NEUROmuscular Training for Enhanced AGE Longevity

The lipid peroxidation will be assessed via blood collections of 8-isoprostaglandin (nmol /mmol)

The antioxidants enzymes will be assessed via blood collections of glutathione peroxidase (IU/g Hb).

The brain neuroplasticity will be assessed via blood collections of brain-derived neurotrophic factor (BDNF) (ng/ml)

The interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) (pg/mL) will be assessed via blood collections.

The bone metabolism will be assessed via blood collections of Type I procollagen N-terminal propeptide (P1NP) are marker of bone formation.

The bone metabolism will be assessed via blood collections of terminal telopeptide of collagen type I (β-CTX- 1) which is a marker of bone resoption.

Systolic and diastolic blood pressure (mmHg) will be measured

Glutamic-oxaloacetic transaminase (U/L) will be measured.

Creatine (mg/dL) will be measured.

The lipid profile will be assessed via blood collections: total cholesterol (mg/dL), high-density lipoprotein cholesterol (mg/dL), low-density lipoprotein cholesterol (mg/dL) and triglycerides (mg/dL).

The metabolic profile will be assessed via blood collections of basal glucose (mmol/l).

The metabolic profile will be assessed via blood collections of human ghrelin (GHRL) (pg/ml), human acylated ghrelin (AG) (pg/ml), human glucagon like peptide 1 (GLP1) (pg/ml), and human cholecystokinin (CCK) (pg/ml).

The metabolic profile will be assessed via blood collections of leptin (ng/mL).

The metabolic profile will be assessed via blood collections of human peptide YY (PYY) (10 µCi).

The cognitive function will be assessed by the Trail Making Test (TMT) questionnaire (seconds; mistakes).

It will be assessed through the application of the MMSE. This test evaluates cognitive impairment, it contains 28 closed questions that measure 8 of the 11 main aspects of cognitive status.

It will be measured by Wisconsin Card Sorting Test (WCST). This test measures abstraction ability, concept formation, and cognitive strategy change in response to changes in environmental contingencies.

It will be measured by Vienna Test System (VTS). Specifically, will be use the Determination Test (DT), and Reaction Test (RT). This tests are used to check the capacity of observation and visual control, as well as the visual capacity of orientation and the speed of comprehension.

The dynamic maximal concentric muscle strength of the dominant side of knee, elbow, and shoulder (flexion and extension muscle groups) will be measured with an isokinetic dynamometer (Biodex System 4 Pro; Biodex, Shirley, NY) at two angular velocities, 60°/s and 180°/s. Maximal voluntary concentric isokinetic torque will be assessed in Newton-meters (N-m).

The isometric maximal muscle hand grip strength of both hands will be measured with a dynamometer (kg).

Will be assessed via the FRAX ® tool (%).

T-Score (SD) will be assessed for whole body, proximal femur on the non-dominant side (femoral nech, trochanter, intertrochanter, Wards triangle and total hip) and lumbar spine segments (L1-L4) (L2-L4) (L1-L3) and single vertebrae (L1, L2, L3, L4) will be assessed using a dual-energy-X ray fan beam absorptiometry (QDR® Hologic Discovery Wi, Hologic Inc., Waltham, MA, USA) equipped with APEX software (APEX Corp., version 12.4, Waltham, MA, USA).

BMD (g/cm2) will be assessed for whole body, proximal femur on the non-dominant side (femoral nech, trochanter, intertrochanter, Wards triangle and total hip) and lumbar spine segments (L1-L4) (L2-L4) (L1-L3) and single vertebrae (L1, L2, L3, L4) will be assessed using a dual-energy-X ray fan beam absorptiometry (QDR® Hologic Discovery Wi, Hologic Inc., Waltham, MA, USA) equipped with APEX software (APEX Corp., version 12.4, Waltham, MA, USA).

