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| ID | Type | Description | Link |
|---|---|---|---|
| R01AR072328 | U.S. NIH Grant/Contract | View source |
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| Name | Class |
|---|---|
| National Institutes of Health (NIH) | NIH |
| University of Arizona | OTHER |
| National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) | NIH |
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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.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Stimulation | Experimental | Electrical stimulation of hemidiaphragm |
|
| Control | No Intervention | No stimulation of hemidiaphragm |
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Electrical stimulation of hemidiaphragm | Other | Electrical impulses |
|
| Measure | Description | Time Frame |
|---|---|---|
| Mitochondrial Respiration | High-resolution respirometry will be used to assess mitochondrial respiration of permeablilized diaphragm bundles. Addition of substrate medium to the Oroboros O2K respirometry instrument enables quantification of leak respiration and peak uncoupled respiration, expressed as pmol oxygen/sec/mg wet weight. | Up to eight hours |
| Aconitase Activity | In order to evaluate mitochondrial damage, actonitase activity will be measured spectrophotometrically. It will be quantified as units/mg protein. | Up to eight hours |
| Lipid Peroxidation | Lipid peroxidation will be assessed by measuring 4-hydroxy-2-nonenal-modified proteins. It will be quantified as arbitrary optical density units. | Up to eight hours |
| Citrate Cynthase Activity | Changes in electron transport chain will be assessed by measuring citrate cynthase activity. It will be quantified as nmol/mg protein/min. | Up to eight hours |
| Single Diaphragm Fiber, Specific Force | Specific force of single diaphragm fibers represents the force generated per unit area. | Up to eight hours |
| Single Diaphragm Fiber, Rate of Tension Redevelopment | Single diaphragm fiber mechanical force properties will be measured. The rate of tension redevelopment is quantified as s^(-1). | Up to eight hours |
| Calcium Sensitivity (pCa50) |
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| Measure | Description | Time Frame |
|---|---|---|
| Mitochondrial Reactive Oxygen Species Production | 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. | Up to eight hours |
Inclusion Criteria:
Exclusion Criteria:
12 females and 12 males (based on gender identity) will be recruited for the study
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| Name | Affiliation | Role |
|---|---|---|
| Anatole D Martin, PhD | University of Florida | Principal Investigator |
| Thomas M Beaver, MD | University of Florida | Principal Investigator |
| Barbara Smith, PhD, PT | University of Florida | Principal Investigator |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| University of Florida | Gainesville | Florida | 32610 | United States |
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| ID | Title | Description |
|---|---|---|
| FG000 | Participants Undergoing Elective, Open Cardiothoracic Surgical Procedures Lasting 4 Hours or Greater | Patients undergoing complex, elective prolonged surgeries, usually lasting 5-8 hours or longer, including lung transplants (e.g. valvuloplasty, coronary artery bypass and/or aortic repairs) |
| Title | Milestones | Reasons Not Completed | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Overall Study |
|
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| ID | Title | Description |
|---|---|---|
| BG000 | Participants Undergoing Elective, Open Cardiothoracic Surgical Procedures Lasting 4 Hours or Greater | Patients undergoing complex, elective prolonged surgeries, usually lasting 5-8 hours or longer, including lung transplants (e.g. valvuloplasty, coronary artery bypass and/or aortic repairs) |
| Units | Counts |
|---|---|
| Participants |
|
| Title | Description | Population Description | Parameter Type | Dispersion Type | Unit of Measure | Calculate Percentage | Denominator Units Selected | Denominators | Classes |
|---|---|---|---|---|---|---|---|---|---|
| Age, Categorical | Count of Participants |
| Type | Title | Description | Population Description | Reporting Status | Anticipated Posting Date | Parameter Type | Dispersion Type | Unit of Measure | Calculate Percentage | Time Frame | Units Analyzed | Denominator Units Selected | Arm/Group Information | Denominators | Classes | Analyses | |||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Primary | Mitochondrial Respiration | High-resolution respirometry will be used to assess mitochondrial respiration of permeablilized diaphragm bundles. Addition of substrate medium to the Oroboros O2K respirometry instrument enables quantification of leak respiration and peak uncoupled respiration, expressed as pmol oxygen/sec/mg wet weight. | Mitochondrial respiration was measured in the stimulated (n=19) and unstimulated (n=19) hemidiaphragms from subjects who completed the entire intra-operative stimulation protocol. Biopsy tissue was specifically reserved for analysis. From the full patient sample, insufficient contractile tissue was available to measure mitochondrial respiration in two unstimulated and two stimulated hemidiaphragms. | Posted | Mean | Standard Deviation | pmol/s/mg wwt | Up to eight hours |
|
From enrollment of each patient until their hospital discharge, which was typically within 7-10 days post-op.
