Protein and Polycystic Ovary Syndrome (PCOS)

Total body fat and body fat distribution will measured using dual x-ray absorptiometry.

Sleep quality and duration will be assessed objectively via an ActiGraph triaxial wrist accelerometer GT3X+, ActiGraph, LLC, Pensacola, FL, USA), a validated method of sleep assessment. Each participant will wear an ActiGraph monitor on the non-dominant wrist for 24-hour per day for seven days (except when bathing or involved in water activities) prior to the start of the intervention, 8-, and 16-weeks. Actigraphs will be fitted securely on each participants wrist. Participants will receive sleep diaries to define "time in bed" and "time out of bed". Researchers will use the indicated "start" and "end" points to define a sleep region to be analyzed within the ActiGraph software. Sleep outcomes will be calculated based on epoch-to-epoch sleep/wake algorithms within the defined sleep period. Data were processed by using the ActiLife Version 6.9.2 software (Pensacola, FL, USA) and sleep will be scored via the Cole-Kripke algorithm.

The Profile of Mood States (POMS) questionnaire consists of 65 questions containing a one-word adjective of mood to measure and identify six affective states. The six identifiable mood/affective states are tension-anxiety, depression-dejection, anger-hostility, vigor-activity, fatigue/-inertia, and confusion-bewilderment. Participants will be instructed to define their mood on a 5-point Likert scale ranging from 0 to 4. The numbers refer to the following descriptive phrases: 0 = Not at all, 1 = A little, 2 = Moderately, 3 = Quite a bit, 4 = Extremely. To obtain the score for reach identifiable mood/affective state subscale, the sum of the responses for each adjective is calculated. The subscale scores range from 0 up to 36, 60, 48, 32, 28, and 28 for tension-anxiety depression-dejection, anger-hostility, vigor-activity, fatigue/-inertia, and confusion-bewilderment respectively. Higher subscale scores for all affect states, but the vigor domain represent poorer mood.

Health-related quality of life will be assessed using the 36-Item Short Form Health Survey (SF-36). The instrument consists of 36 items grouped into eight domains: physical functioning, role limitations due to physical health, bodily pain, general health perceptions, vitality, social functioning, role limitations due to emotional problems, and mental health. Items will be scored using standard Likert-type response scales and coded according to established procedures. Raw scores for each domain were summed and linearly transformed to a 0-100 scale, with higher scores indicating better health status.

Indirect calorimetry will be used to measure energy expenditure

Plasma amino acids will be measured using UPL.

Inflammatory status will be evaluated using established circulating biomarkers that reflect systemic, low-grade inflammation. The primary analytes include high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). hs-CRP is an acute-phase reactant produced by the liver and is widely used as a sensitive indicator of chronic inflammation and cardiovascular risk. IL-6 is a pro-inflammatory cytokine that plays a central role in immune regulation and metabolic function, while TNF-α is a key mediator of systemic inflammation and tissue catabolism. Together, these biomarkers provide complementary information on both acute-phase and cytokine-mediated pathways of inflammation and are commonly used in clinical and epidemiological studies to quantify inflammatory burden.

Subjective measure of sleep quality will be assessed via the Pittsburgh Sleep Quality Index (PSQI) . The 19-item PSQI questionnaire addresses seven components of subjective sleep quality: subjective sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbances, use of sleep medications, and daytime dysfunction. In scoring the PSQI, seven component scores are derived, each scored 0 (no difficulty) to 3 (severe difficulty). The component scores are summed to produce a global sleeping score (GSS) with a range of 0 to 21. Higher scores indicate worse sleep quality. A compiled global score of the seven scored components distinguishes good sleepers (≤ 5) from poor sleepers (>5). According to a test-retest reliability and validity experiment, as a marker for sleep disturbances, a PSQI GSS > 5 has a sensitivity of 98.7 and specificity of 84.4.