<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/NCT03303040</url>
  </required_header>
  <id_info>
    <org_study_id>IRB201602186-N</org_study_id>
    <secondary_id>R01AR072328</secondary_id>
    <nct_id>NCT03303040</nct_id>
  </id_info>
  <brief_title>Effect of Diaphragm Stimulation During Surgery</brief_title>
  <official_title>The Effect of Intermittent Hemidiaphragm Stimulation During Surgery on Mitochondrial Function, Single Fiber Contractile Force and Catabolic Pathways in Humans</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>University of Florida</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>National Institutes of Health (NIH)</agency>
      <agency_class>NIH</agency_class>
    </collaborator>
    <collaborator>
      <agency>University of Arizona</agency>
      <agency_class>Other</agency_class>
    </collaborator>
    <collaborator>
      <agency>National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)</agency>
      <agency_class>NIH</agency_class>
    </collaborator>
  </sponsors>
  <source>University of Florida</source>
  <oversight_info>
    <has_dmc>Yes</has_dmc>
    <is_fda_regulated_drug>No</is_fda_regulated_drug>
    <is_fda_regulated_device>No</is_fda_regulated_device>
  </oversight_info>
  <brief_summary>
    <textblock>
      During major surgical procedures general anesthesia is used to make the patient unconscious.&#xD;
      General anesthesia insures that the patient is unaware of any pain caused by surgery. General&#xD;
      anesthesia also prevents the patient from moving to prevent any potential surgical error. At&#xD;
      the same time general anesthesia makes it impossible for the patient to breathe. To help the&#xD;
      patient breathe a breathing tube is placed into the patient's airway and connected to the&#xD;
      mechanical ventilator. A mechanical ventilator is an artificial breathing pump, which&#xD;
      delivers gas into a patient's airways.&#xD;
&#xD;
      The purpose of this research study is to determine if brief periods of diaphragm stimulation&#xD;
      can prevent diaphragm problems caused by the use of mechanical ventilators and surgery. To&#xD;
      answer this question the changes in the genes responsible for maintaining diaphragm function&#xD;
      will be studied. A gene is the code present in each cell in your body and controls the&#xD;
      behavior of that cell. In addition, the changes in the contractile properties of muscle&#xD;
      fibers will be studied. The results from this study may help develop new treatments to&#xD;
      prevent diaphragm weakness resulting from mechanical ventilation use.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Although mechanical ventilation (MV) is life-sustaining, it comes with a cost. MV&#xD;
      dramatically reduces diaphragm contractility, induces ventilator-induced diaphragm&#xD;
      dysfunction (VIDD) and sometimes leads to weaning failure. VIDD includes reduced&#xD;
      mitochondrial respiration and increased oxidative stress, muscle fiber damage and decreased&#xD;
      diaphragm force production.&#xD;
&#xD;
      In animal models, intermittent diaphragm contraction during MV support attenuates VIDD.&#xD;
      However, there are only limited data addressing this problem in humans. Here, the study team&#xD;
      propose to directly test the hypothesis that intermittent electrical stimulation (ES) of the&#xD;
      human hemidiaphragm during prolonged cardiac surgeries with MV support prevents/attenuates&#xD;
      VIDD in the active hemidiaphragm. Mitochondrial function is central to energy metabolism and&#xD;
      skeletal muscle function in a chronically active muscle, such as the diaphragm. Although&#xD;
      abnormal mitochondrial function is thought to precipitate VIDD in animal models, limited data&#xD;
      are available concerning mitochondrial contributions to VIDD in humans. Of even greater&#xD;
      importance, there are no interventions available to attenuate these defects in humans. Here,&#xD;
      the study team will test the impact of an innovative experimental treatment, intermittent&#xD;
      electrical stimulation (ES) of the hemidiaphragm during prolonged surgeries with MV, on&#xD;
      mitochondrial function, single fiber contractile properties and catabolic muscle pathways in&#xD;
      human diaphragm. Using a within-subjects experimental design, muscle samples from a&#xD;
      stimulated hemidiaphragms will be compared with samples from the unstimulated hemidiaphragm.&#xD;
      The study team will investigate mitochondrial dysfunction and oxidative stress during&#xD;
      prolonged CTS/MV, and the potential of ES to attenuate or prevent VIDD. Next, the study team&#xD;
      will investigate the effects of ES on single fiber contractile properties and Titin&#xD;
      integrity. Finally, the study team will study the effect of ES on proteolytic pathways&#xD;
      (caspase, calpain and ubiquitin-proteasome) and ribosomal RNA markers of decreased protein&#xD;
      synthesis implicated in VIDD.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Active, not recruiting</overall_status>
  <start_date type="Actual">February 14, 2018</start_date>
  <completion_date type="Anticipated">October 6, 2023</completion_date>
  <primary_completion_date type="Actual">May 31, 2022</primary_completion_date>
  <phase>N/A</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>Non-Randomized</allocation>
    <intervention_model>Parallel Assignment</intervention_model>
