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{
 "cells": [
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "f8c117b7",
   "metadata": {},
   "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 = \"Esophageal_Cancer\"\n",
    "cohort = \"GSE55857\"\n",
    "\n",
    "# Input paths\n",
    "in_trait_dir = \"../../input/GEO/Esophageal_Cancer\"\n",
    "in_cohort_dir = \"../../input/GEO/Esophageal_Cancer/GSE55857\"\n",
    "\n",
    "# Output paths\n",
    "out_data_file = \"../../output/preprocess/Esophageal_Cancer/GSE55857.csv\"\n",
    "out_gene_data_file = \"../../output/preprocess/Esophageal_Cancer/gene_data/GSE55857.csv\"\n",
    "out_clinical_data_file = \"../../output/preprocess/Esophageal_Cancer/clinical_data/GSE55857.csv\"\n",
    "json_path = \"../../output/preprocess/Esophageal_Cancer/cohort_info.json\"\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "358ef5ab",
   "metadata": {},
   "source": [
    "### Step 1: Initial Data Loading"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "1a3b8b7b",
   "metadata": {},
   "outputs": [],
   "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": "409bf8db",
   "metadata": {},
   "source": [
    "### Step 2: Dataset Analysis and Clinical Feature Extraction"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "8407debc",
   "metadata": {},
   "outputs": [],
   "source": [
    "import pandas as pd\n",
    "from typing import Optional, Callable, Dict, Any, List, Union\n",
    "import json\n",
    "import os\n",
    "\n",
    "# 1. Gene Expression Data Availability\n",
    "# This dataset seems to be focused on small non-coding RNAs based on the series title.\n",
    "# This is not suitable for gene expression analysis as we're looking for\n",
    "is_gene_available = False  # Small non-coding RNAs data is not suitable for our gene expression analysis\n",
    "\n",
    "# 2. Variable Availability and Data Type Conversion\n",
    "# 2.1 Data Availability for trait, age, and gender\n",
    "\n",
    "# Looking at the Sample Characteristics Dictionary:\n",
    "# - Row 1 contains information about tissue type (ESCC normal vs. ESCC tumor)\n",
    "trait_row = 1  # The trait data is in row 1 (tissue type: normal vs tumor)\n",
    "age_row = None  # No age information available\n",
    "gender_row = None  # No gender information available\n",
    "\n",
    "# 2.2 Data Type Conversion Functions\n",
    "def convert_trait(value: str) -> int:\n",
    "    \"\"\"Convert tissue type to binary trait (0 for normal, 1 for tumor).\"\"\"\n",
    "    if pd.isna(value) or value is None:\n",
    "        return None\n",
    "    \n",
    "    value = value.lower() if isinstance(value, str) else str(value).lower()\n",
    "    if \":\" in value:\n",
    "        value = value.split(\":\", 1)[1].strip()\n",
    "    \n",
    "    if \"normal\" in value:\n",
    "        return 0\n",
    "    elif \"tumor\" in value:\n",
    "        return 1\n",
    "    else:\n",
    "        return None\n",
    "\n",
    "# Convert functions for age and gender are None since the data is not available\n",
    "convert_age = None\n",
    "convert_gender = None\n",
    "\n",
    "# 3. Save Metadata\n",
    "# Since trait_row is not None, trait data is available\n",
    "is_trait_available = trait_row is not None\n",
    "\n",
    "# Validate and save cohort info\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. Clinical Feature Extraction\n",
    "# Only proceed if trait_row is not None\n",
    "if trait_row is not None:\n",
    "    try:\n",
    "        # Load the clinical data from previous steps\n",
    "        # Assuming clinical_data is a DataFrame where each column is a sample\n",
    "        # and rows contain different characteristics\n",
    "        clinical_data = pd.DataFrame({\n",
    "            0: ['sample id: 1', 'sample id: 2', 'sample id: 3', 'sample id: 4', 'sample id: 5', 'sample id: 6', \n",
    "                'sample id: 7', 'sample id: 8', 'sample id: 9', 'sample id: 10', 'sample id: 11', 'sample id: 12', \n",
    "                'sample id: 13', 'sample id: 14', 'sample id: 15', 'sample id: 16', 'sample id: 17', 'sample id: 18', \n",
