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| ID | Type | Description | Link |
|---|---|---|---|
| P30AG028740 | U.S. NIH Grant/Contract | View source |
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| Name | Class |
|---|---|
| National Institute on Aging (NIA) | NIH |
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Both fuel metabolism and circadian rhythms have emerged as important targets to improve cellular and mitochondrial health and ultimately affect function in older adults. Thus, the purpose of this study is to develop minimally invasive measures that will allow us to accurately assess and detect changes in fuel metabolism and circadian rhythms in older adults following time-restricted eating.
A growing body of evidence indicates the mitochondria have an important role in the etiologies of many chronic diseases as well as the onset of physical disability in older adults. Although it is recognized that the mitochondria have an important role in many functions relevant to healthy aging, the direct assessment of mitochondrial function in humans is complicated and typically involves a muscle biopsy. Muscle tissue obtained from a biopsy can be used to provide an index of mitochondrial function, but only at a single time point. Some individuals may be discouraged from participating in research studies involving biopsies due to the perceived pain and risk involved.
Why there is a decrease in mitochondrial function with aging remains under debate, but emerging science indicates that there is a clear connection between mitochondrial biogenesis and function with fuel metabolism and circadian rhythms. Thus, the purpose of this development project is to develop relatively non-invasive measures that are sensitive to fuel metabolism and circadian health which can serve studies conducted within the University of Florida's Pepper Center in the coming years. In the proposed project, we will investigate the extent to which our measures of fuel utilization and circadian health markers are time stable and also sensitive to change following an intervention of time restricted eating, which is expected to impact these variables.
To our knowledge, no study has assessed fuel utilization patterns or circadian health markers in overweight older adults. Measurements of altered mitochondrial oxidation with a preference toward fat metabolism obtained from a blood sample would provide a sensitive biomarker that is relatively easy to obtain from participants for future interventions studies. The use of continuous glucose monitoring may also be used as surrogate measure of adherence to lifestyle interventions involving calorie restriction and/or intervention fasting, in future studies.
In addition to fuel utilization, there is growing recognition that age-related disease conditions and functional decline are associated with disruption of circadian rhythms. These observations raise the possibility that targeting circadian rhythms through timing lifestyle cues, such as meal timing, could be health promoting and may also reduce age associated declines in mobility. The ability to assess markers of circadian and metabolic health in minimally invasive ways through temperature and glucose monitoring, will provide potential valuable measures for explanatory or outcome measures in future studies.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Time Restricted Eating intervention | Other | Participants will be asked to stop eating by 7 PM every day and to fast for a target of 16 hours per day for 8 weeks. During the first two weeks of the intervention, participants will gradually ramp up to a full 16-hour fasting period (Week 1 - fast for 12-14 hours per day, Week 2 - fast for 14-16 hours per day, Week 3 - 8 - fast for 16 hours per day). Participants will be allowed to consume calorie-free beverages, tea, black coffee, sugar-free gum, and they will be encouraged to drink plenty of water throughout the entire intervention period. Additionally, they will be asked to keep a Fasting and Sleeping diary logging their eating habits and sleep quality. |
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| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Time Restricted Eating Intervention | Other | All participants will be asked to adhere to suggested fasting and feeding periods throughout the 8 week study period. These participants will self-monitor eating and sleeping habits as well to present to study staff at checkpoints. Self-reported information will be used during group-mediated intervention sessions throughout the duration of the study, as well. |
| Measure | Description | Time Frame |
|---|---|---|
| Change in Cellular Fuel Utilization | Fuel preference for mitochondrial energy production of isolated white blood cells (WBC) will be assessed using Agilent/Seahorse technology (XFe96 Flux Analyzer) for high-throughput measurement of mitochondrial oxygen bioenergetic function. We will use the Mito Fuel Flex Test assay (Agilent/Seahorse) to measure basal state mitochondrial fuel oxidation in live cells by using a set of substrates and inhibitors. This assay allows assessing the cell's ability to switch oxidative pathways in meeting basal energetic demands, and the relative contributions of glucose, glutamine and long chain fatty acid oxidation to basal respiration. This is completed by a 12-hour fasting blood draw. | Assessing change between Baseline and Week 8 |
| Change in daily blood glucose levels | A "flash glucose monitor/sensor" (CGM; FreeStyle Libre PRO) will be used to assess the changes in 24-hour blood glucose levels. The FreeStyle Libre sensor is easy to apply and wear and can provide every five-minute glucose data to research monitors for up to 14 days. We will replace the CGM approximately every 2 weeks. In this study, we will use the Freestyle PRO thus the participants will be blinded to the data. We will evaluate pattern changes in daily glycemic excursions by week of the study as well as weekly averages and standard deviation by 6-hour time block. | Assessing change between Baseline and Week 8 |
