<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT01710618</url>
  </required_header>
  <id_info>
    <org_study_id>359367-12</org_study_id>
    <nct_id>NCT01710618</nct_id>
  </id_info>
  <brief_title>SEXI: Sciatic Expression and Identification of Differential Proteins in Traumatized Versus Nontraumatized Nerves</brief_title>
  <acronym>SEXI</acronym>
  <official_title>SEXI: Sciatic Expression and Identification of Differential Proteins in Traumatized Versus Non-traumatized Nerves</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Defense and Veterans Center for Integrative Pain Management</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Duke University</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Defense and Veterans Center for Integrative Pain Management</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      The purpose of the study is to elucidate the different proteins that may appear on the&#xD;
      &quot;injured&quot; end of the sciatic nerve and compare it to the other end of the cut nerve, which is&#xD;
      &quot;uninjured&quot;. By comparing the &quot;normal&quot; versus &quot;traumatized&quot; ends of the cut sciatic nerve,&#xD;
      the investigators hope to identify proteins that may be specifically related to pain. For&#xD;
      instance, if the non-traumatized end of the nerve has none of Protein X but the traumatized&#xD;
      end of the nerve has a lot of Protein X, then Protein X would be a protein that the&#xD;
      investigators will want to study further and see if it plays any role in the sensation of&#xD;
      pain.&#xD;
&#xD;
      Other studies have shown that there are specific pain-related proteins and they may be&#xD;
      increased or decreased after traumatic injury. If the investigators are able to identify&#xD;
      specific proteins that are related to pain, then that may eventually lead to the development&#xD;
      of medications that will block those proteins and decrease the sensation of pain.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Subjects scheduled for final amputation revision and closure(s) will be approached to&#xD;
      participate in the study. On the day of the study participant's planned amputation revision,&#xD;
      research staff will liaise with surgical staff to coordinate nerve specimen retrieval. If a&#xD;
      patient is to have an above the knee amputation, a sciatic nerve specimen will be obtained,&#xD;
      if a patient is to have a below the knee or through knee amputation, a tibial nerve specimen&#xD;
      will be obtained (the major component of the sciatic nerve after it divides into the tibial&#xD;
      and peroneal nerves in the popliteal fossa of the knee). The surgeon will place a suture in&#xD;
      the proximal end of the specimen to indicate the non-traumatized end of the nerve, as well as&#xD;
      provide an estimate of the longitudinal location of the nerve sampled (i.e. 10cm distal to&#xD;
      the knee joint line, 15 cm proximal to the knee at the knee joint level). Each sample will be&#xD;
      excised in the same manner - using a scalpel. Once the tissue is excised by the surgeon, it&#xD;
      will be placed in a plastic test tube that is labeled with the subjects' study identification&#xD;
      number (with the suffix &quot;A&quot; or &quot;B&quot; if two samples are being retrieved from that subject),&#xD;
      transferred to a member of the research staff and immediately frozen in liquid nitrogen. The&#xD;
      frozen specimen will then be delivered to the Department of Research Programs laboratory&#xD;
      where it will be stored at -80°C prior to analysis. The promixal, middle, and distal samples&#xD;
      will be obtained at the DRP lab and the remaining segments of nerve will be replaced in the&#xD;
      -80 freezer for future analysis. Samples will then be sent to Duke University for processing&#xD;
      and analysis via a secure air mail carrier. The task of processing samples will be performed&#xD;
      immediately prior to mass spectroscopy analysis by colleagues at Duke University.&#xD;
      Collaborator Dr Moseley's lab responsibilities include both protein extraction and assay. The&#xD;
      number of extractions per sample will vary depending on its length, from 2 to about 50.&#xD;
      Collaborator Dr Van de ven's lab responsibilities include sample handling, pathologic&#xD;
      examination, removing thin slices for histologic examination and segment homogenization for&#xD;
      RNA,DNA and protein extraction.&#xD;
&#xD;
      Individual segments will be thawed on ice and homogenized immediately upon thawing.&#xD;
      Homogenization methods will be optimized to produce the highest dissolved protein content.&#xD;
      Methods to be trialed will include sonication, mechanical disruption by laboratory blender,&#xD;
      mechanical disruption by manual chopping followed by Potter device, and freeze thawing.&#xD;
&#xD;
      Buffer and solvent system variables to be trialed include salt concentration, pH and buffer&#xD;
      type, chaotrope concentration (urea and thiourea), presence of organic co-solvents such as&#xD;
      DMSO, and the presence of nonionic detergent. All homogenization will be performed between 0&#xD;
      and 4°C and in the presence of protease inhibitors.&#xD;
&#xD;
      For proteomic analysis, High-performance liquid chromatography/electrospray mass spectrometry&#xD;
      will be employed as a complementary route to discovering differences in protein expression&#xD;
      between healthy and traumatized tissue. Following solid-phase extraction of soluble and&#xD;
      solubilized proteins, we will optimize conditions for separation of protein peaks detected by&#xD;
      UV absorbance and/or total ion current and may employ more than one set of conditions in&#xD;
      order to focus on different sets of extracted proteins. We will use high-porosity C-18&#xD;
      reversed-phase columns to separate cytosolic proteins, and C-4 for membrane proteins.&#xD;
      Standard methods of ionization, fragmentation, and detection will be used that are consistent&#xD;
      with the Shimadzu LCMS-IT-TOF mass spectrometer at the Research Operations Service (ROS).&#xD;
&#xD;
      For RNA analysis, extracted RNA will be brought to the Duke Center for Genomic and&#xD;
      Computational Biology Genomic Technologies core. There, RNA quality will be determined using&#xD;
      an Agilent 2100bioanalyzer followed by Affymetrix GeneChip analysis. David Corcoran, the&#xD;
      biostatistician who works at the Duke Center for Genomic and Computational Biology, will&#xD;
      perform data analysis.&#xD;
&#xD;
      In this study, no changes to the standard of care will be implemented. The sciatic nerve&#xD;
      specimens would otherwise be discarded as human tissue; there are no other ongoing&#xD;
      investigations using these specimens.&#xD;
&#xD;
      Following the analysis of the sciatic nerve, we will collect data - demographic, injury, and&#xD;
      pain treatment information, as well as pre-operative pain scores - from subjects' charts to&#xD;
      determine secondary outcome variables. (See Appendix B - Case Report Forms)&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Active, not recruiting</overall_status>
  <start_date type="Actual">July 2011</start_date>
  <completion_date type="Anticipated">March 2025</completion_date>
  <primary_completion_date type="Anticipated">March 2025</primary_completion_date>
  <study_type>Observational</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <observational_model>Cohort</observational_model>
    <time_perspective>Prospective</time_perspective>
  </study_design_info>
  <enrollment type="Anticipated">100</enrollment>
  <condition>Pain</condition>
  <biospec_retention>Samples Without DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      sciatic nerve specimen will be obtained&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Male or female Operation Iraqi Freedom (OIF)/(Operation New Dawn (OND) or Operation&#xD;
        Enduring Freedom (OEF) veterans who meet eligibility criteria who require an amputation&#xD;
        revision occurring during their initial hospital admission.&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Non-Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Males and females 18 years and older, capable of providing informed consent indicating&#xD;
             awareness of the investigational nature of the study, in keeping with institutional&#xD;
             policy&#xD;
&#xD;
          -  Written informed consent must be obtained from each subject prior to entering the&#xD;
             study&#xD;
&#xD;
          -  Traumatic lower extremity (above knee or below knee) amputation that occurred as a&#xD;
             result of exposure to improvised explosive device (IED) blast, gunshot, or vehicle&#xD;
             rollover during combat in support of OIF/OND or OEF.&#xD;
&#xD;
          -  Scheduled final amputation revision to occur during the initial hospital admission at&#xD;
             Walter Reed National Military Medical Center&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Inability to provide informed consent&#xD;
&#xD;
          -  Traumatic or surgical foot amputations&#xD;
&#xD;
          -  Amputation occurring at Walter Reed Army Medical Center or another continental United&#xD;
             States (CONUS) hospital after failed limb salvage&#xD;
&#xD;
          -  Nerve specimen less than 5cm in length&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>89 Years</maximum_age>
    <healthy_volunteers>No</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Chester c Buckenmaier</last_name>
    <role>Principal Investigator</role>
    <affiliation>Defense and Veterans Center for Integrative Pain Management</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Walter Reed National Military Medical Center</name>
      <address>
        <city>Bethesda</city>
        <state>Maryland</state>
        <zip>20889</zip>
        <country>United States</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>United States</country>
  </location_countries>
  <verification_date>April 2023</verification_date>
  <study_first_submitted>October 17, 2012</study_first_submitted>
  <study_first_submitted_qc>October 18, 2012</study_first_submitted_qc>
  <study_first_posted type="Estimate">October 19, 2012</study_first_posted>
  <last_update_submitted>April 5, 2023</last_update_submitted>
  <last_update_submitted_qc>April 5, 2023</last_update_submitted_qc>
  <last_update_posted type="Actual">April 6, 2023</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>Protein Expression of Sciatic Nerve</keyword>
  <keyword>traumatized sciatic nerve</keyword>
  <patient_data>
    <sharing_ipd>No</sharing_ipd>
  </patient_data>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT01710618
org study id: 359367-12
nct id: NCT01710618
lead sponsor: 
collaborator: 
has dmc: No
The purpose of the study is to elucidate the different proteins that may appear on the
      "injured" end of the sciatic nerve and compare it to the other end of the cut nerve, which is
      "uninjured". By comparing the "normal" versus "traumatized" ends of the cut sciatic nerve,
      the investigators hope to identify proteins that may be specifically related to pain. For
      instance, if the non-traumatized end of the nerve has none of Protein X but the traumatized
      end of the nerve has a lot of Protein X, then Protein X would be a protein that the
      investigators will want to study further and see if it plays any role in the sensation of
      pain.

      Other studies have shown that there are specific pain-related proteins and they may be
      increased or decreased after traumatic injury. If the investigators are able to identify
      specific proteins that are related to pain, then that may eventually lead to the development
      of medications that will block those proteins and decrease the sensation of pain.
Subjects scheduled for final amputation revision and closure(s) will be approached to
      participate in the study. On the day of the study participant's planned amputation revision,
      research staff will liaise with surgical staff to coordinate nerve specimen retrieval. If a
      patient is to have an above the knee amputation, a sciatic nerve specimen will be obtained,
      if a patient is to have a below the knee or through knee amputation, a tibial nerve specimen
      will be obtained (the major component of the sciatic nerve after it divides into the tibial
      and peroneal nerves in the popliteal fossa of the knee). The surgeon will place a suture in
      the proximal end of the specimen to indicate the non-traumatized end of the nerve, as well as
      provide an estimate of the longitudinal location of the nerve sampled (i.e. 10cm distal to
      the knee joint line, 15 cm proximal to the knee at the knee joint level). Each sample will be
      excised in the same manner - using a scalpel. Once the tissue is excised by the surgeon, it
      will be placed in a plastic test tube that is labeled with the subjects' study identification
      number (with the suffix "A" or "B" if two samples are being retrieved from that subject),
      transferred to a member of the research staff and immediately frozen in liquid nitrogen. The
      frozen specimen will then be delivered to the Department of Research Programs laboratory
      where it will be stored at -80°C prior to analysis. The promixal, middle, and distal samples
      will be obtained at the DRP lab and the remaining segments of nerve will be replaced in the
      -80 freezer for future analysis. Samples will then be sent to Duke University for processing
      and analysis via a secure air mail carrier. The task of processing samples will be performed
      immediately prior to mass spectroscopy analysis by colleagues at Duke University.
      Collaborator Dr Moseley's lab responsibilities include both protein extraction and assay. The
      number of extractions per sample will vary depending on its length, from 2 to about 50.
      Collaborator Dr Van de ven's lab responsibilities include sample handling, pathologic
      examination, removing thin slices for histologic examination and segment homogenization for
      RNA,DNA and protein extraction.

