WEEK 5 isotopes Flashcards
What are isotopes and what makes them different?
Certain elements are composed of atoms that are chemically identical but of a slightly different weight due to different numbers of neutrons
How is isotopic enrichment expressed?
expressed as the tracer:tracee ratio (TTR), corrected for the baseline TTR or as atom % excess (APE) or mole % excess (MPE)
What is very important when working with isotopes?
Measuring the background levels is very important
Nomenclature:
[2H7] = ?
[U-13C] = ?
7 atoms of 2H
Every 13C has been replaced by a stable isotope (U = uniformly labelled)
Why is it important that when working with stable isotopes, the quantity is low?
Because stable isotopes tracers have mass:
You dont want the concentration of the molecules (or elements) being measured to significantly change
What is tracer vs tracee?
Tracer = labelled compound
Tracee = original molecule
What is the most widely used method for measuring molecules labelled with a stable isotope?
Gas chromatography mass spectronemy
What is the principle and application of isotope dilution?
Add known amount of tracer -> mixes in body pools -> reach steady state (tracer same loss rate as tracee = flux) -> sampling body pools -> measure dilution = measure of size of body pool
Application: measurement of body composition
What are assumptions for isotope dilution?
- Stable tracee
- Assumed tracer dilutes in a single body pool (e.g. plasma)
When is a priming dose added?
When you have a prolonged duration of tracer infusion due to a large body pool size relative to flux.
What is the principle of the dual isotope method?
Tracer of the molecule/element of interest is given orally with a meal and a tracer that is distinguishable from the oral tracer is given itravenously (or both meal), isotopes are analyzed: different in appearence reflects absorption
Application of dual isotope method? Requirements?
Measuring nutrient absorption. Or bioavailability
Mathematical models required
Application of precursor product method?
Application: measuring macromolecule synthesis rates.
Dual tracer technique for measuring mineral absorption (calcium) :
Method: Stable isotope of calcium in meal -> second isotope iv -> oral tracer mixes with centrum Ca pool, iv Ca normalises for variations in calcium pool mass between individuals -> urine collection: analysis of isotopic enrichment with the two tracers.
How is fractional calcium absorption calculated?
as the ratio of the oral to the iv tracer recovered in urine
Tracers are also used to measure..
Vit A bioavailability
How is muscle protein synthesis measured?
- can be measured by the incorporation of isotopically labelled amino acids into muscle protein, sampled by needle biopsy
Leucine
When is a biomarker sensitive?
> Sensitivity is high, if a biomarker captures true intake, or if a biomarker identifies an abnormal nutritional status (e.g. deficiency) accurately
When is a biomarker specific?
> Specificity is high, if biomarker is not influenced by other factors, or if the biomarker accurately
identifies a normal nutritional status
Pros of biomarkers of intake vs interview/record methods?
▪ Objective: not dependent on memory, truthfulness, or social desirable answers
▪ May be less time consuming for subject and researcher
▪ Easier quality control
▪ Not just a reflection of intake; includes bioavailability
Cons of biomarkers of intake vs interview/record methods?
▪ Several important nutrients lack good biomarkers
▪ No data on food distribution over the day or on eating behaviour
▪ Effect of food intake is mixed with effects of genome and metabolism
True or false: Biomarker is not restricted to nutrient intake. They may be affected by other factors, e.g. environment or genes
True