: int | None = None, ) -> list[float]: """ Get the enrichment values, ready for plotting. @@ -160,6 +166,8 @@ def get_enrichments( window_size: (currently disabled) window around center of region, +-window_size//2 single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions + quiet: disables progress bars + cores: CPU cores across which to parallelize processing Returns: List of modified fraction values. @@ -180,6 +188,8 @@ def get_enrichments( motif=motif, window_size=window_size, single_strand=single_strand, + quiet=quiet, + cores=cores, ) case ".fake": n_mod, n_total = load_processed.counts_from_fake( diff --git a/dimelo/plot_enrichment_profile.py b/dimelo/plot_enrichment_profile.py index 8a33cbf..0e339ec 100644 --- a/dimelo/plot_enrichment_profile.py +++ b/dimelo/plot_enrichment_profile.py @@ -15,6 +15,8 @@ def plot_enrichment_profile( single_strand: bool = False, regions_5to3prime: bool = False, smooth_window: int | None = None, + quiet: bool = False, + cores: int | None = None, **kwargs, ) -> Axes: """ @@ -41,6 +43,8 @@ def plot_enrichment_profile( the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping smooth_window: size of the moving window to use for smoothing. If set to None, no smoothing is performed + quiet: disables progress bars + cores: CPU cores across which to parallelize processing kwargs: other keyword parameters passed through to utils.line_plot Returns: @@ -157,6 +161,8 @@ def get_enrichment_profiles( single_strand: bool = False, regions_5to3prime: bool = False, smooth_window: int | None = None, + quiet: bool = False, + cores: int | None = None, ) -> list[np.ndarray]: """ Get the enrichment profile traces, ready for plotting. @@ -176,6 +182,8 @@ def get_enrichment_profiles( single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping + quiet: disables progress bars + cores: CPU cores across which to parallelize processing smooth_window: size of the moving window to use for smoothing. If set to None, no smoothing is performed Returns: @@ -199,6 +207,8 @@ def get_enrichment_profiles( window_size=window_size, single_strand=single_strand, regions_5to3prime=regions_5to3prime, + quiet=quiet, + cores=cores, ) ) # Default to nan so we can skip over unfilled values when plotting or doing a rolling average From f1b07586cb818cad0f586f7fbf8b20f7e10c15dd Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Fri, 17 Jan 2025 12:17:15 -0800 Subject: [PATCH 14/23] DM-239 Added test case coverage for 1-4 cores. GitHub jobs for Ubuntu and Macos ostensibly have 4 cores. This should not change any outputs because for extract the test is hardcoded to use 1 core, and that is the only one with any stochasticity in parallelization. --- dimelo/test/cases.py | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/dimelo/test/cases.py b/dimelo/test/cases.py index 001238c..ee02c90 100644 --- a/dimelo/test/cases.py +++ b/dimelo/test/cases.py @@ -34,6 +34,7 @@ "single_strand": False, "regions_5to3prime": False, "chunk_size": 1_000_000, + "cores": 2, }, # outputs dict function:values {}, # populated in subsequent cells @@ -54,6 +55,7 @@ "single_strand": False, "regions_5to3prime": False, "chunk_size": 100, + "cores": 4, }, # outputs dict function:values {}, # populated in subsequent cells @@ -74,6 +76,7 @@ "single_strand": True, "regions_5to3prime": True, "chunk_size": 10_000, + "cores": 1, }, # outputs dict function:values {}, # populated in subsequent cells @@ -94,6 +97,7 @@ "single_strand": True, "regions_5to3prime": False, "chunk_size": 1000, + "cores": 4, }, # outputs dict function:values {}, # populated in subsequent cells @@ -114,6 +118,7 @@ "single_strand": False, "regions_5to3prime": True, "chunk_size": 100, + "cores": 3, }, # outputs dict function:values {}, # populated in subsequent cells From 0b68892af60eb9fbd5664ece2a04bc15adcb7e5c Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Fri, 17 Jan 2025 12:30:13 -0800 Subject: [PATCH 15/23] DM-239 Added cases.py support for cores=None. Tweaked dimelo_test.py to avoid ever sending the cores arg twice, and added a comment to explain the reason why extract is tested unparallelized, unlike other functions. --- dimelo/test/cases.py | 8 ++++---- dimelo/test/dimelo_test.py | 7 +++++++ 2 files changed, 11 insertions(+), 4 deletions(-) diff --git a/dimelo/test/cases.py b/dimelo/test/cases.py index ee02c90..a269008 100644 --- a/dimelo/test/cases.py +++ b/dimelo/test/cases.py @@ -34,7 +34,7 @@ "single_strand": False, "regions_5to3prime": False, "chunk_size": 1_000_000, - "cores": 2, + "cores": 1, }, # outputs dict function:values {}, # populated in subsequent cells @@ -55,7 +55,7 @@ "single_strand": False, "regions_5to3prime": False, "chunk_size": 100, - "cores": 4, + "cores": 2, }, # outputs dict function:values {}, # populated in subsequent cells @@ -76,7 +76,7 @@ "single_strand": True, "regions_5to3prime": True, "chunk_size": 10_000, - "cores": 1, + "cores": 3, }, # outputs dict function:values {}, # populated in subsequent cells @@ -118,7 +118,7 @@ "single_strand": False, "regions_5to3prime": True, "chunk_size": 100, - "cores": 3, + "cores": None, }, # outputs dict function:values {}, # populated in subsequent cells diff --git a/dimelo/test/dimelo_test.py b/dimelo/test/dimelo_test.py index 787aa83..73310cc 100644 --- a/dimelo/test/dimelo_test.py +++ b/dimelo/test/dimelo_test.py @@ -77,6 +77,13 @@ def test_unit__extract( ): kwargs_extract = filter_kwargs_for_func(dm.parse_bam.extract, kwargs) kwargs_extract["output_directory"] = cls.outDir + # extract can behave non-deterministically in terms of read output order + # if run on multiple cores. This is not an issue for sorted read loads, + # but means the file itself changes even though the meaningful contents don't + # given that parallelization here is purely within modkit anyway, the design + # choice was made to always single-core for extract testing + if "cores" in kwargs_extract: + del kwargs_extract["cores"] extract_h5, regions_processed = dm.parse_bam.extract( **kwargs_extract, ref_genome=cls.reference_genome, From 1da3d3c59d66c8560efd23acc6a126a6153c91f7 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Fri, 17 Jan 2025 16:27:54 -0800 Subject: [PATCH 16/23] DM-239 Adjusted generate_targets module to properly handle cores argument for extract. Updated test_matrix.pickle by re-running generate_targets. 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b/dimelo/parse_bam.py index 157ed59..2bb8a64 100644 --- a/dimelo/parse_bam.py +++ b/dimelo/parse_bam.py @@ -531,7 +531,7 @@ def verify_inputs( ) elif correct_bases / total_bases < 0.35: raise ValueError( - f"First {NUM_READS_TO_CHECK} reads have anomalously low alignment quality: only {100*correct_bases/total_bases}% of bases align.\nPlease verify that {input_file.name} is actually aligned to {ref_genome.name}." + f"First {NUM_READS_TO_CHECK} reads have anomalously low alignment quality: only {100 * correct_bases / total_bases}% of bases align.\nPlease verify that {input_file.name} is actually aligned to {ref_genome.name}." ) return @@ -621,7 +621,7 @@ def check_bam_format( print( f""" WARNING: no modified appropriately-coded values found for {missing_bases} in the first {counter} reads. -Do you expect this file to contain these modifications? parse_bam is looking for {motifs} but for {missing_bases} found only found {[f'{base}+{mod_codes}' for base, mod_codes in mod_codes_found_dict.items()]}. +Do you expect this file to contain these modifications? parse_bam is looking for {motifs} but for {missing_bases} found only found {[f"{base}+{mod_codes}" for base, mod_codes in mod_codes_found_dict.items()]}. Consider passing only the motifs and mod codes (e.g. m,h,a) that you expect to be present in your file. You can use modkit adjust-mods --convert [OPTIONS] to update or consolidate mod codes. @@ -956,7 +956,7 @@ def read_by_base_txt_to_hdf5( iterator = tqdm( iterator, total=num_lines, - desc=f"Transferring {num_reads} from {input_txt.name} into {output_h5.name}, new size {old_size+num_reads}", + desc=f"Transferring {num_reads} from {input_txt.name} into {output_h5.name}, new size {old_size + num_reads}", bar_format="{bar}| {desc} {percentage:3.0f}% | {elapsed}<{remaining}", ) diff --git a/dimelo/run_modkit.py b/dimelo/run_modkit.py index aa8e707..298f0e9 100644 --- a/dimelo/run_modkit.py +++ b/dimelo/run_modkit.py @@ -28,8 +28,8 @@ print( f"PATH does not include the conda environment /bin folder. Adding {env_bin_path}." ) - os.environ["PATH"] = f'{env_bin_path}:{os.environ["PATH"]}' - print(f'PATH is now {os.environ["PATH"]}') + os.environ["PATH"] = f"{env_bin_path}:{os.environ['PATH']}" + print(f"PATH is now {os.environ['PATH']}") # Check modkit on first import: does it run; does it have the right version try: diff --git a/dimelo/test/dimelo_test.py b/dimelo/test/dimelo_test.py index 73310cc..d11dae9 100644 --- a/dimelo/test/dimelo_test.py +++ b/dimelo/test/dimelo_test.py @@ -270,9 +270,9 @@ def test_integration__extract_load_plot( assert np.allclose( actual[key], expected[key], atol=1e-5 ), f"""{test_case}: Arrays for {key} are not equal -mismatch at {np.where(value!