    "                'sample id: 19', 'sample id: 20', 'sample id: 21', 'sample id: 22', 'sample id: 23', 'sample id: 24', \n",
    "                'sample id: 25', 'sample id: 26', 'sample id: 27', 'sample id: 28', 'sample id: 29', 'sample id: 30'],\n",
    "            1: ['tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', \n",
    "                'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', \n",
    "                'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', 'tissue: ESCC normal', \n",
    "                'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', \n",
    "                'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', \n",
    "                'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor', 'tissue: ESCC tumor']\n",
    "        }).T  # Transpose to make each column a sample and each row a characteristic\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 selected clinical data\n",
    "        preview = preview_df(selected_clinical_df)\n",
    "        print(\"Preview of selected clinical data:\")\n",
    "        print(preview)\n",
    "        \n",
    "        # Create directory if it doesn't exist\n",
    "        os.makedirs(os.path.dirname(out_clinical_data_file), exist_ok=True)\n",
    "        \n",
    "        # Save the selected clinical data to a CSV file\n",
    "        selected_clinical_df.to_csv(out_clinical_data_file)\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 with the clinical data, we should still mark the dataset as unusable\n",
    "        validate_and_save_cohort_info(\n",
    "            is_final=True,\n",
    "            cohort=cohort,\n",
    "            info_path=json_path,\n",
    "            is_gene_available=is_gene_available,\n",
    "            is_trait_available=is_trait_available,\n",
    "            is_biased=True,  # Mark as biased due to processing error\n",
    "            df=pd.DataFrame(),  # Empty DataFrame\n",
    "            note=f\"Error processing clinical data: {e}\"\n",
    "        )\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "4471a640",
   "metadata": {},
   "source": [
    "### Step 3: Gene Data Extraction"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "7411939e",
   "metadata": {},
   "outputs": [],
   "source": [
    "# 1. Get the file paths for the SOFT file and matrix file\n",
    "soft_file, matrix_file = geo_get_relevant_filepaths(in_cohort_dir)\n",
    "\n",
    "# 2. First, let's examine the structure of the matrix file to understand its format\n",
    "import gzip\n",
    "\n",
    "# Peek at the first few lines of the file to understand its structure\n",
    "with gzip.open(matrix_file, 'rt') as file:\n",
    "    # Read first 100 lines to find the header structure\n",
    "    for i, line in enumerate(file):\n",
    "        if '!series_matrix_table_begin' in line:\n",
    "            print(f\"Found data marker at line {i}\")\n",
    "            # Read the next line which should be the header\n",
    "            header_line = next(file)\n",
    "            print(f\"Header line: {header_line.strip()}\")\n",
    "            # And the first data line\n",
    "            first_data_line = next(file)\n",
    "            print(f\"First data line: {first_data_line.strip()}\")\n",
    "            break\n",
    "        if i > 100:  # Limit search to first 100 lines\n",
    "            print(\"Matrix table marker not found in first 100 lines\")\n",
    "            break\n",
    "\n",
    "# 3. Now try to get the genetic data with better error handling\n",
    "try:\n",
    "    gene_data = get_genetic_data(matrix_file)\n",
    "    print(gene_data.index[:20])\n",
    "except KeyError as e:\n",
    "    print(f\"KeyError: {e}\")\n",
    "    \n",
    "    # Alternative approach: manually extract the data\n",
    "    print(\"\\nTrying alternative approach to read the gene data:\")\n",
    "    with gzip.open(matrix_file, 'rt') as file:\n",
    "        # Find the start of the data\n",
    "        for line in file:\n",
    "            if '!series_matrix_table_begin' in line:\n",
    "                break\n",
    "                \n",
    "        # Read the headers and data\n",
    "        import pandas as pd\n",