Standard CT.gov definitions were used, and assessments were conducted systematically through the medical record and consultation with the study surgeons.
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| ID | Title | Description | Deaths (Affected) | Deaths (At Risk) | Serious Events (Affected) | Serious Events (At Risk) | Other Events (Affected) | Other Events (At Risk) |
|---|---|---|---|---|---|---|---|---|
| EG000 | Participants Undergoing Elective, Open Cardiothoracic Surgical Procedures Lasting 4 Hours or Greater | Patients undergoing complex, elective prolonged surgeries, usually lasting 5-8 hours or longer, including lung transplants (e.g. valvuloplasty, coronary artery bypass and/or aortic repairs) |
| Term | Organ System | Source Vocabulary | Assessment Type | Notes | Statistical Information |
|---|---|---|---|---|---|
| Arrhythmias | Cardiac disorders | Systematic Assessment |
| Term | Organ System | Source Vocabulary | Assessment Type | Notes | Statistical Information |
|---|---|---|---|---|---|
| Postoperative Pain | General disorders | Systematic Assessment |
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| Title | Organization | Phone | Extension | |
|---|---|---|---|---|
| Dr. Barbara K. Smith | University of Florida | 352-294-5315 | bksmith@ufl.edu |
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| Type | Includes Protocol | Includes SAP | Includes ICF | Document Label | Document Date | Document Uploaded Date | Document File Name |
|---|---|---|---|---|---|---|---|
| Prot_SAP | Yes | Yes | No | Study Protocol and Statistical Analysis Plan | Sep 3, 2020 | Jul 31, 2023 | Prot_SAP_000.pdf |
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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.
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The pCa50 value is the logarithmic scale of pCa (sensitivity of Ca+2) at which half-maximal force generation was obtained. The pCa value is calculated as the -log10[Ca (nm)]; the pCa50 is the -log10[Ca (nm)] at which half-maximal force is generated. |
| Up to eight hours |
| Difference in Total Titin to Myosin Heavy Chain Ratio | The quantities of total titin protein and myosin heavy chain protein content in homogenized diaphragm fiber specimens were measured and then calculated as a ratio of total titin to myosin heavy chain content (unitless value). The statistical approach was selected apriori as the difference of the ratio between the stimulated and unstimulated sides. | Up to eight hours |
| Difference in Titin Exon Composition | The composition of titin exons will be assessed and quantified via real-time polymerase chain reaction (qPCR). The N2A and tT2 will be calculated as a percentage of total titin. | Up to eight hours |
| Difference in Titin Binding Protein Content | The content of titin binding proteins will be quantified via Western blot. It will be normalized to a reference protein (GAPDH) and presented as optical intensity (AU). | Up to eight hours |
| Difference in Calpain 1 Protein Content | Calpain 1 (mu-calpain) will be measured with Western Blot analysis and will be presented as percent of total intensity in stimulated and unstimulated hemidiaphragms | Up to eight hours |
| Difference in Calpain 2 Protein Content | Calpain 2 will be measured with automated, capillary-based immunoassay using a Jess System, normalized to total protein, and will be presented as an area of corrected peak (AU) in stimulated and unstimulated hemidiaphragms. | Up to eight hours |
| Difference in Calpain 3 Protein Content | Calpain 3 will be measured with Western Blot analysis and will be presented as a ratio of cleaved to total calpain 3 (unitless value) in stimulated and unstimulated hemidiaphragms. | Up to eight hours |
| Difference in Caspase-3 Protein Content | Caspase-3 will be measured with Western Blot analysis, normalized to total protein loaded in each lane, and will be presented as an area of corrected peak (AU) in stimulated and unstimulated hemidiaphragm muscle fibers. | Up to eight hours |
| Atrogin 1 | Atrogin 1 will be measured with Jess protein immunoassay analysis, normalized to total protein, and will be presented as the corrected peak area (AU) in stimulated and unstimulated hemidiaphragm muscle fibers. | Up to eight hours |
| Cytochrome c Oxidase (COX) Activity |
Changes in electron transport chain will be assessed by measuring cytochrome c oxidase (COX) activity. It will be quantifed as Units/mcg protein. |
