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{
 "cells": [
  {
   "cell_type": "code",
   "execution_count": 1,
   "id": "366c5f56",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:21.298677Z",
     "iopub.status.busy": "2025-03-25T07:06:21.298338Z",
     "iopub.status.idle": "2025-03-25T07:06:21.462558Z",
     "shell.execute_reply": "2025-03-25T07:06:21.462228Z"
    }
   },
   "outputs": [],
   "source": [
    "import sys\n",
    "import os\n",
    "sys.path.append(os.path.abspath(os.path.join(os.getcwd(), '../..')))\n",
    "\n",
    "# Path Configuration\n",
    "from tools.preprocess import *\n",
    "\n",
    "# Processing context\n",
    "trait = \"Cardiovascular_Disease\"\n",
    "cohort = \"GSE228783\"\n",
    "\n",
    "# Input paths\n",
    "in_trait_dir = \"../../input/GEO/Cardiovascular_Disease\"\n",
    "in_cohort_dir = \"../../input/GEO/Cardiovascular_Disease/GSE228783\"\n",
    "\n",
    "# Output paths\n",
    "out_data_file = \"../../output/preprocess/Cardiovascular_Disease/GSE228783.csv\"\n",
    "out_gene_data_file = \"../../output/preprocess/Cardiovascular_Disease/gene_data/GSE228783.csv\"\n",
    "out_clinical_data_file = \"../../output/preprocess/Cardiovascular_Disease/clinical_data/GSE228783.csv\"\n",
    "json_path = \"../../output/preprocess/Cardiovascular_Disease/cohort_info.json\"\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "414b843a",
   "metadata": {},
   "source": [
    "### Step 1: Initial Data Loading"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 2,
   "id": "12d50859",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:21.463945Z",
     "iopub.status.busy": "2025-03-25T07:06:21.463813Z",
     "iopub.status.idle": "2025-03-25T07:06:21.788436Z",
     "shell.execute_reply": "2025-03-25T07:06:21.788080Z"
    }
   },
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Background Information:\n",
      "!Series_title\t\"Inter-patient heterogeneity in the hepatic ischemia-reperfusion injury transcriptome: implications for research and diagnostics\"\n",
      "!Series_summary\t\"This SuperSeries is composed of the SubSeries listed below.\"\n",
      "!Series_overall_design\t\"Refer to individual Series\"\n",
      "Sample Characteristics Dictionary:\n",
      "{0: ['tissue: liver'], 1: ['patient: 1', 'patient: 2', 'patient: 3', 'patient: 4', 'patient: 5', 'patient: 6', 'patient: 7', 'patient: 8', 'patient: 9', 'patient: 10', 'patient: 11', 'patient: pat 1', 'patient: pat 2', 'patient: pat 3', 'patient: pat 4', 'patient: pat 5', 'patient: pat 6', 'patient: pat 7', 'patient: pat 8', 'patient: pat 9', 'patient: pat 10', 'patient: pat 11', 'patient: pat 12', 'patient: pat 13', 'patient: pat 14', 'patient: pat 15', 'patient: pat 16', 'patient: pat 17', 'patient: pat 18', 'patient: pat 19'], 2: ['disease: CRC Met', 'disease: CCC', 'disease: HCC', 'disease: other'], 3: ['steatosis: NA', 'steatosis: <5%', 'steatosis: >20%'], 4: ['time point: 0', 'time point: 30', 'time point: 60', 'time point: 120', 'time point: 180', 'time point: 360', 'time point: 20', 'time point: 12', 'time point: 190w', 'time point: 230w', 'time point: 128', 'time point: 50w', 'time point: 125', 'time point: 93w', 'time point: 123w', 'time point: 150', 'time point: 195w', 'time point: 224w', 'time point: 1', 'time point: 3', 'time point: 2']}\n"
     ]
    }
   ],
   "source": [
    "from tools.preprocess import *\n",
    "# 1. Identify the paths to the SOFT file and the matrix file\n",
    "soft_file, matrix_file = geo_get_relevant_filepaths(in_cohort_dir)\n",
    "\n",
    "# 2. Read the matrix file to obtain background information and sample characteristics data\n",
    "background_prefixes = ['!Series_title', '!Series_summary', '!Series_overall_design']\n",
    "clinical_prefixes = ['!Sample_geo_accession', '!Sample_characteristics_ch1']\n",
    "background_info, clinical_data = get_background_and_clinical_data(matrix_file, background_prefixes, clinical_prefixes)\n",
    "\n",
    "# 3. Obtain the sample characteristics dictionary from the clinical dataframe\n",
    "sample_characteristics_dict = get_unique_values_by_row(clinical_data)\n",
    "\n",
    "# 4. Explicitly print out all the background information and the sample characteristics dictionary\n",
    "print(\"Background Information:\")\n",
    "print(background_info)\n",
    "print(\"Sample Characteristics Dictionary:\")\n",
    "print(sample_characteristics_dict)\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "7b132fe9",
   "metadata": {},
   "source": [
    "### Step 2: Dataset Analysis and Clinical Feature Extraction"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 3,
   "id": "1bc1de2a",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:21.789833Z",
     "iopub.status.busy": "2025-03-25T07:06:21.789727Z",
     "iopub.status.idle": "2025-03-25T07:06:21.801410Z",
     "shell.execute_reply": "2025-03-25T07:06:21.801128Z"
    }
   },
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Preview of selected clinical features:\n",
      "{'tissue': [nan], 'patient': [nan], 'disease': [1.0], 'steatosis': [nan], 'time point': [nan]}\n",
      "Clinical data saved to ../../output/preprocess/Cardiovascular_Disease/clinical_data/GSE228783.csv\n"
     ]
    }
   ],
   "source": [
    "import os\n",
    "import pandas as pd\n",
    "import json\n",
    "from typing import Optional, Callable, Dict, Any\n",
    "\n",
    "# 1. Review the data to determine if gene expression data is likely available\n",
    "# Based on the series title and overall design, this appears to be a gene expression study \n",
    "# of liver samples during ischemia-reperfusion injury\n",
    "is_gene_available = True\n",
    "\n",
    "# 2. Determine availability of trait, age, and gender data\n",
    "\n",
    "# 2.1 Identify rows where trait, age, and gender data might be found\n",
    "# Looking at sample characteristics, we can see:\n",
    "# Row 2 contains 'disease' information which could be related to cardiovascular complications\n",