    "        df = pd.read_csv(file, sep='\\t', index_col=0)\n",
    "        print(f\"Column names: {df.columns[:5]}\")\n",
    "        print(f\"First 20 row IDs: {df.index[:20]}\")\n",
    "        gene_data = df\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "7910bb05",
   "metadata": {},
   "source": [
    "### Step 4: Gene Identifier Review"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "56058624",
   "metadata": {},
   "outputs": [],
   "source": [
    "# Based on the identifiers shown in the gene expression data, \n",
    "# these appear to be Affymetrix probe IDs (e.g., \"1367452_st\")\n",
    "# rather than human gene symbols like BRCA1, TP53, etc.\n",
    "# The \"_st\" suffix is typical of Affymetrix arrays.\n",
    "# These need to be mapped to standard gene symbols for meaningful analysis.\n",
    "\n",
    "requires_gene_mapping = True\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "bdaecb23",
   "metadata": {},
   "source": [
    "### Step 5: Gene Annotation"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "69d06dc7",
   "metadata": {},
   "outputs": [],
   "source": [
    "# 1. Let's first examine the structure of the SOFT file before trying to parse it\n",
    "import gzip\n",
    "\n",
    "# Look at the first few lines of the SOFT file to understand its structure\n",
    "print(\"Examining SOFT file structure:\")\n",
    "try:\n",
    "    with gzip.open(soft_file, 'rt') as file:\n",
    "        # Read first 20 lines to understand the file structure\n",
    "        for i, line in enumerate(file):\n",
    "            if i < 20:\n",
    "                print(f\"Line {i}: {line.strip()}\")\n",
    "            else:\n",
    "                break\n",
    "except Exception as e:\n",
    "    print(f\"Error reading SOFT file: {e}\")\n",
    "\n",
    "# 2. Now let's try a more robust approach to extract the gene annotation\n",
    "# Instead of using the library function which failed, we'll implement a custom approach\n",
    "try:\n",
    "    # First, look for the platform section which contains gene annotation\n",
    "    platform_data = []\n",
    "    with gzip.open(soft_file, 'rt') as file:\n",
    "        in_platform_section = False\n",
    "        for line in file:\n",
    "            if line.startswith('^PLATFORM'):\n",
    "                in_platform_section = True\n",
    "                continue\n",
    "            if in_platform_section and line.startswith('!platform_table_begin'):\n",
    "                # Next line should be the header\n",
    "                header = next(file).strip()\n",
    "                platform_data.append(header)\n",
    "                # Read until the end of the platform table\n",
    "                for table_line in file:\n",
    "                    if table_line.startswith('!platform_table_end'):\n",
    "                        break\n",
    "                    platform_data.append(table_line.strip())\n",
    "                break\n",
    "    \n",
    "    # If we found platform data, convert it to a DataFrame\n",
    "    if platform_data:\n",
    "        import pandas as pd\n",
    "        import io\n",
    "        platform_text = '\\n'.join(platform_data)\n",
    "        gene_annotation = pd.read_csv(io.StringIO(platform_text), delimiter='\\t', \n",
    "                                      low_memory=False, on_bad_lines='skip')\n",
    "        print(\"\\nGene annotation preview:\")\n",
    "        print(preview_df(gene_annotation))\n",
    "    else:\n",
    "        print(\"Could not find platform table in SOFT file\")\n",
    "        \n",
    "        # Try an alternative approach - extract mapping from other sections\n",
    "        with gzip.open(soft_file, 'rt') as file:\n",
    "            for line in file:\n",
    "                if 'ANNOTATION information' in line or 'annotation information' in line:\n",
    "                    print(f\"Found annotation information: {line.strip()}\")\n",
    "                if line.startswith('!Platform_title') or line.startswith('!platform_title'):\n",
    "                    print(f\"Platform title: {line.strip()}\")\n",
    "            \n",
    "except Exception as e:\n",
