{ "cells": [ { "cell_type": "markdown", "metadata": {}, "source": [ "# PMA Treatment\n", "Here we will visualize the PMA treatment data separately from the Plasma treated data. We will generate heatmaps using subsampling and downsampling." ] }, { "cell_type": "code", "execution_count": 1, "metadata": { "collapsed": true }, "outputs": [], "source": [ "from clustergrammer_widget import *\n", "net = Network(clustergrammer_widget)" ] }, { "cell_type": "code", "execution_count": 2, "metadata": { "collapsed": true }, "outputs": [], "source": [ "net.load_file('../cytof_data/PMA_UCT.txt')\n", "pma_df = net.export_df()" ] }, { "cell_type": "code", "execution_count": 3, "metadata": { "collapsed": true }, "outputs": [], "source": [ "# manually set treatment colors\n", "net.set_cat_color('col', 1, 'Marker-type: phospho marker', 'red')\n", "net.set_cat_color('col', 1, 'Marker-type: surface marker', 'blue')\n", "\n", "# manually set row colors: downsample\n", "net.set_cat_color('row', 2, 'Majority-Category: B cells', '#22316C')\n", "net.set_cat_color('row', 2, 'Majority-Category: Basophils', '#000033')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD14hi monocytes', 'yellow')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD14low monocytes', '#93b8bf')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD1c DCs', '#3636e2')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells', 'blue')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells_CD127hi', '#FF6347')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells CD161hi', '#F87531')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells_Tregs', '#8B4513')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells+CD27hi', '#330303')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD8 Tcells', '#ffb247')\n", "net.set_cat_color('row', 2, 'Majority-Category: Neutrophils', 'purple')\n", "net.set_cat_color('row', 2, 'Majority-Category: NK cells_CD16hi', 'red')\n", "net.set_cat_color('row', 2, 'Majority-Category: NK cells_CD16hi_CD57hi', 'orange')\n", "net.set_cat_color('row', 2, 'Majority-Category: NK cells_CD56hi', '#e052e5')\n", "net.set_cat_color('row', 2, 'Majority-Category: Undefined', 'gray')\n", "\n", "# manually set row colors: subsample\n", "net.set_cat_color('row', 2, 'B cells', '#22316C')\n", "net.set_cat_color('row', 2, 'Basophils', '#000033')\n", "net.set_cat_color('row', 2, 'CD14hi monocytes', 'yellow')\n", "net.set_cat_color('row', 2, 'CD14low monocytes', '#93b8bf')\n", "net.set_cat_color('row', 2, 'CD1c DCs', '#3636e2')\n", "net.set_cat_color('row', 2, 'CD4 Tcells', 'blue')\n", "net.set_cat_color('row', 2, 'CD4 Tcells_CD127hi', '#FF6347')\n", "net.set_cat_color('row', 2, 'CD4 Tcells CD161hi', '#F87531')\n", "net.set_cat_color('row', 2, 'CD4 Tcells_Tregs', '#8B4513')\n", "net.set_cat_color('row', 2, 'CD4 Tcells+CD27hi', '#330303')\n", "net.set_cat_color('row', 2, 'CD8 Tcells', '#ffb247')\n", "net.set_cat_color('row', 2, 'Neutrophils', 'purple')\n", "net.set_cat_color('row', 2, 'NK cells_CD16hi', 'red')\n", "net.set_cat_color('row', 2, 'NK cells_CD16hi_CD57hi', 'orange')\n", "net.set_cat_color('row', 2, 'NK cells_CD56hi', '#e052e5')\n", "net.set_cat_color('row', 2, 'Undefined', 'gray')\n", "\n", "# manually set treatment colors\n", "net.set_cat_color('row', 1, 'Majority-Treatment: Plasma', 'blue')\n", "net.set_cat_color('row', 1, 'Majority-Treatment: PMA', 'red')\n", "\n", "net.set_cat_color('row', 1, 'Treatment: Plasma', 'blue')\n", "net.set_cat_color('row', 1, 'Treatment: PMA', 'red')" ] }, { "cell_type": "markdown", "metadata": { "collapsed": true }, "source": [ "# PMA Subsample All Markers" ] }, { "cell_type": "code", "execution_count": 4, "metadata": {}, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "aee9d8a0ba3548e096a8317c3a6045ba" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(pma_df)\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "\n", "# subsample the data so that both treatments have the same number of cells\n", "net.random_sample(axis='row', num_samples=1000, random_state=99)\n", "\n", "# clip z-scores since we do not care about extreme outliers\n", "net.clip(-10,10)\n", "\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "# PMA Downsample All Markers" ] }, { "cell_type": "code", "execution_count": 5, "metadata": {}, "outputs": [ { "name": "stderr", "output_type": "stream", "text": [ "/Users/nickfernandez/anaconda/lib/python2.7/site-packages/sklearn/cluster/k_means_.py:1382: RuntimeWarning: init_size=300 should be larger than k=1000. Setting it to 3*k\n", " init_size=init_size)\n" ] }, { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "87582131c83e49cf9bc83fc954aab3c5" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(pma_df)\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "# downsample to 1000 clusters\n", "ds_data = net.downsample(ds_type='kmeans', axis='row', num_samples=1000)\n", "# clip z-scores since we do not care about extreme outliers\n", "net.clip(-10,10)\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": { "collapsed": true }, "source": [ "# PMA Phopho Markers Subsample" ] }, { "cell_type": "code", "execution_count": 6, "metadata": {}, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "31fb2e4e4f1441f3999bb7bd355e3002" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(pma_df)\n", "net.filter_cat('col', 1, 'Marker-type: phospho marker')\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "net.random_sample(axis='row', num_samples=1000, random_state=1000)\n", "net.clip(-10,10)\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "# PMA Phospho Markers Downsample" ] }, { "cell_type": "code", "execution_count": 7, "metadata": {}, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "93abc04e6c09445e80998b2cf3695b67" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(pma_df)\n", "net.filter_cat('col', 1, 'Marker-type: phospho marker')\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "ds_data = net.downsample(ds_type='kmeans', axis='row', num_samples=1000)\n", "net.clip(-10,10)\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "Similarly to our results clustering the combined Plasma and PMA treated cells, we again find a prominent cluster of CD14hi monocytes with high phosphorylation across several markers." ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": true }, "outputs": [], "source": [] } ], "metadata": { "anaconda-cloud": {}, "kernelspec": { "display_name": "Python [Root]", "language": "python", "name": "Python [Root]" }, "language_info": { "codemirror_mode": { "name": "ipython", "version": 2 }, "file_extension": ".py", "mimetype": "text/x-python", "name": "python", "nbconvert_exporter": "python", "pygments_lexer": "ipython2", "version": "2.7.12" }, "widgets": { "application/vnd.jupyter.widget-state+json": { "state": { "1674220b2b23436fa6c1d030ae3f3a0a": { "model_module": "jupyter-js-widgets", "model_module_version": "~2.1.4", "model_name": "LayoutModel", "state": { "_model_module_version": "~2.1.4", "_view_module_version": "~2.1.4" } }, "2ff4da4e7d144aa98a956c8c72623288": { "model_module": "clustergrammer_widget", "model_module_version": "*", 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1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-8\", \"clust\": 342, \"cat_2_index\": 647, \"cat_1_index\": 188, \"cat_0_index\": 8, \"cat-2\": \"number in clust: 122\", \"rank\": 426, \"cat-1\": \"Majority-Category: CD4 Tcells\", \"ini\": 992, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 187}, {\"group\": [147.0, 77.0, 27.0, 10.0, 5.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-9\", \"clust\": 147, \"cat_2_index\": 966, \"cat_1_index\": 403, \"cat_0_index\": 9, \"cat-2\": \"number in clust: 319\", \"rank\": 344, \"cat-1\": \"Majority-Category: CD4 Tcells+CD27hi\", \"ini\": 991, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 205}, {\"group\": [801.0, 360.0, 115.0, 35.0, 17.0, 6.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-10\", \"clust\": 801, \"cat_2_index\": 318, \"cat_1_index\": 55, \"cat_0_index\": 10, \"cat-2\": \"number in clust: 59\", \"rank\": 567, \"cat-1\": \"Majority-Category: Basophils\", \"ini\": 990, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 965}, {\"group\": [443.0, 221.0, 70.0, 21.0, 10.0, 2.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-11\", \"clust\": 442, \"cat_2_index\": 987, \"cat_1_index\": 633, \"cat_0_index\": 11, \"cat-2\": \"number in clust: 386\", \"rank\": 89, \"cat-1\": \"Majority-Category: CD8 Tcells\", \"ini\": 989, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 12}, {\"group\": [739.0, 345.0, 111.0, 34.0, 16.0, 5.