Collaborative Research Group for Necrotizing Enterocolitis

Study Overview
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Brief Summary
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This proposal will test the hypothesis that synthesis and catabolism of epidermal growth factor (EGF), the genotype of the EGF gene, and the microbiome interact to influence EGF expression in infants at risk for necrotizing enterocolitis (NEC).

Detailed Description
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Preterm infants will receive a six hour intravenous infusion of [5,5,5-2H3]leucine (2H3) through an existing intravenous line (IV) to measure EGF synthesis rate. Two blood samples will be obtained, one prior to the start of infusion, and one during the infusion. The enrichment of the stable isotope labeled leucine will be measured in the plasma from these samples; DNA will be extracted from the residual cell pellets. The EGF and EGF receptor genes will be sequenced. Saliva and urine will be obtained for 5 days following infusion to measure EGF and the rate of incorporation of leucine into EGF using liquid chromatography (LC)/mass spectroscopy (MS)/MS technology, as well as enzyme-linked immunosorbent assay (ELISA) . Saliva will be obtained by a Q tip swab and urine and stool obtained from the diaper. Stool will be obtained every 3 to 7 days through 5 weeks to evaluate inflammatory markers and the microbiome. If breastfeeding, a single sample of mother's milk will be obtained for measurement of EGF after adequate volumes for infant feeds are achieved.

Official Title
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Collaborative Research Group for Necrotizing Enterocolitis

Conditions
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Prematurity, Necrotizing Enterocolitis

Intervention / Treatment
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* Biological: [5,5,5-2H3]leucine (stable isotope labeled leucine)


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: gestation 32 weeks or less 1 week of age or less intravenous line in place for clinical purposes Exclusion Criteria: imminent death active infection pre-existing diagnosis of NEC fluid or electrolyte imbalance

Ages Eligible for Study
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Minimum Age: 1 Day
Maximum Age: 7 Days

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Preterm Infants for EGF Profiles<br>Premature infants born at < 32 weeks gestation who are 7 days old or less. Infants received and intravenous infusion of [5,5,5-2H3]leucine (stable isotope labeled leucine) with sampling of blood, urine and saliva. | Biological: [5,5,5-2H3]leucine (stable isotope labeled leucine)<br>* intravenous infusion of labeled leucine dissolved in 5% glucose water: priming dose of 18 micromoles (1.8 ml)/kg over 5 minutes, then 18 micromoles (1.8 ml)/hr for 6 hours; one infusion total<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Salivary EGF (Epidermal Growth Factor) Protein Levels | Salivary EGF protein levels obtained from oral swabs were analyzed by commercially available EGF ELISA kit (R &D systems Inc). EGF protein levels were normalized to micrograms of protein in saliva, and expressed as picogram of EGF protein per microgram of total salivary protein. | Sampling occurred on average day of life 9 with a range from day of life 7 to 21 | 
| Urinary EGF Protein Levels | Urinary EGF protein levels obtained from free flowing urine samples retrieved from subject diaper were analyzed by commercially available EGF ELISA kit (R &D systems Inc). EGF protein levels were normalized to milligrams of creatinine in urine, and expressed as nanograms of EGF protein per milligram of urinary creatinine. | Sampling occurred on average day of life 9 with a range from day of life 7 to 21 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| EGF Gene Sequencing | Identification of computationally predicted functional variants in EGF gene | Sampling occurred on average day of life 9 with a range from day of life 7 to 21 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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newborn, necrotizing enterocolitis, genetics, growth factors, metabolism