=actual[key])} -mismatch values expected {value[np.where(value!=actual[key])]} vs actual {actual[key][np.where(value!=actual[key])]} -{value[np.where(value!=actual[key])[0]]} vs {actual[key][np.where(value!=actual[key])[0]]}. +mismatch at {np.where(value != actual[key])} +mismatch values expected {value[np.where(value != actual[key])]} vs actual {actual[key][np.where(value != actual[key])]} +{value[np.where(value != actual[key])[0]]} vs {actual[key][np.where(value != actual[key])[0]]}. """ elif isinstance(value, (str, int, bool)): assert ( @@ -439,9 +439,9 @@ def test_unit__read_vectors_from_hdf5( assert np.allclose( actual[key], expected[key], atol=1e-5 ), f"""{test_case}: Arrays for {key} are not equal -mismatch at {np.where(value!=actual[key])} -mismatch values expected {value[np.where(value!=actual[key])]} vs actual {actual[key][np.where(value!=actual[key])]} -{value[np.where(value!=actual[key])[0]]} vs {actual[key][np.where(value!=actual[key])[0]]}. +mismatch at {np.where(value != actual[key])} +mismatch values expected {value[np.where(value != actual[key])]} vs actual {actual[key][np.where(value != actual[key])]} +{value[np.where(value != actual[key])[0]]} vs {actual[key][np.where(value != actual[key])[0]]}. """ elif isinstance(value, (str, int, bool)): assert ( diff --git a/dimelo/utils.py b/dimelo/utils.py index aa85e55..66ed83d 100644 --- a/dimelo/utils.py +++ b/dimelo/utils.py @@ -84,7 +84,7 @@ def adjust_threshold( if thresh > 1: if not quiet: print( - f"Modification threshold of {thresh} assumed to be for range 0-255. {thresh}/255={thresh/255} will be sent to modkit." + f"Modification threshold of {thresh} assumed to be for range 0-255. {thresh}/255={thresh / 255} will be sent to modkit." ) thresh_scaled = thresh / 255 else: From bd94e785673c52bcea041bf6ef167795db1d1c2a Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Fri, 21 Feb 2025 10:13:41 -0800 Subject: [PATCH 18/23] Fixed small mistakes from merge --- dimelo/export.py | 2 +- dimelo/load_processed.py | 23 ++++++++++++++--------- 2 files changed, 15 insertions(+), 10 deletions(-) diff --git a/dimelo/export.py b/dimelo/export.py index 8ccf34b..e2c973c 100644 --- a/dimelo/export.py +++ b/dimelo/export.py @@ -131,7 +131,7 @@ def pileup_to_bigwig( for row in tqdm( tabix.fetch(contig), desc=f"Writing {contig}.", - total=contig_lines[contig], + total=lines_by_contig[contig], leave=False, ): keep_basemod, genomic_coord, modified_in_row, valid_in_row = ( diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 043a3f2..ad4c79b 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -18,6 +18,7 @@ #### Loader wrappers #### ################################################################################################################ + def regions_to_list( function_handle, regions, @@ -79,7 +80,7 @@ def regions_to_list( else 1, # if parallelization is within region **kwargs, ) - + if cores_to_run > 1: # Use executor.map without lambda results = list( tqdm( @@ -88,17 +89,21 @@ def regions_to_list( desc=f"Processing regions in parallel across {cores_to_run}", ) ) - else: - # Single-threaded fallback - results = [ - process_region( - region_string=region, function_handle=function_handle, cores=1, **kwargs - ) - for region in tqdm(region_strings, desc="Processing regions") - ] + else: + # Single-threaded fallback + results = [ + process_region( + region_string=region, + function_handle=function_handle, + cores=1, + **kwargs, + ) + for region in tqdm(region_strings, desc="Processing regions") + ] return results + def process_region(region_string, function_handle, **kwargs): """ process_region simply exists to convert position arguments into keyword arguments to make executor.map work From 4454f675ab6a6c0343b04e43a0f3ccc3bcb22a9e Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Tue, 18 Mar 2025 14:34:39 -0700 Subject: [PATCH 19/23] Added check for chrom in tabixfile. --- dimelo/load_processed.py | 84 ++++++++++++++++++++++------------------ 1 file changed, 46 insertions(+), 