The purpose of this clinical trial is to explore the effects of either a 2-week high-intensity interval training (HIIT) or breath training intervention on measures of overall health, circulating biomarkers of stress, and immune function. Specific aims include: - Does a 2-week HIIT or breath training intervention improve measurements of overall health, including heart rate variability, physical activity, sleep quality, and severity of depression, anxiety, and stress? -Does a 2-week HIIT or breath training intervention improve circulating concentrations of stress-related biomarkers? Does a 2-week HIIT or breath training intervention improve immune function? Researchers will compare HIIT and breath training to see if equivalent immune improvements are observed. Participants will: -Undergo 2 weeks of HIIT or breath training interventions at a frequency of 3 times per week for 30 minutes or 5 times per week for 5 minutes if placed into an intervention group. -Undergo testing measures at the two pre- and post-intervention time points, if placed in the intervention groups or the healthy control group.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
HEALTH_SERVICES_RESEARCH
Masking
NONE
Enrollment
36
Participants assigned to this intervention group will complete 6 supervised HIIT sessions over 2 weeks.
Participants of this intervention group will complete a total of 10 remote guided breathing sessions on Zoom over 2 weeks.
University of Northern Colorado, 1610 Gunter Hall, 1828 10th Ave, Greeley, CO 80631
Greeley, Colorado, United States
Serum Analysis: C-reactive protein (CRP)
Approximately 60 mL of whole blood will be collected into serum separator tubes (Beckton Dickinson, East Rutherford, NJ, USA), allowed to clot for 30-min at room temperature, and then centrifuged at 2000 RPM for 15 min. The serum will be pipetted into 1.5 mL microcentrifuge tubes (Eppendorf AG, Hamburg, Germany) and immediately stored in a -80 °C freezer. Serum concentrations of CRP will be determined with a commercially available enzyme-linked immunosorbent assay (ALPCO Diagnostics, Salem, NH, USA). Microplates will be read with an ELx800 BioTek microplate reader (BioTek Instruments, Inc., Winooski, VT, USA) at the recommended wavelength of 450 nanometers.
Time frame: Visit 1(Week 1) and Visit 3 (Week 4): 15 minutes for blood draw. The assay itself takes 3 hours.
Serum Analysis: Brain Derived Neurotrophic Factor (BDNF)
Approximately 40 mL of whole blood will be collected into serum separator tubes (Beckton Dickinson, East Rutherford, NJ, USA), allowed to clot for 30 minutes at room temperature, and then centrifuged at 2000 RPM for 15 min. The serum will be pipetted into 1.5 mL microcentrifuge tubes (Eppendorf AG, Hamburg, Germany) and immediately stored in a -80 °C freezer. Serum concentrations of BDNF (Sigma-Aldrich, St. Louis, MO, USA) will be determined with commercially available enzyme-linked immunosorbent assays. Microplates will be read with an ELx800 BioTek microplate reader (BioTek Instruments, Inc., Winooski, VT, USA) at the recommended wavelength of 450 nanometers.
Time frame: Visit 1A (Week 1) and Visit 2A (Week 4): 15 minutes for blood draw. The assay itself takes 3 hours.
Natural Killer cell quantity
15mL of whole blood will be obtained using sodium heparin tubes and a peripheral venous puncture. Blood samples will be mixed with Dulbecco's phosphate buffer solution (DPBS) at a 1:1 ratio and layered over 20 mL of Ficoll gradient matrix. Blood samples will be centrifugation at 111 x g for 40 minutes, acceleration 7, brake 0 (Eppendorf 5810R centrifuge, Hamburg, Germany). Samples will be washed 3 times, then resuspended at 1.0 \* 10\^6 cells/mL and incubated for 4 hours at 37 degrees Celsius, with 5% CO2. Next, samples will be pelleted, resuspended, and antibody-receiving samples will each receive 500uL DPBS and 1uL of Fragment crystabillize blocker (FcB)/1.0\*10\^6 cells and incubate on ice for 20 minutes. Samples will be pelleted, resuspended in 500uL DPBS and receive 500uL of DPBS and 2uL of CD56 (NCAM) monoclonal antibodies, incubate on ice for 20 minutes, pelleted, and resuspended in 1mL of DPBS, then analyzed using an Attune NXT flow cytometer (Thermo Fischer, Waltham, MA).
Time frame: Visit 1(Week 1) and Visit 3 (Week 4): 15 minutes for blood draw. The assay itself takes ~10-12 hours.
Natural Killer Cell Cytotoxicity
Chronic human myelogenous leukemia cells (K562; ATCC, Manassas, VA) are used to evaluate NKC effector function. K562 cells are stained with Alexa-Fluor 647-conjugated anti-CD71. K562 cells are further stained with Calcein-AM viability dye (Thermo Fisher Scientific, Waltham, MA), then incubated on ice for 20 min. K562 cells are washed and resuspended at 1.0 × 106 cells/mL in complete RPMI 1640 medium. Human PBMCs containing effector NK and NKT cells are co-cultured with K562 cells in 5 mL Eppendorf tubes at a 40:1 PBMC: K562 effector: target (E: T) ratio and incubated for 4 h at 37 °C in a cell incubator. Following co-incubation, samples are centrifuged at 400 x g for 5 min and resuspended in ice-cold DPBS for analysis. After the 4-hour co-culture, these samples are analyzed via flow cytometry to determine the total proportion of alive (Calcein-AM+/CD71+) versus estimated K562 cells. Changes in proportion and MFI of these populations were further analyzed using Floreada.io.