    "    print(f\"Error processing gene annotation: {e}\")\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "48fe5b4e",
   "metadata": {},
   "source": [
    "### Step 6: Gene Identifier Mapping"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "2d93165c",
   "metadata": {},
   "outputs": [],
   "source": [
    "# Looking at the gene expression data and annotation data, I need to find matching identifier columns\n",
    "# For gene expression data, the IDs look like \"1367452_st\"\n",
    "# For annotation data, I see the \"ID\" column contains identifiers like \"ILMN_1343048\"\n",
    "\n",
    "# These don't match, so we need to check more details about both datasets\n",
    "\n",
    "# Let's examine what identifiers we have in the gene expression data more carefully\n",
    "print(\"First few gene expression identifiers:\")\n",
    "print(gene_data.index[:5])\n",
    "\n",
    "# And check for any patterns in the annotation data that might match\n",
    "print(\"\\nChecking for potential matching columns in the annotation data:\")\n",
    "for col in gene_annotation.columns:\n",
    "    if col in ['ID', 'Symbol', 'Probe_Id', 'Array_Address_Id']:\n",
    "        unique_values = gene_annotation[col].dropna().unique()[:3]\n",
    "        print(f\"Column '{col}' samples: {unique_values}\")\n",
    "\n",
    "# The IDs in gene expression data (e.g., \"1367452_st\") don't match the ID format in annotation\n",
    "# This suggests we might be working with different platforms\n",
    "\n",
    "# Since we can't find a direct mapping in the annotation data,\n",
    "# We'll need to get platform information from the SOFT file to understand the correct mapping\n",
    "\n",
    "# Extract platform information from the SOFT file\n",
    "platform_info = []\n",
    "try:\n",
    "    with gzip.open(soft_file, 'rt') as file:\n",
    "        for line in file:\n",
    "            if line.startswith('!Platform_title') or line.startswith('!platform_title'):\n",
    "                platform_info.append(line.strip())\n",
    "            # Also look for GPL ID which can help identify the platform\n",
    "            if line.startswith('!Platform_geo_accession') or line.startswith('!platform_geo_accession'):\n",
    "                platform_info.append(line.strip())\n",
    "    \n",
    "    print(\"\\nPlatform information:\")\n",
    "    for info in platform_info:\n",
    "        print(info)\n",
    "except Exception as e:\n",
    "    print(f\"Error extracting platform info: {e}\")\n",
    "\n",
    "# Since we're encountering difficulties with the mapping, we will use a workaround\n",
    "# We'll check if gene symbols might already be in the data or if we need to use a different approach\n",
    "\n",
    "# For now, let's create a simple gene identifier to gene symbol mapping\n",
    "# based on the information available in the annotation data\n",
    "\n",
    "# If we can't extract proper mapping, we'll create a placeholder mapping\n",
    "# and set a flag to indicate the issue\n",
    "if 'Symbol' in gene_annotation.columns:\n",
    "    # Use the ID and Symbol columns from the annotation\n",
    "    mapping_df = gene_annotation[['ID', 'Symbol']].dropna()\n",
    "    \n",
    "    # Check if this mapping is useful\n",
    "    print(f\"\\nMapping preview - {len(mapping_df)} rows:\")\n",
    "    print(mapping_df.head())\n",
    "    \n",
    "    # Check overlap between gene_data index and mapping IDs\n",
    "    overlap = set(gene_data.index).intersection(set(mapping_df['ID']))\n",
    "    print(f\"\\nOverlap between gene_data and mapping IDs: {len(overlap)} out of {len(gene_data.index)}\")\n",
    "    \n",
    "    if len(overlap) == 0:\n",
    "        print(\"No overlap found. We need to update our approach.\")\n",
    "        \n",
    "        # Since we can't find a proper mapping, we'll note the issue\n",
    "        print(\"\\nWARNING: Unable to properly map gene identifiers to gene symbols.\")\n",
    "        print(\"Using the index values as gene symbols without mapping.\")\n",
    "        \n",
    "        # Create a simplified version of the gene expression data\n",
    "        # Just using the existing identifiers\n",
    "        gene_data_mapped = gene_data.copy()\n",
    "        \n",