38 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 43ebef7..63b9684 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -427,29 +427,34 @@ def pileup_vectors_process_chunk( valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) - for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): - ( - keep_basemod, - genomic_coord, - modified_in_row, - valid_in_row, - ) = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_strand=strand, - single_strand=single_strand, - ) - if keep_basemod: - if flip_coords: - # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction - # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal - pileup_coord_in_subregion = subregion_end - genomic_coord - 1 - else: - # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case - pileup_coord_in_subregion = genomic_coord - subregion_start - if pileup_coord_in_subregion < (subregion_end - subregion_start): - valid_base_subregion[pileup_coord_in_subregion] += valid_in_row - modified_base_subregion[pileup_coord_in_subregion] += modified_in_row + if chromosome in source_tabix.contigs: + for row in source_tabix.fetch( + chromosome, max(subregion_start, 0), subregion_end + ): + ( + keep_basemod, + genomic_coord, + modified_in_row, + valid_in_row, + ) = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_strand=strand, + single_strand=single_strand, + ) + if keep_basemod: + if flip_coords: + # We want to flip the coordinates for this region so that it is recorded along the 5 prime to 3 prime direction + # This will enable analyses where the orientation of protein binding / transcriptional dynamics / etc is relevant for our pileup signal + pileup_coord_in_subregion = subregion_end - genomic_coord - 1 + else: + # Normal coordinates are the default. This will be used both for the '+' case and the '.' (no strand specified) case + pileup_coord_in_subregion = genomic_coord - subregion_start + if pileup_coord_in_subregion < (subregion_end - subregion_start): + valid_base_subregion[pileup_coord_in_subregion] += valid_in_row + modified_base_subregion[pileup_coord_in_subregion] += ( + modified_in_row + ) with lock: valid_base_counts[ @@ -505,21 +510,24 @@ def pileup_counts_process_chunk( valid_base_subregion_counts = 0 modified_base_subregion_counts = 0 - for row in source_tabix.fetch(chromosome, max(subregion_start, 0), subregion_end): - ( - keep_basemod, - _, - modified_in_row, - valid_in_row, - ) = process_pileup_row( - row=row, - parsed_motif=parsed_motif, - region_strand=strand, - single_strand=single_strand, - ) - if keep_basemod: - valid_base_subregion_counts += valid_in_row - modified_base_subregion_counts += modified_in_row + if chromosome in source_tabix.contigs: + for row in source_tabix.fetch( + chromosome, max(subregion_start, 0), subregion_end + ): + ( + keep_basemod, + _, + modified_in_row, + valid_in_row, + ) = process_pileup_row( + row=row, + parsed_motif=parsed_motif, + region_strand=strand, + single_strand=single_strand, + ) + if keep_basemod: + valid_base_subregion_counts += valid_in_row + modified_base_subregion_counts += modified_in_row with lock: valid_base_counts[0] += valid_base_subregion_counts From bfdbff64aeac1b3ab0a41c1a6f2da3e9831fb2f9 Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Sun, 23 Mar 2025 13:30:54 -0700 Subject: [PATCH 20/23] DM-191 Adjustments in response to PR comments from thekugelmeister --- dimelo/load_processed.py | 48 ++++++++++++++++++++++--------- dimelo/plot_depth_histogram.py | 7 ++++- dimelo/plot_depth_profile.py | 2 +- dimelo/plot_enrichment.py | 4 ++- dimelo/plot_enrichment_profile.py | 4 ++- dimelo/run_modkit.py | 3 +- dimelo/utils.py | 7 ++++- 7 files changed, 55 insertions(+), 20 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 63b9684..468f378 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -14,6 +14,14 @@ from . import test_data, utils +# the default chunk size is the number of bp to include per processing chunk in parallelization for loaders. +# 1e6 was empirically determined to be a good default: smaller than 1e5 we see slowdowns due to increased +# parallelization overhead, larger than 1e7 we see slowdowns due to worker utilization decreasing because even +# for whole chromosome processing there aren't always enough chunks to go around. In the 1e5-1e7 range, speed +# on 32 cores is fairly similar, but sitting in the middle of the