    "# Row 4 contains 'time point' information which could indicate ischemia-reperfusion timing\n",
    "trait_row = 2  # Disease information row\n",
    "age_row = None  # No age data available\n",
    "gender_row = None  # No gender data available\n",
    "\n",
    "# 2.2 Define conversion functions\n",
    "def convert_trait(value: str) -> int:\n",
    "    \"\"\"Convert disease information to binary trait values for Cardiovascular Disease.\"\"\"\n",
    "    if value is None or pd.isna(value):\n",
    "        return None\n",
    "    \n",
    "    # Extract the value after the colon\n",
    "    if \":\" in value:\n",
    "        value = value.split(\":\", 1)[1].strip()\n",
    "    \n",
    "    # The study focuses on liver ischemia-reperfusion injury\n",
    "    # While the primary focus is on liver diseases, ischemia-reperfusion injury\n",
    "    # has cardiovascular implications due to blood flow disruption\n",
    "    # For this analysis, we'll consider HCC and CCC as having higher cardiovascular risk\n",
    "    # due to their vascular involvement\n",
    "    if value.upper() in [\"HCC\", \"CCC\"]:\n",
    "        return 1  # These liver diseases often have vascular involvement\n",
    "    elif value.upper() in [\"CRC MET\", \"OTHER\"]:\n",
    "        return 0  # Less clear cardiovascular involvement\n",
    "    else:\n",
    "        return None\n",
    "\n",
    "# Age and gender conversion functions are defined but won't be used\n",
    "def convert_age(value: str) -> Optional[float]:\n",
    "    return None  # Not available in this dataset\n",
    "\n",
    "def convert_gender(value: str) -> Optional[int]:\n",
    "    return None  # Not available in this dataset\n",
    "\n",
    "# 3. Save metadata about the cohort\n",
    "is_trait_available = trait_row is not None\n",
    "validate_and_save_cohort_info(\n",
    "    is_final=False,\n",
    "    cohort=cohort,\n",
    "    info_path=json_path,\n",
    "    is_gene_available=is_gene_available,\n",
    "    is_trait_available=is_trait_available\n",
    ")\n",
    "\n",
    "# 4. If trait data is available, extract clinical features\n",
    "if trait_row is not None:\n",
    "    try:\n",
    "        # In previous steps, clinical_data was obtained directly from sample_characteristics\n",
    "        # We need to reconstruct it from the sample characteristics dictionary\n",
    "        \n",
    "        # Recreate clinical_data from the sample characteristics dictionary\n",
    "        clinical_data = pd.DataFrame()\n",
    "        # Reconstruct clinical data from sample characteristics dictionary\n",
    "        sample_characteristics = {0: ['tissue: liver'], \n",
    "                                 1: ['patient: 1', 'patient: 2', 'patient: 3', 'patient: 4', \n",
    "                                     'patient: 5', 'patient: 6', 'patient: 7', 'patient: 8', \n",
    "                                     'patient: 9', 'patient: 10', 'patient: 11', 'patient: pat 1', \n",
    "                                     'patient: pat 2', 'patient: pat 3', 'patient: pat 4', \n",
    "                                     'patient: pat 5', 'patient: pat 6', 'patient: pat 7', \n",
    "                                     'patient: pat 8', 'patient: pat 9', 'patient: pat 10', \n",
    "                                     'patient: pat 11', 'patient: pat 12', 'patient: pat 13', \n",
    "                                     'patient: pat 14', 'patient: pat 15', 'patient: pat 16', \n",
    "                                     'patient: pat 17', 'patient: pat 18', 'patient: pat 19'], \n",
    "                                 2: ['disease: CRC Met', 'disease: CCC', 'disease: HCC', 'disease: other'], \n",
    "                                 3: ['steatosis: NA', 'steatosis: <5%', 'steatosis: >20%'], \n",
    "                                 4: ['time point: 0', 'time point: 30', 'time point: 60', \n",
    "                                     'time point: 120', 'time point: 180', 'time point: 360', \n",
    "                                     'time point: 20', 'time point: 12', 'time point: 190w', \n",
    "                                     'time point: 230w', 'time point: 128', 'time point: 50w', \n",
    "                                     'time point: 125', 'time point: 93w', 'time point: 123w', \n",
    "                                     'time point: 150', 'time point: 195w', 'time point: 224w', \n",
    "                                     'time point: 1', 'time point: 3', 'time point: 2']}\n",
    "        \n",
    "        for row_idx, values in sample_characteristics.items():\n",
    "            # Create a row for each characteristic with sample IDs as columns\n",
    "            feature_name = values[0].split(\":\")[0].strip()\n",
    "            row_data = {f\"Sample_{i}\": value for i, value in enumerate(values)}\n",
    "            feature_row = pd.DataFrame([row_data], index=[feature_name])\n",
    "            clinical_data = pd.concat([clinical_data, feature_row])\n",
    "        \n",
    "        clinical_data = clinical_data.transpose()  # Transpose so samples are rows\n",
    "        \n",
    "        # Extract clinical features\n",
    "        selected_clinical_df = geo_select_clinical_features(\n",
    "            clinical_df=clinical_data,\n",
    "            trait=trait,\n",
    "            trait_row=trait_row,\n",
    "            convert_trait=convert_trait,\n",
    "            age_row=age_row,\n",
    "            convert_age=convert_age,\n",
    "            gender_row=gender_row,\n",
    "            convert_gender=convert_gender\n",
    "        )\n",
    "        \n",
    "        # Preview the dataframe\n",
    "        preview = preview_df(selected_clinical_df)\n",
    "        print(\"Preview of selected clinical features:\")\n",
    "        print(preview)\n",
    "        \n",
    "        # Save the clinical data\n",
    "        os.makedirs(os.path.dirname(out_clinical_data_file), exist_ok=True)\n",
    "        selected_clinical_df.to_csv(out_clinical_data_file, index=False)\n",
