Tocotrienol Supplementa as A Senolytic Agent in Middle-aged Adults

Blood pressure in mmHg using Sphygmomanometer

Changes in blood AGEs concentration measured by ELISA will be quantified as circulating glycoxidation markers and expressed in arbitrary units (AU) or µg/mL

Change in blood protein carbonyl concentration measured by ELISA , will be quantified as nmol carbonyl/mg protein

Change in blood malondialdehyde concentration measured by high-performance liquid chromatography (HPLC)

Change in blood DNA damage levels measured using validated laboratory assays

Measured using Inbody 770 Analyzer at Physiology Department; BMI (kg/m²) calculated from weight (kg) and height (m); Body fat percentage (%); visceral fate (cm²); basal metabolic rate (kcal); waist to hip ratio

Food frequency questionnaires (FFQ) will be completed by the participants, and analyse through Diet Information Management System; Result reported for carbohydrate (g), Cholesterol (mg), Energy (kcal), Fat (g), Fibre (g), Protein (g), vitamin E (mg), water (g)

Change in senescence-associated secretory phenotype (SASP) gene expression levels in peripheral blood measured by quantitative real-time polymerase chain reaction (qRT-PCR)

Change in senescence-associated secretory phenotype (SASP) protein expression levels in peripheral blood measured using protein array analysis

Change in blood TNF-α concentration measured by enzyme-linked immunosorbent assay (ELISA), qill be quantified in picograms per millilitre (pg/mL).

Change in blood IL-6 concentration measured by enzyme-linked immunosorbent assay (ELISA)

Change in mitochondrial ATP production in peripheral blood measured using Seahorse analysis

Change in mitochondrial Complex V enzyme activity measured in peripheral blood samples

Changes in mitochondrial membrane potential measured in peripheral blood samples

Change in plasma α-tocotrienol concentration measured by HPLC will be quantified in micromoles per litre (µmol/L) or micrograms per millilitre (µg/mL).

Change in plasma γ-tocotrienol concentration measured by HPLC will be quantified in micromoles per litre (µmol/L) or micrograms per millilitre (µg/mL).

Change in total plasma tocotrienol concentration measured by HPLC will be quantified in micromoles per litre (µmol/L) or micrograms per millilitre (µg/mL).

Appendicular lean mass in kilograms, kg will be measured using Dual-Energy X-ray Absorptiometry, (DEXA)

Fat mass in gram, g will be measured using Dual-Energy X-ray Absorptiometry, (DEXA)

Bone mineral content in gram, g will be measured using Dual-Energy X-ray Absorptiometry, (DEXA)

The assessment instrument, the Montreal Cognitive Assessment (MoCA) test will be administered. The evaluation involves assessing participants through questions and instructions covering executive and spatial cognitive domains. The maximum score is 30. A score of 26 and above indicates no cognitive impairment.

The Rey Auditory Verbal Learning Test (RAVLT) will be utilized. the RAVLT includes two distinct word lists (A and B). Participants will recall the items from list A over five trials (Memory A1 to A5). Following this, an interference list (list B), comprising 15 unrelated nouns, will be introduced, and participants will attempt to recall as many words as possible from it. Subsequently, participants will be asked to recall the words from list A (Delayed recall/memory A6) without the examiner repeating the list. The number of correctly recalled words for each trial will be totaled to generate a score.

Total Learning Score (Sum of Trials A1-A5) to indicate normal performance: 45-65, Mild Cognitive impairment: 30-45, dementia: below 30. For delayed recall score (M6), to indicate normal performance: ≥8-12 words recalled, Mild Cognitive Impairment: 4-7 words recalled, and dementia: ≤3 words recalled

The Digit Span Test involves recalling numbers in the same order (forward), reverse order (backward), or ascending order (sequencing). The test starts with short sequences and increases in length to assess memory capacity. A higher score (20 - 30) indicates better cognitive function, while a lower score (0 - 19) may suggest attention or memory issues. The total score is 30.

Change in arterial stiffness assessed by pulse wave velocity using an Arteriograph device, will be recorded in metres per second (m/s)

Change in arterial wave reflection assessed by augmentation index using an Arteriograph device will be expressed as a percentage (%).

Change in central blood pressure measured using an Arteriograph device will be measured in millimetres of mercury (mmHg)

Change in skin elasticity measured using a Cutometer, quantify in Arbitary unit

Change in skin hydration measured using a Corneometer, quantify in Arbitary unit

Change in skin barrier function measured as transepidermal water loss using a Tewameter quantify in Arbitary unit

Change in skin pigmentation measured using a mexameter quantify in Arbitary unit

Change in skin sebum secretion measured using a Sebumeter quantify in Arbitary unit

Change in wrinkle and skin roughness parameters measured using a visiocan quantify in Arbitary unit