BIOANCIENT: Cohort of Centenarians in Navarra (1926-1927)

Cognitive Function assessed with Mini-Mental State Examination (MMSE) is a 30-point tool designed to assess global cognitive function and screen for cognitive impairment, particularly in older adults. It evaluates several core areas of cognitive functioning, with higher scores indicating better cognitive performance, i.e. 30 (best) and 0 (worst).

The 11 tasks in the MMSE are generally categorized into the following areas of cognitive functioning: Orientation (10 points), Registration (3 points), Attention and Calculation (5 points); Recall (3 points): .Language and Praxis (9 points).

Barthel Index of independence during activities of daily living (ADLs) ranges from 0 (severe functional dependence) to 100 (functional independence).

Physical Function: Assessed using the Short Physical Performance Battery (SPPB), which combines balance, gait velocity, and leg strength as a single score on a 0 (worst) to 12 (best scale).

The Fried Frailty Phenotype is a widely used clinical tool that defines frailty as a distinct syndrome in older adults, based on five key physical markers: unintentional weight loss, self-reported exhaustion, low physical activity, weak grip strength, and slow walking speed. It identifies individuals as frail if they meet 3 or more criteria, and pre-frail if they meet 1-2. Namely, 0 (best) and 5 (worst).

The Lawton Instrumental Activities of Daily Living (IADL) scale is a tool used to evaluate older adult's ability to perform complex, daily tasks necessary for independent community living. It focuses on eight functional domains, typically scoring 0-8 to indicate independence levels. Scale range of 0 (worst) to 8 (best).

Nutritional Status assessed using the Mini Nutritional Assessment - Short Form (MNA-SF), a rapid, validated 6-item screening tool designed to identify older adults (aged 65+) who are malnourished or at risk of malnutrition. It is widely used in hospitals, nursing homes, and community settings to initiate early nutritional interventions. Scoring from14 points (better) and 0 points (worse):12-14 points: Normal nutritional status; 8-11 points: At risk of malnutrition and; 0-7 points: Malnourished.

Muscle Strength assessed by handgrip strength using a dynamometer (Takey Physical Fitness Test) measured in kilograms. The minimum recorded measurement is 5 kg and the maximum is 100 kg. Grip strength is a powerful biomarker of aging and overall muscle strength. Low handgrip strength is associated with increased risks of cardiovascular disease, stroke, type 2 diabetes, and premature mortality.

Lower Limb Muscle Strength. Isometric lower limb muscle strength assessed using a Hoggan dynamometer. Knee extensor strength and hip flexor strength will be measured with participants in a standardized seated position, starting with the knee flexed (90º). Force will be recorded in kilograms (kg). Higher values indicate greater muscle strength.

Risk of sarcopenia assessed using the SARC-F questionnaire, a five-item self-reported screening tool evaluating strength, assistance in walking, rising from a chair, climbing stairs, and falls. Total scores range from 0 to 10, with higher scores indicating greater risk of sarcopenia and functional decline.

Cognitive function assessed using the Clock Drawing Test (CDT), in which participants are asked to draw a clock showing a specified time. The test evaluates executive function, visuospatial ability, and planning. Performance will be scored using a standardized scoring system, with higher scores indicating better cognitive performance. Results range from 0 (worst) to 10 (best).

Depressive symptoms assessed using the Geriatric Depression Scale (GDS-15), a 15-item self-reported questionnaire designed to screen for depression in older adults. Scores range from 0 to 15, with higher scores indicating more severe depressive symptoms. Scores of 0-4 are considered normal, 5-9 suggest mild depression, and 10-15 indicate moderate to severe depression.

Health-related quality of life assessed using the EQ-5D-5L instrument, which includes five dimensions (mobility, self-care, usual activities, pain/discomfort, and anxiety/depression) each with five levels of severity, and a visual analogue scale (EQ VAS). The EQ-5D-5L index value typically ranges from values below 0 (health states worse than death) to 1 (full health), depending on the country-specific value set. The EQ VAS ranges from 0 (worst imaginable health) to 100 (best imaginable health). Higher scores indicate better health status.

Life satisfaction assessed using the Satisfaction With Life Scale (SWLS), a 5-item self-reported questionnaire developed by Diener et al. to evaluate global cognitive judgments of one's life satisfaction. Each item is rated on a 7-point Likert scale. Total scores range from 5 to 35, with higher scores indicating greater life satisfaction. Scores of 5-9 indicate extreme dissatisfaction, 10-14 dissatisfaction, 15-19 slightly below average, 20-24 average satisfaction, 25-29 high satisfaction, and 30-35 very high satisfaction.