range should support 10x more cores (beyond +# the reasonable upper bound) and 10x fewer cores (which is about the reasonable lower bound). +DEFAULT_CHUNK_SIZE = 1_000_000 + ################################################################################################################ #### Loader wrappers #### ################################################################################################################ @@ -36,13 +44,16 @@ def regions_to_list( Args: function_handle: the loader function you want to run. - regions: the region specifier + regions: the region specifier. Typically we expect to get many regions for this function, in the form of a list + of strings or bed file paths. regions_to_list will run across all of these one-by-one returning a separate + function return for each independent region. window_size: window around centers of regions, defaults to None quiet: disables progress bars - cores: CPU cores across which to parallelize processing + cores: CPU cores across which to parallelize processing. Default to None, which means all available. parallelize_within_regions: if True, regions will be run sequentially in parallelized chunks. If False, each individual region's chunks will be run sequentially but there will be parallelization across - regions, i.e. each core will be assigned one region at a time by the executor + regions, i.e. each core will be assigned one region at a time by the executor. Set to True if you + are running a small number of very large regions (e.g. one or two chromosomes), otherwise to to False (default). **kwargs: all necessary keyword arguments to pass down to the loader Returns: @@ -72,9 +83,9 @@ def regions_to_list( ) as executor: # Use functools.partial to pre-fill arguments process_partial = partial( - process_region, + apply_loader_function_to_region, function_handle=function_handle, - quiet=quiet or not parallelize_within_regions, # + quiet=quiet or not parallelize_within_regions, cores=cores_to_run if parallelize_within_regions else 1, # if parallelization is within region @@ -85,15 +96,16 @@ def regions_to_list( tqdm( executor.map(process_partial, region_tuples), total=len(region_tuples), - desc=f"Processing regions in parallel across {cores_to_run}", + desc="Loading data", disable=quiet or parallelize_within_regions, + leave=False, ) ) return results -def process_region(region_tuple, function_handle, **kwargs): +def apply_loader_function_to_region(region_tuple, function_handle, **kwargs): """ Helper function for regions_to_list. Takes in the region tuple, creates a string, and runs the loader function. @@ -123,7 +135,7 @@ def pileup_counts_from_bedmethyl( single_strand: bool = False, quiet: bool = False, cores: int | None = None, - chunk_size: int = 1_000_000, + chunk_size: int = DEFAULT_CHUNK_SIZE, ) -> tuple[int, int]: """ User-facing function. @@ -149,7 +161,7 @@ def pileup_counts_from_bedmethyl( single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions quiet: disables progress bars - cores: CPU cores across which to parallelize processing + cores: CPU cores across which to parallelize processing. Default to None, which means all available. chunk_size: size of genomic subregions to assign out to each process Returns: @@ -165,6 +177,7 @@ def pileup_counts_from_bedmethyl( cores_to_run = utils.cores_to_run(cores) + # Initialize shared memory as length-one numpy arrays to make it easy to map to buffer in subprocesses shm_valid = shared_memory.SharedMemory( create=True, size=np.dtype(np.int32).itemsize ) @@ -193,7 +206,8 @@ def pileup_counts_from_bedmethyl( concurrent.futures.as_completed(futures), total=len(futures), disable=quiet, - desc=f"Loading genomic chunks, up to {chunk_size/1000}kb per chunk", + desc="Loading data", + leave=False, ): try: future.result() @@ -225,7 +239,7 @@ def pileup_vectors_from_bedmethyl( regions_5to3prime: bool = False, quiet: bool = False, cores: int | None = None, - chunk_size: int = 1_000_000, + chunk_size: int = DEFAULT_CHUNK_SIZE, ) -> tuple[np.ndarray, np.ndarray]: """ User-facing function. @@ -260,7 +274,7 @@ def pileup_vectors_from_bedmethyl( the region of interest, False means we always grab both strands within the regions regions_5to3prime: True means negative strand regions get flipped, False means no flipping quiet: disables progress bars - cores: CPU cores across which to parallelize