MedPath

PRE-Pregnancy Weight Loss And the Reducing Effect on CHILDhood Overweight - Aarhus

Not Applicable
Recruiting
Conditions
Weight Loss
Pregnancy Related
Overweight and Obesity
Child Nutrition Sciences
Body Composition
Interventions
Behavioral: Weight loss intervention
Registration Number
NCT05798494
Lead Sponsor
Ulla Kampmann Opstrup
Brief Summary

The study is a single site parallel randomized controlled study. The study will be assessing the effect of a 10% pre-conceptional weight loss intervention vs a control group among healthy couples where the prospective mother is pregnant and overweight or obese (BMI 27-44.9 kg/m\^2). The couples in intervention group will receive dietitian counseling and participate in physical activity sessions to attain the 10% weight loss. The overall objective is to test whether a comprehensive pre-conceptional parental weight loss intervention effectively reduces the risk of offspring overweight, and adiposity and its complications compared to a control group.

Detailed Description

The study is a single site parallel randomized controlled study. The study will be assessing the effect of a pre-conceptional weight loss intervention vs a control group among healthy couples where the prospective mother is pregnant (recruited at gestational age 18-36), overweight or obese (BMI 27-44.9 kg/m\^2), between 18-38 years and planning an additional pregnancy within the following 3 years.

The investigators will recruit a total of 140 healthy pregnant couples who will be randomized 1:1 to either intervention or control, stratified according to maternal pre-pregnancy BMI and previous or present diagnosis of gestational diabetes.

The couples in intervention group will receive dietary counseling by a dietitian from 3 months post-partum of the 1st child until birth of the 2nd child. The goal is to achieve a healthy lifestyle and a moderate weight loss of 10% according to pre-pregnancy weight during the first year. Hereafter, the couples will be guided in weight maintenance until pregnancy of the second child. During pregnancy the goal is to achieve a healthy weight gain. Couples in intervention group will also participate in physical activity sessions every 2nd week and advised to walk with moderate intensity at least 2-4 times a week for at least half an hour.

The overall objective is to test whether a comprehensive pre-conceptional parental weight loss intervention effectively reduces the risk of offspring overweight, and adiposity and its complications compared to a control group. The 1st baby born before intervention is started will be the "index child" and the metabolic and anthropometric parameters of the 1st child will be compared to parameters of the second child to assess the effect of the parent's healthy lifestyle between the two pregnancies.

The investigators hypothesize that parental weight loss intervention, initiated before conception, will facilitate lower parental insulin resistance, inflammation, body weight and adiposity, incretin responses and energy expenditure compared to usual care. For the offspring the investigators hypothesize that the intervention will reduce adverse pregnancy outcomes with a short-term reduction in the risk of being born large for gestational age with excessive fat mass and a long-term reduction in the risk of developing overweight or obesity as a consequence of both social (family approach) and biological (epigenetic modifications) pathways.

Recruitment & Eligibility

Status
RECRUITING
Sex
All
Target Recruitment
140
Inclusion Criteria
  • Paternal pre-pregnancy BMI 18.5-44.9 kg/m^2
  • Paternal age range 18-55 years

Parental Inclusion Criteria:

  • Couples (male and female)
  • Planning pregnancy within 3 years
  • Provided voluntary informed consent
  • Danish or English speaking
  • Intention to permit the planned offspring to participate in the follow-up study
  • Able to store biological samples from the offspring at home in the freezer in a box given by the study personnel

Parental

Exclusion Criteria
  • Diabetes mellitus (Type 1 or 2)
  • Previous or present eating disorder
  • Allergy towards ingredients in the very low calorie diet products
  • Severe heart, liver or kidney disease
  • Conception by in vitro fertilization
  • Any medical condition or concomitant medication as judged by the medical responsible
  • Adherence to vegan diets or other diets interfering with the dietary guidelines in the study
  • Participation in other clinical trial that can affect the results of the current study
  • Engagement in elite sports or similar strenuous exercise ≥5 h/week at inclusion
  • Blood donation or transfusion within the past month before screening
  • Blood donation during the study
  • Inability or unwillingness to follow the study protocol and instructions given by the study personnel