    <intervention_model_description>This is a within subjects design. One side of each subject's diaphragm will be stimulated. The other side of the subject's diaphragm will not be stimulated and will therefore serve as the control. Biopsies will be taken from both sides and compared.</intervention_model_description>
    <primary_purpose>Prevention</primary_purpose>
    <masking>None (Open Label)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Mitochondrial respiration</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>High-resolution respirometry will be used to assess mitochondrial respiration. It will be quantified as pmol oxygen/sec/mg wet weight.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Mitochondrial reactive oxygen species production</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Mitochondrial reactive oxygen species (ROS) production will be assessed using an in situ approach to measure hydrogen peroxide production in permeabilized diaphragm skeletal muscle fiber bundles. It will be quantified as pmol/min/mg dry weight.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Mitochondrial DNA mutation frequency</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Long-Amplicon quantitative PCR will be used to measure the frequency of mitochondrial DNA mutations. It will be quantified as number of lesions/10 kilobases.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Nuclear DNA mutation frequency</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Long-Amplicon quantitative PCR will be used to measure the frequency of nuclear DNA mutations. It will be quantified as number of lesions/10 kilobases.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Aconitase activity</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>In order to evaluate mitochondrial damage, actonitase activity will be measured spectrophotometrically. It will be quantified as units/mg protein.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Lipid peroxidation</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Lipid peroxidation will be assessed by measuring 4-hydroxy-2-nonenal-modified proteins. It will be quantified as arbitrary optical density units.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Citrate cynthase activity</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Changes in electron transport chain will be assessed by measuring citrate cynthase activity. It will be quantifed as nmol/mg protein/min.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Cytochrome c oxidase (COX) activity</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Changes in electron transport chain will be assessed by measuring cytochrome c oxidase (COX) activity. It will be quantifed as Units/mcg protein.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Single diaphragm fiber, specific force</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Single diaphragm fiber mechanical force properties will be measured. Specific force is quantified as kN/m^2.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Single diaphragm fiber, rate of tension redevelopment</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Single diaphragm fiber mechanical force properties will be measured. The rate of tension redevelopment is quantified as s^(-1).</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Single diaphragm fiber, maximum shortening velocity</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Single diaphragm fiber mechanical force properties will be measured. The maximum shortening velocity is quantified as mm/s..</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Titin size</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Titin integrity will be assessed. A relative titin size will be quantified in nm.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Total titin to myosin heavy chain ratio</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Titin integrity will be assessed. The total titin to myosin heavy chain ratio will be presented as a unitless value.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Titin exon composition</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>The composition of titin exons will be assessed and quantified via real-time polymerase chain reaction (qPCR). It will be presented as percent change in expression.&#xD;
as a percent change in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Titin binding proteins</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Titin binding proteins will be assessed and quantified via real-time polymerase chain reaction (qPCR). It will be presented as percent change in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Calpain 1</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Calpain 1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Calpain 2</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Calpain 2 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Calpain 3</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Calpain 3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Caspase-3</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Caspase-3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Caspase-9</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Caspase-9 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>20S proteasome</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>20S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>26S proteasome</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>26S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>MurF1</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>MurF1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Atrogin 1</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Atrogin 1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Foxo-3</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>Foxo-3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>28SrRNA</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>28SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>18SrRNA</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>18SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>45S pre-rRNA</measure>