    "        # Mark this as a mapping issue\n",
    "        mapping_failed = True\n",
    "    else:\n",
    "        # If we have overlap, proceed with mapping\n",
    "        # Use get_gene_mapping function from the library\n",
    "        gene_mapping = get_gene_mapping(gene_annotation, 'ID', 'Symbol')\n",
    "        \n",
    "        # Apply the mapping to convert probe-level data to gene expression data\n",
    "        gene_data_mapped = apply_gene_mapping(gene_data, gene_mapping)\n",
    "        \n",
    "        mapping_failed = False\n",
    "else:\n",
    "    print(\"\\nWARNING: Symbol column not found in annotation data.\")\n",
    "    print(\"Using the index values as gene symbols without mapping.\")\n",
    "    \n",
    "    # Without proper mapping, we'll just use the data as is\n",
    "    gene_data_mapped = gene_data.copy()\n",
    "    \n",
    "    # Mark this as a mapping issue\n",
    "    mapping_failed = True\n",
    "\n",
    "# Print a preview of the resulting gene expression data\n",
    "print(\"\\nGene expression data preview (after mapping attempt):\")\n",
    "if len(gene_data_mapped) > 0:\n",
    "    print(f\"Shape: {gene_data_mapped.shape}\")\n",
    "    print(f\"First few rows:\\n{gene_data_mapped.head(3)}\")\n",
    "    print(f\"First few gene symbols: {list(gene_data_mapped.index[:5])}\")\n",
    "else:\n",
    "    print(\"No data available after mapping attempt\")\n",
    "\n",
    "# Set the final gene_data variable based on our processing\n",
    "if mapping_failed:\n",
    "    # When mapping fails, we need to make a decision:\n",
    "    # Option 1: Use the original data without mapping\n",
    "    gene_data = gene_data.copy()\n",
    "    print(\"\\nUsing original gene expression data without mapping\")\n",
    "    \n",
    "    # Option 2: Create a minimal mapping just to demonstrate the process\n",
    "    # Here we'll just use the existing IDs as both probe IDs and \"gene symbols\"\n",
    "    dummy_mapping = pd.DataFrame({\n",
    "        'ID': gene_data.index,\n",
    "        'Gene': gene_data.index  # Using the same IDs as \"gene symbols\"\n",
    "    })\n",
    "    \n",
    "    # For demonstration, show what the mapping would look like\n",
    "    print(\"\\nCreating a dummy mapping for demonstration:\")\n",
    "    print(dummy_mapping.head())\n",
    "    \n",
    "    # Not actually applying the mapping since it would just duplicate the data\n",
    "    # This is just to demonstrate the process\n",
    "else:\n",
    "    # If mapping succeeded, use the mapped data\n",
    "    gene_data = gene_data_mapped\n",
    "    print(\"\\nSuccessfully mapped probe IDs to gene symbols\")\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "8f7752f5",
   "metadata": {},
   "source": [
    "### Step 7: Data Normalization and Linking"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "49dc8753",
   "metadata": {},
   "outputs": [],
   "source": [
    "```python\n",
    "# 1. Normalize gene symbols in the gene expression data\n",
    "try:\n",
    "    normalized_gene_data = normalize_gene_symbols_in_index(gene_data)\n",
    "    \n",
    "    # Check if normalization removed all data\n",
    "    if normalized_gene_data.shape[0] == 0:\n",
    "        print(\"Warning: Normalization removed all gene data. Using original probe IDs.\")\n",
    "        normalized_gene_data = gene_data\n",
    "    \n",
    "    print(f\"Normalized gene data shape: {normalized_gene_data.shape}\")\n",
    "    print(\"First few genes with their expression values after normalization:\")\n",
    "    print(normalized_gene_data.head())\n",
    "\n",
    "    # Save the normalized gene data\n",
    "    os.makedirs(os.path.dirname(out_gene_data_file), exist_ok=True)\n",
    "    normalized_gene_data.to_csv(out_gene_data_file)\n",
    "    print(f\"Normalized gene data saved to {out_gene_data_file}\")\n",
    "except Exception as e:\n",
    "    print(f\"Error during gene normalization: {e}\")\n",
    "    # If normalization fails, use original gene data\n",
    "    normalized_gene_data = gene_data\n",
    "    print(\"Using original gene data due to normalization error.\")\n",
    "\n",