A detailed description of the exclusion criteria is given below:

Medical conditions as known by the participant:

  • Diabetes mellitus (type 1 and 2)
  • History or diagnosis of eating disorder (e.g. restrained eating, disinhibition, emotional eating)
  • Any significant medical condition as assessed by the investigator (e.g. dysregulated thyroid disease or reproductive diseases)
  • Any significant psychiatric disorder (i.e. schizophrenia, bipolar disease or depression) as assessed by the investigator
  • Severe chronic heart, liver and kidney disease
  • Polycystic ovary syndrome with irregular cycle

Medication:

  • Current use of medication or use within the previous three months with a potential to affect body weight or pregnancy as judged by the investigator (e.g. systemic corticoids)

Personal/other:

  • Severe food allergies, food intolerances or dislike expected to interfere with the study, including allergy towards ingredients in the VLCD products
  • Engagement in elite sports or similar strenuous exercise ≥5 h/week
  • Blood donation or transfusion within the past month before screening
  • Planned blood donation for other purpose than this study during participation
  • Alcohol abuse, as judged by the investigator, within the previous 12 months
  • Drug abuse, as judged by the investigator, within the previous 12 months
  • Psychological or behavioral problems which, in the judgement of the investigator, would lead to difficulty in complying with the study protocol
  • Participation in other clinical trials within the past three months or intention to do so during the study, which are judged by the investigator to affect the present study
  • Unable to consume the interventional product for religious reasons, swallowing disorders, other physiological reasons or any other reasons for not being able to follow the recommended diet
  • Inability or unwillingness to give written informed consent or communicate with study personnel
  • Inability or unwillingness to follow the study protocol and instructions given by the study personnel
  • Illiteracy or inadequate understanding of Danish or English language.
  • Any other condition that judged by the investigator may interfere with the adherence to the study protocol

Study & Design

Study Type
INTERVENTIONAL
Study Design
PARALLEL
Arm && Interventions
GroupInterventionDescription
InterventionWeight loss interventionIntervention group will receive dietary counseling by a trained dietitian from 3 months post-partum of the 1st child until birth of the 2nd child. The mother will receive advice to provide a moderate caloric restriction (-500 to 1000 kcal/d) and attend physical activity sessions to attain a 10 % weight loss according to pre-pregnancy weight. If the father is also overweight or obese, he will receive the same advice.
Primary Outcome Measures
NameTimeMethod
Neonatal fat massCollected within the first week of birth (strived <48 hours)

Offspring's neonatal fat mass (grams) assessed by air displacement plethysmography (Pea POD)

Difference in epigenetic changes in child 1 and 2Assessed at birth

Cord blood from offspring will be collected at birth and stored at -80 °C until use. At the Department of Molecular Medicine AUH genomic DNA will be extracted from peripheral blood using standard methods e.g., QIAmp® Mini Kit (Qiagen, Germany). 1 μg of genomic DNA will be bisulfide converted using Illumina iScan (platform). The methylation level will be measured using Infinium MethylationEPIC DNA Analysis BeadChip (Array type) at Eurofins Genomics AS (Denmark) or similar methods.

The stated required sample size (140 couples) is calculated based on the neonatal fat mass. As no other studies have performed methylation analysis using WGBS in similar cohorts, precise power calculations on epigenetic changes are impractical. However, since the EPICOM study found differentially methylated regions in only 20 cases and 20 controls using Illumina's 450K methylation assay, the power to elucidate epigenetic changes is considered sufficient in the planned study.