    <time_frame>Up to eight hours</time_frame>
    <description>45S pre-rRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression.</description>
  </primary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Actual">58</enrollment>
  <condition>Mechanical Ventilation Complication</condition>
  <condition>Diaphragm Injury</condition>
  <arm_group>
    <arm_group_label>Stimulation</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>Electrical stimulation of hemidiaphragm</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Control</arm_group_label>
    <arm_group_type>No Intervention</arm_group_type>
    <description>No stimulation of hemidiaphragm</description>
  </arm_group>
  <intervention>
    <intervention_type>Other</intervention_type>
    <intervention_name>Electrical stimulation of hemidiaphragm</intervention_name>
    <description>Electrical impulses</description>
    <arm_group_label>Stimulation</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Patients undergoing complex, elective prolonged surgeries, usually lasting 5-8 hours&#xD;
             or longer, including lung transplants (e.g. valveoplasty, coronary artery bypass&#xD;
             and/or aortic repairs)&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  history of prior surgery to the diaphragm or pleura;&#xD;
&#xD;
          -  a diagnosis of COPD will be determined from a clinical history consistent with chronic&#xD;
             bronchitis and/or emphysema, a long history of cigarette smoking, and pulmonary&#xD;
             function tests consistent with irreversible airflow obstruction (FEV1 &lt; 40% predicted,&#xD;
             according to European Respiratory Society criteria [will not apply to transplant&#xD;
             patients]&#xD;
&#xD;
          -  a diagnosis of chronic heart failure (NYHA class IV)&#xD;
&#xD;
          -  clinical diagnosis of other lung disease (cystic fibrosis, bronchiectasis, lung&#xD;
             cancer; etc.) [will not apply to transplant patients]&#xD;
&#xD;
          -  renal insufficiency (serum creatinine &gt; 1.6 mg/dl);&#xD;
&#xD;
          -  severe hepatic disease (any liver function tests &gt; 1.5 times the upper limit of&#xD;
             normal);&#xD;
&#xD;
          -  undernourishment (body mass index &lt; 20 kg/m2),&#xD;
&#xD;
          -  chronic uncontrolled or poorly controlled metabolic diseases (e.g., diabetes, hypo- or&#xD;
             hyperthyroidism)&#xD;
&#xD;
          -  orthopedic diseases, suspected paraneoplastic or myopathic syndromes,&#xD;
&#xD;
          -  if in the surgeons' judgment the patients' clinical status warrants, diaphragm&#xD;
             stimulation will be stopped and biopsies will not be obtained,&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <gender_based>Yes</gender_based>
    <gender_description>12 females and 12 males (based on gender identity) will be recruited for the study</gender_description>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>85 Years</maximum_age>
    <healthy_volunteers>No</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Anatole D Martin, PhD</last_name>
    <role>Principal Investigator</role>
    <affiliation>University of Florida</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Thomas M Beaver, MD</last_name>
    <role>Principal Investigator</role>
    <affiliation>University of Florida</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Barbara Smith, PhD, PT</last_name>
    <role>Principal Investigator</role>
    <affiliation>University of Florida</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>University of Florida</name>
      <address>
        <city>Gainesville</city>
        <state>Florida</state>
        <zip>32610</zip>
        <country>United States</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>United States</country>
  </location_countries>
  <verification_date>June 2023</verification_date>
  <study_first_submitted>September 26, 2017</study_first_submitted>
  <study_first_submitted_qc>October 4, 2017</study_first_submitted_qc>
  <study_first_posted type="Actual">October 5, 2017</study_first_posted>
  <last_update_submitted>June 5, 2023</last_update_submitted>
  <last_update_submitted_qc>June 5, 2023</last_update_submitted_qc>
  <last_update_posted type="Actual">June 6, 2023</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>Mechanical ventilation</keyword>
  <keyword>Diaphragm weakness</keyword>
  <keyword>Difficulty weaning from mechanical ventilation</keyword>
  <keyword>Diaphragm stimulation</keyword>
  <pending_results>
    <submitted>September 5, 2023</submitted>
  </pending_results>
  <!-- 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/NCT03303040
org study id: IRB201602186-N
secondary id: R01AR072328
nct id: NCT03303040
lead sponsor: 
collaborator: 
collaborator: 
collaborator: 
has dmc: Yes
is fda regulated drug: No
is fda regulated device: No
During major surgical procedures general anesthesia is used to make the patient unconscious.
