Asprosin and miRNA Expression in Young Women With Metabolic Syndrome (ASPRO-MetS)

Women aged 18-35 years meeting NCEP ATP III criteria for metabolic syndrome (at least three of five components: triglycerides greater than 150 mg/dL; HDL cholesterol below 50 mg/dL; blood pressure greater than 130/85 mmHg or antihypertensive treatment; fasting glucose greater than 100 mg/dL or previously diagnosed diabetes; waist circumference greater than 88 cm), with no other comorbidity requiring continuous pharmacotherapy. Target n=55.

Fasting venous blood collection for measurement of serum asprosin, BDNF and cortisol concentrations, and expression of six circulating microRNAs (miR-15a-5p, miR-197-3p, miR-23a-3p, miR-21-5p, miR-146a-5p, miR-223-3p), in addition to complete blood count, glucose, glycated hemoglobin, insulin, and lipid profile.

Blood pressure measurement, anthropometric measurements and bioelectrical impedance body composition analysis, and completion of validated questionnaires (IPAQ, 7day-PAR, PSQI, PSS-10, FFQ/KomPAN with the Fitatu application, SF-36).

Healthy women aged 18-35 years not meeting NCEP ATP III criteria for metabolic syndrome, matched for age, not receiving continuous pharmacotherapy. Target n=55.

Fasting venous blood collection for measurement of serum asprosin, BDNF and cortisol concentrations, and expression of six circulating microRNAs (miR-15a-5p, miR-197-3p, miR-23a-3p, miR-21-5p, miR-146a-5p, miR-223-3p), in addition to complete blood count, glucose, glycated hemoglobin, insulin, and lipid profile.

Blood pressure measurement, anthropometric measurements and bioelectrical impedance body composition analysis, and completion of validated questionnaires (IPAQ, 7day-PAR, PSQI, PSS-10, FFQ/KomPAN with the Fitatu application, SF-36).

Fasting serum asprosin concentration measured by enzyme-linked immunosorbent assay, compared between women with metabolic syndrome and healthy controls. Reported in ng/mL.

Relative expression of six circulating microRNAs (miR-15a-5p, miR-197-3p, miR-23a-3p, miR-21-5p, miR-146a-5p, miR-223-3p) quantified in plasma by reverse-transcription quantitative PCR. Expression is normalized using ath-miR159a as an exogenous spike-in control and hsa-miR-186-5p as a reference. All six analytes are measured by the same method and reported in the same unit of measure, relative fold change, with results reported separately for each microRNA.

Proportion of participants in each group meeting each of the five NCEP ATP III components: elevated triglycerides, reduced HDL cholesterol, elevated blood pressure, elevated fasting glucose, and abdominal obesity. Reported as percentage of participants.

HOMA-IR calculated from fasting glucose and fasting insulin as (fasting insulin in microU/mL x fasting glucose in mg/dL) / 405, compared between the metabolic syndrome group and the control group. Reported as an index score (unitless). Higher values indicate greater insulin resistance.

Ratio of total cholesterol to HDL cholesterol, calculated from the fasting lipid profile and compared between the metabolic syndrome group and the control group. Reported as a ratio (unitless). Higher values indicate greater atherogenic risk.

Body fat percentage (PBF) measured by bioelectrical impedance analysis, compared between the metabolic syndrome group and the control group. Reported as a percentage of total body mass.

Fasting serum BDNF concentration measured by enzyme-linked immunosorbent assay, compared between groups and correlated with metabolic syndrome components. Reported in ng/mL.

Fasting serum cortisol concentration measured by immunoassay, compared between groups and correlated with perceived stress and sleep quality scores. Reported in micrograms/dL.

QUICKI calculated from fasting glucose and fasting insulin as 1 / (log fasting insulin in microU/mL + log fasting glucose in mg/dL), compared between the metabolic syndrome group and the control group. Reported as an index score (unitless). Higher values indicate greater insulin sensitivity.

METS-IR calculated from fasting glucose, fasting triglycerides, body mass index and HDL cholesterol, compared between the metabolic syndrome group and the control group. Reported as an index score (unitless). Higher values indicate greater insulin resistance.

TyG index calculated as the natural logarithm of (fasting triglycerides in mg/dL x fasting glucose in mg/dL) / 2, compared between the metabolic syndrome group and the control group. Reported as an index score (unitless). Higher values indicate greater insulin resistance.

Ratio of LDL cholesterol to HDL cholesterol, calculated from the fasting lipid profile and compared between the metabolic syndrome group and the control group. Reported as a ratio (unitless). Higher values indicate greater atherogenic risk.