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-12\", \"clust\": 736, \"cat_2_index\": 915, \"cat_1_index\": 961, \"cat_0_index\": 12, \"cat-2\": \"number in clust: 243\", \"rank\": 44, \"cat-1\": \"Majority-Category: Undefined\", \"ini\": 988, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 220}, {\"group\": [614.0, 302.0, 98.0, 29.0, 15.0, 5.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-13\", \"clust\": 613, \"cat_2_index\": 89, \"cat_1_index\": 957, \"cat_0_index\": 13, \"cat-2\": \"number in clust: 23\", 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{\"group\": [9.0, 6.0, 2.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-888\", \"clust\": 6, \"cat_2_index\": 146, \"cat_1_index\": 374, \"cat_0_index\": 888, \"cat-2\": \"number in clust: 29\", \"rank\": 412, \"cat-1\": \"Majority-Category: CD4 Tcells\", \"ini\": 112, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 196}, {\"group\": [434.0, 214.0, 67.0, 21.0, 10.0, 2.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-889\", \"clust\": 434, \"cat_2_index\": 293, \"cat_1_index\": 739, \"cat_0_index\": 889, \"cat-2\": \"number in clust: 53\", \"rank\": 420, \"cat-1\": \"Majority-Category: CD8 Tcells\", \"ini\": 111, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 504}, {\"group\": [859.0, 372.0, 118.0, 36.0, 18.0, 6.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-890\", \"clust\": 842, \"cat_2_index\": 414, \"cat_1_index\": 52, \"cat_0_index\": 890, \"cat-2\": \"number in clust: 74\", \"rank\": 618, \"cat-1\": \"Majority-Category: B 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\"Majority-Treatment: PMA\", \"rankvar\": 146}, {\"group\": [606.0, 298.0, 96.0, 28.0, 14.0, 5.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-910\", \"clust\": 606, \"cat_2_index\": 882, \"cat_1_index\": 850, \"cat_0_index\": 910, \"cat-2\": \"number in clust: 214\", \"rank\": 108, \"cat-1\": \"Majority-Category: NK cells_CD16hi\", \"ini\": 90, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 288}, {\"group\": [959.0, 405.0, 126.0, 39.0, 19.0, 6.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-911\", \"clust\": 965, \"cat_2_index\": 371, \"cat_1_index\": 150, \"cat_0_index\": 911, \"cat-2\": \"number in clust: 66\", \"rank\": 925, \"cat-1\": \"Majority-Category: CD14hi monocytes\", \"ini\": 89, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 899}, {\"group\": [957.0, 404.0, 126.0, 39.0, 19.0, 6.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-912\", \"clust\": 960, \"cat_2_index\": 372, \"cat_1_index\": 151, \"cat_0_index\": 912, \"cat-2\": 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\"Majority-Treatment: PMA\", \"rankvar\": 798}, {\"group\": [446.0, 222.0, 70.0, 21.0, 10.0, 2.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-948\", \"clust\": 446, \"cat_2_index\": 926, \"cat_1_index\": 748, \"cat_0_index\": 948, \"cat-2\": \"number in clust: 262\", \"rank\": 158, \"cat-1\": \"Majority-Category: CD8 Tcells\", \"ini\": 52, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 74}, {\"group\": [240.0, 123.0, 45.0, 16.0, 7.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-949\", \"clust\": 242, \"cat_2_index\": 202, \"cat_1_index\": 629, \"cat_0_index\": 949, \"cat-2\": \"number in clust: 37\", \"rank\": 844, \"cat-1\": \"Majority-Category: CD4 Tcells_Tregs\", \"ini\": 51, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 583}, {\"group\": [686.0, 322.0, 103.0, 30.0, 16.0, 5.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-950\", \"clust\": 687, \"cat_2_index\": 800, \"cat_1_index\": 937, \"cat_0_index\": 950, \"cat-2\": \"number in 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\"Majority-Category: CD4 Tcells\", \"ini\": 42, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 16}, {\"group\": [749.0, 352.0, 113.0, 34.0, 16.0, 5.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-959\", \"clust\": 748, \"cat_2_index\": 604, \"cat_1_index\": 857, \"cat_0_index\": 959, \"cat-2\": \"number in clust: 112\", \"rank\": 455, \"cat-1\": \"Majority-Category: NK cells_CD16hi\", \"ini\": 41, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 453}, {\"group\": [631.0, 312.0, 100.0, 30.0, 16.0, 5.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-960\", \"clust\": 631, \"cat_2_index\": 615, \"cat_1_index\": 999, \"cat_0_index\": 960, \"cat-2\": \"number in clust: 114\", \"rank\": 76, \"cat-1\": \"Majority-Category: Undefined\", \"ini\": 40, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 171}, {\"group\": [318.0, 171.0, 60.0, 19.0, 9.