| Up to eight hours |
| Nuclear DNA Mutation Frequency | 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. | Up to eight hours |
| Titin Size | Titin integrity will be assessed. A relative titin size will be quantified in nm. | Up to eight hours |
| Caspase-9 | Caspase-9 will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| 20S Proteasome | 20S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| 26S Proteasome | 26S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| 28SrRNA | 28SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| 18SrRNA | 18SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| Foxo-3 | Foxo-3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| 45S Pre-rRNA | 45S pre-rRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| Mitochondrial DNA Mutation Frequency | 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. | Up to eight hours |
| MurF1 | MurF1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Up to eight hours |
| Participants |
|
| Age, Continuous | Mean | Standard Deviation | Number |
|
| Sex: Female, Male | Count of Participants | Participants |
|
| Ethnicity (NIH/OMB) | Count of Participants | Participants |
|
| Race (NIH/OMB) | Count of Participants | Participants |
|
| Region of Enrollment | Number | participants |
|
| Number of stimulations | Mean | Standard Deviation | Number of Stimulations |
|
| Stimulation Current Intensity | Mean | Standard Deviation | mA |
|
| %Force Vital Capacity | Mean | Standard Deviation | Percentage of predicted value |
|
| % Forced Expiratory Volume | Mean | Standard Deviation | Percentage of predicted |
|
| Maximal Inspiratory Pressure | Maximal voluntary static contractions of the inspiratory muscles against a closed valve, measured at the mouth. | Mean | Standard Deviation | cm H2O |
|
| Body Mass Index | Mean | Standard Deviation | Kg/m^2 |
|
| Intubation to Biopsy | Measurement of the amount of time in minutes between intubation and muscle biopsy acquisition | Mean | Standard Deviation | Minutes |
|
| Intubation to Frist Stimulation | Mean | Standard Deviation | Minute |
|
| Cardiopulmonary Bypass | Mean | Standard Deviation | Minutes |
|
| Minimum Core Temperature | Mean | Standard Deviation | Degrees Celsius |
|
| Core Temperature at Biopsy | Mean | Standard Deviation | Degrees Celsius |
|
| OG001 | Stimulation | Electrical stimulation of hemidiaphragm |
|
|
| Primary | Aconitase Activity | In order to evaluate mitochondrial damage, actonitase activity will be measured spectrophotometrically. It will be quantified as units/mg protein. | Aconitase activity was measured in the stimulated (n=20) and unstimulated (n=18) hemidiaphragms from subjects who completed the entire intra-operative stimulation protocol. Insufficient tissue was available in two unstimulated hemidiaphragms. Tissue obtained from one participant contained large proportions of intramuscular fat that prevented a measurement. | Posted | Mean | Standard Deviation | mU/mg protein | Up to eight hours |
|
|
|
| Primary | Lipid Peroxidation | Lipid peroxidation will be assessed by measuring 4-hydroxy-2-nonenal-modified proteins. It will be quantified as arbitrary optical density units. | Protein content was measured by traditional Western blot. Samples were analyzed from the stimulated and unstimulated hemidiaphragms of the first 11 participants. Protein quantification was normalized to total protein and optical density units of the stimulated and unstimulated hemidiaphragm were reported. | Posted | Mean | Standard Deviation | Arbitrary optical density units (U) | Up to eight hours |
|
|
|
| Primary | Citrate Cynthase Activity | Changes in electron transport chain will be assessed by measuring citrate cynthase activity. It will be quantified as nmol/mg protein/min. | Citrate synthase activity was measured in the stimulated (n=20) and unstimulated (n=18) hemidiaphragms from subjects who completed the entire intra-operative stimulation protocol. Insufficient tissue was available in two unstimulated hemidiaphragms. Tissue obtained from one participant contained large proportions of intramuscular fat that prevented a measurement. | Posted | Mean | Standard Deviation | nmol/mg protein/min | Up to eight hours |
|
|
|