Secondary Outcome Measures
NameTimeMethod
Adult reproduction - Time to pregnancyRegistration of 2nd. pregnancy (up to tree years from birth of 1.st. child)

Duration of time to conception defined from selfreported time point they started to try get pregnant to registration of pregnancy. The time of conception will be calculated from the ultrasound scan in week 12

VO2-max adult - cardiorespiratory fitnessCollected at study visit 3, 12 and 24 months pp after birth of 1st child

The VO2-max test will be performed as an incremental exercise test on a bicycle ergometer.

Cardiac function offspring - TAPSE and RV fractional area changeCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of TAPSE and RV fractional area change will be done.

Offspring blood sample - CD163Collected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for CD163

Offspring blood sample - HgbCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Hgb

Offspring blood sample - TriglycerideCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Triglyceride

Offspring blood sample - Cholesterol LDLCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Cholesterol LDL

Offspring blood sample - FGF-21Collected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for FGF-21

Cord blood -InsulinCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for Insulin

Cord blood -IGF-1Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for IGF-1

Cord blood -FGF21Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for FGF21

Adult female blood sample - ProlactinCollected at GA34-GA36 in both pregnancies

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will only be collected from the female participant during pregnancy and be analysed for Prolactin

Adult blood sample -CortisolCollected at GA34-GA36 in both pregnancies

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Cortisol

Offspring blood sample - IGF-1Collected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for IGF-1

Offspring blood sample - IGFBP-1Collected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for IGFBP-1

Adult Oral Glucose Tolerance Test (OGTT) during pregnancy (only female)Collected in GA28 of 1st and 2nd pregnancy (performed after risk score at gestational age~20 and ~28 or only ~28)

OGTT performed during pregnancy according to the screening guidelines for GDM will be obtained from the mother's medical journal in both pregnancies.

Obstetric - pregnancy complicationsCollected at gestational age (GA) 34-36 and after birth of both offsprings

Pregnancy complications e.g. miscarriage, abortion (induced and indication for abortion), hydramnios, gestational hypertension, preeclampsia, need for maternal corticosteroid treatment for fetal lung maturation, urinary tract infection) will be collected from medical record.

Obstetrics - birth defects/abnormalitiesCollected at birth of both offsprings

Birth defects/abnormalities

Offspring growth - small for gestational ageCollected at birth of both offsprings

Small for gestational age. Prevalence of offspring being small for gestational age (SGA).

Adult weightIntervention: each visit throughout study (GA34-36 of 1st child to 18mo. after birth of 2nd child). Control: In GA34-36 in both pregnancies, at study visits (3, 12 and 24mo. pp) and follow up visits (3, 12 and 18 months pp of 2nd child)

Adult body weight is measured to the nearest 0.1 kg using a calibrated scale

Obstetrics outcome - length of gestational ageAssessed at birth of both offsprings

Length of gestational age

Fetal and offspring growth - femur lengthCollected from medical record during pregnancy, measured in GA34-GA36 via ultrasound in both pregnancies,

Femur length will be measured with ultrasound during pregnancy

Offspring blood pressureCollected 12 and 18 months after birth of both offsprings

Offspring blood pressure (diastolic and systolic blood pressure) will be measured three times after 10 minutes rest using an automated device placed on the arm and in direct contact with the skin.

Adult medication useCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st. child

Use of medication will be collected throughout the study and for each medication the start date, stop date or continuing, form and indication will be noted in appropriate form.

Adult glucose metabolism (OGTT)Performed at study visits 3, 12 and 24 months pp of 1st. child

3-h oral glucose tolerance test (OGTT) will be performed at with 75g of glucose dissolved in 250 ml of water and added 1 gram of paracetamol. Blood will collected for measurements of blood glucose, hormones (insulin, C-Peptide, glucagon), paracetamol, glucagon-like-peptide-1 (GLP-1), Glucose insulinotropic polypeptide (GIP) and metabolites (FFA). Blood samples will be drawn at time 0, 15, 30, 45, 60, 90, 120 and 180 min

Placenta - weight (gram)Collected at birth of both offsprings

Placenta will be weighted after birth

Obstetric - delivery modeCollected at birth of both offsprings

Delivery mode (rupture of membranes or premature contractions, vaginal, cesarean section, instrumental delivery, miscarriage and stillbirth).