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-961\", \"clust\": 312, \"cat_2_index\": 927, 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\"cat-1\": \"Majority-Category: CD4 Tcells\", \"ini\": 176, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 154}, {\"group\": [91.0, 41.0, 14.0, 7.0, 3.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-825\", \"clust\": 94, \"cat_2_index\": 356, \"cat_1_index\": 924, \"cat_0_index\": 825, \"cat-2\": \"number in clust: 66\", \"rank\": 453, \"cat-1\": \"Majority-Category: NK cells_CD16hi\", \"ini\": 175, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 690}, {\"group\": [532.0, 137.0, 42.0, 13.0, 7.0, 3.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-826\", \"clust\": 530, \"cat_2_index\": 8, \"cat_1_index\": 802, \"cat_0_index\": 826, \"cat-2\": \"number in clust: 6\", \"rank\": 378, \"cat-1\": \"Majority-Category: CD8 Tcells\", \"ini\": 174, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 721}, {\"group\": [552.0, 146.0, 45.0, 14.0, 7.0, 3.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-827\", \"clust\": 553, \"cat_2_index\": 678, 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\"Majority-Treatment: PMA\", \"rankvar\": 269}, {\"group\": [265.0, 78.0, 25.0, 8.0, 3.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-833\", \"clust\": 266, \"cat_2_index\": 840, \"cat_1_index\": 634, \"cat_0_index\": 833, \"cat-2\": \"number in clust: 185\", \"rank\": 95, \"cat-1\": \"Majority-Category: CD4 Tcells\", \"ini\": 167, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 99}, {\"group\": [546.0, 145.0, 45.0, 14.0, 7.0, 3.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-834\", \"clust\": 546, \"cat_2_index\": 606, \"cat_1_index\": 926, \"cat_0_index\": 834, \"cat-2\": \"number in clust: 109\", \"rank\": 439, \"cat-1\": \"Majority-Category: NK cells_CD16hi\", \"ini\": 166, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 603}, {\"group\": [795.0, 211.0, 59.0, 17.0, 9.0, 4.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-835\", \"clust\": 794, \"cat_2_index\": 451, \"cat_1_index\": 235, \"cat_0_index\": 835, \"cat-2\": \"number in clust: 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\"Majority-Treatment: PMA\", \"rankvar\": 569}, {\"group\": [948.0, 229.0, 63.0, 18.0, 10.0, 4.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-929\", \"clust\": 948, \"cat_2_index\": 428, \"cat_1_index\": 678, \"cat_0_index\": 929, \"cat-2\": \"number in clust: 77\", \"rank\": 618, \"cat-1\": \"Majority-Category: CD4 Tcells\", \"ini\": 71, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 539}, {\"group\": [378.0, 104.0, 32.0, 10.0, 5.0, 2.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-930\", \"clust\": 374, \"cat_2_index\": 611, \"cat_1_index\": 811, \"cat_0_index\": 930, \"cat-2\": \"number in clust: 110\", \"rank\": 683, \"cat-1\": \"Majority-Category: CD8 Tcells\", \"ini\": 70, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 634}, {\"group\": [125.0, 50.0, 17.0, 7.0, 3.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-931\", \"clust\": 95, \"cat_2_index\": 313, \"cat_1_index\": 679, \"cat_0_index\": 931, \"cat-2\": \"number in clust: 58\", 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\"Majority-Treatment: PMA\", \"rankvar\": 658}, {\"group\": [473.0, 122.0, 37.0, 11.0, 6.0, 3.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-940\", \"clust\": 470, \"cat_2_index\": 151, \"cat_1_index\": 813, \"cat_0_index\": 940, \"cat-2\": \"number in clust: 33\", \"rank\": 710, \"cat-1\": \"Majority-Category: CD8 Tcells\", \"ini\": 60, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 542}, {\"group\": [688.0, 181.0, 53.0, 16.0, 9.0, 4.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-941\", \"clust\": 687, \"cat_2_index\": 297, \"cat_1_index\": 249, \"cat_0_index\": 941, \"cat-2\": \"number in clust: 55\", \"rank\": 966, \"cat-1\": \"Majority-Category: CD14hi monocytes\", \"ini\": 59, \"cat-0\": \"Majority-Treatment: PMA\", \"rankvar\": 911}, {\"group\": [671.0, 179.0, 53.0, 16.0, 9.0, 4.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-942\", \"clust\": 670, \"cat_2_index\": 224, \"cat_1_index\": 250, \"cat_0_index\": 942, \"cat-2\": \"number in clust: 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