| Primary | Single Diaphragm Fiber, Specific Force | Specific force of single diaphragm fibers represents the force generated per unit area. | Specific force represents force generation per cross sectional area of single diaphragm fibers and was calculated from an average of 10 slow and 10 fast single fibers obtained from each hemidiaphragm, per subject. Of the 21 patients who completed intraoperative stimulation procedures, single fiber measurements were not obtained in two participants due to unavailability of study personnel (n=2) and non-cardiac surgery (n=1). | Posted | Mean | Standard Error | kN/m2 | Up to eight hours | Muscle Fibers | Muscle Fibers |
|
|
|
| Primary | Single Diaphragm Fiber, Rate of Tension Redevelopment | Single diaphragm fiber mechanical force properties will be measured. The rate of tension redevelopment is quantified as s^(-1). | Rate constant of tension development (ktr) represents the half-time of force recovery after release of a standardized stretch. ktr was calculated from an average of 10 slow and 10 fast single fibers obtained from each hemidiaphragm, per subject. Of the 21 patients who completed intraoperative stimulation procedures, single fiber measurements were not obtained in two participants due to unavailability of study personnel (n=2) and non-cardiac surgery (n=1). | Posted | Mean | Standard Error | per second (force recovery rate) | Up to eight hours | Muscle Fibers | Muscle Fibers |
|
|
|
| Primary | Calcium Sensitivity (pCa50) | The pCa50 value is the logarithmic scale of pCa (sensitivity of Ca+2) at which half-maximal force generation was obtained. The pCa value is calculated as the -log10[Ca (nm)]; the pCa50 is the -log10[Ca (nm)] at which half-maximal force is generated. | pCa50 represents the sensitivity of pCa (concentration of Ca+2: -log10[Ca+2]) at which half-maximal force generation was obtained. It was calculated from an average of 10 slow and 10 fast single fibers obtained from each hemidiaphragm, per subject. Of the 21 patients who completed intraoperative stimulation procedures, single fiber measurements were not obtained in two participants due to unavailability of study personnel (n=2) and non-cardiac surgery (n=1). | Posted | Mean | Standard Error | -log10[Ca2+] | Up to eight hours | Muscle Fibers | Muscle Fibers |
|
|
|
| Primary | Difference in Total Titin to Myosin Heavy Chain Ratio | The quantities of total titin protein and myosin heavy chain protein content in homogenized diaphragm fiber specimens were measured and then calculated as a ratio of total titin to myosin heavy chain content (unitless value). The statistical approach was selected apriori as the difference of the ratio between the stimulated and unstimulated sides. | The quantities of total titin protein and myosin heavy chain protein content in homogenized diaphragm fiber specimens were measured and then calculated as a ratio of total titin to myosin heavy chain content. The statistical approach selected to test the effect of stimulation on titin:myosin heavy chain ratio was selected apriori as the difference between the stimulated and unstimulated sides. | Posted | Mean | Standard Deviation | Total titan/MHC Ratio | Up to eight hours |
|
|
|
| Primary | Difference in Titin Exon Composition | The composition of titin exons will be assessed and quantified via real-time polymerase chain reaction (qPCR). The N2A and tT2 will be calculated as a percentage of total titin. | The composition of the N2A and tT2 exons of the titin protein was obtained from homogenized tissue of the unstimulated and stimulated hemidiaphragms of the first 11 subjects. The relative abundance of each exon was calculated as a percentage of total titin. | Posted | Mean | Standard Deviation | Percent of total titin | Up to eight hours |
|
|
|
| Primary | Difference in Titin Binding Protein Content | The content of titin binding proteins will be quantified via Western blot. It will be normalized to a reference protein (GAPDH) and presented as optical intensity (AU). | Content of titin binding proteins including MARP1 (ANKRD-1) and MARP2 (ANKRD-2) were measured in homogenized tissue obtained from the stimulated and unstimulated hemidiaphragms of the first 11 subjects and quantified using western blot. The results were normalized to the optical intensity of GAPDH (AU). | Posted | Mean | Standard Deviation | AU | Up to eight hours |
|
|
|