Obstetric - obstetrics complicationsCollected at birth of both offsprings

Obstetrics complications (e.g. preterm prelabour rupture of the membranes, shoulder dystocia, birth trauma (perinatal rupture), postpartum hemorrhage, maternal infections, maternal hospitalization).

Fetal and offspring growth - body weightCollected from medical record during pregnancy, measured in gestational age (GA) GA34-GA36 via ultrasound in both pregnancies, at birth and 3, 12 and 18 months after birth of both offsprings

Fetal body weight will be measured with ultrasound during pregnancy and offspring body weight will be measured using an electronic scale.

Fetal and offspring growth - lengthCollected from medical record during pregnancy, measured in GA34-GA36 via ultrasound in both pregnancies, and measured at birth and 3, 12 and 18 months after birth of both offsprings

Fetal length will be measured with ultrasound during pregnancy and offspring length will be measured using an infantometer as appropriate for measuring infant, baby and toddler length.

Fetal and offspring growth - crown circumferenceCollected from medical record during pregnancy, measured in GA34-GA36 via ultrasound in both pregnancies

Crown circumference will be measured with ultrasound during pregnancy

Offspring growth - weight for gestational ageCollected at birth and 3, 12 and 18 months after birth of both offsprings

Weight for gestational age. Measured as percent of normal weight related to gestational age.

Offspring growthCollected at birth and 3, 12 and 18 months after birth of offspring.

Offspring circumference (thigh and upper arm) and skinfold (triceps, subscapularis and suprailiac). Circumferences will be measured to the nearest 0.1 cm using a stretch-resistant measuring tape and skinfolds are measured to the nearest 0.02 cm using a caliper to lightly pinch the skin and the underlying fat.

Offspring growth - body composition by the POD systemCollected within the first week of birth and 3 months post partum in both offsprings

Offspring fat free mass (g) and fat mass (g) will be assessed by air displacement plethysmography using the POD system (Pea POD for 1-8kg)

Offspring heart rateCollected 12 and 18 months after birth of both offsprings

Offspring heart rate will be measured three times after 10 minutes rest using an automated device placed on the arm and in direct contact with the skin.

Offspring growth - self-reported body weight18 months after birth the study staff will make an annual follow-up phone call to the parents to collect information about the offspring's self-reported body weight until the offspring is 18 of age

Self-reported body weight

Adult blood pressureCollected at GA34-36 in 1st pregnancy, study visits 3, 12 and 24 months pp. of 1st child, GA34-36 in 2nd pregnancy and 3 months pp after 2nd child

Blood pressure (diastolic and systolic blood pressure) will be measured three times with two minutes apart in right arm after 15 minutes rest using a validated automatic device. The reading is recorded to the nearest 1 mmHg.

Offspring medication useBirth, after 3, 12, and 18 months after birth of both offsprings

Use of medication will be collected throughout the study and for each medication the start date, stop date or continuing, form and indication will be noted in appropriate form.

Neonatal morbiditiesCollected at birth of both offsprings

Neonatal morbidity (e.g. major congenital malformations (ICD10 Q00-Q99 or requiring medical or surgical treatment), birth injury, respiratory distress, transient tachypnoea, neonatal hypoglycemia, systemic infections, admission to neonatal intensive care unit, number of admission days, hyperbilirubinemia), deaths.