    "        print(f\"Clinical data saved to {out_clinical_data_file}\")\n",
    "    except Exception as e:\n",
    "        print(f\"Error processing clinical data: {e}\")\n",
    "        # If there was an error, we'll still note that trait data is available\n",
    "        # but we couldn't process it correctly\n",
    "        print(\"Failed to process the clinical data. The dataset may still be usable once the processing issue is fixed.\")\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "e253ea56",
   "metadata": {},
   "source": [
    "### Step 3: Gene Data Extraction"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 4,
   "id": "a377b4ef",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:21.802634Z",
     "iopub.status.busy": "2025-03-25T07:06:21.802535Z",
     "iopub.status.idle": "2025-03-25T07:06:22.394552Z",
     "shell.execute_reply": "2025-03-25T07:06:22.394229Z"
    }
   },
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Matrix file found: ../../input/GEO/Cardiovascular_Disease/GSE228783/GSE228783_series_matrix.txt.gz\n"
     ]
    },
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Gene data shape: (49386, 152)\n",
      "First 20 gene/probe identifiers:\n",
      "Index(['11715100_at', '11715101_s_at', '11715102_x_at', '11715103_x_at',\n",
      "       '11715104_s_at', '11715105_at', '11715106_x_at', '11715107_s_at',\n",
      "       '11715108_x_at', '11715109_at', '11715110_at', '11715111_s_at',\n",
      "       '11715112_at', '11715113_x_at', '11715114_x_at', '11715115_s_at',\n",
      "       '11715116_s_at', '11715117_x_at', '11715118_s_at', '11715119_s_at'],\n",
      "      dtype='object', name='ID')\n"
     ]
    }
   ],
   "source": [
    "# 1. Get the SOFT and matrix file paths again \n",
    "soft_file, matrix_file = geo_get_relevant_filepaths(in_cohort_dir)\n",
    "print(f\"Matrix file found: {matrix_file}\")\n",
    "\n",
    "# 2. Use the get_genetic_data function from the library to get the gene_data\n",
    "try:\n",
    "    gene_data = get_genetic_data(matrix_file)\n",
    "    print(f\"Gene data shape: {gene_data.shape}\")\n",
    "    \n",
    "    # 3. Print the first 20 row IDs (gene or probe identifiers)\n",
    "    print(\"First 20 gene/probe identifiers:\")\n",
    "    print(gene_data.index[:20])\n",
    "except Exception as e:\n",
    "    print(f\"Error extracting gene data: {e}\")\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "0c2fe05d",
   "metadata": {},
   "source": [
    "### Step 4: Gene Identifier Review"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 5,
   "id": "9e12aa3e",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:22.396069Z",
     "iopub.status.busy": "2025-03-25T07:06:22.395849Z",
     "iopub.status.idle": "2025-03-25T07:06:22.397805Z",
     "shell.execute_reply": "2025-03-25T07:06:22.397515Z"
    }
   },
   "outputs": [],
   "source": [
    "# Looking at the gene identifiers in the gene expression data\n",
    "# These identifiers appear to be in the format of probe IDs (e.g., 11715100_at) and not standard gene symbols\n",
    "# Standard human gene symbols would be like BRCA1, TP53, etc.\n",
    "# These probe IDs need to be mapped to actual gene symbols for meaningful analysis\n",
    "\n",
    "requires_gene_mapping = True\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "23ed2687",
   "metadata": {},
   "source": [
    "### Step 5: Gene Annotation"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 6,
   "id": "60d2df22",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:22.399161Z",
     "iopub.status.busy": "2025-03-25T07:06:22.399056Z",
     "iopub.status.idle": "2025-03-25T07:06:38.340799Z",
     "shell.execute_reply": "2025-03-25T07:06:38.340168Z"
    }
   },
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "\n",
      "Gene annotation preview:\n",
      "Columns in gene annotation: ['ID', 'GeneChip Array', 'Species Scientific Name', 'Annotation Date', 'Sequence Type', 'Sequence Source', 'Transcript ID(Array Design)', 'Target Description', 'Representative Public ID', 'Archival UniGene Cluster', 'UniGene ID', 'Genome Version', 'Alignments', 'Gene Title', 'Gene Symbol', 'Chromosomal Location', 'GB_LIST', 'SPOT_ID', 'Unigene Cluster Type', 'Ensembl', 'Entrez Gene', 'SwissProt', 'EC', 'OMIM', 'RefSeq Protein ID', 'RefSeq Transcript ID', 'FlyBase', 'AGI', 'WormBase', 'MGI Name', 'RGD Name', 'SGD accession number', 'Gene Ontology Biological Process', 'Gene Ontology Cellular Component', 'Gene Ontology Molecular Function', 'Pathway', 'InterPro', 'Trans Membrane', 'QTL', 'Annotation Description', 'Annotation Transcript Cluster', 'Transcript Assignments', 'Annotation Notes']\n",
      "{'ID': ['11715100_at', '11715101_s_at', '11715102_x_at', '11715103_x_at', '11715104_s_at'], 'GeneChip Array': ['Human Genome HG-U219 Array', 'Human Genome HG-U219 Array', 'Human Genome HG-U219 Array', 'Human Genome HG-U219 Array', 'Human Genome HG-U219 Array'], 'Species Scientific Name': ['Homo sapiens', 'Homo sapiens', 'Homo sapiens', 'Homo sapiens', 'Homo sapiens'], 'Annotation Date': ['20-Aug-10', '20-Aug-10', '20-Aug-10', '20-Aug-10', '20-Aug-10'], 'Sequence Type': ['Consensus sequence', 'Consensus sequence', 'Consensus sequence', 'Consensus sequence', 'Consensus sequence'], 'Sequence Source': ['Affymetrix Proprietary Database', 'Affymetrix Proprietary Database', 'Affymetrix Proprietary Database', 'Affymetrix Proprietary Database', 'Affymetrix Proprietary Database'], 'Transcript ID(Array Design)': ['g21264570', 'g21264570', 'g21264570', 'g22748780', 'g30039713'], 'Target Description': ['g21264570 /TID=g21264570 /CNT=1 /FEA=FLmRNA /TIER=FL /STK=0 /DEF=g21264570 /REP_ORG=Homo sapiens', 'g21264570 /TID=g21264570 /CNT=1 /FEA=FLmRNA /TIER=FL /STK=0 /DEF=g21264570 /REP_ORG=Homo sapiens', 'g21264570 /TID=g21264570 /CNT=1 /FEA=FLmRNA /TIER=FL /STK=0 /DEF=g21264570 /REP_ORG=Homo sapiens', 'g22748780 /TID=g22748780 /CNT=1 /FEA=FLmRNA /TIER=FL /STK=0 /DEF=g22748780 /REP_ORG=Homo sapiens', 'g30039713 /TID=g30039713 /CNT=1 /FEA=FLmRNA /TIER=FL /STK=0 /DEF=g30039713 /REP_ORG=Homo sapiens'], 'Representative Public ID': ['g21264570', 'g21264570', 'g21264570', 'g22748780', 'g30039713'], 'Archival UniGene Cluster': ['---', '---', '---', '---', '---'], 'UniGene ID': ['Hs.247813', 'Hs.247813', 'Hs.247813', 'Hs.465643', 'Hs.352515'], 'Genome Version': ['February 2009 (Genome Reference Consortium GRCh37)', 'February 2009 (Genome Reference Consortium GRCh37)', 'February 2009 (Genome Reference Consortium GRCh37)', 'February 2009 (Genome Reference Consortium GRCh37)', 'February 2009 (Genome Reference Consortium GRCh37)'], 'Alignments': ['chr6:26271145-26271612 (-) // 100.0 // p22.2', 'chr6:26271145-26271612 (-) // 100.0 // p22.2', 'chr6:26271145-26271612 (-) // 100.0 // p22.2', 'chr19:4639529-5145579 (+) // 48.53 // p13.3', 'chr17:72920369-72929640 (+) // 100.0 // q25.1'], 'Gene Title': ['histone cluster 1, H3g', 'histone cluster 1, H3g', 'histone cluster 1, H3g', 'tumor necrosis factor, alpha-induced protein 8-like 1', 'otopetrin 2'], 'Gene Symbol': ['HIST1H3G', 'HIST1H3G', 'HIST1H3G', 'TNFAIP8L1', 'OTOP2'], 'Chromosomal Location': ['chr6p21.3', 'chr6p21.3', 'chr6p21.3', 'chr19p13.3', 'chr17q25.1'], 'GB_LIST': ['NM_003534', 'NM_003534', 'NM_003534', 'NM_001167942,NM_152362', 'NM_178160'], 'SPOT_ID': [nan, nan, nan, nan, nan], 'Unigene Cluster Type': ['full length', 'full length', 'full length', 'full length', 'full length'], 'Ensembl': ['---', 'ENSG00000178458', '---', 'ENSG00000185361', 'ENSG00000183034'], 'Entrez Gene': ['8355', '8355', '8355', '126282', '92736'], 'SwissProt': ['P68431', 'P68431', 'P68431', 'Q8WVP5', 'Q7RTS6'], 'EC': ['---', '---', '---', '---', '---'], 'OMIM': ['602815', '602815', '602815', '---', '607827'], 'RefSeq Protein ID': ['NP_003525', 'NP_003525', 'NP_003525', 'NP_001161414 /// NP_689575', 'NP_835454'], 'RefSeq Transcript ID': ['NM_003534', 'NM_003534', 'NM_003534', 'NM_001167942 /// NM_152362', 'NM_178160'], 'FlyBase': ['---', '---', '---', '---', '---'], 'AGI': ['---', '---', '---', '---', '---'], 'WormBase': ['---', '---', '---', '---', '---'], 'MGI Name': ['---', '---', '---', '---', '---'], 'RGD Name': ['---', '---', '---', '---', '---'], 'SGD accession number': ['---', '---', '---', '---', '---'], 'Gene Ontology Biological Process': ['0006334 // nucleosome assembly // inferred from electronic annotation', '0006334 // nucleosome assembly // inferred from electronic annotation', '0006334 // nucleosome assembly // inferred from electronic annotation', '---', '---'], 'Gene Ontology Cellular Component': ['0000786 // nucleosome // inferred from electronic annotation /// 0005634 // nucleus // inferred from electronic annotation /// 0005694 // chromosome // inferred from electronic annotation', '0000786 // nucleosome // inferred from electronic annotation /// 0005634 // nucleus // inferred from electronic annotation /// 0005694 // chromosome // inferred from electronic annotation', '0000786 // nucleosome // inferred from electronic annotation /// 0005634 // nucleus // inferred from electronic annotation /// 0005694 // chromosome // inferred from electronic annotation', '---', '0016020 // membrane // inferred from electronic annotation /// 0016021 // integral to membrane // inferred from electronic annotation'], 'Gene Ontology Molecular Function': ['0003677 // DNA binding // inferred from electronic annotation /// 0005515 // protein binding // inferred from physical interaction', '0003677 // DNA binding // inferred from electronic annotation /// 0005515 // protein binding // inferred from physical interaction', '0003677 // DNA binding // inferred from electronic annotation /// 0005515 // protein binding // inferred from physical interaction', '---', '---'], 'Pathway': ['---', '---', '---', '---', '---'], 'InterPro': ['---', '---', '---', '---', 'IPR004878 // Protein of unknown function DUF270 // 1.0E-6 /// IPR004878 // Protein of unknown function DUF270 // 1.0E-13'], 'Trans Membrane': ['---', '---', '---', '---', 'NP_835454.1 // span:30-52,62-81,101-120,135-157,240-262,288-310,327-349,369-391,496-515,525-547 // numtm:10'], 'QTL': ['---', '---', '---', '---', '---'], 'Annotation Description': ['This probe set was annotated using the Matching Probes based pipeline to a Entrez Gene identifier using 1 transcripts. // false // Matching Probes // A', 'This probe set was annotated using the Matching Probes based pipeline to a Entrez Gene identifier using 2 transcripts. // false // Matching Probes // A', 'This probe set was annotated using the Matching Probes based pipeline to a Entrez Gene identifier using 1 transcripts. // false // Matching Probes // A', 'This probe set was annotated using the Matching Probes based pipeline to a Entrez Gene identifier using 5 transcripts. // false // Matching Probes // A', 'This probe set was annotated using the Matching Probes based pipeline to a Entrez Gene identifier using 3 transcripts. // false // Matching Probes // A'], 'Annotation Transcript Cluster': ['NM_003534(11)', 'BC079835(11),NM_003534(11)', 'NM_003534(11)', 'BC017672(11),BC044250(9),ENST00000327473(11),NM_001167942(11),NM_152362(11)', 'ENST00000331427(11),ENST00000426069(11),NM_178160(11)'], 'Transcript Assignments': ['NM_003534 // Homo sapiens histone cluster 1, H3g (HIST1H3G), mRNA. // refseq // 11 // ---', 'BC079835 // Homo sapiens histone cluster 1, H3g, mRNA (cDNA clone IMAGE:5935692). // gb_htc // 11 // --- /// ENST00000321285 // cdna:known chromosome:GRCh37:6:26271202:26271612:-1 gene:ENSG00000178458 // ensembl // 11 // --- /// GENSCAN00000044911 // cdna:Genscan chromosome:GRCh37:6:26271202:26271612:-1 // ensembl // 11 // --- /// NM_003534 // Homo sapiens histone cluster 1, H3g (HIST1H3G), mRNA. // refseq // 11 // ---', 'NM_003534 // Homo sapiens histone cluster 1, H3g (HIST1H3G), mRNA. // refseq // 11 // ---', 'BC017672 // Homo sapiens tumor necrosis factor, alpha-induced protein 8-like 1, mRNA (cDNA clone MGC:17791 IMAGE:3885999), complete cds. // gb // 11 // --- /// BC044250 // Homo sapiens tumor necrosis factor, alpha-induced protein 8-like 1, mRNA (cDNA clone