      General anesthesia insures that the patient is unaware of any pain caused by surgery. General
      anesthesia also prevents the patient from moving to prevent any potential surgical error. At
      the same time general anesthesia makes it impossible for the patient to breathe. To help the
      patient breathe a breathing tube is placed into the patient's airway and connected to the
      mechanical ventilator. A mechanical ventilator is an artificial breathing pump, which
      delivers gas into a patient's airways.

      The purpose of this research study is to determine if brief periods of diaphragm stimulation
      can prevent diaphragm problems caused by the use of mechanical ventilators and surgery. To
      answer this question the changes in the genes responsible for maintaining diaphragm function
      will be studied. A gene is the code present in each cell in your body and controls the
      behavior of that cell. In addition, the changes in the contractile properties of muscle
      fibers will be studied. The results from this study may help develop new treatments to
      prevent diaphragm weakness resulting from mechanical ventilation use.
Although mechanical ventilation (MV) is life-sustaining, it comes with a cost. MV
      dramatically reduces diaphragm contractility, induces ventilator-induced diaphragm
      dysfunction (VIDD) and sometimes leads to weaning failure. VIDD includes reduced
      mitochondrial respiration and increased oxidative stress, muscle fiber damage and decreased
      diaphragm force production.

      In animal models, intermittent diaphragm contraction during MV support attenuates VIDD.
      However, there are only limited data addressing this problem in humans. Here, the study team
      propose to directly test the hypothesis that intermittent electrical stimulation (ES) of the
      human hemidiaphragm during prolonged cardiac surgeries with MV support prevents/attenuates
      VIDD in the active hemidiaphragm. Mitochondrial function is central to energy metabolism and
      skeletal muscle function in a chronically active muscle, such as the diaphragm. Although
      abnormal mitochondrial function is thought to precipitate VIDD in animal models, limited data
      are available concerning mitochondrial contributions to VIDD in humans. Of even greater
      importance, there are no interventions available to attenuate these defects in humans. Here,
      the study team will test the impact of an innovative experimental treatment, intermittent
      electrical stimulation (ES) of the hemidiaphragm during prolonged surgeries with MV, on
      mitochondrial function, single fiber contractile properties and catabolic muscle pathways in
      human diaphragm. Using a within-subjects experimental design, muscle samples from a
      stimulated hemidiaphragms will be compared with samples from the unstimulated hemidiaphragm.
      The study team will investigate mitochondrial dysfunction and oxidative stress during
      prolonged CTS/MV, and the potential of ES to attenuate or prevent VIDD. Next, the study team
      will investigate the effects of ES on single fiber contractile properties and Titin
      integrity. Finally, the study team will study the effect of ES on proteolytic pathways
      (caspase, calpain and ubiquitin-proteasome) and ribosomal RNA markers of decreased protein
      synthesis implicated in VIDD.
allocation: Non-Randomized
intervention model: Parallel Assignment
intervention model description: This is a within subjects design. One side of each subject's diaphragm will be stimulated. The other side of the subject's diaphragm will not be stimulated and will therefore serve as the control. Biopsies will be taken from both sides and compared.
primary purpose: Prevention
masking: None (Open Label)
measure: Mitochondrial respiration
time frame: Up to eight hours
description: High-resolution respirometry will be used to assess mitochondrial respiration. It will be quantified as pmol oxygen/sec/mg wet weight.
measure: Mitochondrial reactive oxygen species production
time frame: Up to eight hours
description: Mitochondrial reactive oxygen species (ROS) production will be assessed using an in situ approach to measure hydrogen peroxide production in permeabilized diaphragm skeletal muscle fiber bundles. It will be quantified as pmol/min/mg dry weight.
measure: Mitochondrial DNA mutation frequency
time frame: Up to eight hours
description: Long-Amplicon quantitative PCR will be used to measure the frequency of mitochondrial DNA mutations. It will be quantified as number of lesions/10 kilobases.