      Individual segments will be thawed on ice and homogenized immediately upon thawing.
      Homogenization methods will be optimized to produce the highest dissolved protein content.
      Methods to be trialed will include sonication, mechanical disruption by laboratory blender,
      mechanical disruption by manual chopping followed by Potter device, and freeze thawing.

      Buffer and solvent system variables to be trialed include salt concentration, pH and buffer
      type, chaotrope concentration (urea and thiourea), presence of organic co-solvents such as
      DMSO, and the presence of nonionic detergent. All homogenization will be performed between 0
      and 4°C and in the presence of protease inhibitors.

      For proteomic analysis, High-performance liquid chromatography/electrospray mass spectrometry
      will be employed as a complementary route to discovering differences in protein expression
      between healthy and traumatized tissue. Following solid-phase extraction of soluble and
      solubilized proteins, we will optimize conditions for separation of protein peaks detected by
      UV absorbance and/or total ion current and may employ more than one set of conditions in
      order to focus on different sets of extracted proteins. We will use high-porosity C-18
      reversed-phase columns to separate cytosolic proteins, and C-4 for membrane proteins.
      Standard methods of ionization, fragmentation, and detection will be used that are consistent
      with the Shimadzu LCMS-IT-TOF mass spectrometer at the Research Operations Service (ROS).

      For RNA analysis, extracted RNA will be brought to the Duke Center for Genomic and
      Computational Biology Genomic Technologies core. There, RNA quality will be determined using
      an Agilent 2100bioanalyzer followed by Affymetrix GeneChip analysis. David Corcoran, the
      biostatistician who works at the Duke Center for Genomic and Computational Biology, will
      perform data analysis.

      In this study, no changes to the standard of care will be implemented. The sciatic nerve
      specimens would otherwise be discarded as human tissue; there are no other ongoing
      investigations using these specimens.

      Following the analysis of the sciatic nerve, we will collect data - demographic, injury, and
      pain treatment information, as well as pre-operative pain scores - from subjects' charts to
      determine secondary outcome variables. (See Appendix B - Case Report Forms)
observational model: Cohort
time perspective: Prospective
sciatic nerve specimen will be obtained
study pop: 
sampling method: Non-Probability Sample
criteria: 
gender: All
minimum age: 18 Years
maximum age: 89 Years
healthy volunteers: No
last name: Chester c Buckenmaier
role: Principal Investigator
affiliation: Defense and Veterans Center for Integrative Pain Management
facility: 
country: United States
responsible party type: Sponsor
sharing ipd: No