Digital Health and Exercise for Autonomous Longevity Program

Primary outcomes

1. Speed, agility and dynamic balance

   Time frame: 3 months

   The following physical fitness and health parameters will be evaluated:

   6-Minute Walk Test (Distance)

   At the start signal, the subjects were instructed to walk as fast as possible (without running) for 6 minutes. During the test, the supervisor walked slightly behind the participant, avoiding accompanying them side by side so as not to influence the self-selected walking pace.

2. Speed, agility and dynamic balance

   Time frame: 3 months

   Eight-Foot Up and Go Test (Seconds)

   A chair was placed at the starting point with a cone positioned 8 feet (2.44 m) away. Participants, starting seated with hands on knees, stood up, walked quickly around the cone, and returned to the chair, with timing starting on "go" and stopping when they sat back down.

3. Strength assessment

   Time frame: 3 months

   The following physical fitness and health parameters will be evaluated:

   Seated Medicine Ball Throw (Meters)

   The seated medicine ball throw test measured upper body strength and power. Participants were seated on a chair, with the trunk against the chair back, holding a 3 kg medicine ball in front of the trunk with both hands. Rotation of the trunk and hips was restricted during the test. Three throws were performed with one-minute rest intervals to prevent fatigue. The distance from the point of release to where the ball contacted the ground was measured using a metric tape. The mean distance of the throws was used for subsequent analysis.

4. Strength assessment

   Time frame: 3 months

   The following physical fitness and health parameters will be evaluated:

   Arm Curl Test (n. Rep)

   In the arm curl test, participants sat on a chair and were asked to flex the elbow of the strongest arm as many times as possible in 30 seconds while holding a 3 kg dumbbell. Participants were instructed to maintain a normal breathing pattern and keep the elbow at the side of the body throughout the test. A single repetition consisted of complete elbow flexion from maximum extension.

5. Strength assessment

   Time frame: 3 months

   The following physical fitness and health parameters will be evaluated:

   Handgrip Strength Test (Kg)

   Handgrip strength test, indicative of upper limb muscle function, was measured using an adjustable digital hand dynamometer (Saehan, Model DHD-1, South Korea). Participants were seated with hips, knees, and elbows flexed at 90º, gripping the dynamometer in a neutral forearm position. Three measurements of grip strength were taken for each hand, with a two-minute rest period between attempts. The average of these measurements was used for data analysis.

6. Strength assessment

   Time frame: 3 months

   The following physical fitness and health parameters will be evaluated:

   Sit-to-Stand Test (n. Rep)

   The sit-stand test was employed to indirectly assess lower limb muscle strength and functionality. Participants used a chair with a 40 cm height and a straight backrest [29]. With arms folded across the chest and plantar support on the floor throughout, participants were instructed to stand up and sit down as many times as possible within 30 seconds. The outcome of this test was determined by counting the number of times the participant successfully completed the sit-stand cycle.

7. Flexibility assessment

   Time frame: 3 months

   The following physical fitness and health parameters will be evaluated:

   Chair Sit-and-Reach Test (Cm)

   Starting in a seated position, the participant moves their body forwards until they are sitting on the edge of the chair seat. The fold between the top of the leg and the buttocks should be level with the edge of the seat. With one leg bent and the foot fully on the ground, the other leg (the preferred leg) is extended towards the thigh, with the heel on the ground and the foot bent (approx. 90º). The participant should be encouraged to exhale as they flex forwards, avoiding sudden, rapid and strong movements, never reaching the limit of pain.

   With the leg extended (but not hyper-extended), the participant flexes slowly forwards as far as the hip joint (the spine should remain as straight as possible, with the head in the extension of the spine, therefore not flexed), sliding the hands (one over the other, with the fingertips overlapping) along the extended leg, trying to touch the t

8. Flexibility assessment

   Time frame: 3 months

   The following physical fitness and health parameters will be evaluated:

   Back Scratch Test (Cm)

   In the standing position, the participant places their dominant hand on top of it and reaches as low as possible towards the middle of their back, palm down and fingers extended (elbow pointing upwards). The hand of the other arm is placed underneath and behind, palm facing upwards, trying to reach as far as possible in an attempt to touch (or overlap) the middle fingers of both hands.

9. Cognitive assessment

   Time frame: 3 months

   The cognitive assessment protocol aims to evaluate multiple dimensions of cognition, including memory, concentration, attention, and executive cognitive functions, as well as their severity. The assessment will include the following validated tests:

   Mini-Mental State Illiterate < 15

   1 to 11 years of schooling < 22 With more than 11 years of schooling < 27

10. Salivary assessment

    Time frame: 3 months

    Additionally, saliva samples will be analyzed for inflammatory markers such as IL-6 and IL-10, which have been reported in the literature as altered in individuals with dementia

    The assessment of biomarkers was conducted at baseline, after 6 weeks, and post-intervention. To ensure the quality of the samples collected, the experimental collection protocol adhered to strict guidelines. Sampling was consistently performed at the same time (9-11 a.m.) to account for the circadian rhythm of hormones and salivary proteins. The "passive drool" method (unstimulated saliva) was used, where participants sat comfortably, rinsed their mouth with water to remove any residue, and waited 10 minutes before collection began. Saliva was accumulated under the tongue and collected in Falcon tubes. The samples were then transferred to cryotubes and stored at -80 °C.

11. Salivary assessment

    Time frame: 3 months

    Additionally, saliva samples will be analyzed for inflammatory markers such as TNF-α, which have been reported in the literature as altered in individuals with dementia.

    The assessment of biomarkers was conducted at baseline, after 6 weeks, and post-intervention. To ensure the quality of the samples collected, the experimental collection protocol adhered to strict guidelines. Sampling was consistently performed at the same time (9-11 a.m.) to account for the circadian rhythm of hormones and salivary proteins. The "passive drool" method (unstimulated saliva) was used, where participants sat comfortably, rinsed their mouth with water to remove any residue, and waited 10 minutes before collection began. Saliva was accumulated under the tongue and collected in Falcon tubes. The samples were then transferred to cryotubes and stored at -80 °C.

12. Body Composition Assessment

    Time frame: 3 months

    Bodyweight (kilogram) BMI The percentage of fat mass was assessed using bioelectrical impedance analysis (BC-BC-613; TANITA, United Kingdom). Body mass index (BMI) was calculated as weight (kg) divided by height squared (m^2). Height measurements were taken using a stadiometer (Cabral, model 14). Participants removed their shoes and stood upright with feet flat on the floor, heels in contact, and their trunk and head positioned together against the stadiometer. The head position was neutral with the chin neither lifted nor tucked. Height measurements were recorded after participants exhaled.

13. Body Composition Assessment

    Time frame: 3 months

    Free Fat mass (kilogram) Fat mass (%)

    The percentage of fat mass and free fat mass was assessed using bioelectrical impedance analysis (BC-BC-613; TANITA, United Kingdom).

14. Assessment of memory, concentration, attention and executive cognitive functions

    Time frame: 3 months

    Rey Auditory-Verbal Learning Test scores of ≤10 for more advanced dementia and ≤ 12 for mild/moderate

15. Assessment of memory, concentration, attention and executive cognitive functions

    Time frame: 3 months

    Verbal Fluency Test (e.g., Units on a Scale) The Verbal Fluency Test is scored based on the total number of words or items generated by the individual.

16. Quality of Life

    Time frame: 3 months

    Qality of Life PHQ-9 0 to 4: No depression. 5 to 9: Mild depression. 10 to 14: Moderate depression. 15 to 19: Moderately severe depression. 20 to 27: Severe depression.