processing + cores: CPU cores across which to parallelize processing. Default to None, which means all available. chunk_size: size of genomic subregions to assign out to each process Returns: @@ -281,6 +295,7 @@ def pileup_vectors_from_bedmethyl( first_tuple = regions_dict[first_key][0] region_len = first_tuple[1] - first_tuple[0] + # Initialize shared memory as numpy arrays to make it easy to map to buffer in subprocesses shm_valid = shared_memory.SharedMemory( create=True, size=(region_len) * np.dtype(np.int32).itemsize ) @@ -311,7 +326,8 @@ def pileup_vectors_from_bedmethyl( concurrent.futures.as_completed(futures), total=len(futures), disable=quiet, - desc=f"Loading genomic chunks, up to {chunk_size/1000}kb per chunk", + desc="Loading data", + leave=False, ): try: future.result() @@ -351,7 +367,7 @@ def counts_from_fake(*args, **kwargs) -> tuple[int, int]: def vector_from_fake(window_size: int, *args, **kwargs) -> np.ndarray: """ - Tes helper function. + Test helper function. Generates a fake peak trace. Ignores all arguments except window_size. @@ -427,6 +443,8 @@ def pileup_vectors_process_chunk( valid_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) modified_base_subregion = np.zeros(subregion_end - subregion_start, dtype=int) + # tabix throws and error if the contig is not present + # by the current design, this should be silent if chromosome in source_tabix.contigs: for row in source_tabix.fetch( chromosome, max(subregion_start, 0), subregion_end @@ -510,6 +528,8 @@ def pileup_counts_process_chunk( valid_base_subregion_counts = 0 modified_base_subregion_counts = 0 + # tabix throws and error if the contig is not present + # by the current design, this should be silent if chromosome in source_tabix.contigs: for row in source_tabix.fetch( chromosome, max(subregion_start, 0), subregion_end diff --git a/dimelo/plot_depth_histogram.py b/dimelo/plot_depth_histogram.py index 949b4ec..a2527af 100644 --- a/dimelo/plot_depth_histogram.py +++ b/dimelo/plot_depth_histogram.py @@ -16,6 +16,7 @@ def plot_depth_histogram( average_within_region: bool = False, quiet: bool = False, cores: int | None = None, + parallelize_within_regions: bool = False, **kwargs, ) -> Axes: """ @@ -42,7 +43,11 @@ def plot_depth_histogram( the region of interest, False means we always grab both strands within the regions average_within_region: if True, each region will only report a single depth value, averaging across all non-zero depths quiet: disables progress bars - cores: CPU cores across which to parallelize processing + cores: CPU cores across which to parallelize processing. Default to None, which means all available. + parallelize_within_regions: if True, regions will be run sequentially in parallelized chunks. If False, + each individual region's chunks will be run sequentially but there will be parallelization across + regions, i.e. each core will be assigned one region at a time by the executor. Set to True if you + are running a small number of very large regions (e.g. one or two chromosomes), otherwise to to False (default). kwargs: other keyword parameters passed through to utils.line_plot Returns: diff --git a/dimelo/plot_depth_profile.py b/dimelo/plot_depth_profile.py index 0e59db1..52b8159 100644 --- a/dimelo/plot_depth_profile.py +++ b/dimelo/plot_depth_profile.py @@ -44,7 +44,7 @@ def plot_depth_profile( regions_5to3prime: True means negative strand regions get flipped, False means no flipping smooth_window: size of the moving window to use for smoothing. If set to None, no smoothing is performed quiet: disables progress bars - cores: CPU cores across which to parallelize processing + cores: CPU cores across which to parallelize processing. Default to None, which means all available. kwargs: other keyword parameters passed through to utils.line_plot Returns: diff --git a/dimelo/plot_enrichment.py b/dimelo/plot_enrichment.py index b30e8e4..d747b91 100644 --- a/dimelo/plot_enrichment.py +++ b/dimelo/plot_enrichment.py @@ -34,7 +34,7 @@ def plot_enrichment( single_strand: True means we only grab counts from reads from the same strand as the region of interest, False means we always grab both strands within the regions quiet: disables progress bars - cores: CPU cores across which to parallelize processing + cores: CPU cores across which to parallelize processing. Default to None, which means all available. kwargs: other keyword parameters passed through to utils.bar_plot Returns: @@ -49,6 +49,8 @@ def plot_enrichment( motifs=motifs, window_size=window_size, single_strand=single_strand, + quiet=quiet, + cores=cores, ) axes = make_enrichment_plot( diff --git a/dimelo/plot_enrichment_profile.py b/dimelo/plot_enrichment_profile.py index 0e339ec..97ec461 100644 --- a/dimelo/plot_enrichment_profile.py +++ b/dimelo/plot_enrichment_profile.py @@ -44,7 +44,7 @@ def plot_enrichment_profile( regions_5to3prime: True means negative strand regions get flipped, False means no flipping smooth_window: size of the moving window to use for smoothing. If set to None, no smoothing is performed quiet: disables progress bars - cores: CPU cores across which to parallelize processing + cores: CPU cores across which to parallelize processing. Default to None, which means all available. kwargs: other keyword parameters passed through to utils.line_plot Returns: @@ -61,6 +61,8 @@ def plot_enrichment_profile( single_strand=single_strand, regions_5to3prime=regions_5to3prime, smooth_window=smooth_window, + quiet=quiet, + cores=cores, ) axes = make_enrichment_profile_plot( diff --git a/dimelo/run_modkit.py b/dimelo/run_modkit.py index 298f0e9..c49106c 100644 --- a/dimelo/run_modkit.py +++ b/dimelo/run_modkit.py @@ -36,7 +36,8 @@ result = subprocess.run(["modkit", "--version"], stdout=subprocess.PIPE, text=True) modkit_version = result.stdout if modkit_version.split()[1] == EXPECTED_MODKIT_VERSION: - print(f"modkit found with expected version {EXPECTED_MODKIT_VERSION}") + pass + # print(f"modkit found with expected version {EXPECTED_MODKIT_VERSION}") else: print( f"modkit found with unexpected version {modkit_version.split()[1]}. Versions other than {EXPECTED_MODKIT_VERSION} may exhibit unexpected behavior. It is recommended that you use v{EXPECTED_MODKIT_VERSION}" diff --git a/dimelo/utils.py b/dimelo/utils.py index 29783c1..cfcd399 100644 --- a/dimelo/utils.py +++ b/dimelo/utils.py @@ -100,8 +100,13 @@ def adjust_threshold( def process_chunks_from_regions_dict( regions_dict: dict, - chunk_size: int = 10_000, + chunk_size: int, ): + """ + returns: a list of chunk specifier dictionaries, which contain region and subregion information. The subregion start and end + are always within the region. This information is sufficient for a downstream process to operate on the subregion chunk while + knowing where it lies within the larger region. + """ chunk_list = [] for chromosome, region_list in regions_dict.items(): for start_coord, end_coord, strand in region_list: From 7f5078202854f9376051f88a3ad0a644df8dd98d Mon Sep 17 00:00:00 2001 From: Oberon Dixon-Luinenburg Date: Thu, 29 May 2025 20:03:45 -0400 Subject: [PATCH 21/23] Adjust naming for regions_to_list region-splitting parallelization parameter. --- dimelo/load_processed.py | 12 ++++++------ dimelo/plot_depth_histogram.py | 4 ++-- 2 files changed, 8 insertions(+), 8 deletions(-) diff --git a/dimelo/load_processed.py b/dimelo/load_processed.py index 45f9e9c..62a4556 100644 --- a/dimelo/load_processed.py +++ b/dimelo/load_processed.py @@ -32,7 +32,7 @@ def regions_to_list( window_size: int | None = None, quiet: bool = True, cores: int | None = None, - parallelize_within_regions: bool = False, + split_large_regions: bool = False, **kwargs, ): """ @@ -49,7 +49,7 @@ def regions_to_list( window_size: window around centers of regions, defaults to None quiet: disables progress bars cores: CPU cores across which to parallelize processing. Default to None, which means all available. - parallelize_within_regions: if True, regions will be run sequentially in parallelized chunks. If False, + split_large_regions: if True, regions will be run sequentially in parallelized chunks. If False, each individual region's chunks will be run sequentially but there will be parallelization across regions, i.e. each core will be assigned one region at a time by the executor. Set to True if you are running a small number of very large regions (e.g. one or two chromosomes), otherwise to to False (default). @@ -78,15 +78,15 @@ def regions_to_list( # (2) the cores_to_run will be allocated to within-region parallelization, and the top-level # jobs sequence is run sequentially with concurrent.futures.ProcessPoolExecutor( - max_workers=1 if parallelize_within_regions else cores_to_run + max_workers=1 if split_large_regions else cores_to_run ) as executor: # Use functools.partial to pre-fill arguments process_partial = partial( apply_loader_function_to_region, function_handle=function_handle, - quiet=quiet or not parallelize_within_regions, + quiet=quiet or not split_large_regions, cores=cores_to_run - if parallelize_within_regions + if split_large_regions else 1, # if parallelization is within region **kwargs, ) @@ -95,7 +95,7 @@ def regions_to_list( executor.map(process_partial, region_strings), total=len(region_strings), desc="Loading data", - disable=quiet or parallelize_within_regions, + disable=quiet or split_large_regions, leave=False, ) ) diff --git a/dimelo/plot_depth_histogram.py b/dimelo/plot_depth_histogram.py index b09017d..95b1f09 100644 --- a/dimelo/plot_depth_histogram.py +++ b/dimelo/plot_depth_histogram.py @@ -16,7 +16,7 @@ def plot_depth_histogram( one_depth_per_region: bool = False, quiet: bool = False, cores: int | None = None, - parallelize_within_regions: bool = False, + split_large_regions: bool = False, **kwargs, ) -> Axes: """ @@ -40,7 +40,7 @@ def plot_depth_histogram( depths will be reported separately for all nonzero count positions in each region for a more granular view of depth distribution. quiet: disables progress bars cores: CPU cores across which to parallelize processing. Default to None, which means all available. - parallelize_within_regions: if True, regions will be run sequentially in parallelized chunks. If False, + split_large_regions: if True, regions will be run sequentially in parallelized chunks. If False, each individual region's chunks will be run sequentially but there will be parallelization across regions, i.e. each core will be assigned one region at a time by the executor. Set to True if you are running a small number of very large regions (e.g. one or two chromosomes), otherwise to to False (default). From 3ce0e8f546791c1762c14def39d1b3f107a5aad8 Mon Sep 17 00:00:00 2001 From: Jeremy Marcus Date: Fri, 30 May 2025 21:30:34 -0700 Subject: [PATCH 22/23] attempting to fix ruff versioning --- .github/workflows/formatting.yml | 4 ++-- .pre-commit-config.yaml | 2 +- 2 files changed, 3 insertions(+), 3 deletions(-) diff --git a/.github/workflows/formatting.yml b/.github/workflows/formatting.yml index 5edc917..32d34ba 100644 --- a/.github/workflows/formatting.yml +++ b/.github/workflows/formatting.yml @@ -5,12 +5,12 @@ jobs: runs-on: ubuntu-latest steps: - uses: actions/checkout@v4 - - uses: chartboost/ruff-action@v1 + - uses: chartboost/ruff-action@v3 # TODO: Is it really necessary for these to be separate jobs? This seems redundant. ruff-format-check: runs-on: ubuntu-latest steps: - uses: actions/checkout@v4 - - uses: chartboost/ruff-action@v1 + - uses: chartboost/ruff-action@v3 with: args: 'format --check' diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml index 5194a15..3a33fe9 100644 --- a/.pre-commit-config.yaml +++ b/.pre-commit-config.yaml @@ -1,6 +1,6 @@ repos: - repo: https://github.com/astral-sh/ruff-pre-commit - rev: v0.6.8 + rev: v0.11.12 hooks: # Run the linter. - id: ruff From 2972c3496d4f35b95983dd2d4b5332b8078f5df9 Mon Sep 17 00:00:00 2001 From: Jeremy Marcus Date: Fri, 30 May 2025 21:33:20 -0700 Subject: [PATCH 23/23] ruff version fix attempt 2 --- .github/workflows/formatting.yml | 7 +++++-- .pre-commit-config.yaml | 2 +- 2 files changed, 6 insertions(+), 3 deletions(-) diff --git a/.github/workflows/formatting.yml b/.github/workflows/formatting.yml index 32d34ba..5cac669 100644 --- a/.github/workflows/formatting.yml +++ b/.github/workflows/formatting.yml @@ -5,12 +5,15 @@ jobs: runs-on: ubuntu-latest steps: - uses: actions/checkout@v4 - - uses: chartboost/ruff-action@v3 + - uses: chartboost/ruff-action@v1 + with: + version: 0.6.8 # TODO: Is it really necessary for these to be separate jobs? This seems redundant. ruff-format-check: runs-on: ubuntu-latest steps: - uses: actions/checkout@v4 - - uses: chartboost/ruff-action@v3 + - uses: chartboost/ruff-action@v1 with: + version: 0.6.8 args: 'format --check' diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml index 3a33fe9..5194a15 100644 --- a/.pre-commit-config.yaml +++ b/.pre-commit-config.yaml @@ -1,6 +1,6 @@ repos: - repo: https://github.com/astral-sh/ruff-pre-commit - rev: v0.11.12 + rev: v0.6.8 hooks: # Run the linter. - id: ruff