Fetal and offspring growth - abdominal circumferenceCollected from medical record during pregnancy, measured in GA34-GA36 via ultrasound in both pregnancies, and measured at birth and 3, 12 and 18 months after birth of both offsprings

Abdominal circumference will be measured with ultrasound during pregnancy and offspring abdominal circumference will be measured three times to the nearest 0.1 cm using a stretch-resistant measuring tape

Fetal growth - fractional limb volumeCollected from medical record during pregnancy, measured in GA34-GA36 via ultrasound in both pregnancies,

Fractional limb volume will be measured with ultrasound during pregnancy

Offspring growth - BMI z-scoreCollected at birth and 3, 12 and 18 months after birth of both offsprings

Offspring body mass index (BMI) z-score (BMI adjusted for age and sex).

Offspring growth - large for gestational ageCollected at birth of both offsprings

Large for gestational age. Prevalence of infants being large for gestational age (LGA).

Offspring growth - self-reported height18 months after birth the study staff will make an annual follow-up phone call to the parents to collect information about the offspring's self-reported body weight until the offspring is 18 of age

Self-reported height

Obstetric - medication useCollected from initiation of labor until completed birth of both offsprings

Medication use during labor

Obstetric - Neonatal unit admissionsCollected at birth and until 12 months after birth of both offsprings

Neonatal unit admissions

Offspring growth - apgar scoreCollected at birth of offspring of both offsprings

Apgar score (0-10). The higher the score, the better health condition of the offspring)

Adult HeightCollected in GA34-GA36 in index pregnancy

Adult height will be measured to the nearest 0.5 cm using a wall-mounted stadiometer.

Adult BMICollected in GA34-36 in 1st pregnancy, study visits 3, 12 and 24mo. pp after 1st child, GA34-36 in 2nd pregnancy and at follow up vist (3, 12 and 18mo. pp of 2nd child).

Calculated as weight (kg)/height(m)\^2

Adult anthropometricIntervention: each visit throughout study (GA34-36 of 1st child to 18mo. after birth of 2nd child). Control: In GA34-36 in both pregnancies, at study visits (3, 12 and 24mo. pp) and follow up visits (3, 12 and 18 months pp of 2nd child)

Waist- and hip circumference will be measured to the nearest 0.1 cm. During pregnancy and at birth the waist of the female will not be measured.

Adult body composition by DEXA scanCollected at study visits 3, 12 and 24 months pp. of 1st child

Lean mass (g), fat mass (g), abdominal fat (g), visceral fat (g) assessed by a Dual-energy X-ray absorptiometry (DEXA) -scan .

Adult bone markers by DEXA scan - bone mineral contentCollected at study visits 3, 12 and 24 months pp. of 1st child

Bone mineral content (g) assessed by a Dual-energy X-ray absorptiometry-scan.

Adult resting metabolic rateCollected at study visit 3, 12, and 24 months pp after 1st child.

Resting metabolic rate (calories/24-h) will be measured by indirect calorimetry using a ventilated hood system. Participants will lie in the supine position and rest for at least 30 min before measurements commence. The measurement will last for 20 min.

Cardiac function offspring - Diastolic function, left ventricular E/A-ratioCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of left ventricular E/A-ratio will be done.

Cardiac function offspring - Diastolic function, tissue velocities (E', A', S')Collected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of tissue velocities (E', A', S') will be done.

Cardiac function offspring - Diastolic function, strainCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of strain will be done.

Cardiac function offspring - Pulmonary artery acceleration timeCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of Pulmonary artery acceleration time will be done.

Cord blood - pHCollected at birth

Cord blood will be drawn immediately after birth. pH will be assessed.

Adult heart rateCollected at GA34-36 in 1st pregnancy, study visits 3, 12 and 24 months pp. of 1st child, GA34-36 in 2nd pregnancy and 3 months pp after 2nd child

Heart rate will be measured three times with two minutes apart in right arm after 5-10 minutes rest using a validated automatic device. The reading is recorded to the nearest 1 beats/minute for heart rate

Cardiac function offspring - Left ventricular and right ventricular morphologyCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of Interventricular septal thickness diastole and valve- and chamber dimensions will be done.