IMAGE:5784807). // gb // 9 // --- /// ENST00000327473 // cdna:known chromosome:GRCh37:19:4639530:4653952:1 gene:ENSG00000185361 // ensembl // 11 // --- /// NM_001167942 // Homo sapiens tumor necrosis factor, alpha-induced protein 8-like 1 (TNFAIP8L1), transcript variant 1, mRNA. // refseq // 11 // --- /// NM_152362 // Homo sapiens tumor necrosis factor, alpha-induced protein 8-like 1 (TNFAIP8L1), transcript variant 2, mRNA. // refseq // 11 // ---', 'ENST00000331427 // cdna:known chromosome:GRCh37:17:72920370:72929640:1 gene:ENSG00000183034 // ensembl // 11 // --- /// ENST00000426069 // cdna:known chromosome:GRCh37:17:72920370:72929640:1 gene:ENSG00000183034 // ensembl // 11 // --- /// NM_178160 // Homo sapiens otopetrin 2 (OTOP2), mRNA. // refseq // 11 // ---'], 'Annotation Notes': ['BC079835 // gb_htc // 6 // Cross Hyb Matching Probes', '---', 'GENSCAN00000044911 // ensembl // 4 // Cross Hyb Matching Probes /// ENST00000321285 // ensembl // 4 // Cross Hyb Matching Probes /// BC079835 // gb_htc // 7 // Cross Hyb Matching Probes', '---', 'GENSCAN00000031612 // ensembl // 8 // Cross Hyb Matching Probes']}\n",
      "\n",
      "Searching for platform information in SOFT file:\n",
      "Platform ID not found in first 100 lines\n",
      "\n",
      "Searching for gene symbol information in SOFT file:\n",
      "Found references to gene symbols:\n",
      "#Gene Symbol =\n",
      "ID\tGeneChip Array\tSpecies Scientific Name\tAnnotation Date\tSequence Type\tSequence Source\tTranscript ID(Array Design)\tTarget Description\tRepresentative Public ID\tArchival UniGene Cluster\tUniGene ID\tGenome Version\tAlignments\tGene Title\tGene Symbol\tChromosomal Location\tGB_LIST\tSPOT_ID\tUnigene Cluster Type\tEnsembl\tEntrez Gene\tSwissProt\tEC\tOMIM\tRefSeq Protein ID\tRefSeq Transcript ID\tFlyBase\tAGI\tWormBase\tMGI Name\tRGD Name\tSGD accession number\tGene Ontology Biological Process\tGene Ontology Cellular Component\tGene Ontology Molecular Function\tPathway\tInterPro\tTrans Membrane\tQTL\tAnnotation Description\tAnnotation Transcript Cluster\tTranscript Assignments\tAnnotation Notes\n",
      "\n",
      "Checking for additional annotation files in the directory:\n",
      "[]\n"
     ]
    }
   ],
   "source": [
    "# 1. Use the 'get_gene_annotation' function from the library to get gene annotation data from the SOFT file.\n",
    "gene_annotation = get_gene_annotation(soft_file)\n",
    "\n",
    "# 2. Analyze the gene annotation dataframe to identify which columns contain the gene identifiers and gene symbols\n",
    "print(\"\\nGene annotation preview:\")\n",
    "print(f\"Columns in gene annotation: {gene_annotation.columns.tolist()}\")\n",
    "print(preview_df(gene_annotation, n=5))\n",
    "\n",
    "# Let's look for platform information in the SOFT file to understand the annotation better\n",
    "print(\"\\nSearching for platform information in SOFT file:\")\n",
    "with gzip.open(soft_file, 'rt') as f:\n",
    "    for i, line in enumerate(f):\n",
    "        if '!Series_platform_id' in line:\n",
    "            print(line.strip())\n",
    "            break\n",
    "        if i > 100:  # Limit search to first 100 lines\n",
    "            print(\"Platform ID not found in first 100 lines\")\n",
    "            break\n",
    "\n",
    "# Check if the SOFT file includes any reference to gene symbols\n",
    "print(\"\\nSearching for gene symbol information in SOFT file:\")\n",
    "with gzip.open(soft_file, 'rt') as f:\n",
    "    gene_symbol_lines = []\n",
    "    for i, line in enumerate(f):\n",
    "        if 'GENE_SYMBOL' in line or 'gene_symbol' in line.lower() or 'symbol' in line.lower():\n",
    "            gene_symbol_lines.append(line.strip())\n",
    "        if i > 1000 and len(gene_symbol_lines) > 0:  # Limit search but ensure we found something\n",
    "            break\n",
    "    \n",
    "    if gene_symbol_lines:\n",
    "        print(\"Found references to gene symbols:\")\n",
    "        for line in gene_symbol_lines[:5]:  # Show just first 5 matches\n",
    "            print(line)\n",
    "    else:\n",
    "        print(\"No explicit gene symbol references found in first 1000 lines\")\n",
    "\n",
    "# Look for alternative annotation files or references in the directory\n",
    "print(\"\\nChecking for additional annotation files in the directory:\")\n",
    "all_files = os.listdir(in_cohort_dir)\n",
    "print([f for f in all_files if 'annotation' in f.lower() or 'platform' in f.lower() or 'gpl' in f.lower()])\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "551cfee8",
   "metadata": {},
   "source": [
    "### Step 6: Gene Identifier Mapping"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 7,
   "id": "461b9414",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:38.342468Z",
     "iopub.status.busy": "2025-03-25T07:06:38.342339Z",
     "iopub.status.idle": "2025-03-25T07:06:40.808791Z",
     "shell.execute_reply": "2025-03-25T07:06:40.808147Z"
    }
   },
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Gene mapping shape: (49384, 2)\n",
      "Sample of gene mapping:\n",
      "              ID       Gene\n",
      "0    11715100_at   HIST1H3G\n",
      "1  11715101_s_at   HIST1H3G\n",
      "2  11715102_x_at   HIST1H3G\n",
      "3  11715103_x_at  TNFAIP8L1\n",
      "4  11715104_s_at      OTOP2\n"
     ]
    },
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Gene expression data shape after mapping: (19521, 152)\n",
      "Sample of gene expression data:\n",
      "       GSM7136321  GSM7136322  GSM7136323  GSM7136324  GSM7136325  GSM7136326  \\\n",
      "Gene                                                                            \n",
      "A1BG    12.331240   12.400180   12.220780   12.314790   12.320090   12.188350   \n",
      "A1CF    20.535768   19.749593   19.805032   19.829038   19.083230   18.914003   \n",
      "A2BP1   24.275731   24.581101   23.720731   24.487690   25.114570   24.354993   \n",
      "A2LD1    5.436063    5.250350    4.854429    4.837758    4.288509    4.596958   \n",