Comorbidity burden assessed using the Cumulative Illness Rating Scale for Geriatrics (CIRS-G), which evaluates the severity of chronic medical conditions across 14 organ systems. Each system is rated on a 5-point scale (0 = no problem to 4 = extremely severe impairment). The total score ranges from 0 to 56, with higher scores indicating greater comorbidity burden.

Proteomic biomarkers assessed through the identification and quantification of circulating proteins associated with aging and frailty using high-throughput proteomic techniques. Protein expression levels will be measured in biological samples (e.g., plasma or serum), and relative or absolute concentrations will be analyzed.

Mobility and functional performance assessed using the Timed Up and Go (TUG) test. Participants are instructed to stand up from a standard chair, walk a distance of 3 meters, turn around, walk back to the chair, and sit down. The total time to complete the task is recorded in seconds. Lower times indicate better mobility and functional performance. Times greater than or equal to 12 seconds are commonly associated with increased risk of falls in older adults.

Polypharmacy assessed as the concurrent use of multiple medications. The total number of regularly prescribed medications will be recorded for each participant. Polypharmacy will be defined as the use of five or more medications (≥5). The outcome will be analyzed both as a continuous variable (number of medications) and as a categorical variable (presence vs absence of polypharmacy). Higher values indicate greater medication burden.

Prior and Concomitant Medication History; Baseline assessment of chronic pharmacological exposure. Participants report lifetime use (lasting >1 year) of 27 key drug classes (Yes/No), including cardiovascular, endocrine, respiratory, analgesic, anti-infective, neuropsychiatric, gastrointestinal, bone, and immune therapies. Long-term historical medications are also captured via open text. Reported as counts and percentages.

Bioimpedance Analysis (BIVA - Bioelectrical Impedance Vector Analysis): Whole-body bioelectrical impedance vector analysis (BIVA) assessed using a phase-sensitive bioimpedance device (e.g., BIVA Pro). Raw bioelectrical parameters including resistance (R, ohms) and reactance (Xc, ohms) will be measured at 50 kHz and normalized by height (R/H, Xc/H).

BIVA provides a qualitative and semi-quantitative assessment of body composition, including hydration status and body cell mass, without relying on predictive equations. Vector displacement patterns will be interpreted as follows:

Vector shortening (↓R/H, ↓Xc/H): fluid overload or increased hydration Vector lengthening (↑R/H, ↑Xc/H): dehydration or reduced fluid content Increased phase angle: better cellular integrity and nutritional status Decreased phase angle: impaired cellular health or frailty

Total number of unintentional falls experienced by the participant during the 12 months prior to the assessment. A fall is defined as an unexpected event in which the participant comes to rest on the ground, floor, or lower level. Data will be collected via self-report calendar.

Assessment of the participant's fear of falling, categorized as a dichotomous outcome (Yes or No). Participants are asked the single standard question: "Are you afraid of falling?". A response of "Yes" indicates the presence of fear, while "No" indicates its absence.

Pain intensity will be assessed using a standard Visual Analog Scale (VAS). The scale consists of a 10 cm horizontal line, where the left endpoint (0) represents "no pain" and the right endpoint (10) represents "worst imaginable pain". Participants mark the point on the line that best represents their current pain level. Higher scores indicate greater pain severity.

Assessment of shoulder external rotation and abduction flexibility using a simplified functional reach test. Participants are asked to place the palm of their hand flat on the back of their neck (nuca). The outcome is categorized as a dichotomous variable: "Successful" if the participant can touch the back of the neck with the palm without compensatory movements, or "Unsuccessful" if they are unable to complete the movement. Both shoulders will be evaluated separately.

Assessment of shoulder internal rotation and adduction flexibility using a simplified functional reach test. Participants are asked to place the back of their hand (dorso) on their lower back, specifically above the sacrum. The outcome is categorized as a dichotomous variable: "Successful" if the participant can touch the area above the sacrum with the back of the hand without compensatory movements, or "Unsuccessful" if they are unable to complete the movement. Both shoulders will be evaluated separately.

Adherence to the Mediterranean diet will be assessed using the 14-item Mediterranean Diet Adherence Screener (MEDAS). The questionnaire evaluates the frequency of consumption of traditional Mediterranean food groups (such as olive oil, nuts, fruits, vegetables, and fish) and negative items (such as sugar-sweetened beverages and commercial sweets). Each of the 14 items is scored as 0 (does not meet the criteria) or 1 (meets the criteria). The total score ranges from 0 to 14, where a score of 9 or higher indicates high adherence to the Mediterranean diet. Higher scores represent greater adherence.