Time frame: Visit 1(Week 1) and Visit 3 (Week 4): 15 minutes for blood draw. The assay itself takes ~10-12 hours.
Heart Rate Variability (HRV)
Heart rate variability (HRV) will be analyzed to determine the fluctuation in the time intervals between adjacent heartbeats to assess autonomic regulation of the heart. Resting HR and HRV were measured using the electrocardiograph (GE CASE Exercise Testing System Version 6.0, Chicago, Illinois). To measure HRV, the standard deviation of normal to normal beats (SDNN) was used with the 10-second electrocardiogram reading. Based on the outcome of this assessment, participants with SDNN values below 50 ms will be classified as unhealthy, 50-100 ms will be classified as having compromised health, and above 100 ms will be classified as healthy.
Time frame: Visit 2 (week 1) and Vist 4 (week 4)
Physical Activity Readiness Questionnaire for Everyone (PAR-Q+)
PAR-Q+ utilizes yes or no generalized health questions to screen participants interested in engaging in an exercise program for the need of medical clearance and physician release. The PAR-Q+ has 7 general health questions ranging from Does the participant feel chest pain at rest or during activities of daily living? Individuals who are eligible for this investigation require no further medical clearance before participating in an exercise program and should truthfully answer NO to every question. Participants who answer "Yes" to any questions will be required to answer follow-up questions to determine the need for medical clearance. If medical clearance is suggested, the individual will not be eligible to participate.
Time frame: Visit 1 (week 1) 10 minutes
Depression Anxiety and Stress Scale (DASS-21)
The DASS-21 contains 3 subscales assessing Depression, Anxiety, and Stress consisting of 7 questions for each subscale, where a high score of 28+ indicates "Extremely Severe" levels of Depression, a high score of 20+ indicates "Extremely Severe" levels of Anxiety, and a high score of 34+ indicates "Extremely Severe" levels of Stress, while low scores of 0-9 indicates "Normal" levels of Depression, scores of 0-7 indicates "Normal" levels of Anxiety, and scores of 0-14 indicates "Normal" levels of Stress. Raw scores for each subcategory will be recorded and multiplied by 2 to calculate the subcategory score.
Time frame: Visit 1 (week 1) and Visit 3 (week 4) 10 minutes each
Pittsburg Sleep Quality Index (PSQI)
Participants will be asked to complete a paper version of the Pittsburgh Sleep Quality Index (PSQI). The PSQI is a 19-question self-reported sleep quality tool divided into 7 subcategories for scoring: subjective sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbances, use of sleeping medication, and daytime dysfunction. Five additional questions directed to the respondent's roommate or bed partner are not used in the scoring but are utilized for clinical purposes.
Time frame: Visit 1 (week 1) and Visit 3 (week 4) 10 minutes each
Wisconsin Upper Respiratory Symptom Survey (WURSS-21)
The WURSS is an illness-specific quality of life measurement designed to assess the impact of the common cold (acute upper respiratory infection). It consists of 21 questions that investigate the perception of sickness, the severity of sickness-related symptoms over the last 24 hours, the extent to which sickness has hindered the ability to think clearly and accomplish daily tasks over the past 24 hours, and the comparison of sickness to the day before.
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Time frame: Visit 1 (week 1) and Visit 3 (week 4) 10 minutes each
Physical Activity and Sleep Tracking
Heart rate, sleep duration, and steps will be measured and obtained via the WHOOP Strap 4. Measures obtained from the WHOOP strap include continuous heart rate monitoring, sleep cycle, sleep stage, and total step data. The HRV measurement from the WHOOP strap will then be compared to the HRV analysis performed using the standard deviation of the normal-to-normal beat method.
Time frame: 4 weeks
VO2Max
Maximal oxygen consumption will be assessed on a cycle ergometer using a TrueOne 2400 Metabolic Measurement System. A Polar Heart Rate Sensor H1 will be outfitted for each participant before starting the assessment. Participants will rest for 5 minutes in a seated position when heart rate, blood pressure, and blood lactate are measured. Next, the participant will be fitted to the cycle ergometer and perform a brief 5-minute warm-up at 0.5 kg and 60 rpm. Next, the pedal cadence increases to 75-85 rpm, and the participant starts stage 1. Every stage lasts 3 minutes, at the end of every stage, heart rate, blood lactate, blood pressure, and RPE are recorded, and 0.5 kg of resistance is added to the weight basket. Once blood lactate concentration is above 4.0 mmol/L, an incremental increase of 0.2 kg occurs. Assessment is terminated after volitional fatigue, failure to maintain pedal cadence, failure to increase heart rate or VO2 in subsequent stages, and the participant's request.
Time frame: Visit 2 (week 1) 30-45 minutes