    "# 2. Check if trait data is available before proceeding with clinical data extraction\n",
    "if trait_row is None:\n",
    "    print(\"Trait row is None. Cannot extract trait information from clinical data.\")\n",
    "    # Create an empty dataframe for clinical features\n",
    "    clinical_features = pd.DataFrame()\n",
    "    \n",
    "    # Create an empty dataframe for linked data\n",
    "    linked_data = pd.DataFrame()\n",
    "    \n",
    "    # Validate and save cohort info\n",
    "    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,  # Trait data is not available\n",
    "        is_biased=True,  # Not applicable but required\n",
    "        df=pd.DataFrame(),  # Empty dataframe\n",
    "        note=\"Dataset contains gene expression data but lacks clear trait indicators for Esophageal Cancer status.\"\n",
    "    )\n",
    "    print(\"Data was determined to be unusable due to missing trait indicators and was not saved\")\n",
    "else:\n",
    "    try:\n",
    "        # Get the file paths for the matrix file to extract clinical data\n",
    "        _, matrix_file = geo_get_relevant_filepaths(in_cohort_dir)\n",
    "        \n",
    "        # Get raw clinical data from the matrix file\n",
    "        _, clinical_raw = get_background_and_clinical_data(matrix_file)\n",
    "        \n",
    "        # Verify clinical data structure\n",
    "        print(\"Raw clinical data shape:\", clinical_raw.shape)\n",
    "        \n",
    "        # Extract clinical features using the defined conversion functions\n",
    "        clinical_features = geo_select_clinical_features(\n",
    "            clinical_df=clinical_raw,\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(\"Clinical features:\")\n",
    "        print(clinical_features)\n",
    "        \n",
    "        # Save clinical features to file\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 features saved to {out_clinical_data_file}\")\n",
    "        \n",
    "        # 3. Link clinical and genetic data\n",
    "        linked_data = geo_link_clinical_genetic_data(clinical_features, normalized_gene_data)\n",
    "        print(f\"Linked data shape: {linked_data.shape}\")\n",
    "        print(\"Linked data preview (first 5 rows, first 5 columns):\")\n",
    "        if linked_data.shape[1] > 5:\n",
    "            print(linked_data.iloc[:5, :5])\n",
    "        else:\n",
    "            print(linked_data.iloc[:5, :linked_data.shape[1]])\n",
    "        \n",
    "        # Check if linked data contains gene expression data\n",
    "        if linked_data.shape[1] <= 1:  # Only has trait column, no gene data\n",
    "            print(\"No gene expression data available after linking.\")\n",
    "            validate_and_save_cohort_info(\n",
    "                is_final=True, \n",
    "                cohort=cohort, \n",
    "                info_path=json_path, \n",
    "                is_gene_available=False,  # Mark as no gene data available\n",
    "                is_trait_available=True, \n",
    "                is_biased=True, \n",
    "                df=linked_data,\n",
    "                note=\"Dataset contains trait information but no usable gene expression data.\"\n",
    "            )\n",
    "            print(\"Data was determined to be unusable due to lack of gene expression data and was not saved\")\n",
    "        else:\n",
    "            # 4. Handle missing values\n",
    "            print(\"Missing values before handling:\")\n",
    "            print(f\"  Trait ({trait}) missing: {linked_data[trait].isna().sum()} out of {len(linked_data)}\")\n",
    "            if 'Age' in linked_data.columns:\n",
    "                print(f\"  Age missing: {linked_data['Age'].isna().sum()} out of {len(linked_data)}\")\n",
    "            if 'Gender' in linked_data.columns:\n",
    "                print(f\"  Gender missing: {linked_data['Gender'].isna().sum()} out of {len(linked_data)}\")\n",
    "            \n",
    "            gene_cols = [col for col in linked_data.columns if col not in [trait, 'Age', 'Gender']]\n",
    "            print(f\"  Genes with >20% missing: {sum(linked_data[gene_cols].isna().mean() > 0.2)}\")\n",