Evaluation of participant medical history and current health status classified according to the International Classification of Diseases, 10th Revision (ICD-10). Diagnoses will be extracted from electronic medical records or clinical interviews and mapped to their corresponding ICD-10 codes. The analysis will focus on the total number of coexisting chronic conditions per participant (disease burden) and the prevalence of major ICD-10 diagnostic chapters (e.g., cardiovascular, metabolic, musculoskeletal diseases).

Evaluation of laboratory blood markers prior to baseline to characterize the participants' physiological status. The panels include: 1) Hematology: Hemoglobin, hematocrit, white blood cells, and neutrophils; 2) Glycemic Control: Fasting glucose and HbA1c; 3) Lipid Profile: Total cholesterol, LDL, HDL, and triglycerides; 4) Renal Function: Creatinine, eGFR, and urea; 5) Hepatic Function: AST, ALT, and bilirubin; 6) Inflammation \& Nutrition: C-reactive protein (CRP), albumin, and total proteins; 7) Endocrine \& Bone Metabolism: Vitamin D, TSH, and calcium. All analytes are measured from fasting venous blood samples using automated laboratory assays.

Systolic and diastolic blood pressure measured in mmHg. Reported as mean/median descriptive baseline value.

Heart rate measured in beats per minute (bpm). Reported as mean/median descriptive baseline value.

Peripheral oxygen saturation measured as percentage (%). Reported as mean/median descriptive baseline value.

Body temperature measured in degrees Celsius (°C). Reported as mean/median descriptive baseline value.

Body weight measured in kilograms (kg). Reported as mean/median descriptive baseline value.

Height measured in meters (m). Reported as mean/median descriptive baseline value.

Standing height (cm) estimated using knee height measurement according to the validated Chumlea equations. Knee height is measured as the distance (cm) from the heel (plantar surface) to the anterior surface of the thigh just proximal to the knee (femoral condyle), with the knee and ankle positioned at 90° angles.

Sex-specific equations incorporating age are applied to estimate stature. Measurement Units: Centimeters (cm). This method is validated for populations in whom direct standing height measurement is not feasible (e.g., bedridden, frail, or with postural deformities). Women: Height (cm) = 84.88 + (1.83 × knee height [cm]) - (0.24 × age [years]). Men: Height (cm) = 64.19 + (2.02 × knee height [cm]) - (0.04 × age [years])

Body mass index calculated as kg/m². Reported as mean/median descriptive baseline value.

Calf circumference measured in centimeters (cm) at the point of maximum circumference of the calf. Measurements are obtained with the participant in a seated or supine position using a non-stretchable measuring tape.

Method of Assessment: Anthropometric measurement performed using a calibrated measuring tape. The measurement is taken on the non-dominant leg when feasible. Reported as descriptive statistics (mean and standard deviation or median and interquartile range, as appropriate).

Mid-upper arm circumference measured in centimeters (cm) at the midpoint between the acromion and the olecranon process on the upper arm.

Method of Assessment: Anthropometric measurement using a non-stretchable measuring tape. The measurement is performed on the non-dominant arm when feasible, with the arm relaxed. Reported as descriptive statistics (mean and standard deviation or median and interquartile range, as appropriate).

Composite outcome based on clinical ocular screening findings assessed through direct examination. The presence (Yes/No) of the following signs will be recorded for each participant: Eyelid crusting or pus; Excessive ocular discharge; Inverted eyelashes (trichiasis); Abnormal eyelid closure; Scleral redness; Iris opacity and/or redness.

Each component will be assessed as a binary variable (present/absent). The composite outcome may be defined as the presence of ≥1 abnormal finding.

Measurement Units: Categorical (Yes/No) Standardized clinical ocular examination performed by trained healthcare personnel using visual inspection under appropriate lighting conditions.

Individual components will be reported as participant counts and percentages The composite outcome will be reported as the proportion of participants with ≥1 abnormal finding.Total number of abnormalities per participant may be summarized (e.g., mean or median count)

Waist-to-hip ratio (WHR) calculated as the ratio of waist circumference to hip circumference. Waist circumference is measured (cm) at the midpoint between the lower margin of the last palpable rib and the top of the iliac crest. Hip circumference is measured (cm) at the widest portion of the buttocks.

WHR is used as an indicator of body fat distribution and central (abdominal) obesity. Measurement Units: Unitless ratio (waist circumference [cm] / hip circumference [cm]). Anthropometric measurements performed using a non-stretchable measuring tape. Measurements are taken with the participant standing, with light clothing, and recorded to the nearest 0.1 cm. WHR is calculated using recorded values and is reported as mean and standard deviation or median and interquartile range, as appropriate. May also be categorized using established risk thresholds (e.g., >0.90 in men, >0.85 in women). It is a validated indicator of central adiposity and is associated with cardiometabolic risk.