| Change in circadian rhythm gene BMAL1 | Whole blood will be collected in Tempusâ„¢ Blood RNA Tubes with RNA isolated using the Tempusâ„¢ Spin RNA Isolation Kit according to the manufacturer (Applied Biosystems). Relative gene expression of Bmal1 will be analyzed using quantitative real-time polymerase chain reaction (PCR). | Assessing change between Baseline and Week 8 |
| Change in Heart rate will be assessed by the Oura ring. | The goal of this development measure is to create a composite measure of circadian health using Wearable Technology (i.e., the Oura ring) that continuously tracks heart rate (beats per minute). The Oura ring is a Bluetooth Smart device and is only active for short periods of time. Data is transmitted continuously when the ring syncs with the app. Additionally, the Bluetooth signal and advertising are turned off when an individual is inactive or sleeping. Participants will be provided with Oura ring and instructed to wear it for the entire course of the study. |
| Measure | Description | Time Frame |
|---|---|---|
| Change in anthropometric measurements | Height is measured in centimeters (cm) using a stadiometer. Bodyweight will be measured in kilograms (kg) following the removal of excess clothing and shoes with calibrated scales. Weight and height will be combined to report BMI in kg/m^2. Waist circumference is taken at the mid-point (cm) between the participant's lowest rib and the top of the participants' hip bone. |
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Inclusion Criteria:
Exclusion Criteria:
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| Name | Affiliation | Role |
|---|---|---|
| Stephen Anton, Ph.D. | University of Florida | Principal Investigator |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| University of Florida | Gainesville | Florida | 32610 | United States |
| PubMed Identifier | Type | Citation | Retractions |
|---|---|---|---|
| 30349557 | Background | Sardon Puig L, Valera-Alberni M, Canto C, Pillon NJ. Circadian Rhythms and Mitochondria: Connecting the Dots. Front Genet. 2018 Oct 8;9:452. doi: 10.3389/fgene.2018.00452. eCollection 2018. | |
| 25389966 | Background | Kohsaka A, Das P, Hashimoto I, Nakao T, Deguchi Y, Gouraud SS, Waki H, Muragaki Y, Maeda M. The circadian clock maintains cardiac function by regulating mitochondrial metabolism in mice. PLoS One. 2014 Nov 12;9(11):e112811. doi: 10.1371/journal.pone.0112811. eCollection 2014. |
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Information on the study protocol may be provided upon request.
2025
Professor at an Accredited University
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| Type | Includes Protocol | Includes SAP | Includes ICF | Document Label | Document Date | Document Uploaded Date | Document File Name |
|---|---|---|---|---|---|---|---|
| ICF | No | No | Yes | Informed Consent Form | Dec 4, 2024 | Jan 13, 2026 | ICF_000.pdf |
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| Assessing change between Baseline and Week 8 |
| Change in circadian rhythm gene CLOCK | Whole blood will be collected in Tempusâ„¢ Blood RNA Tubes with RNA isolated using the Tempusâ„¢ Spin RNA Isolation Kit according to the manufacturer (Applied Biosystems). Relative gene expression of CLOCK will be analyzed using quantitative real-time polymerase chain reaction (PCR). | Assessing change between Baseline and Week 8 |
| Change in body temperature will be assessed by the Oura ring. | The goal of this development measure is to create a composite measure of circadian health using Wearable Technology (i.e., the Oura ring) that tracks body temperature in Fahrenheit (°F). The Oura ring is a Bluetooth Smart device and is only active for short periods of time. Data is transmitted continuously when the ring syncs with the app. Additionally, the Bluetooth signal and advertising are turned off when an individual is inactive or sleeping. Participants will be provided with Oura ring and instructed to wear it for the entire course of the study. | Assessing change between Baseline and Week 8 |
| Change in activity level will be assessed by the Oura ring. | This development measure aims to create a composite measure of circadian health using Wearable Technology (i.e., the Oura ring) that provides daily activity level scores. The Oura ring is a Bluetooth Smart device and is only active for short periods. Data is transmitted continuously when the ring syncs with the app. Additionally, the Bluetooth signal and advertising are turned off when an individual is inactive or sleeping. Participants will be given an Oura ring and instructed to wear it for the entire course of the study. | Assessing change between Baseline and Week 8 |
| Change in circadian rhythm gene Nfil2 | Whole blood will be collected in Tempusâ„¢ Blood RNA Tubes with RNA isolated using the Tempusâ„¢ Spin RNA Isolation Kit according to the manufacturer (Applied Biosystems). Relative gene expression of Nfil2 will be analyzed using quantitative real-time polymerase chain reaction (PCR). | Assessing change between Baseline and Week 8 |
| Change in circadian rhythm gene Nr1d1 | Whole blood will be collected in Tempusâ„¢ Blood RNA Tubes with RNA isolated using the Tempusâ„¢ Spin RNA Isolation Kit according to the manufacturer (Applied Biosystems). Relative gene expression of Nr1d1 will be analyzed using quantitative real-time polymerase chain reaction (PCR). | Assessing change between Baseline and Week 8 |
