scholarly journals High-Sensitivity Chemiluminescence Immunoassays for Detection of Growth Hormone Doping in Sports

2009 ◽  
Vol 55 (3) ◽  
pp. 445-453 ◽  
Author(s):  
Martin Bidlingmaier ◽  
Jennifer Suhr ◽  
Andrea Ernst ◽  
Zida Wu ◽  
Alexandra Keller ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Single Injection ◽  
Recombinant Human Growth Hormone ◽  
Human Growth ◽  
High Sensitivity ◽  
High Dose ◽  
Recreational Athletes ◽  
Before And After ◽  
Sandwich Type ◽  
Preferential Recognition

Abstract Background: Recombinant human growth hormone (rhGH) is abused in sports, but adequate routine doping tests are lacking. Analysis of serum hGH isoform composition has been shown to be effective in detecting rhGH doping. We developed and validated selective immunoassays for isoform analysis with potential utility for screening and confirmation in doping tests. Methods: Monoclonal antibodies with preference for pituitary hGH (phGH) or rhGH were used to establish 2 pairs of sandwich-type chemiluminescence assays with differential recognition of rhGH (recA and recB) and phGH (pitA and pitB). We analyzed specimens from volunteers before and after administration of rhGH and calculated ratios between the respective rec- and pit-assay results. Results: Functional sensitivities were <0.05 μg/L, with intra- and interassay imprecision ≤8.4% and ≤13.7%, respectively. In 2 independent cohorts of healthy subjects, rec/pit ratios (median range) were 0.84 (0.09–1.32)/0.81 (0.27–1.21) (recA/pitA) and 0.68 (0.08–1.20)/0.80 (0.25–1.36) (recB/pitB), with no sex difference. In 20 recreational athletes, ratios (median SD) increased after a single injection of rhGH, reaching 350% (73%) (recA/pitA) and 400% (93%) (recB/pitB) of baseline ratios. At a moderate dose (0.033 mg/kg), mean recA/pitA and recB/pitB ratios remained significantly increased for 18 h (men) and 26 h (women). After high-dose rhGH (0.083 mg/kg), mean rec/pit ratios remained increased for 32 h (recA/pitA) and 34 h (recB/pitB) in men and were still increased after 36 h in women. Conclusions: Using sensitive chemiluminescence assays with preferential recognition of phGH or rhGH, detection of a single injection of rhGH was possible for up to 36 h.

scholarly journals Expression of insulin target genes in skeletal muscle and adipose tissue in adult patients with growth hormone deficiency: effect of one year recombinant human growth hormone therapy

2001 ◽  
Vol 171 (2) ◽  
pp. 285-292 ◽  
Author(s):  
Y Khalfallah ◽  
G Sassolas ◽  
F Borson-Chazot ◽  
N Vega ◽  
H Vidal
Keyword(s):  
Growth Hormone ◽  
Adipose Tissue ◽  
Recombinant Human Growth Hormone ◽  
Human Growth ◽  
Mrna Levels ◽  
Hexokinase Ii ◽  
Gh Therapy ◽  
Rhgh Therapy ◽  
Before And After ◽  
One Year

Our aim was to investigate the effects of one year recombinant human growth hormone (rhGH) therapy on the regulation by insulin of gene expression in muscle and adipose tissue in adults with secondary GH deficiency (GHD). Six GHD subjects without upper-body obesity were submitted to a 3-h euglycemic hyperinsulinemic clamp before and after one year of rhGH therapy. Muscle and abdominal subcutaneous adipose tissue biopsies were taken before and at the end of each clamp. The mRNA levels of insulin receptor, p85 alpha-phosphatidylinositol-3 kinase (p85 alpha PI-3K), insulin dependent glucose transporter (Glut4), hexokinase II, glycogen synthase, lipoprotein lipase (LPL) in muscle and in adipose tissue, hormone sensitive lipase and peroxisome proliferator-activated receptor gamma (PPAR gamma) in adipose tissue were quantified by RT-competitive PCR. One year treatment with rhGH (1.25 IU/day) increased plasma IGF-I concentrations (54+/-7 vs 154+/-11 ng/ml, P<0.01) but did not affect insulin-stimulated glucose disposal rate measured during the hyperinsulinemic clamp (74+/-9 vs 85+/-5 micromol/kg free fat mass/min). Insulin significantly increased p85 alpha PI-3K, hexokinase II and Glut4 mRNA levels in muscle both before and after rhGH treatment. One year of GH therapy increased LPL mRNA levels in muscle (38+/-2 vs 70+/-7 amol/microg total RNA, P<0.05) and in adipose tissue (2490+/-260 vs 4860+/-880 amol/microg total RNA, P<0.05), but did not change the expression of the other mRNAs. We conclude from this study that GH therapy did not alter whole body insulin sensitivity and the response of gene expression to insulin in skeletal muscle of adult GHD patients, but it did increase LPL expression in muscle and adipose tissue. This result could be related to the documented beneficial effect of GH therapy on lipid metabolism.

Effect of long-term rhGH administration in GH-deficient adults on fat cell epinephrine response

1992 ◽  
Vol 263 (3) ◽  
pp. E467-E472 ◽  
Author(s):  
M. Beauville ◽  
I. Harant ◽  
F. Crampes ◽  
D. Riviere ◽  
M. T. Tauber ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Recombinant Human Growth Hormone ◽  
Human Growth ◽  
Similar Response ◽  
Fat Cell ◽  
Double Blind ◽  
Placebo Controls ◽  
Before And After ◽  
Isolated Adipocytes

Besides exerting its own lipolytic effect, growth hormone (GH) has been reported to potentiate the lipolytic response of adipose tissue to epinephrine. It was thought interesting to find out whether long-term recombinant human growth hormone (rhGH) administration modifies epinephrine-induced lipolysis in isolated adipocytes of GH-deficient adults. In a double-blind protocol, GH-deficient subjects received either 6 mo placebo (controls, n = 5) or 6 mo rhGH (treated, n = 5). Biopsies of fat were obtained from the periumbilical region before and after placebo or rhGH administration. The response of the collagenase-isolated fat cells to various concentrations of epinephrine was assessed by glycerol release, measured by bioluminescence. Epinephrine-induced lipolysis was not altered by 6 mo placebo, while it was significantly increased by 6 mo rhGH. A similar response was obtained with isoproterenol, but no significant differences occurred in either group with UK 14304, an alpha 2-adrenoreceptor agonist. Thus, in GH-deficient adults, long-term rhGH administration improves the lipolytic response of isolated adipocytes to epinephrine, essentially by increasing the efficiency of the beta-adrenergic pathway.

scholarly journals High Dose Recombinant Human Growth Hormone (GH) Treatment of GH-Deficient Patients in Puberty Increases Near-Final Height: A Randomized, Multicenter Trial1

2000 ◽  
Vol 85 (10) ◽  
pp. 3653-3660 ◽  
Author(s):  
Nelly Mauras ◽  
Kenneth M. