Macarron is a workflow to systematically annotate and prioritize potentially bioactive (and often unannotated) small molecules in microbial community metabolomic datasets. Macarron prioritizes metabolic features as potentially bioactive in a phenotype/condition of interest using a combination of (a) covariance with annotated metabolites, (b) ecological properties such as abundance with respect to covarying annotated compounds, and (c) differential abundance in the phenotype/condition of interest.
If you have questions, please direct it to: Macarron Forum
Macarron requires R version 4.2.0 or higher. Install Bioconductor and then install Macarron:
if(!requireNamespace("BiocManager", quietly = TRUE))
install.packages("BiocManager")
BiocManager::install("Macarron")
Macarron can be run from the command line or as an R function. Both methods require the same
arguments, have the same options, and use the same default settings. The package includes the
wrapper Macarron()
as well as functions which perform different steps in the Macarron
framework.
Macarron requires 4 comma-separated, appropriately formatted input files. The files and their formatting constraints are described below.
If you do not have the chemical taxonomy file, you can generate this file using the annotation dataframe and Macarron utility decorate_ID
(see Advanced Topics).
By default, all files will be stored in a folder named Macarron_output inside the current working directory. The main prioritization results are stored in prioritized_metabolites_all.csv
. Another file, prioritized_metabolites_characterizable.csv
is a subset of prioritized_metabolites_all.csv
and only contains metabolic features which covary with at least one annotated metabolite.
The columns in these output files are:
Maaslin2
.Example (demo) input files can be found under inst/extdata
folder of the Macarron
source. These files were generated from the PRISM study of stool metabolomes of individuals with inflammatory bowel disease (IBD) and healthy “Control” individuals. Control and IBD are the two phenotypes in this example. Macarron will be applied to prioritize metabolic features with respect to their bioactivity in IBD. Therefore, in this example, the phenotype of interest is “IBD” and the reference phenotype is “Control”. The four input files are demo_abundances.csv
, demo_annotations.csv
, demo_metadata.csv
, and demo_taxonomy.csv
.
library(Macarron)
## Loading required package: SummarizedExperiment
## Loading required package: MatrixGenerics
## Loading required package: matrixStats
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## Attaching package: 'MatrixGenerics'
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## Welcome to Bioconductor
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## Vignettes contain introductory material; view with
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prism_abundances <- system.file(
'extdata','demo_abundances.csv', package="Macarron")
prism_annotations <-system.file(
'extdata','demo_annotations.csv', package="Macarron")
prism_metadata <-system.file(
'extdata','demo_metadata.csv', package="Macarron")
mets_taxonomy <-system.file(
'extdata','demo_taxonomy.csv', package="Macarron")
prism_prioritized <- Macarron::Macarron(input_abundances = prism_abundances,
input_annotations = prism_annotations,
input_metadata = prism_metadata,
input_taxonomy = mets_taxonomy)
## 2024-05-04 20:52:21.267416 INFO::Creating output folder.
## 2024-05-04 20:52:21.283474 INFO::Writing function arguments to log file
## Samples with both abundances and metadata: 102
## 2024-05-04 20:52:21.346835 INFO::Summarized Experiment created.
## 2024-05-04 20:52:21.348851 INFO::Metadata chosen for prevalence filtering: diagnosis
## 869 features pass chosen minimum prevalence threshold of 0.7.
## Calculating pairwise correlations in phenotype: IBD
##
## Calculating pairwise correlations in phenotype: Control
## Distance matrix with 869 features created.
## 2024-05-04 20:52:27.518943 INFO::Distance matrix with 869 metabolic features created.
## Initiating module detection
## 2024-05-04 20:52:27.520661 INFO::Minimum module size used for this dataset: 10
## Tree constructed
## Evaluating measures of success
## ..cutHeight not given, setting it to 0.99 ===> 99% of the (truncated) height range in dendro.
## ..done.
## ..cutHeight not given, setting it to 0.99 ===> 99% of the (truncated) height range in dendro.
## ..done.
## ..cutHeight not given, setting it to 0.99 ===> 99% of the (truncated) height range in dendro.
## ..done.
## ..cutHeight not given, setting it to 0.99 ===> 99% of the (truncated) height range in dendro.
## ..done.
## ..cutHeight not given, setting it to 0.99 ===> 99% of the (truncated) height range in dendro.
## ..done.
## 2024-05-04 20:52:29.905827 INFO::Total number of modules detected: 30
## Initiating AVA calculations
## Finding anchors
## Calculating AVA
## Initiating q-value calculations
## [1] "Creating output folder"
## [1] "Creating output feature tables folder"
## [1] "Creating output fits folder"
## [1] "Creating output figures folder"
## 2024-05-04 20:52:31.978141 INFO::Writing function arguments to log file
## 2024-05-04 20:52:31.984839 INFO::Verifying options selected are valid
## 2024-05-04 20:52:32.039071 INFO::Determining format of input files
## 2024-05-04 20:52:32.040645 INFO::Input format is data samples as columns and metadata samples as rows
## 2024-05-04 20:52:32.05908 INFO::Formula for fixed effects: expr ~ diagnosis + age + antibiotics
## 2024-05-04 20:52:32.061252 INFO::Filter data based on min abundance and min prevalence
## 2024-05-04 20:52:32.062095 INFO::Total samples in data: 102
## 2024-05-04 20:52:32.062812 INFO::Min samples required with min abundance for a feature not to be filtered: 0.000000
## 2024-05-04 20:52:32.075756 INFO::Total filtered features: 0
## 2024-05-04 20:52:32.077113 INFO::Filtered feature names from abundance and prevalence filtering:
## 2024-05-04 20:52:32.097346 INFO::Total filtered features with variance filtering: 0
## 2024-05-04 20:52:32.098733 INFO::Filtered feature names from variance filtering:
## 2024-05-04 20:52:32.099433 INFO::Running selected normalization method: NONE
## 2024-05-04 20:52:32.100248 INFO::Applying z-score to standardize continuous metadata
## 2024-05-04 20:52:32.1324 INFO::Running selected transform method: NONE
## 2024-05-04 20:52:32.133816 INFO::Running selected analysis method: LM
## 2024-05-04 20:52:32.14115 INFO::Fitting model to feature number 1, F1
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## 2024-05-04 20:52:32.878895 INFO::Fitting model to feature number 185, F191
## 2024-05-04 20:52:32.882749 INFO::Fitting model to feature number 186, F192
## 2024-05-04 20:52:32.886724 INFO::Fitting model to feature number 187, F193
## 2024-05-04 20:52:32.890857 INFO::Fitting model to feature number 188, F194
## 2024-05-04 20:52:32.894909 INFO::Fitting model to feature number 189, F195
## 2024-05-04 20:52:32.898903 INFO::Fitting model to feature number 190, F196
## 2024-05-04 20:52:32.902754 INFO::Fitting model to feature number 191, F197
## 2024-05-04 20:52:32.906629 INFO::Fitting model to feature number 192, F198
## 2024-05-04 20:52:32.910683 INFO::Fitting model to feature number 193, F199
## 2024-05-04 20:52:32.914491 INFO::Fitting model to feature number 194, F200
## 2024-05-04 20:52:32.918275 INFO::Fitting model to feature number 195, F201
## 2024-05-04 20:52:32.922094 INFO::Fitting model to feature number 196, F202
## 2024-05-04 20:52:32.925946 INFO::Fitting model to feature number 197, F203
## 2024-05-04 20:52:32.929756 INFO::Fitting model to feature number 198, F204
## 2024-05-04 20:52:32.933584 INFO::Fitting model to feature number 199, F206
## 2024-05-04 20:52:32.937361 INFO::Fitting model to feature number 200, F207
## 2024-05-04 20:52:32.941155 INFO::Fitting model to feature number 201, F208
## 2024-05-04 20:52:32.944962 INFO::Fitting model to feature number 202, F209
## 2024-05-04 20:52:32.94894 INFO::Fitting model to feature number 203, F210
## 2024-05-04 20:52:32.95274 INFO::Fitting model to feature number 204, F211
## 2024-05-04 20:52:32.956528 INFO::Fitting model to feature number 205, F212
## 2024-05-04 20:52:32.960355 INFO::Fitting model to feature number 206, F213
## 2024-05-04 20:52:32.964222 INFO::Fitting model to feature number 207, F214
## 2024-05-04 20:52:32.968067 INFO::Fitting model to feature number 208, F215
## 2024-05-04 20:52:32.971934 INFO::Fitting model to feature number 209, F216
## 2024-05-04 20:52:32.975769 INFO::Fitting model to feature number 210, F217
## 2024-05-04 20:52:32.979626 INFO::Fitting model to feature number 211, F218
## 2024-05-04 20:52:32.983458 INFO::Fitting model to feature number 212, F219
## 2024-05-04 20:52:32.987291 INFO::Fitting model to feature number 213, F220
## 2024-05-04 20:52:32.991086 INFO::Fitting model to feature number 214, F221
## 2024-05-04 20:52:32.99486 INFO::Fitting model to feature number 215, F222
## 2024-05-04 20:52:32.99868 INFO::Fitting model to feature number 216, F223
## 2024-05-04 20:52:33.002485 INFO::Fitting model to feature number 217, F224
## 2024-05-04 20:52:33.006351 INFO::Fitting model to feature number 218, F225
## 2024-05-04 20:52:33.010083 INFO::Fitting model to feature number 219, F226
## 2024-05-04 20:52:33.013825 INFO::Fitting model to feature number 220, F227
## 2024-05-04 20:52:33.01765 INFO::Fitting model to feature number 221, F228
## 2024-05-04 20:52:33.070407 INFO::Fitting model to feature number 222, F229
## 2024-05-04 20:52:33.074632 INFO::Fitting model to feature number 223, F230
## 2024-05-04 20:52:33.078457 INFO::Fitting model to feature number 224, F231
## 2024-05-04 20:52:33.082185 INFO::Fitting model to feature number 225, F232
## 2024-05-04 20:52:33.085891 INFO::Fitting model to feature number 226, F233
## 2024-05-04 20:52:33.089601 INFO::Fitting model to feature number 227, F234
## 2024-05-04 20:52:33.093296 INFO::Fitting model to feature number 228, F235
## 2024-05-04 20:52:33.096941 INFO::Fitting model to feature number 229, F236
## 2024-05-04 20:52:33.100693 INFO::Fitting model to feature number 230, F237
## 2024-05-04 20:52:33.104413 INFO::Fitting model to feature number 231, F238
## 2024-05-04 20:52:33.108141 INFO::Fitting model to feature number 232, F239
## 2024-05-04 20:52:33.111863 INFO::Fitting model to feature number 233, F240
## 2024-05-04 20:52:33.115544 INFO::Fitting model to feature number 234, F241
## 2024-05-04 20:52:33.119241 INFO::Fitting model to feature number 235, F242
## 2024-05-04 20:52:33.122942 INFO::Fitting model to feature number 236, F243
## 2024-05-04 20:52:33.126786 INFO::Fitting model to feature number 237, F244
## 2024-05-04 20:52:33.130448 INFO::Fitting model to feature number 238, F245
## 2024-05-04 20:52:33.134149 INFO::Fitting model to feature number 239, F246
## 2024-05-04 20:52:33.137861 INFO::Fitting model to feature number 240, F247
## 2024-05-04 20:52:33.141585 INFO::Fitting model to feature number 241, F248
## 2024-05-04 20:52:33.145291 INFO::Fitting model to feature number 242, F249
## 2024-05-04 20:52:33.149007 INFO::Fitting model to feature number 243, F250
## 2024-05-04 20:52:33.152733 INFO::Fitting model to feature number 244, F252
## 2024-05-04 20:52:33.156484 INFO::Fitting model to feature number 245, F253
## 2024-05-04 20:52:33.16023 INFO::Fitting model to feature number 246, F254
## 2024-05-04 20:52:33.164123 INFO::Fitting model to feature number 247, F255
## 2024-05-04 20:52:33.16787 INFO::Fitting model to feature number 248, F256
## 2024-05-04 20:52:33.171637 INFO::Fitting model to feature number 249, F257
## 2024-05-04 20:52:33.175468 INFO::Fitting model to feature number 250, F259
## 2024-05-04 20:52:33.179212 INFO::Fitting model to feature number 251, F260
## 2024-05-04 20:52:33.183115 INFO::Fitting model to feature number 252, F261
## 2024-05-04 20:52:33.186833 INFO::Fitting model to feature number 253, F262
## 2024-05-04 20:52:33.190619 INFO::Fitting model to feature number 254, F263
## 2024-05-04 20:52:33.194387 INFO::Fitting model to feature number 255, F264
## 2024-05-04 20:52:33.198109 INFO::Fitting model to feature number 256, F265
## 2024-05-04 20:52:33.201865 INFO::Fitting model to feature number 257, F266
## 2024-05-04 20:52:33.205654 INFO::Fitting model to feature number 258, F267
## 2024-05-04 20:52:33.2094 INFO::Fitting model to feature number 259, F269
## 2024-05-04 20:52:33.213159 INFO::Fitting model to feature number 260, F270
## 2024-05-04 20:52:33.216932 INFO::Fitting model to feature number 261, F271
## 2024-05-04 20:52:33.220706 INFO::Fitting model to feature number 262, F272
## 2024-05-04 20:52:33.224474 INFO::Fitting model to feature number 263, F273
## 2024-05-04 20:52:33.228266 INFO::Fitting model to feature number 264, F274
## 2024-05-04 20:52:33.232079 INFO::Fitting model to feature number 265, F276
## 2024-05-04 20:52:33.235783 INFO::Fitting model to feature number 266, F277
## 2024-05-04 20:52:33.239514 INFO::Fitting model to feature number 267, F278
## 2024-05-04 20:52:33.243241 INFO::Fitting model to feature number 268, F279
## 2024-05-04 20:52:33.246903 INFO::Fitting model to feature number 269, F280
## 2024-05-04 20:52:33.250606 INFO::Fitting model to feature number 270, F281
## 2024-05-04 20:52:33.254296 INFO::Fitting model to feature number 271, F282
## 2024-05-04 20:52:33.257969 INFO::Fitting model to feature number 272, F283
## 2024-05-04 20:52:33.26167 INFO::Fitting model to feature number 273, F284
## 2024-05-04 20:52:33.265476 INFO::Fitting model to feature number 274, F285
## 2024-05-04 20:52:33.269418 INFO::Fitting model to feature number 275, F286
## 2024-05-04 20:52:33.273271 INFO::Fitting model to feature number 276, F287
## 2024-05-04 20:52:33.277045 INFO::Fitting model to feature number 277, F288
## 2024-05-04 20:52:33.281052 INFO::Fitting model to feature number 278, F289
## 2024-05-04 20:52:33.285194 INFO::Fitting model to feature number 279, F290
## 2024-05-04 20:52:33.289051 INFO::Fitting model to feature number 280, F292
## 2024-05-04 20:52:33.292991 INFO::Fitting model to feature number 281, F293
## 2024-05-04 20:52:33.296876 INFO::Fitting model to feature number 282, F294
## 2024-05-04 20:52:33.300779 INFO::Fitting model to feature number 283, F295
## 2024-05-04 20:52:33.304653 INFO::Fitting model to feature number 284, F296
## 2024-05-04 20:52:33.308657 INFO::Fitting model to feature number 285, F297
## 2024-05-04 20:52:33.312529 INFO::Fitting model to feature number 286, F298
## 2024-05-04 20:52:33.316319 INFO::Fitting model to feature number 287, F299
## 2024-05-04 20:52:33.320129 INFO::Fitting model to feature number 288, F300
## 2024-05-04 20:52:33.324027 INFO::Fitting model to feature number 289, F301
## 2024-05-04 20:52:33.327907 INFO::Fitting model to feature number 290, F302
## 2024-05-04 20:52:33.331818 INFO::Fitting model to feature number 291, F303
## 2024-05-04 20:52:33.335626 INFO::Fitting model to feature number 292, F304
## 2024-05-04 20:52:33.339436 INFO::Fitting model to feature number 293, F305
## 2024-05-04 20:52:33.343408 INFO::Fitting model to feature number 294, F306
## 2024-05-04 20:52:33.347345 INFO::Fitting model to feature number 295, F307
## 2024-05-04 20:52:33.351207 INFO::Fitting model to feature number 296, F308
## 2024-05-04 20:52:33.355141 INFO::Fitting model to feature number 297, F309
## 2024-05-04 20:52:33.359197 INFO::Fitting model to feature number 298, F310
## 2024-05-04 20:52:33.363019 INFO::Fitting model to feature number 299, F311
## 2024-05-04 20:52:33.367103 INFO::Fitting model to feature number 300, F312
## 2024-05-04 20:52:33.371018 INFO::Fitting model to feature number 301, F313
## 2024-05-04 20:52:33.375123 INFO::Fitting model to feature number 302, F315
## 2024-05-04 20:52:33.379167 INFO::Fitting model to feature number 303, F316
## 2024-05-04 20:52:33.382987 INFO::Fitting model to feature number 304, F317
## 2024-05-04 20:52:33.387016 INFO::Fitting model to feature number 305, F318
## 2024-05-04 20:52:33.390965 INFO::Fitting model to feature number 306, F319
## 2024-05-04 20:52:33.394906 INFO::Fitting model to feature number 307, F320
## 2024-05-04 20:52:33.398971 INFO::Fitting model to feature number 308, F321
## 2024-05-04 20:52:33.402719 INFO::Fitting model to feature number 309, F322
## 2024-05-04 20:52:33.406458 INFO::Fitting model to feature number 310, F323
## 2024-05-04 20:52:33.41022 INFO::Fitting model to feature number 311, F324
## 2024-05-04 20:52:33.414066 INFO::Fitting model to feature number 312, F325
## 2024-05-04 20:52:33.417864 INFO::Fitting model to feature number 313, F326
## 2024-05-04 20:52:33.421665 INFO::Fitting model to feature number 314, F327
## 2024-05-04 20:52:33.425434 INFO::Fitting model to feature number 315, F328
## 2024-05-04 20:52:33.429264 INFO::Fitting model to feature number 316, F329
## 2024-05-04 20:52:33.433053 INFO::Fitting model to feature number 317, F330
## 2024-05-04 20:52:33.436755 INFO::Fitting model to feature number 318, F331
## 2024-05-04 20:52:33.440514 INFO::Fitting model to feature number 319, F332
## 2024-05-04 20:52:33.444388 INFO::Fitting model to feature number 320, F333
## 2024-05-04 20:52:33.448174 INFO::Fitting model to feature number 321, F334
## 2024-05-04 20:52:33.452009 INFO::Fitting model to feature number 322, F335
## 2024-05-04 20:52:33.455751 INFO::Fitting model to feature number 323, F336
## 2024-05-04 20:52:33.459501 INFO::Fitting model to feature number 324, F337
## 2024-05-04 20:52:33.46333 INFO::Fitting model to feature number 325, F338
## 2024-05-04 20:52:33.467075 INFO::Fitting model to feature number 326, F339
## 2024-05-04 20:52:33.470792 INFO::Fitting model to feature number 327, F340
## 2024-05-04 20:52:33.474453 INFO::Fitting model to feature number 328, F342
## 2024-05-04 20:52:33.478124 INFO::Fitting model to feature number 329, F343
## 2024-05-04 20:52:33.481795 INFO::Fitting model to feature number 330, F344
## 2024-05-04 20:52:33.485471 INFO::Fitting model to feature number 331, F345
## 2024-05-04 20:52:33.489226 INFO::Fitting model to feature number 332, F346
## 2024-05-04 20:52:33.492925 INFO::Fitting model to feature number 333, F347
## 2024-05-04 20:52:33.49661 INFO::Fitting model to feature number 334, F348
## 2024-05-04 20:52:33.500314 INFO::Fitting model to feature number 335, F350
## 2024-05-04 20:52:33.504055 INFO::Fitting model to feature number 336, F351
## 2024-05-04 20:52:33.507851 INFO::Fitting model to feature number 337, F352
## 2024-05-04 20:52:33.511639 INFO::Fitting model to feature number 338, F353
## 2024-05-04 20:52:33.515376 INFO::Fitting model to feature number 339, F355
## 2024-05-04 20:52:33.519234 INFO::Fitting model to feature number 340, F356
## 2024-05-04 20:52:33.522957 INFO::Fitting model to feature number 341, F357
## 2024-05-04 20:52:33.526727 INFO::Fitting model to feature number 342, F358
## 2024-05-04 20:52:33.530471 INFO::Fitting model to feature number 343, F359
## 2024-05-04 20:52:33.534267 INFO::Fitting model to feature number 344, F360
## 2024-05-04 20:52:33.537994 INFO::Fitting model to feature number 345, F361
## 2024-05-04 20:52:33.541751 INFO::Fitting model to feature number 346, F362
## 2024-05-04 20:52:33.545495 INFO::Fitting model to feature number 347, F363
## 2024-05-04 20:52:33.549279 INFO::Fitting model to feature number 348, F364
## 2024-05-04 20:52:33.553063 INFO::Fitting model to feature number 349, F365
## 2024-05-04 20:52:33.55687 INFO::Fitting model to feature number 350, F366
## 2024-05-04 20:52:33.560685 INFO::Fitting model to feature number 351, F367
## 2024-05-04 20:52:33.564517 INFO::Fitting model to feature number 352, F368
## 2024-05-04 20:52:33.568336 INFO::Fitting model to feature number 353, F369
## 2024-05-04 20:52:33.572133 INFO::Fitting model to feature number 354, F370
## 2024-05-04 20:52:33.575847 INFO::Fitting model to feature number 355, F371
## 2024-05-04 20:52:33.579571 INFO::Fitting model to feature number 356, F372
## 2024-05-04 20:52:33.583515 INFO::Fitting model to feature number 357, F373
## 2024-05-04 20:52:33.587381 INFO::Fitting model to feature number 358, F374
## 2024-05-04 20:52:33.591118 INFO::Fitting model to feature number 359, F375
## 2024-05-04 20:52:33.595113 INFO::Fitting model to feature number 360, F376
## 2024-05-04 20:52:33.599037 INFO::Fitting model to feature number 361, F377
## 2024-05-04 20:52:33.602816 INFO::Fitting model to feature number 362, F378
## 2024-05-04 20:52:33.606508 INFO::Fitting model to feature number 363, F379
## 2024-05-04 20:52:33.610258 INFO::Fitting model to feature number 364, F380
## 2024-05-04 20:52:33.614026 INFO::Fitting model to feature number 365, F381
## 2024-05-04 20:52:33.61783 INFO::Fitting model to feature number 366, F382
## 2024-05-04 20:52:33.621593 INFO::Fitting model to feature number 367, F383
## 2024-05-04 20:52:33.625308 INFO::Fitting model to feature number 368, F384
## 2024-05-04 20:52:33.62908 INFO::Fitting model to feature number 369, F386
## 2024-05-04 20:52:33.632857 INFO::Fitting model to feature number 370, F387
## 2024-05-04 20:52:33.636603 INFO::Fitting model to feature number 371, F388
## 2024-05-04 20:52:33.640366 INFO::Fitting model to feature number 372, F389
## 2024-05-04 20:52:33.644428 INFO::Fitting model to feature number 373, F390
## 2024-05-04 20:52:33.648424 INFO::Fitting model to feature number 374, F391
## 2024-05-04 20:52:33.652279 INFO::Fitting model to feature number 375, F392
## 2024-05-04 20:52:33.656189 INFO::Fitting model to feature number 376, F393
## 2024-05-04 20:52:33.660201 INFO::Fitting model to feature number 377, F394
## 2024-05-04 20:52:33.664081 INFO::Fitting model to feature number 378, F395
## 2024-05-04 20:52:33.66813 INFO::Fitting model to feature number 379, F396
## 2024-05-04 20:52:33.672178 INFO::Fitting model to feature number 380, F397
## 2024-05-04 20:52:33.67611 INFO::Fitting model to feature number 381, F398
## 2024-05-04 20:52:33.680193 INFO::Fitting model to feature number 382, F399
## 2024-05-04 20:52:33.684156 INFO::Fitting model to feature number 383, F400
## 2024-05-04 20:52:33.688243 INFO::Fitting model to feature number 384, F401
## 2024-05-04 20:52:33.692161 INFO::Fitting model to feature number 385, F402
## 2024-05-04 20:52:33.696152 INFO::Fitting model to feature number 386, F403
## 2024-05-04 20:52:33.700176 INFO::Fitting model to feature number 387, F404
## 2024-05-04 20:52:33.704092 INFO::Fitting model to feature number 388, F406
## 2024-05-04 20:52:33.708091 INFO::Fitting model to feature number 389, F407
## 2024-05-04 20:52:33.711968 INFO::Fitting model to feature number 390, F408
## 2024-05-04 20:52:33.716003 INFO::Fitting model to feature number 391, F409
## 2024-05-04 20:52:33.719899 INFO::Fitting model to feature number 392, F410
## 2024-05-04 20:52:33.723949 INFO::Fitting model to feature number 393, F411
## 2024-05-04 20:52:33.727785 INFO::Fitting model to feature number 394, F412
## 2024-05-04 20:52:33.731761 INFO::Fitting model to feature number 395, F413
## 2024-05-04 20:52:33.735739 INFO::Fitting model to feature number 396, F414
## 2024-05-04 20:52:33.739671 INFO::Fitting model to feature number 397, F415
## 2024-05-04 20:52:33.743718 INFO::Fitting model to feature number 398, F416
## 2024-05-04 20:52:33.747498 INFO::Fitting model to feature number 399, F417
## 2024-05-04 20:52:33.751425 INFO::Fitting model to feature number 400, F418
## 2024-05-04 20:52:33.755589 INFO::Fitting model to feature number 401, F419
## 2024-05-04 20:52:33.759698 INFO::Fitting model to feature number 402, F420
## 2024-05-04 20:52:33.763426 INFO::Fitting model to feature number 403, F421
## 2024-05-04 20:52:33.767335 INFO::Fitting model to feature number 404, F422
## 2024-05-04 20:52:33.771306 INFO::Fitting model to feature number 405, F423
## 2024-05-04 20:52:33.775222 INFO::Fitting model to feature number 406, F425
## 2024-05-04 20:52:33.779187 INFO::Fitting model to feature number 407, F426
## 2024-05-04 20:52:33.783148 INFO::Fitting model to feature number 408, F428
## 2024-05-04 20:52:33.787093 INFO::Fitting model to feature number 409, F429
## 2024-05-04 20:52:33.791102 INFO::Fitting model to feature number 410, F430
## 2024-05-04 20:52:33.795188 INFO::Fitting model to feature number 411, F431
## 2024-05-04 20:52:33.799182 INFO::Fitting model to feature number 412, F432
## 2024-05-04 20:52:33.803124 INFO::Fitting model to feature number 413, F433
## 2024-05-04 20:52:33.807045 INFO::Fitting model to feature number 414, F434
## 2024-05-04 20:52:33.810976 INFO::Fitting model to feature number 415, F435
## 2024-05-04 20:52:33.814946 INFO::Fitting model to feature number 416, F436
## 2024-05-04 20:52:33.81888 INFO::Fitting model to feature number 417, F437
## 2024-05-04 20:52:33.822838 INFO::Fitting model to feature number 418, F438
## 2024-05-04 20:52:33.826719 INFO::Fitting model to feature number 419, F439
## 2024-05-04 20:52:33.830612 INFO::Fitting model to feature number 420, F440
## 2024-05-04 20:52:33.834512 INFO::Fitting model to feature number 421, F441
## 2024-05-04 20:52:33.838388 INFO::Fitting model to feature number 422, F442
## 2024-05-04 20:52:33.842281 INFO::Fitting model to feature number 423, F443
## 2024-05-04 20:52:33.846183 INFO::Fitting model to feature number 424, F444
## 2024-05-04 20:52:33.850115 INFO::Fitting model to feature number 425, F445
## 2024-05-04 20:52:33.853827 INFO::Fitting model to feature number 426, F446
## 2024-05-04 20:52:33.857743 INFO::Fitting model to feature number 427, F447
## 2024-05-04 20:52:33.861517 INFO::Fitting model to feature number 428, F448
## 2024-05-04 20:52:33.865247 INFO::Fitting model to feature number 429, F449
## 2024-05-04 20:52:33.869011 INFO::Fitting model to feature number 430, F450
## 2024-05-04 20:52:33.872766 INFO::Fitting model to feature number 431, F451
## 2024-05-04 20:52:33.876561 INFO::Fitting model to feature number 432, F452
## 2024-05-04 20:52:33.880475 INFO::Fitting model to feature number 433, F454
## 2024-05-04 20:52:33.88435 INFO::Fitting model to feature number 434, F455
## 2024-05-04 20:52:33.888254 INFO::Fitting model to feature number 435, F456
## 2024-05-04 20:52:33.892179 INFO::Fitting model to feature number 436, F457
## 2024-05-04 20:52:33.896075 INFO::Fitting model to feature number 437, F458
## 2024-05-04 20:52:33.9 INFO::Fitting model to feature number 438, F459
## 2024-05-04 20:52:33.903952 INFO::Fitting model to feature number 439, F461
## 2024-05-04 20:52:33.907918 INFO::Fitting model to feature number 440, F462
## 2024-05-04 20:52:33.911785 INFO::Fitting model to feature number 441, F463
## 2024-05-04 20:52:33.915704 INFO::Fitting model to feature number 442, F464
## 2024-05-04 20:52:33.919603 INFO::Fitting model to feature number 443, F465
## 2024-05-04 20:52:33.923512 INFO::Fitting model to feature number 444, F466
## 2024-05-04 20:52:33.927364 INFO::Fitting model to feature number 445, F467
## 2024-05-04 20:52:33.931188 INFO::Fitting model to feature number 446, F468
## 2024-05-04 20:52:33.935085 INFO::Fitting model to feature number 447, F469
## 2024-05-04 20:52:33.938992 INFO::Fitting model to feature number 448, F470
## 2024-05-04 20:52:33.942896 INFO::Fitting model to feature number 449, F471
## 2024-05-04 20:52:33.946977 INFO::Fitting model to feature number 450, F474
## 2024-05-04 20:52:33.950968 INFO::Fitting model to feature number 451, F475
## 2024-05-04 20:52:33.954834 INFO::Fitting model to feature number 452, F476
## 2024-05-04 20:52:33.958771 INFO::Fitting model to feature number 453, F477
## 2024-05-04 20:52:33.96263 INFO::Fitting model to feature number 454, F478
## 2024-05-04 20:52:33.966538 INFO::Fitting model to feature number 455, F479
## 2024-05-04 20:52:33.970514 INFO::Fitting model to feature number 456, F480
## 2024-05-04 20:52:33.974348 INFO::Fitting model to feature number 457, F481
## 2024-05-04 20:52:33.978177 INFO::Fitting model to feature number 458, F482
## 2024-05-04 20:52:33.982036 INFO::Fitting model to feature number 459, F483
## 2024-05-04 20:52:33.98593 INFO::Fitting model to feature number 460, F484
## 2024-05-04 20:52:33.989797 INFO::Fitting model to feature number 461, F485
## 2024-05-04 20:52:33.993654 INFO::Fitting model to feature number 462, F486
## 2024-05-04 20:52:33.99749 INFO::Fitting model to feature number 463, F487
## 2024-05-04 20:52:34.001377 INFO::Fitting model to feature number 464, F488
## 2024-05-04 20:52:34.005204 INFO::Fitting model to feature number 465, F489
## 2024-05-04 20:52:34.009305 INFO::Fitting model to feature number 466, F490
## 2024-05-04 20:52:34.013151 INFO::Fitting model to feature number 467, F491
## 2024-05-04 20:52:34.017112 INFO::Fitting model to feature number 468, F492
## 2024-05-04 20:52:34.021199 INFO::Fitting model to feature number 469, F493
## 2024-05-04 20:52:34.025222 INFO::Fitting model to feature number 470, F494
## 2024-05-04 20:52:34.029038 INFO::Fitting model to feature number 471, F495
## 2024-05-04 20:52:34.033031 INFO::Fitting model to feature number 472, F496
## 2024-05-04 20:52:34.037126 INFO::Fitting model to feature number 473, F497
## 2024-05-04 20:52:34.041288 INFO::Fitting model to feature number 474, F498
## 2024-05-04 20:52:34.045421 INFO::Fitting model to feature number 475, F499
## 2024-05-04 20:52:34.049584 INFO::Fitting model to feature number 476, F500
## 2024-05-04 20:52:34.05379 INFO::Fitting model to feature number 477, F501
## 2024-05-04 20:52:34.057898 INFO::Fitting model to feature number 478, F502
## 2024-05-04 20:52:34.062061 INFO::Fitting model to feature number 479, F503
## 2024-05-04 20:52:34.066214 INFO::Fitting model to feature number 480, F504
## 2024-05-04 20:52:34.070389 INFO::Fitting model to feature number 481, F505
## 2024-05-04 20:52:34.074574 INFO::Fitting model to feature number 482, F506
## 2024-05-04 20:52:34.078686 INFO::Fitting model to feature number 483, F507
## 2024-05-04 20:52:34.082839 INFO::Fitting model to feature number 484, F508
## 2024-05-04 20:52:34.086928 INFO::Fitting model to feature number 485, F509
## 2024-05-04 20:52:34.091063 INFO::Fitting model to feature number 486, F510
## 2024-05-04 20:52:34.095186 INFO::Fitting model to feature number 487, F511
## 2024-05-04 20:52:34.099315 INFO::Fitting model to feature number 488, F512
## 2024-05-04 20:52:34.103445 INFO::Fitting model to feature number 489, F513
## 2024-05-04 20:52:34.10761 INFO::Fitting model to feature number 490, F514
## 2024-05-04 20:52:34.111764 INFO::Fitting model to feature number 491, F515
## 2024-05-04 20:52:34.115893 INFO::Fitting model to feature number 492, F516
## 2024-05-04 20:52:34.120031 INFO::Fitting model to feature number 493, F517
## 2024-05-04 20:52:34.124167 INFO::Fitting model to feature number 494, F518
## 2024-05-04 20:52:34.128327 INFO::Fitting model to feature number 495, F519
## 2024-05-04 20:52:34.132624 INFO::Fitting model to feature number 496, F520
## 2024-05-04 20:52:34.136775 INFO::Fitting model to feature number 497, F521
## 2024-05-04 20:52:34.141042 INFO::Fitting model to feature number 498, F522
## 2024-05-04 20:52:34.14519 INFO::Fitting model to feature number 499, F523
## 2024-05-04 20:52:34.149245 INFO::Fitting model to feature number 500, F524
## 2024-05-04 20:52:34.153353 INFO::Fitting model to feature number 501, F525
## 2024-05-04 20:52:34.157475 INFO::Fitting model to feature number 502, F526
## 2024-05-04 20:52:34.161558 INFO::Fitting model to feature number 503, F527
## 2024-05-04 20:52:34.165621 INFO::Fitting model to feature number 504, F528
## 2024-05-04 20:52:34.169726 INFO::Fitting model to feature number 505, F529
## 2024-05-04 20:52:34.173783 INFO::Fitting model to feature number 506, F530
## 2024-05-04 20:52:34.177883 INFO::Fitting model to feature number 507, F531
## 2024-05-04 20:52:34.181974 INFO::Fitting model to feature number 508, F532
## 2024-05-04 20:52:34.18605 INFO::Fitting model to feature number 509, F533
## 2024-05-04 20:52:34.190154 INFO::Fitting model to feature number 510, F534
## 2024-05-04 20:52:34.194286 INFO::Fitting model to feature number 511, F535
## 2024-05-04 20:52:34.198436 INFO::Fitting model to feature number 512, F536
## 2024-05-04 20:52:34.20264 INFO::Fitting model to feature number 513, F537
## 2024-05-04 20:52:34.206763 INFO::Fitting model to feature number 514, F539
## 2024-05-04 20:52:34.210918 INFO::Fitting model to feature number 515, F540
## 2024-05-04 20:52:34.214993 INFO::Fitting model to feature number 516, F541
## 2024-05-04 20:52:34.21909 INFO::Fitting model to feature number 517, F543
## 2024-05-04 20:52:34.223302 INFO::Fitting model to feature number 518, F544
## 2024-05-04 20:52:34.227557 INFO::Fitting model to feature number 519, F545
## 2024-05-04 20:52:34.231544 INFO::Fitting model to feature number 520, F546
## 2024-05-04 20:52:34.235652 INFO::Fitting model to feature number 521, F547
## 2024-05-04 20:52:34.239578 INFO::Fitting model to feature number 522, F548
## 2024-05-04 20:52:34.243778 INFO::Fitting model to feature number 523, F549
## 2024-05-04 20:52:34.247862 INFO::Fitting model to feature number 524, F550
## 2024-05-04 20:52:34.251899 INFO::Fitting model to feature number 525, F551
## 2024-05-04 20:52:34.255778 INFO::Fitting model to feature number 526, F552
## 2024-05-04 20:52:34.259655 INFO::Fitting model to feature number 527, F553
## 2024-05-04 20:52:34.263489 INFO::Fitting model to feature number 528, F554
## 2024-05-04 20:52:34.267313 INFO::Fitting model to feature number 529, F555
## 2024-05-04 20:52:34.271082 INFO::Fitting model to feature number 530, F556
## 2024-05-04 20:52:34.274944 INFO::Fitting model to feature number 531, F557
## 2024-05-04 20:52:34.278808 INFO::Fitting model to feature number 532, F558
## 2024-05-04 20:52:34.282685 INFO::Fitting model to feature number 533, F559
## 2024-05-04 20:52:34.286711 INFO::Fitting model to feature number 534, F560
## 2024-05-04 20:52:34.291362 INFO::Fitting model to feature number 535, F561
## 2024-05-04 20:52:34.296402 INFO::Fitting model to feature number 536, F562
## 2024-05-04 20:52:34.301471 INFO::Fitting model to feature number 537, F563
## 2024-05-04 20:52:34.306608 INFO::Fitting model to feature number 538, F564
## 2024-05-04 20:52:34.311834 INFO::Fitting model to feature number 539, F566
## 2024-05-04 20:52:34.317159 INFO::Fitting model to feature number 540, F567
## 2024-05-04 20:52:34.322398 INFO::Fitting model to feature number 541, F568
## 2024-05-04 20:52:34.327659 INFO::Fitting model to feature number 542, F569
## 2024-05-04 20:52:34.332383 INFO::Fitting model to feature number 543, F570
## 2024-05-04 20:52:34.337387 INFO::Fitting model to feature number 544, F571
## 2024-05-04 20:52:34.342408 INFO::Fitting model to feature number 545, F572
## 2024-05-04 20:52:34.347324 INFO::Fitting model to feature number 546, F573
## 2024-05-04 20:52:34.352648 INFO::Fitting model to feature number 547, F574
## 2024-05-04 20:52:34.357607 INFO::Fitting model to feature number 548, F575
## 2024-05-04 20:52:34.361661 INFO::Fitting model to feature number 549, F576
## 2024-05-04 20:52:34.365881 INFO::Fitting model to feature number 550, F577
## 2024-05-04 20:52:34.41635 INFO::Fitting model to feature number 551, F578
## 2024-05-04 20:52:34.421268 INFO::Fitting model to feature number 552, F579
## 2024-05-04 20:52:34.425383 INFO::Fitting model to feature number 553, F580
## 2024-05-04 20:52:34.429522 INFO::Fitting model to feature number 554, F581
## 2024-05-04 20:52:34.433698 INFO::Fitting model to feature number 555, F582
## 2024-05-04 20:52:34.437812 INFO::Fitting model to feature number 556, F583
## 2024-05-04 20:52:34.441822 INFO::Fitting model to feature number 557, F584
## 2024-05-04 20:52:34.445888 INFO::Fitting model to feature number 558, F585
## 2024-05-04 20:52:34.450044 INFO::Fitting model to feature number 559, F586
## 2024-05-04 20:52:34.454153 INFO::Fitting model to feature number 560, F587
## 2024-05-04 20:52:34.458167 INFO::Fitting model to feature number 561, F588
## 2024-05-04 20:52:34.462246 INFO::Fitting model to feature number 562, F589
## 2024-05-04 20:52:34.466402 INFO::Fitting model to feature number 563, F590
## 2024-05-04 20:52:34.470469 INFO::Fitting model to feature number 564, F591
## 2024-05-04 20:52:34.474436 INFO::Fitting model to feature number 565, F592
## 2024-05-04 20:52:34.478497 INFO::Fitting model to feature number 566, F593
## 2024-05-04 20:52:34.48277 INFO::Fitting model to feature number 567, F594
## 2024-05-04 20:52:34.486894 INFO::Fitting model to feature number 568, F595
## 2024-05-04 20:52:34.490882 INFO::Fitting model to feature number 569, F596
## 2024-05-04 20:52:34.494942 INFO::Fitting model to feature number 570, F597
## 2024-05-04 20:52:34.499082 INFO::Fitting model to feature number 571, F598
## 2024-05-04 20:52:34.503127 INFO::Fitting model to feature number 572, F599
## 2024-05-04 20:52:34.507218 INFO::Fitting model to feature number 573, F600
## 2024-05-04 20:52:34.511196 INFO::Fitting model to feature number 574, F601
## 2024-05-04 20:52:34.515386 INFO::Fitting model to feature number 575, F602
## 2024-05-04 20:52:34.520157 INFO::Fitting model to feature number 576, F603
## 2024-05-04 20:52:34.524425 INFO::Fitting model to feature number 577, F605
## 2024-05-04 20:52:34.528537 INFO::Fitting model to feature number 578, F606
## 2024-05-04 20:52:34.533151 INFO::Fitting model to feature number 579, F607
## 2024-05-04 20:52:34.537 INFO::Fitting model to feature number 580, F608
## 2024-05-04 20:52:34.540812 INFO::Fitting model to feature number 581, F609
## 2024-05-04 20:52:34.544505 INFO::Fitting model to feature number 582, F610
## 2024-05-04 20:52:34.548124 INFO::Fitting model to feature number 583, F611
## 2024-05-04 20:52:34.55189 INFO::Fitting model to feature number 584, F612
## 2024-05-04 20:52:34.55557 INFO::Fitting model to feature number 585, F613
## 2024-05-04 20:52:34.559259 INFO::Fitting model to feature number 586, F614
## 2024-05-04 20:52:34.562935 INFO::Fitting model to feature number 587, F615
## 2024-05-04 20:52:34.566599 INFO::Fitting model to feature number 588, F616
## 2024-05-04 20:52:34.570411 INFO::Fitting model to feature number 589, F617
## 2024-05-04 20:52:34.574279 INFO::Fitting model to feature number 590, F618
## 2024-05-04 20:52:34.578099 INFO::Fitting model to feature number 591, F619
## 2024-05-04 20:52:34.581862 INFO::Fitting model to feature number 592, F620
## 2024-05-04 20:52:34.585608 INFO::Fitting model to feature number 593, F621
## 2024-05-04 20:52:34.589414 INFO::Fitting model to feature number 594, F622
## 2024-05-04 20:52:34.593216 INFO::Fitting model to feature number 595, F623
## 2024-05-04 20:52:34.597001 INFO::Fitting model to feature number 596, F624
## 2024-05-04 20:52:34.600892 INFO::Fitting model to feature number 597, F625
## 2024-05-04 20:52:34.604536 INFO::Fitting model to feature number 598, F626
## 2024-05-04 20:52:34.608147 INFO::Fitting model to feature number 599, F627
## 2024-05-04 20:52:34.611824 INFO::Fitting model to feature number 600, F628
## 2024-05-04 20:52:34.615588 INFO::Fitting model to feature number 601, F629
## 2024-05-04 20:52:34.619293 INFO::Fitting model to feature number 602, F630
## 2024-05-04 20:52:34.622981 INFO::Fitting model to feature number 603, F631
## 2024-05-04 20:52:34.626673 INFO::Fitting model to feature number 604, F632
## 2024-05-04 20:52:34.630395 INFO::Fitting model to feature number 605, F633
## 2024-05-04 20:52:34.634132 INFO::Fitting model to feature number 606, F634
## 2024-05-04 20:52:34.637893 INFO::Fitting model to feature number 607, F635
## 2024-05-04 20:52:34.641558 INFO::Fitting model to feature number 608, F636
## 2024-05-04 20:52:34.645228 INFO::Fitting model to feature number 609, F637
## 2024-05-04 20:52:34.64905 INFO::Fitting model to feature number 610, F638
## 2024-05-04 20:52:34.652836 INFO::Fitting model to feature number 611, F639
## 2024-05-04 20:52:34.656562 INFO::Fitting model to feature number 612, F640
## 2024-05-04 20:52:34.66029 INFO::Fitting model to feature number 613, F641
## 2024-05-04 20:52:34.664067 INFO::Fitting model to feature number 614, F642
## 2024-05-04 20:52:34.667806 INFO::Fitting model to feature number 615, F643
## 2024-05-04 20:52:34.671579 INFO::Fitting model to feature number 616, F644
## 2024-05-04 20:52:34.675387 INFO::Fitting model to feature number 617, F645
## 2024-05-04 20:52:34.679237 INFO::Fitting model to feature number 618, F646
## 2024-05-04 20:52:34.683137 INFO::Fitting model to feature number 619, F647
## 2024-05-04 20:52:34.686928 INFO::Fitting model to feature number 620, F648
## 2024-05-04 20:52:34.690732 INFO::Fitting model to feature number 621, F649
## 2024-05-04 20:52:34.694513 INFO::Fitting model to feature number 622, F650
## 2024-05-04 20:52:34.698299 INFO::Fitting model to feature number 623, F651
## 2024-05-04 20:52:34.702077 INFO::Fitting model to feature number 624, F652
## 2024-05-04 20:52:34.705837 INFO::Fitting model to feature number 625, F653
## 2024-05-04 20:52:34.709589 INFO::Fitting model to feature number 626, F654
## 2024-05-04 20:52:34.713352 INFO::Fitting model to feature number 627, F655
## 2024-05-04 20:52:34.717109 INFO::Fitting model to feature number 628, F656
## 2024-05-04 20:52:34.720786 INFO::Fitting model to feature number 629, F657
## 2024-05-04 20:52:34.724723 INFO::Fitting model to feature number 630, F658
## 2024-05-04 20:52:34.728513 INFO::Fitting model to feature number 631, F659
## 2024-05-04 20:52:34.732271 INFO::Fitting model to feature number 632, F660
## 2024-05-04 20:52:34.736021 INFO::Fitting model to feature number 633, F661
## 2024-05-04 20:52:34.739754 INFO::Fitting model to feature number 634, F662
## 2024-05-04 20:52:34.743487 INFO::Fitting model to feature number 635, F663
## 2024-05-04 20:52:34.747152 INFO::Fitting model to feature number 636, F664
## 2024-05-04 20:52:34.75097 INFO::Fitting model to feature number 637, F665
## 2024-05-04 20:52:34.754712 INFO::Fitting model to feature number 638, F666
## 2024-05-04 20:52:34.758469 INFO::Fitting model to feature number 639, F667
## 2024-05-04 20:52:34.76221 INFO::Fitting model to feature number 640, F668
## 2024-05-04 20:52:34.765969 INFO::Fitting model to feature number 641, F669
## 2024-05-04 20:52:34.769699 INFO::Fitting model to feature number 642, F670
## 2024-05-04 20:52:34.773449 INFO::Fitting model to feature number 643, F671
## 2024-05-04 20:52:34.77716 INFO::Fitting model to feature number 644, F672
## 2024-05-04 20:52:34.780924 INFO::Fitting model to feature number 645, F673
## 2024-05-04 20:52:34.784727 INFO::Fitting model to feature number 646, F674
## 2024-05-04 20:52:34.788428 INFO::Fitting model to feature number 647, F675
## 2024-05-04 20:52:34.792118 INFO::Fitting model to feature number 648, F676
## 2024-05-04 20:52:34.795908 INFO::Fitting model to feature number 649, F677
## 2024-05-04 20:52:34.799703 INFO::Fitting model to feature number 650, F678
## 2024-05-04 20:52:34.803437 INFO::Fitting model to feature number 651, F679
## 2024-05-04 20:52:34.807176 INFO::Fitting model to feature number 652, F680
## 2024-05-04 20:52:34.810924 INFO::Fitting model to feature number 653, F681
## 2024-05-04 20:52:34.814719 INFO::Fitting model to feature number 654, F682
## 2024-05-04 20:52:34.818483 INFO::Fitting model to feature number 655, F683
## 2024-05-04 20:52:34.822234 INFO::Fitting model to feature number 656, F684
## 2024-05-04 20:52:34.826005 INFO::Fitting model to feature number 657, F685
## 2024-05-04 20:52:34.829749 INFO::Fitting model to feature number 658, F686
## 2024-05-04 20:52:34.833447 INFO::Fitting model to feature number 659, F687
## 2024-05-04 20:52:34.837149 INFO::Fitting model to feature number 660, F688
## 2024-05-04 20:52:34.840978 INFO::Fitting model to feature number 661, F689
## 2024-05-04 20:52:34.844782 INFO::Fitting model to feature number 662, F690
## 2024-05-04 20:52:34.848556 INFO::Fitting model to feature number 663, F691
## 2024-05-04 20:52:34.852291 INFO::Fitting model to feature number 664, F692
## 2024-05-04 20:52:34.856058 INFO::Fitting model to feature number 665, F693
## 2024-05-04 20:52:34.859668 INFO::Fitting model to feature number 666, F694
## 2024-05-04 20:52:34.863493 INFO::Fitting model to feature number 667, F695
## 2024-05-04 20:52:34.867285 INFO::Fitting model to feature number 668, F696
## 2024-05-04 20:52:34.871065 INFO::Fitting model to feature number 669, F697
## 2024-05-04 20:52:34.874742 INFO::Fitting model to feature number 670, F698
## 2024-05-04 20:52:34.878427 INFO::Fitting model to feature number 671, F699
## 2024-05-04 20:52:34.882053 INFO::Fitting model to feature number 672, F700
## 2024-05-04 20:52:34.885745 INFO::Fitting model to feature number 673, F701
## 2024-05-04 20:52:34.889506 INFO::Fitting model to feature number 674, F702
## 2024-05-04 20:52:34.893296 INFO::Fitting model to feature number 675, F704
## 2024-05-04 20:52:34.89709 INFO::Fitting model to feature number 676, F705
## 2024-05-04 20:52:34.900877 INFO::Fitting model to feature number 677, F706
## 2024-05-04 20:52:34.904715 INFO::Fitting model to feature number 678, F707
## 2024-05-04 20:52:34.908589 INFO::Fitting model to feature number 679, F708
## 2024-05-04 20:52:34.912458 INFO::Fitting model to feature number 680, F709
## 2024-05-04 20:52:34.916201 INFO::Fitting model to feature number 681, F710
## 2024-05-04 20:52:34.920035 INFO::Fitting model to feature number 682, F711
## 2024-05-04 20:52:34.924021 INFO::Fitting model to feature number 683, F712
## 2024-05-04 20:52:34.92772 INFO::Fitting model to feature number 684, F713
## 2024-05-04 20:52:34.931791 INFO::Fitting model to feature number 685, F714
## 2024-05-04 20:52:34.935551 INFO::Fitting model to feature number 686, F715
## 2024-05-04 20:52:34.939306 INFO::Fitting model to feature number 687, F716
## 2024-05-04 20:52:34.942976 INFO::Fitting model to feature number 688, F717
## 2024-05-04 20:52:34.946824 INFO::Fitting model to feature number 689, F718
## 2024-05-04 20:52:34.95073 INFO::Fitting model to feature number 690, F719
## 2024-05-04 20:52:34.954457 INFO::Fitting model to feature number 691, F720
## 2024-05-04 20:52:34.958202 INFO::Fitting model to feature number 692, F721
## 2024-05-04 20:52:34.961989 INFO::Fitting model to feature number 693, F722
## 2024-05-04 20:52:34.965776 INFO::Fitting model to feature number 694, F723
## 2024-05-04 20:52:34.969578 INFO::Fitting model to feature number 695, F724
## 2024-05-04 20:52:34.973466 INFO::Fitting model to feature number 696, F725
## 2024-05-04 20:52:34.977247 INFO::Fitting model to feature number 697, F726
## 2024-05-04 20:52:34.980951 INFO::Fitting model to feature number 698, F727
## 2024-05-04 20:52:34.984704 INFO::Fitting model to feature number 699, F728
## 2024-05-04 20:52:34.988397 INFO::Fitting model to feature number 700, F729
## 2024-05-04 20:52:34.992139 INFO::Fitting model to feature number 701, F730
## 2024-05-04 20:52:34.995904 INFO::Fitting model to feature number 702, F731
## 2024-05-04 20:52:34.999682 INFO::Fitting model to feature number 703, F732
## 2024-05-04 20:52:35.003429 INFO::Fitting model to feature number 704, F733
## 2024-05-04 20:52:35.007202 INFO::Fitting model to feature number 705, F734
## 2024-05-04 20:52:35.010933 INFO::Fitting model to feature number 706, F735
## 2024-05-04 20:52:35.014699 INFO::Fitting model to feature number 707, F736
## 2024-05-04 20:52:35.018451 INFO::Fitting model to feature number 708, F737
## 2024-05-04 20:52:35.022191 INFO::Fitting model to feature number 709, F739
## 2024-05-04 20:52:35.025997 INFO::Fitting model to feature number 710, F740
## 2024-05-04 20:52:35.029797 INFO::Fitting model to feature number 711, F741
## 2024-05-04 20:52:35.033573 INFO::Fitting model to feature number 712, F742
## 2024-05-04 20:52:35.037308 INFO::Fitting model to feature number 713, F743
## 2024-05-04 20:52:35.041077 INFO::Fitting model to feature number 714, F744
## 2024-05-04 20:52:35.044845 INFO::Fitting model to feature number 715, F745
## 2024-05-04 20:52:35.048669 INFO::Fitting model to feature number 716, F746
## 2024-05-04 20:52:35.052479 INFO::Fitting model to feature number 717, F747
## 2024-05-04 20:52:35.05626 INFO::Fitting model to feature number 718, F748
## 2024-05-04 20:52:35.06001 INFO::Fitting model to feature number 719, F749
## 2024-05-04 20:52:35.063786 INFO::Fitting model to feature number 720, F750
## 2024-05-04 20:52:35.06754 INFO::Fitting model to feature number 721, F751
## 2024-05-04 20:52:35.071306 INFO::Fitting model to feature number 722, F752
## 2024-05-04 20:52:35.075145 INFO::Fitting model to feature number 723, F753
## 2024-05-04 20:52:35.078907 INFO::Fitting model to feature number 724, F754
## 2024-05-04 20:52:35.082658 INFO::Fitting model to feature number 725, F755
## 2024-05-04 20:52:35.086412 INFO::Fitting model to feature number 726, F756
## 2024-05-04 20:52:35.090232 INFO::Fitting model to feature number 727, F757
## 2024-05-04 20:52:35.094018 INFO::Fitting model to feature number 728, F758
## 2024-05-04 20:52:35.097775 INFO::Fitting model to feature number 729, F759
## 2024-05-04 20:52:35.101563 INFO::Fitting model to feature number 730, F760
## 2024-05-04 20:52:35.105291 INFO::Fitting model to feature number 731, F761
## 2024-05-04 20:52:35.109019 INFO::Fitting model to feature number 732, F762
## 2024-05-04 20:52:35.1127 INFO::Fitting model to feature number 733, F763
## 2024-05-04 20:52:35.11643 INFO::Fitting model to feature number 734, F764
## 2024-05-04 20:52:35.120174 INFO::Fitting model to feature number 735, F765
## 2024-05-04 20:52:35.123917 INFO::Fitting model to feature number 736, F766
## 2024-05-04 20:52:35.127669 INFO::Fitting model to feature number 737, F767
## 2024-05-04 20:52:35.131388 INFO::Fitting model to feature number 738, F768
## 2024-05-04 20:52:35.135169 INFO::Fitting model to feature number 739, F769
## 2024-05-04 20:52:35.139122 INFO::Fitting model to feature number 740, F770
## 2024-05-04 20:52:35.142841 INFO::Fitting model to feature number 741, F771
## 2024-05-04 20:52:35.146566 INFO::Fitting model to feature number 742, F772
## 2024-05-04 20:52:35.150284 INFO::Fitting model to feature number 743, F773
## 2024-05-04 20:52:35.154017 INFO::Fitting model to feature number 744, F774
## 2024-05-04 20:52:35.157745 INFO::Fitting model to feature number 745, F775
## 2024-05-04 20:52:35.161458 INFO::Fitting model to feature number 746, F776
## 2024-05-04 20:52:35.165123 INFO::Fitting model to feature number 747, F777
## 2024-05-04 20:52:35.16882 INFO::Fitting model to feature number 748, F778
## 2024-05-04 20:52:35.172526 INFO::Fitting model to feature number 749, F779
## 2024-05-04 20:52:35.176229 INFO::Fitting model to feature number 750, F780
## 2024-05-04 20:52:35.179954 INFO::Fitting model to feature number 751, F781
## 2024-05-04 20:52:35.183707 INFO::Fitting model to feature number 752, F782
## 2024-05-04 20:52:35.18748 INFO::Fitting model to feature number 753, F783
## 2024-05-04 20:52:35.191214 INFO::Fitting model to feature number 754, F784
## 2024-05-04 20:52:35.194928 INFO::Fitting model to feature number 755, F785
## 2024-05-04 20:52:35.19863 INFO::Fitting model to feature number 756, F786
## 2024-05-04 20:52:35.202317 INFO::Fitting model to feature number 757, F787
## 2024-05-04 20:52:35.206032 INFO::Fitting model to feature number 758, F788
## 2024-05-04 20:52:35.20978 INFO::Fitting model to feature number 759, F789
## 2024-05-04 20:52:35.213509 INFO::Fitting model to feature number 760, F790
## 2024-05-04 20:52:35.217263 INFO::Fitting model to feature number 761, F791
## 2024-05-04 20:52:35.221021 INFO::Fitting model to feature number 762, F792
## 2024-05-04 20:52:35.224739 INFO::Fitting model to feature number 763, F793
## 2024-05-04 20:52:35.228539 INFO::Fitting model to feature number 764, F794
## 2024-05-04 20:52:35.232314 INFO::Fitting model to feature number 765, F795
## 2024-05-04 20:52:35.236063 INFO::Fitting model to feature number 766, F796
## 2024-05-04 20:52:35.239838 INFO::Fitting model to feature number 767, F797
## 2024-05-04 20:52:35.243599 INFO::Fitting model to feature number 768, F798
## 2024-05-04 20:52:35.247342 INFO::Fitting model to feature number 769, F799
## 2024-05-04 20:52:35.251036 INFO::Fitting model to feature number 770, F800
## 2024-05-04 20:52:35.254711 INFO::Fitting model to feature number 771, F801
## 2024-05-04 20:52:35.258421 INFO::Fitting model to feature number 772, F802
## 2024-05-04 20:52:35.262149 INFO::Fitting model to feature number 773, F803
## 2024-05-04 20:52:35.265942 INFO::Fitting model to feature number 774, F804
## 2024-05-04 20:52:35.269751 INFO::Fitting model to feature number 775, F805
## 2024-05-04 20:52:35.273519 INFO::Fitting model to feature number 776, F806
## 2024-05-04 20:52:35.277347 INFO::Fitting model to feature number 777, F808
## 2024-05-04 20:52:35.281168 INFO::Fitting model to feature number 778, F809
## 2024-05-04 20:52:35.284922 INFO::Fitting model to feature number 779, F810
## 2024-05-04 20:52:35.288661 INFO::Fitting model to feature number 780, F811
## 2024-05-04 20:52:35.292365 INFO::Fitting model to feature number 781, F812
## 2024-05-04 20:52:35.296108 INFO::Fitting model to feature number 782, F813
## 2024-05-04 20:52:35.29985 INFO::Fitting model to feature number 783, F814
## 2024-05-04 20:52:35.303532 INFO::Fitting model to feature number 784, F815
## 2024-05-04 20:52:35.307207 INFO::Fitting model to feature number 785, F816
## 2024-05-04 20:52:35.310956 INFO::Fitting model to feature number 786, F817
## 2024-05-04 20:52:35.314777 INFO::Fitting model to feature number 787, F818
## 2024-05-04 20:52:35.318577 INFO::Fitting model to feature number 788, F819
## 2024-05-04 20:52:35.322417 INFO::Fitting model to feature number 789, F820
## 2024-05-04 20:52:35.326228 INFO::Fitting model to feature number 790, F821
## 2024-05-04 20:52:35.329992 INFO::Fitting model to feature number 791, F822
## 2024-05-04 20:52:35.33379 INFO::Fitting model to feature number 792, F823
## 2024-05-04 20:52:35.337651 INFO::Fitting model to feature number 793, F824
## 2024-05-04 20:52:35.341618 INFO::Fitting model to feature number 794, F825
## 2024-05-04 20:52:35.345369 INFO::Fitting model to feature number 795, F826
## 2024-05-04 20:52:35.349066 INFO::Fitting model to feature number 796, F827
## 2024-05-04 20:52:35.352825 INFO::Fitting model to feature number 797, F828
## 2024-05-04 20:52:35.356574 INFO::Fitting model to feature number 798, F829
## 2024-05-04 20:52:35.360358 INFO::Fitting model to feature number 799, F830
## 2024-05-04 20:52:35.364128 INFO::Fitting model to feature number 800, F831
## 2024-05-04 20:52:35.367935 INFO::Fitting model to feature number 801, F832
## 2024-05-04 20:52:35.371699 INFO::Fitting model to feature number 802, F833
## 2024-05-04 20:52:35.375473 INFO::Fitting model to feature number 803, F834
## 2024-05-04 20:52:35.379297 INFO::Fitting model to feature number 804, F835
## 2024-05-04 20:52:35.383054 INFO::Fitting model to feature number 805, F836
## 2024-05-04 20:52:35.386859 INFO::Fitting model to feature number 806, F837
## 2024-05-04 20:52:35.390626 INFO::Fitting model to feature number 807, F838
## 2024-05-04 20:52:35.394454 INFO::Fitting model to feature number 808, F839
## 2024-05-04 20:52:35.398243 INFO::Fitting model to feature number 809, F840
## 2024-05-04 20:52:35.402024 INFO::Fitting model to feature number 810, F841
## 2024-05-04 20:52:35.405823 INFO::Fitting model to feature number 811, F842
## 2024-05-04 20:52:35.409538 INFO::Fitting model to feature number 812, F843
## 2024-05-04 20:52:35.413279 INFO::Fitting model to feature number 813, F844
## 2024-05-04 20:52:35.41701 INFO::Fitting model to feature number 814, F845
## 2024-05-04 20:52:35.420746 INFO::Fitting model to feature number 815, F846
## 2024-05-04 20:52:35.424469 INFO::Fitting model to feature number 816, F847
## 2024-05-04 20:52:35.428466 INFO::Fitting model to feature number 817, F848
## 2024-05-04 20:52:35.432271 INFO::Fitting model to feature number 818, F849
## 2024-05-04 20:52:35.435993 INFO::Fitting model to feature number 819, F850
## 2024-05-04 20:52:35.439762 INFO::Fitting model to feature number 820, F851
## 2024-05-04 20:52:35.443517 INFO::Fitting model to feature number 821, F852
## 2024-05-04 20:52:35.447216 INFO::Fitting model to feature number 822, F853
## 2024-05-04 20:52:35.450967 INFO::Fitting model to feature number 823, F854
## 2024-05-04 20:52:35.454656 INFO::Fitting model to feature number 824, F855
## 2024-05-04 20:52:35.45845 INFO::Fitting model to feature number 825, F856
## 2024-05-04 20:52:35.462135 INFO::Fitting model to feature number 826, F857
## 2024-05-04 20:52:35.46587 INFO::Fitting model to feature number 827, F858
## 2024-05-04 20:52:35.469679 INFO::Fitting model to feature number 828, F859
## 2024-05-04 20:52:35.473444 INFO::Fitting model to feature number 829, F860
## 2024-05-04 20:52:35.477194 INFO::Fitting model to feature number 830, F861
## 2024-05-04 20:52:35.481021 INFO::Fitting model to feature number 831, F862
## 2024-05-04 20:52:35.484923 INFO::Fitting model to feature number 832, F863
## 2024-05-04 20:52:35.488686 INFO::Fitting model to feature number 833, F864
## 2024-05-04 20:52:35.492346 INFO::Fitting model to feature number 834, F865
## 2024-05-04 20:52:35.496019 INFO::Fitting model to feature number 835, F866
## 2024-05-04 20:52:35.499717 INFO::Fitting model to feature number 836, F867
## 2024-05-04 20:52:35.503537 INFO::Fitting model to feature number 837, F868
## 2024-05-04 20:52:35.507294 INFO::Fitting model to feature number 838, F869
## 2024-05-04 20:52:35.511052 INFO::Fitting model to feature number 839, F870
## 2024-05-04 20:52:35.514874 INFO::Fitting model to feature number 840, F871
## 2024-05-04 20:52:35.518604 INFO::Fitting model to feature number 841, F872
## 2024-05-04 20:52:35.522385 INFO::Fitting model to feature number 842, F873
## 2024-05-04 20:52:35.526226 INFO::Fitting model to feature number 843, F874
## 2024-05-04 20:52:35.530066 INFO::Fitting model to feature number 844, F875
## 2024-05-04 20:52:35.533809 INFO::Fitting model to feature number 845, F876
## 2024-05-04 20:52:35.537576 INFO::Fitting model to feature number 846, F877
## 2024-05-04 20:52:35.541345 INFO::Fitting model to feature number 847, F878
## 2024-05-04 20:52:35.545088 INFO::Fitting model to feature number 848, F879
## 2024-05-04 20:52:35.548989 INFO::Fitting model to feature number 849, F880
## 2024-05-04 20:52:35.552664 INFO::Fitting model to feature number 850, F881
## 2024-05-04 20:52:35.556336 INFO::Fitting model to feature number 851, F882
## 2024-05-04 20:52:35.559992 INFO::Fitting model to feature number 852, F883
## 2024-05-04 20:52:35.563781 INFO::Fitting model to feature number 853, F884
## 2024-05-04 20:52:35.567611 INFO::Fitting model to feature number 854, F885
## 2024-05-04 20:52:35.571308 INFO::Fitting model to feature number 855, F886
## 2024-05-04 20:52:35.575022 INFO::Fitting model to feature number 856, F887
## 2024-05-04 20:52:35.578747 INFO::Fitting model to feature number 857, F888
## 2024-05-04 20:52:35.582494 INFO::Fitting model to feature number 858, F889
## 2024-05-04 20:52:35.586237 INFO::Fitting model to feature number 859, F890
## 2024-05-04 20:52:35.589889 INFO::Fitting model to feature number 860, F891
## 2024-05-04 20:52:35.59359 INFO::Fitting model to feature number 861, F892
## 2024-05-04 20:52:35.597272 INFO::Fitting model to feature number 862, F893
## 2024-05-04 20:52:35.600926 INFO::Fitting model to feature number 863, F894
## 2024-05-04 20:52:35.604617 INFO::Fitting model to feature number 864, F895
## 2024-05-04 20:52:35.608323 INFO::Fitting model to feature number 865, F896
## 2024-05-04 20:52:35.611975 INFO::Fitting model to feature number 866, F897
## 2024-05-04 20:52:35.615724 INFO::Fitting model to feature number 867, F898
## 2024-05-04 20:52:35.619486 INFO::Fitting model to feature number 868, F899
## 2024-05-04 20:52:35.623251 INFO::Fitting model to feature number 869, F900
## 2024-05-04 20:52:35.85007 INFO::Counting total values for each feature
## 2024-05-04 20:52:35.979226 INFO::Writing filtered data to file Macarron_output/maaslin2_results/features/filtered_data.tsv
## 2024-05-04 20:52:36.116389 INFO::Writing filtered, normalized data to file Macarron_output/maaslin2_results/features/filtered_data_norm.tsv
## 2024-05-04 20:52:36.255779 INFO::Writing filtered, normalized, transformed data to file Macarron_output/maaslin2_results/features/filtered_data_norm_transformed.tsv
## 2024-05-04 20:52:36.395947 INFO::Writing residuals to file Macarron_output/maaslin2_results/fits/residuals.rds
## 2024-05-04 20:52:36.467103 INFO::Writing fitted values to file Macarron_output/maaslin2_results/fits/fitted.rds
## 2024-05-04 20:52:36.524526 INFO::Writing all results to file (ordered by increasing q-values): Macarron_output/maaslin2_results/all_results.tsv
## 2024-05-04 20:52:36.552286 INFO::Writing the significant results (those which are less than or equal to the threshold of 0.250000 ) to file (ordered by increasing q-values): Macarron_output/maaslin2_results/significant_results.tsv
## 2024-05-04 20:52:36.565008 INFO::Writing heatmap of significant results to file: Macarron_output/maaslin2_results/heatmap.pdf
## Initiating effect size calculations
## Calculating mean abundance in: IBD
## Calculating mean abundance in: Control
## Calculating effect size in: IBD
## Initiating prioritization
## Calculating meta-rank and prioritizing metabolic features
## 2024-05-04 20:52:36.88223 INFO::Writing all prioritized metabolites to file: Macarron_output/prioritized_metabolites_all.csv
## 2024-05-04 20:52:36.897837 INFO::Writing characterizable prioritized metabolites to file: Macarron_output/prioritized_metabolites_characterizable.csv
## 2024-05-04 20:52:36.925448 INFO::Writing highly prioritized metabolites in IBD to file: Macarron_output/highly_prioritized_per_module_in_IBD.csv
abundances_df = read.csv(file = prism_abundances, row.names = 1) # setting features as rownames
annotations_df = read.csv(file = prism_annotations, row.names = 1) # setting features as rownames
metadata_df = read.csv(file = prism_metadata, row.names = 1) # setting samples as rownames
taxonomy_df = read.csv(file = mets_taxonomy)
# Running Macarron
prism_prioritized <- Macarron::Macarron(input_abundances = abundances_df,
input_annotations = annotations_df,
input_metadata = metadata_df,
input_taxonomy = taxonomy_df)
The Macarron::Macarron()
function is a wrapper for the Macarron framework. Users can also apply individual functions on the input dataframes to achieve same results as the wrapper with the added benefit of storing output from each function for other analyses. There are seven steps:
# Step 1: Storing input data in a summarized experiment object
prism_mbx <- prepInput(input_abundances = abundances_df,
input_annotations = annotations_df,
input_metadata = metadata_df)
# Step 2: Creating a distance matrix from pairwise correlations in abundances of metabolic features
prism_w <- makeDisMat(se = prism_mbx)
# Step 3: Finding covariance modules
prism_modules <- findMacMod(se = prism_mbx,
w = prism_w,
input_taxonomy = taxonomy_df)
# The output is a list containing two dataframes- module assignments and measures of success
# if evaluateMOS=TRUE. To write modules to a separate dataframe, do:
prism_module_assignments <- prism_modules[[1]]
prism_modules_mos <- prism_modules[[2]]
# Step 4: Calculating AVA
prism_ava <- calAVA(se = prism_mbx,
mod.assn = prism_modules)
# Step 5: Calculating q-value
prism_qval <- calQval(se = prism_mbx,
mod.assn = prism_modules)
# Step 6: Calculating effect size
prism_es <- calES(se = prism_mbx,
mac.qval = prism_qval)
# Step 7: Prioritizing metabolic features
prism_prioritized <- prioritize(se = prism_mbx,
mod.assn = prism_modules,
mac.ava = prism_ava,
mac.qval = prism_qval,
mac.es = prism_es)