BMC(g) will be assessed for whole body, proximal femur on the non-dominant side (femoral nech, trochanter, intertrochanter, Wards triangle and total hip) and lumbar spine segments (L1-L4) (L2-L4) (L1-L3) and single vertebrae (L1, L2, L3, L4) will be assessed using a dual-energy-X ray fan beam absorptiometry (QDR® Hologic Discovery Wi, Hologic Inc., Waltham, MA, USA) equipped with APEX software (APEX Corp., version 12.4, Waltham, MA, USA).

Will be assessed via Short form quality of life questionnaire (SF36) (0-100 points).

Will be assessed via Lawton and Brody instrumental activities of daily living scale. There are eight domains of function measured with the Lawton and Brody scale. A summary score ranges from 0 (low function, dependent) to 8 (high function, independent).

Will be assessed via Barthel index of activities of daily living (score 0-100).

Adherence levels (%) will be calculated at the end of the training period. Nonadherence is define as those not attending at follow-up and have not attend for 4 weeks during the training period, and have not given a reason for nonattendance or those who have stated they are dropping out.

The eye health will be measured with Visual Function index (VF14) (0-100 points). This test provides an subjective assessment of visual quality.

The state of eye health will be measured with Functional Acuity Contrast Test (FACT) (log) associated with contrast sensitivity.

The intraocular pressure will be measured using an iCare rebound tonometer (mmHg).

The ocular perfusion pressure (mmHg) will be calculate with a formula that includes intraocular pressure and blood pressure.

Neuropathic pain will be measured with Western Ontario and McMaster Universities Arthritis Index (WOMAC). This index is widely used in the evaluation of Hip and Knee Osteoarthritis. It is a self-administered questionnaire consisting of 24 items divided into 3 subscales: a) pain (0-20 points); b) stiffness (0-8 points); c) physical function (0-68 points).

Neuropathic pain will be measured with Leeds Assessment of Neuropathic Symptoms and Signs (LANSS) scale. This scale can be used to identify pain of predominantly neuropathic origin, thus distinguishing between neuropathic pain and nociceptive pain. The LANSS scale comprises of a 7-item pain scale (5 are symptom related, and 2 are examination items) (more than 12 points on the scale might also be suffering from neuropathic pain; less than 12 points on the scale might not be suffering from neuropathic pain).

Neuropathic pain will be measured with Visual Analogue Scale (VAS) scale. This scale measure the intensity or frequency of various symptoms (0-100 points; A higher score indicates greater pain intensity).

Neuropathic pain will be measured with Neuropathic pain scale (NPS). This scale measures 10 specific qualities associated with neuropathic pain (0-10 points; A higher score indicates greater pain intensity).

Height (cm) will be recorded to the nearest 0.01 cm using a stadiometer (Seca 711, Hamburg, Germany)

Weight (kg) will be measured with a bioelectrical impedance scale (Tanita model Tanita MC 780-P MA, Tokyo, Japan; precision 0.01 kg).

Weight and height will be combined to report body mass index (BMI) (kg/m^2 ).

Fat mass (kg) will be assessed for whole body and appendicular/visceral regions using a dual-energy-X ray fan beam absorptiometry (QDR® Hologic Discovery Wi, Hologic Inc., Waltham, MA, USA) equipped with APEX software (APEX Corp., version 12.4, Waltham, MA, USA). Furthermore, the same variable will be assessed using a bioelectrical impedance scale (Tanita model Tanita MC 780-P MA, Tokyo, Japan; precision 0.01 kg).

Percentage of fat mass (%) will be assessed for whole body and appendicular/visceral regions using a dual-energy-X ray fan beam absorptiometry (QDR® Hologic Discovery Wi, Hologic Inc., Waltham, MA, USA) equipped with APEX software (APEX Corp., version 12.4, Waltham, MA, USA). Furthermore, the same variable will be assessed using a bioelectrical impedance scale (Tanita model Tanita MC 780-P MA, Tokyo, Japan; precision 0.01 kg).

Fat-free mass (kg) will be assessed for whole body and appendicular/visceral regions using a dual-energy-X ray fan beam absorptiometry (QDR® Hologic Discovery Wi, Hologic Inc., Waltham, MA, USA) equipped with APEX software (APEX Corp., version 12.4, Waltham, MA, USA). Furthermore, the same variable will be assessed using a bioelectrical impedance scale (Tanita model MC 780-P MA, Tokyo, Japan; precision 0.01 kg).

Dietary intake will be assessed by applying the Mini Nutritional Assessment-short form (MNA-f1). This brief questionnaire consists of six items, which correspond to questions about decreased food consumption, weight loss, mobility, presence of acute illness, neuropsychological problems and anthropometry.

Balance will be measured with Romberg test (points).

Balance will be measured with Functional Reach Test (FRT) (cm).

The functional performance will be assessed with Senior Fitness tests that involve 3 tests: Chair Stand (stand up from a chair as often as possible within 30s); Up-and-Go (for dynamic balance, time for got up from the chair, walk as quickly as possible around a cone placed 2.4 m from the chair, and resume the seated position); and Six-Minute walking test (for aerobic capacity, distance walked in 6 minutes).

It will be measured by Short Physical Performance Battery (SPPB): This is a performance instrument that includes 3 tests: balance, walking speed and getting up and sitting down from a chair 5 times. Each test is scored from 0 (worst performance) to 4 (best performance). A total score is obtained for the entire battery, which is the sum of the 3 tests and ranges from 0 to 12 and is classified as: low performance (0-6 points); intermediate (7-9 points) and high performance (10-12 points)

It will be measured by 2-Minute Step Test: The test considers the number of times the right knee reaches the individual reference height. If it does not reach the indicated height, the speed will be reduced so that the test is valid without stopping time. The score at the end will be given according to the number of steps taken, where the right knee reached the set height.

It will be measured through the GDS-15 test, the most widely used screening scale for depression in older people, consisting of 15 questions, with a dichotomous yes/no response. Scores from 0 to 4 points are considered normal, 5-10 indicates mild depression, and scores greater than 10 indicate established depression

The level of physical activity (low, medium, high) of the participants will be assessed by Global Physical Activity Questionnaire (GPAQ) (min/week; MET/week).

The level of physical activity (low, medium, high) of the participants will be assessed by International Physical Activity Questionnaires (IPAQ) (min/week; MET/week).

Working heart rate will be assessed with a Polar FT1 (Polar Electro, Tampere, Finland) (bpm). This variable will be measured in the first stage of the project (Validation studies.

Neuromuscular activation will be measured using electromyographic (EMG). The EMG signal will be obtained using two two-channel handheld devices (RealtimeTechnologies Ltd., Dublin, Ireland) with 16-bit analog-to-digital (A/D) conversion. EMGdata will be monitored using validated mDurance software for Android (mDuranceSolutions S.L., Granada, Spain). This variable will be measured in the first stage of the project (Validation studies).

Total repetitions performed will be recorded for later analysis with photogrammetry and video techniques. This variable will be measured in the first stage of the project (Validation studies).

Will be measured using a portable metabolimeter (VO2 Master Health Sensors Inc., Canada) (ml/kg/min).

Blood lactate concentrations will be measured from capillary blood obtained from the fingertips. The samples were analyzed using a portablel actate analyzer (Lactate Pro 2, Arkray Inc., Kyoto, Japan) (mmol/L).

Rating of perceived exertion will be measured with OMNI-Resistance Exercise Scale in the elderly (0-10 points). This variable will be measured in the first stage of the project (Validation studies).

It will be assessed by the Pittsburgh Sleep Quality Questionnaire. This instrument provides an overall score of subjects sleep quality and 7 partial scores belonging to different partial scores pertaining to different components: subjective sleep quality, latency, duration, habitual efficiency, disturbances, use of hypnotic medication and daytime dysfunction. The questions refer to the last month. The score for each of the seven components ranges from 0 (no difficulty) to 3 (severe difficulty); the overall score ranges between 0 (no difficulty) and 21 (difficulties in all areas), with a cut-off point at score 5 to differentiate between good and bad sleepers.