Base-10 logarithm of the ratio of fasting triglycerides to HDL cholesterol, both expressed in mmol/L, compared between the metabolic syndrome group and the control group. Reported as a log-transformed ratio (unitless). Higher values indicate greater atherogenic risk.

Fat-free mass (FFM) measured by bioelectrical impedance analysis, compared between the metabolic syndrome group and the control group. Reported in kilograms.

Body mass index calculated as body mass in kilograms divided by the square of body height in metres, compared between the metabolic syndrome group and the control group. Reported in kg/m^2.

Ratio of waist circumference to hip circumference, both measured in centimetres with an anthropometric tape, compared between the metabolic syndrome group and the control group. Reported as a ratio (unitless).

Ratio of waist circumference to body height, both measured in centimetres, compared between the metabolic syndrome group and the control group. Reported as a ratio (unitless).

ABSI calculated from waist circumference, body mass index and body height according to Krakauer and Krakauer, compared between the metabolic syndrome group and the control group. Reported as an index score. Higher values indicate greater central adiposity relative to body size.

Hip Index calculated from hip circumference adjusted for body height and body mass, compared between the metabolic syndrome group and the control group. Reported as an index score.

BAI calculated from hip circumference and body height, compared between the metabolic syndrome group and the control group. Reported as estimated body fat expressed as a percentage of total body mass.

VAI calculated from waist circumference, body mass index, fasting triglycerides and HDL cholesterol using the sex-specific formula for women, compared between the metabolic syndrome group and the control group. Reported as an index score (unitless).

LAP calculated for women as (waist circumference in cm minus 58) multiplied by fasting triglycerides in mmol/L, compared between the metabolic syndrome group and the control group. Reported in cm x mmol/L.

Habitual physical activity assessed using the International Physical Activity Questionnaire, compared between the metabolic syndrome group and the control group. Reported as total physical activity in MET-minutes per week. Higher values indicate a higher level of physical activity.

Sleep quality assessed using the Pittsburgh Sleep Quality Index (PSQI). Global score range 0 to 21; higher scores indicate worse sleep quality, with a score above 5 indicating poor sleep quality.

Perceived stress assessed using the 10-item Perceived Stress Scale (PSS-10). Score range 0 to 40; higher scores indicate higher perceived stress.

Habitual daily energy intake assessed using a Food Frequency Questionnaire or the KomPAN questionnaire together with the Fitatu dietary application, compared between the metabolic syndrome group and the control group. Reported in kilocalories per day.

Health-related quality of life assessed using the Short Form-36 Health Survey (SF-36). Domain and summary scores range 0 to 100; higher scores indicate better perceived health.

Waist circumference measured with an anthropometric tape according to standard protocol, compared between the metabolic syndrome group and the control group. Reported in centimetres.

Abdominal circumference measured with an anthropometric tape at the level of the umbilicus, compared between the metabolic syndrome group and the control group. Reported in centimetres.

Hip circumference measured with an anthropometric tape at the level of the greatest gluteal protrusion, compared between the metabolic syndrome group and the control group. Reported in centimetres.

Body mass measured on a calibrated scale integrated with the bioelectrical impedance analyser, compared between groups. Reported in kilograms.

Target body mass calculated by the bioelectrical impedance analyser from measured body composition, compared between groups. Reported in kilograms.

Mass of body fat (MBF) measured by bioelectrical impedance analysis, compared between groups. Reported in kilograms.

Soft lean mass (SLM) measured by bioelectrical impedance analysis, compared between groups. Reported in kilograms.

Skeletal muscle mass (SMM) measured by bioelectrical impedance analysis, compared between groups. Reported in kilograms.

Total body water (TBW) measured by bioelectrical impedance analysis, compared between groups. Reported as a percentage of total body mass.

Ratio of extracellular water to total body water (ECW ratio) measured by bioelectrical impedance analysis, compared between groups. Reported as a ratio (unitless).

Body cell mass (BCM) measured by bioelectrical impedance analysis, compared between groups. Reported in kilograms.

Total body protein content measured by bioelectrical impedance analysis, compared between groups. Reported in kilograms.

Total body mineral content measured by bioelectrical impedance analysis, compared between groups. Reported in kilograms.

Physical activity energy expenditure over the preceding seven days assessed using the 7-day Physical Activity Recall interview, compared between the metabolic syndrome group and the control group. Reported as mean daily energy expenditure in kilocalories per day.

Contribution of protein, fat and carbohydrate to total daily energy intake, assessed using a Food Frequency Questionnaire or the KomPAN questionnaire together with the Fitatu dietary application, compared between the metabolic syndrome group and the control group. Reported as a percentage of total daily energy intake for each macronutrient.