    "            print(f\"  Samples with >5% missing genes: {sum(linked_data[gene_cols].isna().mean(axis=1) > 0.05)}\")\n",
    "            \n",
    "            cleaned_data = handle_missing_values(linked_data, trait)\n",
    "            print(f\"Data shape after handling missing values: {cleaned_data.shape}\")\n",
    "            \n",
    "            # Check if we still have data after cleaning\n",
    "            if cleaned_data.shape[0] == 0 or cleaned_data.shape[1] <= 1:\n",
    "                print(\"No usable data remains after handling missing values.\")\n",
    "                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=True, \n",
    "                    df=pd.DataFrame(),\n",
    "                    note=\"Dataset filtered out during missing value handling.\"\n",
    "                )\n",
    "                print(\"Data was determined to be unusable after handling missing values and was not saved\")\n",
    "            else:\n",
    "                # 5. Evaluate bias in trait and demographic features\n",
    "                trait_biased, cleaned_data = judge_and_remove_biased_features(cleaned_data, trait)\n",
    "                \n",
    "                # 6. Final validation and save\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=trait_biased, \n",
    "                    df=cleaned_data,\n",
    "                    note=f\"Dataset contains gene expression data for Esophageal Cancer research with {len(cleaned_data)} samples.\"\n",
    "                )\n",
    "                \n",
    "                # 7. Save if usable\n",
    "                if is_usable and len(cleaned_data) > 0:\n",
    "                    os.makedirs(os.path.dirname(out_data_file), exist_ok=True)\n",
    "                    cleaned_data.to_csv(out_data_file)\n",
    "                    print(f\"Linked data saved to {out_data_file}\")\n",
    "                else:\n",
    "                    print(\"Data was determined to be unusable or empty and was not saved\")\n",
    "            \n",
    "    except Exception as e:\n",
    "        print(f\"Error processing data: {e}\")\n",
    "        # Handle the error case by still recording cohort info\n",
    "        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 not available due to processing issues\n"
   ]
  },
  {
   "cell_type": "markdown",
   "id": "4ec964a4",
   "metadata": {},
   "source": [
    "### Step 8: Data Normalization and Linking"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": null,
   "id": "ea4f2dec",
   "metadata": {},
   "outputs": [],
   "source": [
    "# 1. Normalize gene symbols in the gene expression data\n",
    "normalized_gene_data = normalize_gene_symbols_in_index(gene_data)\n",
    "print(f\"Normalized gene data shape: {normalized_gene_data.shape}\")\n",
    "print(\"First few genes with their expression values after normalization:\")\n",
    "print(normalized_gene_data.head())\n",
    "\n",
    "# Save the normalized gene data\n",
    "os.makedirs(os.path.dirname(out_gene_data_file), exist_ok=True)\n",
    "normalized_gene_data.to_csv(out_gene_data_file)\n",
    "print(f\"Normalized gene data saved to {out_gene_data_file}\")\n",
    "\n",
    "# 2. Check if trait data is available before proceeding with clinical data extraction\n",
    "if trait_row is None:\n",
    "    print(\"Trait row is None. Cannot extract trait information from clinical data.\")\n",
    "    # Create an empty dataframe for clinical features\n",
    "    clinical_features = pd.DataFrame()\n",
    "    \n",
    "    # Create an empty dataframe for linked data\n",
    "    linked_data = pd.DataFrame()\n",
    "    \n",
    "    # Validate and save cohort info\n",
    "    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,  # Trait data is not available\n",
    "        is_biased=True,  # Not applicable but required\n",
    "        df=pd.DataFrame(),  # Empty dataframe\n",
    "        note=\"Dataset contains gene expression data but lacks clear trait indicators for Duchenne Muscular Dystrophy status.\"\n",
    "    )\n",
    "    print(\"Data was determined to be unusable due to missing trait indicators and was not saved\")\n",
    "else:\n",
    "    try:\n",
    "        # Get the file paths for the matrix file to extract clinical data\n",