Cardiac function offspring - Systolic function, strainCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of strain will be done.

Adult bone markers by DEXA scan - bone mineral densityCollected at study visits 3, 12 and 24 months pp. of 1st child

Bone mineral density (g/cm\^2) assessed by a Dual-energy X-ray absorptiometry-scan.

Adult blood sample - CD163Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for CD163

Adult blood sample - eGFRCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for kreatinin and GFR will be estimated

Adult blood sample - FerritinCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Ferritin

Adult blood sample - TriglycerideCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for triglyceride

Adult blood sample - Total CholesterolCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Total Cholesterol

Adult blood sample - vitamin B12Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for B12

Adult resting respiratory quotient + substrateCollected at study visit 3, 12, and 24 months pp after 1st child.

Resting respiratory quotient (RQ), will be measured by indirect calorimetry using a ventilated hood system. Participants will lie in the supine position and rest for at least 30 min before measurements commence. The measurement will last for 20 min. Lipid, glucose and protein oxidation will be estimated.

Cardiac function offspring - left centricular ejection fractionCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of left ventricular ejection fraction will be done.

Cardiac function offspring - fractional shorteningCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of fractional shortening will be done.

Cardiac function offspring - Systolic function, strain rateCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of strain rate will be done.

Cardiac function offspring - Persistent ductus arteriosus (PDA)Collected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of PDA open or closed and PDA shunt will be done.

Cord blood - Telomore lengthCollected at birth of both offsprings

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for Telomore length.

Adult blood sample - C-reaktivt protein (CRP)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for CRP

Adult blood sample - BilirubinCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Bilirubin

Adult blood sample - vitamin D3Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for vitamin D3

Cardiac function offspring - Pulmonary artery pressureCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of Pulmonary artery pressure will be done.

Cord blood - EpigeneticsCollected at birth of both offsprings

Epigenetics analyses from whole blood. Will be analysed for methylation changes. metabolomics and exosome.

Adult blood sample - KreatininCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for kreatinin

Adult blood sample - Basisk fosfatase (BASP)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for BASP

Adult blood sample - plateletsCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for platelets

Adult blood sample - Cholesterol HDLCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Cholesterol HDL

Adult blood sample - Cholesterol LDLCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Cholesterol LDL

Adult blood sample - Fibroblast growth factor 21 (FGF-21)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for FGF-21

Cardiac function offspring - Overall cardiac functionCollected at birth and at 12 months follow up visit after birth of each child

ECG and echocardiography will be performed according to standard imaging protocol. Assessment of Myocardial performance index (MPI) and cardiac torsion will be done.

Adult bone markers by DEXA scan - bone areaCollected at study visits 3, 12 and 24 months pp. of 1st child

Bone area (cm\^2) assessed by a Dual-energy X-ray absorptiometry-scan.

Adult blood sample - HbA1cCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for markers of glucose metabolism HbA1c

Adult blood sample - Alanintransaminase (ALAT)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for ALAT

Adult blood sample - Gamma-GT (GGT)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for GGT

Adult blood sample - c-peptideCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for c-peptide

Adult blood sample - Insulin-like Growth Factor-1 (IGF-1)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for IGF-1

Adult blood sample - glucoseCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for glucose

Adult blood sample - HemoglobinCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Hemoglobin

Adult blood sample - LeukocytterCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Leucocytter

Adult blood sample - Insulin-like growth factor-binding protein-3 (IGFBP-3)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for IGFBP-3

Adult blood sample - GhrelinCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Ghrelin

Adult blood sample - Interleukin 6 (IL-10)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for IL-10

Adult blood sample - Interleukin-1 alpha (IL-1 alpha)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for IL-1 alpha

Adult blood sample - Tumor Necrosis Factor alfa (TNF-α)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for TNF-α