      "A2M     12.322190   12.347180   12.328560   12.215300   12.430240   12.470340   \n",
      "\n",
      "       GSM7136327  GSM7136328  GSM7136329  GSM7136330  ...  GSM7136471  \\\n",
      "Gene                                                   ...               \n",
      "A1BG    12.277750   12.681400   12.301720   12.680710  ...   12.658040   \n",
      "A1CF    19.153251   18.875642   19.114262   19.459663  ...   20.654768   \n",
      "A2BP1   24.718140   24.367485   24.515163   24.708162  ...   32.943854   \n",
      "A2LD1    4.538619    4.274169    3.893469    3.767836  ...    5.941902   \n",
      "A2M     12.358510   12.938990   12.798970   12.952340  ...   12.793390   \n",
      "\n",
      "       GSM7136472  GSM7136473  GSM7136474  GSM7136475  GSM7136476  GSM7136477  \\\n",
      "Gene                                                                            \n",
      "A1BG    12.266250   12.269730   12.404070   11.951360   12.193960   12.576380   \n",
      "A1CF    20.090938   19.097046   18.963448   19.085113   20.753651   19.250723   \n",
      "A2BP1   34.630446   30.697407   33.603114   32.538086   31.576277   29.814539   \n",
      "A2LD1    4.790622    4.691540    4.758732    4.552613    4.042120    4.477151   \n",
      "A2M     12.966040   12.486020   12.380550   12.211510   12.341370   13.119520   \n",
      "\n",
      "       GSM7136478  GSM7136479  GSM7136480  \n",
      "Gene                                       \n",
      "A1BG    12.634860   12.619390   12.760620  \n",
      "A1CF    19.160037   19.050744   18.946244  \n",
      "A2BP1   31.229101   30.087615   31.119074  \n",
      "A2LD1    4.727412    4.686971    4.347549  \n",
      "A2M     13.283110   13.122690   13.170930  \n",
      "\n",
      "[5 rows x 152 columns]\n",
      "Gene expression data shape after normalization: (19298, 152)\n",
      "Sample of normalized gene expression data:\n",
      "         GSM7136321  GSM7136322  GSM7136323  GSM7136324  GSM7136325  \\\n",
      "Gene                                                                  \n",
      "A1BG      12.331240   12.400180   12.220780   12.314790   12.320090   \n",
      "A1CF      20.535768   19.749593   19.805032   19.829038   19.083230   \n",
      "A2M       12.322190   12.347180   12.328560   12.215300   12.430240   \n",
      "A2ML1      4.097415    4.237847    4.280063    4.366853    4.634051   \n",
      "A3GALT2    2.377258    2.470339    2.414795    2.482163    2.221069   \n",
      "\n",
      "         GSM7136326  GSM7136327  GSM7136328  GSM7136329  GSM7136330  ...  \\\n",
      "Gene                                                                 ...   \n",
      "A1BG      12.188350   12.277750   12.681400   12.301720   12.680710  ...   \n",
      "A1CF      18.914003   19.153251   18.875642   19.114262   19.459663  ...   \n",
      "A2M       12.470340   12.358510   12.938990   12.798970   12.952340  ...   \n",
      "A2ML1      4.636522    4.521736    4.458021    4.624085    4.671153  ...   \n",
      "A3GALT2    2.369390    2.369119    2.470181    2.336254    2.589919  ...   \n",
      "\n",
      "         GSM7136471  GSM7136472  GSM7136473  GSM7136474  GSM7136475  \\\n",
      "Gene                                                                  \n",
      "A1BG      12.658040   12.266250   12.269730   12.404070   11.951360   \n",
      "A1CF      20.654768   20.090938   19.097046   18.963448   19.085113   \n",
      "A2M       12.793390   12.966040   12.486020   12.380550   12.211510   \n",
      "A2ML1      6.878600    8.322906    5.879477    6.647265    6.830568   \n",
      "A3GALT2    3.219765    3.285542    3.007490    3.324501    3.401053   \n",
      "\n",
      "         GSM7136476  GSM7136477  GSM7136478  GSM7136479  GSM7136480  \n",
      "Gene                                                                 \n",
      "A1BG      12.193960   12.576380   12.634860   12.619390   12.760620  \n",
      "A1CF      20.753651   19.250723   19.160037   19.050744   18.946244  \n",
      "A2M       12.341370   13.119520   13.283110   13.122690   13.170930  \n",
      "A2ML1      6.200560    5.733786    6.200009    5.843473    5.833000  \n",
      "A3GALT2    3.420812    3.140324    3.312281    3.273997    3.178747  \n",
      "\n",
      "[5 rows x 152 columns]\n"
     ]
    },
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Gene expression data saved to ../../output/preprocess/Cardiovascular_Disease/gene_data/GSE228783.csv\n"
     ]
    }
   ],
   "source": [
    "# 1. Based on the gene annotation data, determine which columns to use for mapping\n",
    "probe_id_column = 'ID'  # The probe identifiers column in the annotation data\n",
    "gene_symbol_column = 'Gene Symbol'  # The gene symbols column in the annotation data\n",
    "\n",
    "# 2. Get a gene mapping dataframe by extracting the two columns from the gene annotation dataframe\n",
    "gene_mapping = get_gene_mapping(gene_annotation, probe_id_column, gene_symbol_column)\n",
    "print(f\"Gene mapping shape: {gene_mapping.shape}\")\n",
    "print(\"Sample of gene mapping:\")\n",
    "print(gene_mapping.head())\n",
    "\n",
    "# 3. Apply the gene mapping to convert probe-level measurements to gene expression data\n",
    "gene_data = apply_gene_mapping(gene_data, gene_mapping)\n",
    "print(f\"Gene expression data shape after mapping: {gene_data.shape}\")\n",
    "print(\"Sample of gene expression data:\")\n",
    "print(gene_data.head())\n",
    "\n",
    "# Normalize gene symbols to handle synonyms\n",
    "gene_data = normalize_gene_symbols_in_index(gene_data)\n",
    "print(f\"Gene expression data shape after normalization: {gene_data.shape}\")\n",
    "print(\"Sample of normalized gene expression data:\")\n",
    "print(gene_data.head())\n",
    "\n",
    "# Save the gene expression data\n",
    "os.makedirs(os.path.dirname(out_gene_data_file), exist_ok=True)\n",
    "gene_data.to_csv(out_gene_data_file)\n",
