Arginine synthesis is a complex, age-dependant process involving multiple precursors and enzymes, and the interorgan transfer of substrates. The objectives of this thesis were to elucidate arginine synthesis from enteral precursors in newborn infants and in healthy adults using stable isotope methodology.
In the first, of four studies, I validated the use of a non-invasive methodology using urinary amino acids for stable isotope studies of arginine synthesis and demonstrated a known, but under-recognised, problem with D-amino acid contamination of tracers. Implementing chiral chromatography, this issue was further investigated using samples from previous studies in our laboratory with several different isotopes. I demonstrated the novel finding that the impact of D-amino acids is dependent on the tracer used, and also on the age of the subject.
In the second study, I used a multi-tracer design to assess arginine synthesis from enteral proline or glutamate in healthy preterm infants. Labeled arginine (M+2), proline (M+1) and glutamate (M+3) were given enterally to fifteen stable, growing preterm infants (gestational age at birth 30-35 weeks) at 1-3 weeks’ postnatal age. I found only arginine synthesis from proline, with no synthesis from glutamate. I conclude that enteral proline is the major contributor to arginine synthesis in vivo in human preterm infants.
In the third study, I measured arginine synthesis from enteral proline in adults. I have demonstrated that enteral proline contributes significantly, ~25%, to newly synthesised arginine.
In the fourth study, I used two glutamine tracers, 1-13C and 2-15N, to determine if glutamine is a carbon or nitrogen donor for arginine. I showed that enteral glutamine contributes ~50% of the carbon skeleton for arginine and that the 2-15N tracer significantly overestimates arginine synthesis, with the labeled N being transferred through transamination from pyrroline-5-carboxylate (to ornithine, rather than directly from glutamate to pyrroline-5-carboxylate.
In conclusion, proline is the sole precursor for arginine in human neonates and combines with glutamaine as the dietary precusor in adults. As a precussor for arginine, though, glutamine’s main role is in provision of nitrogen independent of the carbon skeleton.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/29893 |
| Date | 31 August 2011 |
| Creators | Tomlinson, Robert Christopher Kennedy |
| Contributors | Pencharz, Paul B., Ball, Ron |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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