| Primary | Difference in Calpain 1 Protein Content | Calpain 1 (mu-calpain) will be measured with Western Blot analysis and will be presented as percent of total intensity in stimulated and unstimulated hemidiaphragms | Calpain-1 (µ-calpain) protein content was measured by traditional Western blot. Samples were analyzed from the stimulated and unstimulated hemidiaphragms of the first 9 participants and normalized to total protein. The statistical approach selected to test the effect of stimulation on oxidative stress-related protein content was selected apriori as the difference between the stimulated and unstimulated sides. Full-length (80kDa) and truncated (76 kDa) isoforms are reported. | Posted | Mean | Standard Deviation | Percent of total Intensity | Up to eight hours |
|
|
|
| Primary | Difference in Calpain 2 Protein Content | Calpain 2 will be measured with automated, capillary-based immunoassay using a Jess System, normalized to total protein, and will be presented as an area of corrected peak (AU) in stimulated and unstimulated hemidiaphragms. | Protein content of AKT, p-AKT, calpain-2, calpain-3, caspase-3, atrogin-1 was measured by automated, capillary-based immunoassay using a Jess System. Samples were analyzed from the stimulated and unstimulated hemidiaphragms of the first 9 participants and normalized to total protein. | Posted | Mean | Standard Deviation | AU | Up to eight hours |
|
|
|
| Primary | Difference in Calpain 3 Protein Content | Calpain 3 will be measured with Western Blot analysis and will be presented as a ratio of cleaved to total calpain 3 (unitless value) in stimulated and unstimulated hemidiaphragms. | Protein content of AKT, p-AKT, calpain-2, calpain-3, caspase-3, atrogin-1 was measured by automated, capillary-based immunoassay using a Jess System. Samples were analyzed from the stimulated and unstimulated hemidiaphragms of the first 9 participants and normalized to total protein. The statistical approach selected to test the effect of stimulation on oxidative stress-related protein was selected apriori as the difference between the stimulated and unstimulated sides. | Posted | Mean | Standard Deviation | ratio of cleaved/total calpain 3 | Up to eight hours |
|
|
|
| Primary | Difference in Caspase-3 Protein Content | Caspase-3 will be measured with Western Blot analysis, normalized to total protein loaded in each lane, and will be presented as an area of corrected peak (AU) in stimulated and unstimulated hemidiaphragm muscle fibers. | Protein content of calpain-2, calpain-3, caspase-3, atrogin-1 was measured by automated, capillary-based immunoassay using a Jess System. Samples were analyzed from the stimulated and unstimulated hemidiaphragms of the first 9 participants and normalized to total protein. | Posted | Mean | Standard Deviation | AU | Up to eight hours |
|
|
|
| Primary | Atrogin 1 | Atrogin 1 will be measured with Jess protein immunoassay analysis, normalized to total protein, and will be presented as the corrected peak area (AU) in stimulated and unstimulated hemidiaphragm muscle fibers. | Protein content of AKT, p-AKT, calpain-2, calpain-3, caspase-3, atrogin-1 was measured by automated, capillary-based immunoassay using a Jess System. Samples were analyzed from the stimulated and unstimulated hemidiaphragms of the first 9 participants and normalized to total protein. | Posted | Mean | Standard Deviation | AU | Up to eight hours |
|
|
|
| Other Pre-specified | Mitochondrial Reactive Oxygen Species Production | 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. | Limited tissue quantity did not permit analysis. | Posted | Up to eight hours |
|
|
| Other Pre-specified | Cytochrome c Oxidase (COX) Activity | Changes in electron transport chain will be assessed by measuring cytochrome c oxidase (COX) activity. It will be quantifed as Units/mcg protein. | Limited tissue quantity did not permit analysis. | Posted | Up to eight hours |
|
|
| Other Pre-specified | Nuclear DNA Mutation Frequency | 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. | Limited tissue quantity did not permit analysis. | Posted | Up to eight hours |
|
|
| Other Pre-specified | Titin Size | Titin integrity will be assessed. A relative titin size will be quantified in nm. | Limited tissue quantity did not permit analysis. | Posted | Up to eight hours |
|
|
| Other Pre-specified | Caspase-9 | Caspase-9 will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Changes in Caspase-9 protein were not evaluated due to lack of a reliable antibody with preparatory validation studies. | Posted | Up to eight hours |
|
|