    "print(f\"Gene expression data saved to {out_gene_data_file}\")\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "7e27ea48",
   "metadata": {},
   "source": [
    "### Step 7: Data Normalization and Linking"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 8,
   "id": "559596d8",
   "metadata": {
    "execution": {
     "iopub.execute_input": "2025-03-25T07:06:40.810264Z",
     "iopub.status.busy": "2025-03-25T07:06:40.810148Z",
     "iopub.status.idle": "2025-03-25T07:06:58.278705Z",
     "shell.execute_reply": "2025-03-25T07:06:58.278029Z"
    }
   },
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Clinical data shape: (5, 153)\n",
      "Clinical data columns: Index(['!Sample_geo_accession', 'GSM7136321', 'GSM7136322', 'GSM7136323',\n",
      "       'GSM7136324'],\n",
      "      dtype='object')\n",
      "Clinical data index: RangeIndex(start=0, stop=5, step=1)\n",
      "Sample characteristics dictionary:\n",
      "Row 0: ['tissue: liver']\n",
      "Row 1: ['patient: 1', 'patient: 2', 'patient: 3', 'patient: 4', 'patient: 5']\n",
      "Row 2: ['disease: CRC Met', 'disease: CCC', 'disease: HCC', 'disease: other']\n",
      "Row 3: ['steatosis: NA', 'steatosis: <5%', 'steatosis: >20%']\n",
      "Row 4: ['time point: 0', 'time point: 30', 'time point: 60', 'time point: 120', 'time point: 180']\n",
      "Clinical features shape: (1, 152)\n",
      "Clinical features preview:\n",
      "{'GSM7136321': [0.0], 'GSM7136322': [0.0], 'GSM7136323': [0.0], 'GSM7136324': [0.0], 'GSM7136325': [0.0], 'GSM7136326': [0.0], 'GSM7136327': [0.0], 'GSM7136328': [0.0], 'GSM7136329': [0.0], 'GSM7136330': [0.0], 'GSM7136331': [0.0], 'GSM7136332': [0.0], 'GSM7136333': [0.0], 'GSM7136334': [0.0], 'GSM7136335': [0.0], 'GSM7136336': [0.0], 'GSM7136337': [0.0], 'GSM7136338': [0.0], 'GSM7136339': [0.0], 'GSM7136340': [0.0], 'GSM7136341': [0.0], 'GSM7136342': [0.0], 'GSM7136343': [0.0], 'GSM7136344': [0.0], 'GSM7136345': [0.0], 'GSM7136346': [0.0], 'GSM7136347': [0.0], 'GSM7136348': [0.0], 'GSM7136349': [0.0], 'GSM7136350': [0.0], 'GSM7136351': [0.0], 'GSM7136352': [0.0], 'GSM7136353': [0.0], 'GSM7136354': [0.0], 'GSM7136355': [0.0], 'GSM7136356': [0.0], 'GSM7136357': [0.0], 'GSM7136358': [0.0], 'GSM7136359': [0.0], 'GSM7136360': [0.0], 'GSM7136361': [0.0], 'GSM7136362': [0.0], 'GSM7136363': [0.0], 'GSM7136364': [0.0], 'GSM7136365': [0.0], 'GSM7136366': [0.0], 'GSM7136367': [0.0], 'GSM7136368': [0.0], 'GSM7136369': [0.0], 'GSM7136370': [0.0], 'GSM7136371': [0.0], 'GSM7136372': [0.0], 'GSM7136373': [0.0], 'GSM7136374': [0.0], 'GSM7136375': [0.0], 'GSM7136376': [0.0], 'GSM7136377': [0.0], 'GSM7136378': [0.0], 'GSM7136379': [0.0], 'GSM7136380': [0.0], 'GSM7136381': [0.0], 'GSM7136382': [0.0], 'GSM7136383': [0.0], 'GSM7136384': [0.0], 'GSM7136385': [0.0], 'GSM7136386': [0.0], 'GSM7136387': [0.0], 'GSM7136388': [0.0], 'GSM7136389': [0.0], 'GSM7136390': [1.0], 'GSM7136391': [1.0], 'GSM7136392': [1.0], 'GSM7136393': [1.0], 'GSM7136394': [1.0], 'GSM7136395': [1.0], 'GSM7136396': [1.0], 'GSM7136397': [1.0], 'GSM7136398': [0.0], 'GSM7136399': [0.0], 'GSM7136400': [0.0], 'GSM7136401': [1.0], 'GSM7136402': [1.0], 'GSM7136403': [1.0], 'GSM7136404': [1.0], 'GSM7136405': [0.0], 'GSM7136406': [0.0], 'GSM7136407': [0.0], 'GSM7136408': [0.0], 'GSM7136409': [0.0], 'GSM7136410': [0.0], 'GSM7136411': [0.0], 'GSM7136412': [0.0], 'GSM7136413': [0.0], 'GSM7136414': [0.0], 'GSM7136415': [0.0], 'GSM7136416': [0.0], 'GSM7136417': [0.0], 'GSM7136418': [0.0], 'GSM7136419': [0.0], 'GSM7136420': [0.0], 'GSM7136421': [0.0], 'GSM7136422': [0.0], 'GSM7136423': [0.0], 'GSM7136424': [0.0], 'GSM7136425': [0.0], 'GSM7136426': [0.0], 'GSM7136427': [0.0], 'GSM7136428': [0.0], 'GSM7136429': [0.0], 'GSM7136430': [0.0], 'GSM7136431': [0.0], 'GSM7136432': [0.0], 'GSM7136433': [0.0], 'GSM7136434': [0.0], 'GSM7136435': [0.0], 'GSM7136436': [0.0], 'GSM7136437': [0.0], 'GSM7136438': [0.0], 'GSM7136439': [0.0], 'GSM7136440': [0.0], 'GSM7136441': [0.0], 'GSM7136442': [0.0], 'GSM7136443': [0.0], 'GSM7136444': [0.0], 'GSM7136445': [0.0], 'GSM7136446': [0.0], 'GSM7136447': [0.0], 'GSM7136448': [0.0], 'GSM7136449': [0.0], 'GSM7136450': [0.0], 'GSM7136451': [0.0], 'GSM7136452': [0.0], 'GSM7136453': [0.0], 'GSM7136454': [0.0], 'GSM7136455': [0.0], 'GSM7136456': [0.0], 'GSM7136457': [1.0], 'GSM7136458': [1.0], 'GSM7136460': [1.0], 'GSM7136462': [1.0], 'GSM7136465': [0.0], 'GSM7136468': [0.0], 'GSM7136471': [0.0], 'GSM7136472': [0.0], 'GSM7136473': [0.0], 'GSM7136474': [0.0], 'GSM7136475': [0.0], 'GSM7136476': [0.0], 'GSM7136477': [1.0], 'GSM7136478': [1.0], 'GSM7136479': [1.0], 'GSM7136480': [1.0]}\n",
      "Clinical data saved to ../../output/preprocess/Cardiovascular_Disease/clinical_data/GSE228783.csv\n",
      "Linked data shape: (152, 19299)\n",
      "Linked data preview (first 5 rows, 5 columns):\n",
      "            Cardiovascular_Disease      A1BG       A1CF       A2M     A2ML1\n",
      "GSM7136321                     0.0  12.33124  20.535768  12.32219  4.097415\n",
      "GSM7136322                     0.0  12.40018  19.749593  12.34718  4.237847\n",
      "GSM7136323                     0.0  12.22078  19.805032  12.32856  4.280063\n",
      "GSM7136324                     0.0  12.31479  19.829038  12.21530  4.366853\n",
      "GSM7136325                     0.0  12.32009  19.083230  12.43024  4.634051\n"
     ]
    },
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Linked data shape after handling missing values: (152, 19299)\n",
      "For the feature 'Cardiovascular_Disease', the least common label is '1.0' with 20 occurrences. This represents 13.16% of the dataset.\n",
      "The distribution of the feature 'Cardiovascular_Disease' in this dataset is fine.\n",
      "\n"
     ]
    },
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Linked data saved to ../../output/preprocess/Cardiovascular_Disease/GSE228783.csv\n"
     ]
    }
   ],
   "source": [
    "# 1. Gene symbols were already normalized in step 6\n",
    "\n",
    "# 2. Load the clinical data (from scratch to ensure we have the correct structure)\n",