# The output is a list containing two dataframes- all prioritized metabolic features and
# only characterizable metabolic features.
all_prioritized <- prism_prioritized[[1]]
char_prioritized <- prism_prioritized[[2]]
# Step 8 (optional): View only the highly prioritized metabolic features in each module
prism_highly_prioritized <- showBest(prism_prioritized)
Session info from running the demo in R can be displayed with the following command.
sessionInfo()
## R version 4.4.0 Patched (2024-04-24 r86482)
## Platform: aarch64-apple-darwin20
## Running under: macOS Ventura 13.6.6
##
## Matrix products: default
## BLAS: /Library/Frameworks/R.framework/Versions/4.4-arm64/Resources/lib/libRblas.0.dylib
## LAPACK: /Library/Frameworks/R.framework/Versions/4.4-arm64/Resources/lib/libRlapack.dylib; LAPACK version 3.12.0
##
## locale:
## [1] C/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8
##
## time zone: America/New_York
## tzcode source: internal
##
## attached base packages:
## [1] stats4 stats graphics grDevices utils datasets methods
## [8] base
##
## other attached packages:
## [1] Macarron_1.9.0 SummarizedExperiment_1.35.0
## [3] Biobase_2.65.0 GenomicRanges_1.57.0
## [5] GenomeInfoDb_1.41.0 IRanges_2.39.0
## [7] S4Vectors_0.43.0 BiocGenerics_0.51.0
## [9] MatrixGenerics_1.17.0 matrixStats_1.3.0
## [11] BiocStyle_2.33.0
##
## loaded via a namespace (and not attached):
## [1] mnormt_2.1.1 DBI_1.2.2 pbapply_1.7-2
## [4] gridExtra_2.3 rlang_1.1.3 magrittr_2.0.3
## [7] compiler_4.4.0 RSQLite_2.3.6 png_0.1-8
## [10] vctrs_0.6.5 stringr_1.5.1 pkgconfig_2.0.3
## [13] crayon_1.5.2 fastmap_1.1.1 backports_1.4.1
## [16] XVector_0.45.0 utf8_1.2.4 rmarkdown_2.26
## [19] preprocessCore_1.67.0 UCSC.utils_1.1.0 bit_4.0.5
## [22] xfun_0.43 zlibbioc_1.51.0 cachem_1.0.8
## [25] jsonlite_1.8.8 blob_1.2.4 DelayedArray_0.31.0
## [28] BiocParallel_1.39.0 psych_2.4.3 Maaslin2_1.19.0
## [31] parallel_4.4.0 cluster_2.1.6 biglm_0.9-2.1
## [34] R6_2.5.1 RColorBrewer_1.1-3 stringi_1.8.3
## [37] bslib_0.7.0 rpart_4.1.23 jquerylib_0.1.4
## [40] Rcpp_1.0.12 bookdown_0.39 iterators_1.0.14
## [43] knitr_1.46 WGCNA_1.72-5 base64enc_0.1-3
## [46] Matrix_1.7-0 splines_4.4.0 nnet_7.3-19
## [49] tidyselect_1.2.1 rstudioapi_0.16.0 abind_1.4-5
## [52] yaml_2.3.8 doParallel_1.0.17 codetools_0.2-20
## [55] plyr_1.8.9 lattice_0.22-6 tibble_3.2.1
## [58] withr_3.0.0 KEGGREST_1.45.0 evaluate_0.23
## [61] foreign_0.8-86 survival_3.6-4 getopt_1.20.4
## [64] Biostrings_2.73.0 pillar_1.9.0 BiocManager_1.30.22
## [67] checkmate_2.3.1 foreach_1.5.2 pcaPP_2.0-4
## [70] generics_0.1.3 ggplot2_3.5.1 munsell_0.5.1
## [73] scales_1.3.0 glue_1.7.0 pheatmap_1.0.12
## [76] Hmisc_5.1-2 tools_4.4.0 robustbase_0.99-2
## [79] data.table_1.15.4 mvtnorm_1.2-4 fastcluster_1.2.6
## [82] grid_4.4.0 optparse_1.7.5 impute_1.79.0
## [85] AnnotationDbi_1.67.0 colorspace_2.1-0 nlme_3.1-164
## [88] GenomeInfoDbData_1.2.12 htmlTable_2.4.2 Formula_1.2-5
## [91] cli_3.6.2 fansi_1.0.6 S4Arrays_1.5.0
## [94] dplyr_1.1.4 DEoptimR_1.1-3 gtable_0.3.5
## [97] hash_2.2.6.3 logging_0.10-108 dynamicTreeCut_1.63-1
## [100] sass_0.4.9 digest_0.6.35 SparseArray_1.5.0
## [103] htmlwidgets_1.6.4 memoise_2.0.1 htmltools_0.5.8.1
## [106] lifecycle_1.0.4 httr_1.4.7 GO.db_3.19.1
## [109] bit64_4.0.5
The input taxonomy dataframe can be generated using the input metabolic features annotation dataframe using Macarron::decorateID()
. This function annotates an HMDB ID or a PubChem CID with the chemical class and subclass of the metabolite.
taxonomy_df <- decorateID(input_annotations = annotations_df)
write.csv(taxonomy_df, file="demo_taxonomy.csv", row.names = FALSE)
A record of all chosen parameters and steps that were followed during execution.
This file provides information about the properties of covariance modules used in the analysis. By default, modules are generated using a minimum module size (MMS) (argument: min_module_size
) equal to cube root of the total number of prevalent metabolic features. Macarron evaluates 9 measures of success (MOS) that collectively capture the “correctness” and chemical homogeneity of the modules. The MOS are as follows:
This folder contains the Maaslin2 log file (maaslin2.log), significant associations found by Maaslin2 (significant_results.tsv) and the linear model residuals file (residuals.rds). For more information, see Maaslin2.
Ideally, at least 50% metabolic features must be retained after prevalence filtering. By default, Macarron uses the union of metabolic features observed (non-zero abundance) in at least 70% samples of any phenotype for further analysis. This prevalence threshold may be high for some metabolomics datasets and can be changed using the min_prevalence
argument.
prism_prioritized <- Macarron::Macarron(input_abundances = abundances_df,
input_annotations = annotations_df,
input_metadata = metadata_df,
input_taxonomy = taxonomy_df,
min_prevalence = 0.5)
# or
prism_w <- makeDisMat(se = prism_mbx,
min_prevalence = 0.5)
By default, cube root of the total number of prevalent features is used as the minimum module size (MMS) (argument: min_module_size
) for module detection and generation. We expect this to work for most real world datasets. To determine if the modules are optimal for further analysis, Macarron evaluates several measures of success (MOS) as described above. In addition to evaluating MOS for modules generated using the default MMS, Macarron also evaluates MOS for MMS values that are larger (MMS+5, MMS+10) and smaller (MMS-5, MMS-10) than the default MMS. If you find that the MOS improve with larger or smaller MMS, you may change the default accordingly. For more details about module detection, please see WGCNA
and dynamicTreeCut
.
# See MOS of modules generated using default
prism_modules <- findMacMod(se = prism_mbx,
w = prism_w,
input_taxonomy = taxonomy_df)
prism_modules_mos <- prism_modules[[2]]
View(prism_modules_mos)
# Change MMS
prism_modules <- findMacMod(se = prism_mbx,
w = prism_w,
input_taxonomy = taxonomy_df,
min_module_size = 10)
Macarron uses Maaslin2 for determining the q-value of differential abundance in a phenotype of interest. For default execution, the phenotype of interest must be a category in column 1 of the metadata dataframe e.g. IBD in diagnosis in the demo. This is also the column that is picked by the metadata_variable
argument for identifying the main phenotypes/conditions in any dataset (see Macarron.log file). Further, in the default execution, all columns in the metadata table are considered as fixed effects and the alphabetically first categorical variable in each covariate with two categories is considered as the reference. Maaslin2 requires reference categories to be explicitly defined for all categorical metadata with more than two categories.
Defaults can be changed with the arguments fixed_effects
, random_effects
and reference
. In the demo example, fixed effects
and reference
can be defined as follows:
prism_qval <- calQval(se = prism_mbx,
mod.assn = prism_modules,
metadata_variable = "diagnosis",
fixed_effects = c("diagnosis","age","antibiotics"),
reference = c("diagnosis,Control";"antibiotics,No"))
The package source contains a script MacarronCMD.R
in inst/scripts
to invoke Macarron in the command line using Rscript.
The inst/scripts
folder also contains a README file that comprehensively documents the usage of the script.