| Other Pre-specified | 20S Proteasome | 20S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Changes in 20S proteasome protein were not evaluated due to lack of a reliable antibody with preparatory validation studies. | Posted | Up to eight hours |
|
|
| Other Pre-specified | 26S Proteasome | 26S proteasome will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Changes in 26S Proteasome protein were not evaluated due to lack of a reliable antibody with preparatory validation studies. | Posted | Up to eight hours |
|
|
| Other Pre-specified | 28SrRNA | 28SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Differences in 28SrRNA were not completed due to challenges with cross-species validation of reagents and reference values. | Posted | Up to eight hours |
|
|
| Other Pre-specified | 18SrRNA | 18SrRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Differences in 18SrRNA were not completed due to challenges with cross-species validation of reagents and reference values. | Posted | Up to eight hours |
|
|
| Other Pre-specified | Foxo-3 | Foxo-3 will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Changes in Foxo-3 protein were not evaluated due to lack of a reliable antibody with preparatory validation studies. | Posted | Up to eight hours |
|
|
| Other Pre-specified | 45S Pre-rRNA | 45S pre-rRNA will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Changes in 45S pre-rRNA protein were not evaluated due to lack of a reliable antibody with preparatory validation studies. | Posted | Up to eight hours |
|
|
| Other Pre-specified | Mitochondrial DNA Mutation Frequency | 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. | Limited tissue quantity did not permit analysis. | Posted | Up to eight hours |
|
|
| Other Pre-specified | MurF1 | MurF1 will be measured with Western Blot anaylsis and will be presented as percent difference in expression. | Changes in MuRF1 protein were not evaluated due to lack of a reliable antibody with preparatory validation studies. | Posted | Up to eight hours |
|
|
| 0 |
| 21 |
| 14 |
| 21 |
| 21 |
| 21 |
| Anemia | Blood and lymphatic system disorders | Systematic Assessment |
|
| Dysphagia | General disorders | Systematic Assessment |
|
| Acute respiratory failure | Respiratory, thoracic and mediastinal disorders | Systematic Assessment |
|
| Methycillin sensitive staph aureus infection | Infections and infestations | Systematic Assessment |
|
| Acute Kidney Injury | Renal and urinary disorders | Systematic Assessment |
|
| Acute respiratory insufficiency | Respiratory, thoracic and mediastinal disorders | Systematic Assessment |
|
| Near-syncope | Cardiac disorders | Systematic Assessment |
|
| Low cardiac index | Cardiac disorders | Systematic Assessment |
|
| Sternal wound infection | Infections and infestations | Systematic Assessment |
|
| Tachycardia | Cardiac disorders | Systematic Assessment |
|
| Acute MCA stroke | Vascular disorders | Systematic Assessment |
|
| Pulmonary embolism | Vascular disorders | Systematic Assessment |
|
| Urinary Tract Infection | Renal and urinary disorders | Systematic Assessment |
|
| Sternal malunion | Musculoskeletal and connective tissue disorders | Systematic Assessment |
|
| Cellulitis | Infections and infestations | Systematic Assessment |
|
| Anemia | Blood and lymphatic system disorders | Systematic Assessment |
|
| Pleural effusion | Respiratory, thoracic and mediastinal disorders | Systematic Assessment |
|
| Hyperglycemia | Metabolism and nutrition disorders | Systematic Assessment |
|
| Atelectasis | Respiratory, thoracic and mediastinal disorders | Systematic Assessment |
|
| Leukocytosis | Blood and lymphatic system disorders | Systematic Assessment |
|
| Acute kidney injury | Renal and urinary disorders | Systematic Assessment |
|
| Hypotension | Vascular disorders | Systematic Assessment |
|
| Bradycardia | Cardiac disorders | Systematic Assessment |
|
| Thrombocytopenia | Blood and lymphatic system disorders | Systematic Assessment |
|
| Pericardial effusion | Cardiac disorders | Systematic Assessment |
|
| Pulmonary edema | Respiratory, thoracic and mediastinal disorders | Systematic Assessment |
|
| Pneumonia | Respiratory, thoracic and mediastinal disorders | Systematic Assessment |
|
| Bilateral lower extremity edema | Blood and lymphatic system disorders | Systematic Assessment |
|
Not provided
Not provided
|
| Fast Fibers |
|
|
|
| Fast Fibers |
|
|
|
| Fast Muscle Fibers |
|
|