Adult blood sample - Insulin-like growth factor-binding protein-1 (IGFBP-1)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for IGFBP-1

Adult blood sample - LeptinCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Leptin

Adult blood sample - AdiponectinCollected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for Adiponectin

Adult blood sample - Free fatty Acids (FFA)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for FFA

Adult blood sample - Interleukin 6 (IL-6)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for IL-6

Offspring blood sample - insulinCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Insulin

Offspring blood sample - IGFBP-3Collected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for IGFBP-3

Cord blood -FFACollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for FFA

Cord blood -CRPCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for CRP

Adult blood sample - Growth Hormone (GH)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for GH

Adult blood sample - Interferon gamma (IFN-γ)Collected at GA34-GA36 in 1st pregnancy, study visit 3, 12 and 24 months pp of 1st child, at GA34 in 2nd pregnancy and at follow visit 3 months pp after 2nd child

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will be analysed for IFN-γ

Adult female blood sample - ProgesteroneCollected at GA34-GA36 in both pregnancies

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will only be collected from the female participant during pregnancy and be analysed for Progesterone

Adult female blood sample - sex hormone binding globulin (SHBG)Collected at GA34-GA36 in both pregnancies

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will only be collected from the female participant during pregnancy and be analysed for SHBG

Adult female blood sample -EstradiolCollected at GA34-GA36 in both pregnancies

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will only be collected from the female participant during pregnancy and be analysed for Estradiol

Offspring blood sample - glucoseCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for glucose

Offspring blood sample - cTroponin I (cTnI)Collected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for cTnI

Cord blood - c-peptideCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for C-peptide

Cord blood -IGFBP-1Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for IGFBP-1

Cord blood -FerritinCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for Ferritin

Adult female blood sample - Humant choriogonadotropin (hCG)Collected at GA34-GA36 in both pregnancies

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will only be collected from the female participant during pregnancy and be analysed for hCG

Offspring blood sample - c-peptideCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for c-peptide

Offspring blood sample - HgbA1cCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for HgbA1c

Offspring blood sample - AdiponectinCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Adiponectin

Offspring blood sample - FFACollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for FFA

Offspring blood sample - Growth Hormone (GH)Collected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for GH

Cord blood -IGFBP-3Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for IGFBP-3

Cord blood -cTnICollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for cTnI

Adult female blood sample - Proinsulin C-peptideCollected at GA34-GA36 in both pregnancies

All blood samples will be collected by venous puncture in the antecubital vein after an overnight fast from at least 10 pm the night before. Samples will only be collected from the female participant during pregnancy and be analysed for Proinsulin C-peptide

Offspring blood sample - FerritinCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Ferritin

Offspring blood sample - CRPCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for CRP

Offspring blood sample - Cholesterol HDLCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Cholesterol HDL

Offspring blood sample - Total CholesterolCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Total Cholesterol

Offspring blood sample - LeptinCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for Leptin

Offspring blood sample - BNPCollected 12 months after birth in both offsprings

A non-fasting venous blood will be drawn. Parents will be instructed that the offspring should not have eaten two hours prior to the assessment. Samples will be analysed for BNP

Cord blood -BNPCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for BNP

Cord blood -CD163Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for CD163

Cord blood -LeptinCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for Leptin

Cord blood - HbA1cCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for HbA1c

Cord blood - IL-10Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for IL-10

Cord blood - IL-1 alphaCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for IL-1 alpha

Cord blood -IFN- gammaCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for IFN- gamma

Cord blood -AdiponectinCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for Adiponectin

Cord blood - IL-6Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for IL-6

Cord blood -TNF- alphaCollected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for-TNF- alpha

Cord blood - Vitamin D3Collected at birth of both offspring

Cord blood will be drawn immediately after birth and analysed. Blood will be analyzed for Vitamin D3

Trial Locations

Locations (1)

Aarhus University Hospital

🇩🇰

Aarhus, Denmark

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