Optimisation of Exercise Intensity During High-Intensity Interval Training for Glucose Control in Prediabetes
Optimisation of Exercise Intensity During High-Intensity Interval Training for Glucose Control in Prediabetes
The aim of the current study is to identify the optimal exercise intensity of the
 high-intensity phase during a bout of High-Intensity Interval Training (HIIT) to increase
 glucose clearance in a prediabetic population.

 The study is a within-subjects randomised crossover design with participants attending the
 laboratory on 11 occasions over a five to eight week period. The first visit will be to
 perform medical screening and to obtain informed consent. The second visit will be to collect
 baseline measures, select meal plans, to collect an activity tracker and to complete a ramped
 exercise test to establish exercise intensity thresholds. The third visit will be to fit an
 interstitial glucose sensor and to collect the standardised meals which will be provided to
 each participant. Visits four, five, six, eight, nine and ten will be the experimental
 trials, with visit seven being to change the interstitial glucose sensor. The 11th visit will
 be to remove the interstitial glucose sensor and return the activity tracker.
Recruitment Recruitment Eight men will be recruited through Church and Borth GP Surgeries in
 Aberystwyth. Patients attending GP appointments who are considered to be at risk of
 developing diabetes will have a blood sample taken as part of usual care, this sample will be
 analysed for HbA1c. If the HbA1c value is between 42 and 47 mmol·mol-1 the patient is
 considered to be prediabetic. Usual care for this group would be to inform the newly
 diagnosed patient by letter of the test result and provide information on diet and exercise
 to reduce the risk of developing T2DM. Patients who have had an HbA1c value of between 42 and
 47 mmol·mol-1 during the previous 12 months will be identified by the surgery who will then
 contact the patient by post with details of the study, including contact information for the
 research team. The research team will have no contact with the patient unless they make
 contact to request more information. Additionally, posters will be placed around Aberystwyth
 University campus, Aberystwyth town, and an email will be sent to IBERS and university staff,
 as well as using social media networks including the WARU (https://waru.org.uk/cms/) Facebook
 and Twitter accounts. Potential participants, not recruited through their GP surgery, but who
 have been diagnosed with prediabetes can contact the research team to express their interest.

 Following contact expressing an interest in participation, the research team will provide the
 individual with the participant information sheet via email or post. If having received, and
 read, the participant information the individual is still interested in becoming involved in
 the study there contact details will also be passed onto the Cardiology Department of
 Bronglais General Hospital. Consent will be obtained prior to contact information being
 shared. An appointment will then be arranged between the cardiologist and participant for the
 first visit, the chief investigator will also attend this first visit.

 Visit One On arrival at the local hospital, participants will be given a detailed verbal
 explanation of the study protocol by the chief investigator, which will explain the purpose
 of the study and explicitly state what is required of them. The participant will also have
 the opportunity to ask questions about the study and will be informed of their right to
 withdraw without the need of an explanation. They will then give written informed consent to
 participate in the study. Once consent has been given, participants will then be asked to
 complete a health screening questionnaire. Following this, the participant will complete an
 exercise stress test with continuous electrocardiogram (ECG) monitoring, using the 'Bruce
 protocol', which will be examined for an indication of cardiovascular disease. If the
 participant is excluded from the study based on their ECG results, they will be informed by
 the cardiologist who will then inform their GP. If the participant is not excluded at this
 stage, a date and time for the second visit will be arranged.

 Visit Two On arrival the participant will be given the opportunity to ask for clarification
 on any aspect of the study and their involvement, they will also be reminded of the right to
 withdraw from the study. The participant will sit for 10 minutes before blood pressure (Omron
 M2 Basic, Automatic Blood Pressure Monitor, Milton Keynes, UK) height (Harpenden Stadiometer,
 Holtain Ltd; Crymych, UK) body mass (Tanita, Body Composition Analyzer, DC-430MA, Amsterdam,
 Netherlands) and waist circumference (Rollfix, Hoechstmass, West Germany) are recorded and
 Body Mass Index (BMI) calculated. A fingertip blood sample will be taken and analysed for
 glycated hemoglobin (HbA1c) and blood lipid profile (Alere Afinion AS100, Abbott, Cheshire,
 UK). Participants will then complete a maximal exercise test which will involve completion of
 a 'ramp' test performed on a cycle ergometer (Lode Excaliber sport V2, Lode BV, Gronigen,
 Netherlands) to determine VO2peak and Gas Exchange Threshold (GET). The ergometer will be
 adjusted so that each subject is comfortable, and the settings will be recorded and
 replicated during all subsequent exercise tests. The test will involve three minutes of
 pedaling at 0 W, followed by a continuous ramped increase in work rate of 15 W·min-1.
 Participants will be instructed to maintain a cadence of 65 ± 5 r.p.m. throughout the test.
 When the participant can no longer maintain ≥ 55 r.p.m., despite verbal encouragement, the
 test will be terminated. Throughout the test, heart rate (HR) will be recorded via short wave
 telemetry (Polar, S610i, Kempele, Finland). Pulmonary gas exchange and ventilation will be
 measured breath-by-breath, (COSMED Quark CPET, COSMED srl, Rome; Italy). The gas analysers
 will be calibrated before each test with gases of known concentration, and the turbine volume
 transducer will be calibrated using a three-liter syringe. VO2peak will be determined as the
 highest VO2 measured over 30 seconds and GET will be determined using the V-slope method
 (Beaver et al. 1986) as the first disproportionate increase in carbon dioxide output (VCO2)
 relative to VO2. The work rate corresponding to 105% of VO2peak, and 50%∆ of GET, which is
 50% of the difference between GET and VO2peak, will be calculated using linear regression of
 VO2 versus work rate with account taken of the lag in VO2 relative to work rate that exists
 during ramp incremental exercise (Whipp et al. 1981). On the completion of exercise, the
 participants will be provided with a menu from which they will choose five meals (two
 breakfasts, one lunch and two dinners) to eat prior to and following each of the six
 experimental trials.

 Finally, participants will be given an ActiGraph activity tracker and instructions on its
 use. The tracker will be worn by the participant for a two and a half day period during each
 of the six experimental trials; the day before a trial from waking, the day of the trial
 until retiring to bed, and the day following the trial from waking until two-hour post second
 carbohydrate (CHO) load.

 Visit Three Visit three will take place the day before the first experimental trial. The
 participant will arrive prior to 2:00 pm to collect their meals. While participants can vary
 the content of the meals either side of a trial, the meals will be duplicated in content and
 time of consumption for each of the six experimental trials. To aid the participant a
 schedule of meals and trials will be completed and provided with the meals. This schedule
 will be used for participants to record any additional food intake, including snacks, that
 are consumed prior to and following the first experimental trial so that they can be
 replicated for the subsequent trials.

 During this visit the participant will also be fitted with a FreeStyle Libre sensor
 (FreeStyle Libre Sensor, Flash Glucose Monitoring System, Abbott Laboratories LTD.,
 Maidenhead; Berkshire), and will be shown how to use the reader. The FreeStyle Libre sensor
 is a 14-day continuous personal glucose monitoring system which logs interstitial glucose
 levels, the sensor is fitted to the non-dominant upper arm for the duration of the study. The
 sensor consists of a small disc, approximately the size of a £2 coin, 5 mm in height and 35
 mm in diameter (Abbott, 2018), with an adhesive to attach the sensor to the participant,
 there is also a small filament, approximately 0.4 mm thick and 5mm in height, which measures
 the interstitial glucose levels (FreeStyle, 2016). Participants will be instructed to scan
 the sensor either before each meal or every eight-hours, and also before they fall asleep and
 as soon as they wake up. Once the sensor is scanned, the data are transferred onto the reader
 and shows the trend of the interstitial glucose levels for the past eight-hours, and will
 store these data for up to 90 days.

 Visits Four - Six and Eight - Ten Participants will attend the laboratory on each of these
 occasions at the same time of day to minimise diurnal variation. The participants will have
 consumed the provided breakfast at least an hour prior to arriving at the laboratory. On
 arrival participants will be seated for 10 min before blood pressure is recorded and a venous
 blood sample drawn from the antecubital vein using a 21-gauge precision needle
 (Becton-Dickinson, Oxford, UK). Blood will be collected into a 6 mL K2EDTA-treated vacutainer
 tube (BD Vacutainer Systems, Plymouth, UK). 1.5 µl of whole blood will be transferred
 directly from the vacutainer into a lipid test cartridge and assayed immediately on the Alere
 Afinion AS100 POC analyser (Abbott, Illinois, United States). A further 1.0 µl of whole blood
 will be assayed immediately for glucose and lactate (2300 Stat Plus, YSI, Yellow Springs,
 USA). The remaining blood will be centrifuged at 1500xg for 10 min at 4°C, the plasma will be
 pipetted into polypropylene microcentrifuge tubes (Eppendorf, Hamburg, Germany) before being
 stored at -80°C for later analysis (insulin).

 Participants will perform each of the three different conditions twice over the duration of
 the study. The experimental conditions will be performed in random order, determined by
 WinPepi for Windows, with each of the experimental trials being separated by five days. The
 three conditions are; completing ten intervals at a work rate which equates to 105% of
 VO2peak with a 1:1 work rest ratio, completing ten intervals at a work rate which equates to
 50% ∆ GET, with a 1:1 work rest ratio, and a Control (CON) session with no exercise. Recovery
 in each of the exercise trials will involve free cycling against no resistance. The duration
 of the interval and recovery periods will vary between trials so that the total work done
 during each trial, excluding CON, will be the same, the duration of intervals will be
 determined by the 50% ∆ trial which will be set at 60 s for work and recovery intervals. Each
 of the exercise trials will be preceded and followed by a five-minute warm-up and cool-down
 at a workload of 30 W. Participants will be able to drink tap water ad libitum before, during
 and after the trial.

 During the exercise trials, the participants will be asked to report their Rating Perceived
 Exertion (RPE) from Borg's RPE scale (Borg, 1982) during each of the recovery intervals.
 Throughout the trial pulmonary gas exchange and ventilation will be measured breath-by-breath
 and hear rate recorded.

 On completion of the cool-down participants will be seated and a post-exercise blood sample
 taken and blood pressure recorded. The participants will be required to consume a CHO load of
 75 g of CHO in the form of glucose monohydrate in 200 mL of water following the five-minute
 recovery period. Participants are then free to shower and change before a final blood sample
 and blood pressure measurement is taken, one hour post completion of the exercise.
 Participants will be provided with a bottle containing 75 g of CHO in the form of glucose
 monohydrate in 200 mL of water, they will be asked to drink this the following day of the
 trial, at the same time as the first CHO load.

 Visits Seven Due to the interstitial glucose sensor only being able to log data for a 14-day
 period, at least one day before visit eight the participants will visit the laboratory and
 the interstitial glucose sensor will be replaced.

 Visit 11 Participants will attend the laboratory for the removal the glucose sensor, and to
 return the interstitial glucose reader and activity tracker.

 Adverse Effects Any adverse events (AE), serious adverse events (SAE), or suspected
 unexpected serious adverse reactions (SUSAR) will be reported to the study sponsor via
 standard Aberystwyth University reporting mechanisms. Potential AE would include; muscle
 soreness/damage, nausea, and cardiovascular events. The risk of each of these will be
 minimised by appropriate warm-up and cool-down exercises, and cardiovascular screening.
Inclusion Criteria:

 -  Males

 -  Aged 18 years and over

 -  Diagnosed with prediabetes within the last 12-months (HbA1c of between 42 and 47
 mmol·mol-1)

 -  Inactive, participating in less than 30-minutes of moderate activity on three days of
 the week, over the previous three months

 -  Mental capacity to understand the study purpose and give informed consent to
 participate

 Exclusion Criteria:

 -  Existing cardiovascular or respiratory disease, or if the individual has any symptoms
 or signs of cardiovascular disease:

 -  pain, discomfort in the chest, neck, jaw, arms, or other areas that may result
 from ischaemia

 -  shortness of breath at rest or with mild exertion

 -  dizziness or syncope

 -  orthopnea or paroxysmal nocturnal dyspnea

 -  ankle edema

 -  palpitations or tachycardia

 -  intermittent claudication

 -  known heart murmur

 -  unusual fatigue or shortness of breath with usual activities

 -  Any musculoskeletal conditions/injuries which could be exacerbated by exercise.