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1
Section of Clinical Biochemistry, Department of Medical Diagnostic Sciences and Special Therapy, University of Padua, 35100 Padua, Italy.
2
Ludwig Boltzmann Institut für Experimentelle
Endokrinologie, A-1100 Wien, Austria.
3
Department of Anesthesia, University of Copenhagen,
DK-2100 Copenhagen, Denmark.
a Author for correspondence. Fax 039-49-657391;
Background: Atrial natriuretic peptide (ANP) is synthesized and
stored in myocytes as prohormone(1-126), which upon release is cleaved
into proANP(1-98) and
-ANP(99-126). In addition, cleavage of
proANP(1-98) produces proANP(1-30), proANP(31-67), and
proANP(79-98) fragments. ProANP(1-30) and proANP(31-67) have roles
in fluid and electrolyte homeostasis. The aim of the present study was
to develop a plasma assay for proANP(1-30) and proANP(31-67) and to
compare results in trained athletes and sedentary subjects.
Methods: Two competitive enzyme immunoassays were established with affinity-purified sheep antiserum against synthetic ANP fragments. The immunoreactivity (ir) of proANP(1-30) and proANP(31-67) was measured in 10-µL plasma samples without extraction in a microwell-based assay. Plasma concentrations in sedentary male subjects (n = 22) and male endurance athletes (n = 14) were examined.
Results: In the assay for ir-proANP(1-30) and ir-proANP(31-67), the concentrations at 95% B/B0 were 4.7 and 14.2 pmol/L, respectively. Within-run CVs were 46% and 56%, and between-run CVs were 9% for both assays. Both assays were linear on dilution (y = 0.9945x - 0.7291 and y = 1.0001x - 3.428), and the recoveries were 102112% and 102106%, respectively. In the sedentary and athletic groups, the ir-proANP(1-30) concentrations were similar: 318 ± 38 pmol/L and 312 ± 25 pmol/L (mean ± SE), respectively, whereas the ir-proANP(31-67) was higher in the rowers (713 ± 81 pmol/L) than in the sedentary subjects (387 ± 71 pmol/L; P <0.005).
Conclusions: The proANP fragment assays are precise (CV <10%) and exhibit nearly quantitative recovery (102112%). Only ir-proANP(31-67) responds to physical training.
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