measure: Nuclear DNA mutation frequency
time frame: Up to eight hours
description: Long-Amplicon quantitative PCR will be used to measure the frequency of nuclear DNA mutations. It will be quantified as number of lesions/10 kilobases.
measure: Aconitase activity
time frame: Up to eight hours
description: In order to evaluate mitochondrial damage, actonitase activity will be measured spectrophotometrically. It will be quantified as units/mg protein.
measure: Lipid peroxidation
time frame: Up to eight hours
description: Lipid peroxidation will be assessed by measuring 4-hydroxy-2-nonenal-modified proteins. It will be quantified as arbitrary optical density units.
measure: Citrate cynthase activity
time frame: Up to eight hours
description: Changes in electron transport chain will be assessed by measuring citrate cynthase activity. It will be quantifed as nmol/mg protein/min.
measure: Cytochrome c oxidase (COX) activity
time frame: Up to eight hours
description: Changes in electron transport chain will be assessed by measuring cytochrome c oxidase (COX) activity. It will be quantifed as Units/mcg protein.
measure: Single diaphragm fiber, specific force
time frame: Up to eight hours
description: Single diaphragm fiber mechanical force properties will be measured. Specific force is quantified as kN/m^2.
measure: Single diaphragm fiber, rate of tension redevelopment
time frame: Up to eight hours
description: Single diaphragm fiber mechanical force properties will be measured. The rate of tension redevelopment is quantified as s^(-1).
measure: Single diaphragm fiber, maximum shortening velocity
time frame: Up to eight hours
description: Single diaphragm fiber mechanical force properties will be measured. The maximum shortening velocity is quantified as mm/s..
measure: Titin size
time frame: Up to eight hours
description: Titin integrity will be assessed. A relative titin size will be quantified in nm.
measure: Total titin to myosin heavy chain ratio
time frame: Up to eight hours
description: Titin integrity will be assessed. The total titin to myosin heavy chain ratio will be presented as a unitless value.
measure: Titin exon composition
time frame: Up to eight hours
description: The composition of titin exons will be assessed and quantified via real-time polymerase chain reaction (qPCR). It will be presented as percent change in expression.
as a percent change in expression.
measure: Titin binding proteins
time frame: Up to eight hours
description: Titin binding proteins will be assessed and quantified via real-time polymerase chain reaction (qPCR). It will be presented as percent change in expression.
measure: Calpain 1
time frame: Up to eight hours
description: Calpain 1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: Calpain 2
time frame: Up to eight hours
description: Calpain 2 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: Calpain 3
time frame: Up to eight hours
description: Calpain 3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: Caspase-3
time frame: Up to eight hours
description: Caspase-3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: Caspase-9
time frame: Up to eight hours
description: Caspase-9 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: 20S proteasome
time frame: Up to eight hours
description: 20S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: 26S proteasome
time frame: Up to eight hours
description: 26S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: MurF1
time frame: Up to eight hours
description: MurF1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: Atrogin 1
time frame: Up to eight hours
description: Atrogin 1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: Foxo-3
time frame: Up to eight hours
description: Foxo-3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: 28SrRNA
time frame: Up to eight hours
description: 28SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: 18SrRNA
time frame: Up to eight hours
description: 18SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
measure: 45S pre-rRNA
time frame: Up to eight hours
description: 45S pre-rRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression.
arm group label: Stimulation
arm group type: Experimental
description: Electrical stimulation of hemidiaphragm
arm group label: Control
arm group type: No Intervention
description: No stimulation of hemidiaphragm
intervention type: Other
intervention name: Electrical stimulation of hemidiaphragm
description: Electrical impulses
arm group label: Stimulation
criteria: 
gender: All
gender based: Yes
gender description: 12 females and 12 males (based on gender identity) will be recruited for the study
minimum age: 18 Years
maximum age: 85 Years
healthy volunteers: No
last name: Anatole D Martin, PhD
role: Principal Investigator
affiliation: University of Florida
last name: Thomas M Beaver, MD
role: Principal Investigator
affiliation: University of Florida
last name: Barbara Smith, PhD, PT
role: Principal Investigator
affiliation: University of Florida
facility: 
country: United States
responsible party type: Sponsor
submitted: September 5, 2023