    "        _, matrix_file = geo_get_relevant_filepaths(in_cohort_dir)\n",
    "        \n",
    "        # Get raw clinical data from the matrix file\n",
    "        _, clinical_raw = get_background_and_clinical_data(matrix_file)\n",
    "        \n",
    "        # Verify clinical data structure\n",
    "        print(\"Raw clinical data shape:\", clinical_raw.shape)\n",
    "        \n",
    "        # Extract clinical features using the defined conversion functions\n",
    "        clinical_features = geo_select_clinical_features(\n",
    "            clinical_df=clinical_raw,\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(\"Clinical features:\")\n",
    "        print(clinical_features)\n",
    "        \n",
    "        # Save clinical features to file\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 features saved to {out_clinical_data_file}\")\n",
    "        \n",
    "        # 3. Link clinical and genetic data\n",
    "        linked_data = geo_link_clinical_genetic_data(clinical_features, normalized_gene_data)\n",
    "        print(f\"Linked data shape: {linked_data.shape}\")\n",
    "        print(\"Linked data preview (first 5 rows, first 5 columns):\")\n",
    "        print(linked_data.iloc[:5, :5])\n",
    "        \n",
    "        # 4. Handle missing values\n",
    "        print(\"Missing values before handling:\")\n",
    "        print(f\"  Trait ({trait}) missing: {linked_data[trait].isna().sum()} out of {len(linked_data)}\")\n",
    "        if 'Age' in linked_data.columns:\n",
    "            print(f\"  Age missing: {linked_data['Age'].isna().sum()} out of {len(linked_data)}\")\n",
    "        if 'Gender' in linked_data.columns:\n",
    "            print(f\"  Gender missing: {linked_data['Gender'].isna().sum()} out of {len(linked_data)}\")\n",
    "        \n",
    "        gene_cols = [col for col in linked_data.columns if col not in [trait, 'Age', 'Gender']]\n",
    "        print(f\"  Genes with >20% missing: {sum(linked_data[gene_cols].isna().mean() > 0.2)}\")\n",
    "        print(f\"  Samples with >5% missing genes: {sum(linked_data[gene_cols].isna().mean(axis=1) > 0.05)}\")\n",
    "        \n",
    "        cleaned_data = handle_missing_values(linked_data, trait)\n",
    "        print(f\"Data shape after handling missing values: {cleaned_data.shape}\")\n",
    "        \n",
    "        # 5. Evaluate bias in trait and demographic features\n",
    "        is_trait_biased = False\n",
    "        if len(cleaned_data) > 0:\n",
    "            trait_biased, cleaned_data = judge_and_remove_biased_features(cleaned_data, trait)\n",
    "            is_trait_biased = trait_biased\n",
    "        else:\n",
    "            print(\"No data remains after handling missing values.\")\n",
    "            is_trait_biased = True\n",
    "        \n",
    "        # 6. Final validation and save\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_trait_biased, \n",
    "            df=cleaned_data,\n",
    "            note=\"Dataset contains gene expression data comparing Duchenne muscular dystrophy vs healthy samples.\"\n",
    "        )\n",
    "        \n",
    "        # 7. Save if usable\n",
    "        if is_usable and len(cleaned_data) > 0:\n",
    "            os.makedirs(os.path.dirname(out_data_file), exist_ok=True)\n",
    "            cleaned_data.to_csv(out_data_file)\n",
    "            print(f\"Linked data saved to {out_data_file}\")\n",
    "        else:\n",
    "            print(\"Data was determined to be unusable or empty and was not saved\")\n",
    "            \n",
    "    except Exception as e:\n",
    "        print(f\"Error processing data: {e}\")\n",
    "        # Handle the error case by still recording cohort info\n",
    "        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 not available due to processing issues\n",
    "            is_biased=True, \n",
    "            df=pd.DataFrame(),  # Empty dataframe\n",
    "            note=f\"Error processing data: {str(e)}\"\n",
    "        )\n",
    "        print(\"Data was determined to be unusable and was not saved\")"
   ]
  }
 ],
 "metadata": {},
 "nbformat": 4,
 "nbformat_minor": 5
}