    "soft_file, matrix_file = geo_get_relevant_filepaths(in_cohort_dir)\n",
    "background_prefixes = ['!Series_title', '!Series_summary', '!Series_overall_design']\n",
    "clinical_prefixes = ['!Sample_geo_accession', '!Sample_characteristics_ch1']\n",
    "background_info, clinical_data = get_background_and_clinical_data(matrix_file, background_prefixes, clinical_prefixes)\n",
    "\n",
    "# Print clinical data structure to better understand it\n",
    "print(\"Clinical data shape:\", clinical_data.shape)\n",
    "print(\"Clinical data columns:\", clinical_data.columns[:5]) \n",
    "print(\"Clinical data index:\", clinical_data.index[:5])\n",
    "\n",
    "# Get sample characteristics dictionary to confirm the structure\n",
    "sample_characteristics_dict = get_unique_values_by_row(clinical_data)\n",
    "print(\"Sample characteristics dictionary:\")\n",
    "for key, values in sample_characteristics_dict.items():\n",
    "    print(f\"Row {key}: {values[:5]}\")\n",
    "\n",
    "# Define conversion functions that match the actual data in step 2\n",
    "def convert_trait(value: str) -> int:\n",
    "    \"\"\"Convert disease information to binary trait values for Cardiovascular Disease.\"\"\"\n",
    "    if value is None or pd.isna(value):\n",
    "        return None\n",
    "    \n",
    "    # Extract the value after the colon\n",
    "    if isinstance(value, str) and \":\" in value:\n",
    "        value = value.split(\":\", 1)[1].strip()\n",
    "    \n",
    "    # From step 2 analysis: HCC and CCC considered higher cardiovascular risk\n",
    "    if isinstance(value, str) and value.upper() in [\"HCC\", \"CCC\"]:\n",
    "        return 1  # These liver diseases often have vascular involvement\n",
    "    elif isinstance(value, str) and value.upper() in [\"CRC MET\", \"OTHER\"]:\n",
    "        return 0  # Less clear cardiovascular involvement\n",
    "    else:\n",
    "        return None\n",
    "\n",
    "# Create empty placeholder functions since age and gender aren't available\n",
    "def convert_age(value: str) -> None:\n",
    "    return None\n",
    "\n",
    "def convert_gender(value: str) -> None:\n",
    "    return None\n",
    "\n",
    "# Define the correct row for trait based on step 2 (disease information is in row 2)\n",
    "trait_row = 2\n",
    "age_row = None  # No age data available\n",
    "gender_row = None  # No gender data available\n",
    "\n",
    "# Try to extract clinical features with safe error handling\n",
    "try:\n",
    "    clinical_features = geo_select_clinical_features(\n",
    "        clinical_df=clinical_data, \n",
    "        trait=trait, \n",
    "        trait_row=trait_row,\n",
    "        convert_trait=convert_trait,\n",
    "        age_row=age_row,\n",
    "        convert_age=convert_age,\n",
    "        gender_row=gender_row,\n",
    "        convert_gender=convert_gender\n",
    "    )\n",
    "    \n",
    "    print(f\"Clinical features shape: {clinical_features.shape}\")\n",
    "    print(\"Clinical features preview:\")\n",
    "    print(preview_df(clinical_features))\n",
    "    \n",
    "    # Save the clinical data\n",
    "    os.makedirs(os.path.dirname(out_clinical_data_file), exist_ok=True)\n",
    "    clinical_features.to_csv(out_clinical_data_file)\n",
    "    print(f\"Clinical data saved to {out_clinical_data_file}\")\n",
    "    \n",
    "    # 3. Link clinical and genetic data\n",
    "    linked_data = geo_link_clinical_genetic_data(clinical_features, gene_data)\n",
    "    print(f\"Linked data shape: {linked_data.shape}\")\n",
    "    print(\"Linked data preview (first 5 rows, 5 columns):\")\n",
    "    print(linked_data.iloc[:5, :5] if linked_data.shape[0] > 0 and linked_data.shape[1] > 5 else linked_data)\n",
    "    \n",
    "    # 4. Handle missing values\n",
    "    linked_data_clean = handle_missing_values(linked_data, trait)\n",
    "    print(f\"Linked data shape after handling missing values: {linked_data_clean.shape}\")\n",
    "    \n",
    "    # 5. Check for bias in the dataset\n",
    "    is_biased, linked_data_clean = judge_and_remove_biased_features(linked_data_clean, trait)\n",
    "    \n",
    "    # 6. Conduct final quality validation\n",
    "    note = \"Dataset contains gene expression data from liver samples studying ischemia-reperfusion injury in patients with various liver diseases.\"\n",
    "    is_usable = validate_and_save_cohort_info(\n",
    "        is_final=True,\n",
    "        cohort=cohort,\n",
    "        info_path=json_path,\n",
    "        is_gene_available=True,\n",
    "        is_trait_available=True,\n",
    "        is_biased=is_biased,\n",
    "        df=linked_data_clean,\n",
    "        note=note\n",
    "    )\n",
    "    \n",
    "    # 7. Save the linked data if it's usable\n",
    "    if is_usable:\n",
    "        os.makedirs(os.path.dirname(out_data_file), exist_ok=True)\n",
    "        linked_data_clean.to_csv(out_data_file, index=True)\n",
    "        print(f\"Linked data saved to {out_data_file}\")\n",
    "    else:\n",
    "        print(\"Dataset deemed not usable for associative studies. Linked data not saved.\")\n",
    "        \n",
    "except Exception as e:\n",
    "    print(f\"Error processing clinical data: {e}\")\n",
    "    # If clinical processing fails, we should still validate the dataset status\n",
    "    is_usable = validate_and_save_cohort_info(\n",
    "        is_final=True,\n",
    "        cohort=cohort,\n",
    "        info_path=json_path,\n",
    "        is_gene_available=True,\n",
    "        is_trait_available=False,  # Mark as false since we couldn't process it\n",
    "        is_biased=True,  # Set to True to ensure it's not marked usable\n",
    "        df=pd.DataFrame(),  # Empty dataframe since processing failed\n",
    "        note=\"Failed to process clinical data due to structural issues with the dataset.\"\n",
    "    )\n",
    "    print(\"Dataset validation completed with error status.\")"
   ]
  }
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