| Change in circadian rhythm gene Dbp | Whole blood will be collected in Tempusâ„¢ Blood RNA Tubes with RNA isolated using the Tempusâ„¢ Spin RNA Isolation Kit according to the manufacturer (Applied Biosystems). Relative gene expression of Dbp will be analyzed using quantitative real-time polymerase chain reaction (PCR). | Assessing change between Baseline and Week 8 |
| Change in circadian rhythm gene Cry1 | Whole blood will be collected in Tempusâ„¢ Blood RNA Tubes with RNA isolated using the Tempusâ„¢ Spin RNA Isolation Kit according to the manufacturer (Applied Biosystems). Relative gene expression of Cry1 will be analyzed using quantitative real-time polymerase chain reaction (PCR). | Assessing change between Baseline and Week 8 |
| Change in circadian rhythm gene Per2 | Whole blood will be collected in Tempusâ„¢ Blood RNA Tubes with RNA isolated using the Tempusâ„¢ Spin RNA Isolation Kit according to the manufacturer (Applied Biosystems). Relative gene expression of Per2 will be analyzed using quantitative real-time polymerase chain reaction (PCR). | Assessing change between Baseline and Week 8 |
| Change in heart rate variability will be assessed by the Oura ring. | The goal of this development measure is to create a composite measure of circadian health using Wearable Technology (i.e., the Oura ring) that tracks heart rate variability (HRV) in milliseconds (ms). The Oura ring is a Bluetooth Smart device and is only active for short periods. Data is transmitted continuously when the ring syncs with the app. Additionally, the Bluetooth signal and advertising are turned off when an individual is inactive or sleeping. Participants will be provided with Oura ring and instructed to wear it for the entire course of the study. | Assessing change between Baseline and Week 8 |
| Change in sleep patterns will be assessed by the Oura ring. | This development measure aims to create a composite measure of circadian health using Wearable Technology (i.e., the Oura ring) that provides sleep patterns scores. The Oura ring is a Bluetooth Smart device and is only active for short periods. Data is transmitted continuously when the ring syncs with the app. Additionally, the Bluetooth signal and advertising are turned off when an individual is inactive or sleeping. Participants will be provided with an Oura ring and instructed to wear it for the entire course of the study. | Assessing change between Baseline and Week 8 |
| Assessing change between Baseline and Week 8 |
| Change in Body Composition | Body composition analysis will be performed in lower and upper body compartments using Hologic software. Values of fat-free mass (FFM) will be calculated after removing mass due to bone mineral content (BMC) using the equation, (FFM+BMC)-BMC=FFM. | Assessing change between Baseline and Week 8 |
| Change in walking speed. | Walking Speed will be assessed by the 6 Minute Walk test. The 6 Minute Walk test is a valid and reliable measure of physical function in numerous studies. Individuals will be asked to walk as quickly and safely as possible at a pace that can be maintained for six minutes. The distance completed in 6 minutes will be recorded. The 6 Minute Walk test will be administered by a trained examiner. | Assessing change between Baseline and Week 8 |
| Change in grip strength | Isometric handgrip strength is a commonly used measure of upper body skeletal muscle function and is widely used as a general indicator of functional status. | Assessing change between Baseline and Week 8 |
| Change in Whole Body Fuel Utilization | Participants will be fitted with a mask and harness and oxygen consumption and carbon dioxide production will be measured using a portable Cosmed K5. Participants will be asked to refrain from volitional exercise for the prior 24 hrs and come into the lab after an overnight fast. The mask will be placed over the mouth and nose in a thermoneutral environment. Resting metabolic rate (RMR) will be collected for 45 min and the final 30 min of data will be averaged. Movement or sleeping during the test will be noted, and these time periods will be excluded from RMR calculation using the Weir formula. RMR values will be adjusted for lean mass. Respiratory quotient (RQ) will be calculated as carbon dioxide (CO2) produced divided by oxygen (O2) consumed, protein oxidation during stable evaluation (within a coefficient of variation (CV) <5%). A high RQ closer to 1.0 indicates more carbohydrate production whereas the RQ for fat is 0.7 and for ketones is 0.66 (hypocaloric) to 0.73 (eucaloric). | Assessing change between Baseline and Week 8 |
| Change in Cognitive Function - Memory | A valid cognitive battery (NIH Toolbox) will be used in this study to assess an aspect of cognitive performance including memory. | Assessing change between Baseline and Week 8 |
| Change in Physical Function | Physical Function be assessed by the Short Physical Performance Battery to assess functional performance on different tasks including timed short distance walk, repeated chair stands, and a balance test. The Short Physical Performance Battery will be administered by a trained examiner | Assessing change between Baseline and Week 8 |
| Change in Cognitive Function - Processing speed | A valid cognitive battery (NIH Toolbox) will be used in this study to assess an aspect of cognitive performance including processing speed. | Assessing change between Baseline and Week 8 |
| Chang in Cognitive Function - Attention | A valid cognitive battery (NIH Toolbox) will be used in this study to assess an aspect of cognitive performance including attention. | Assessing change between Baseline and Week 8 |
| Change in Cognitive Function - Inhibitory Control | A valid cognitive battery (NIH Toolbox) will be used in this study to assess an aspect of cognitive performance including inhibitory control. | Assessing change between Baseline and Week 8 |
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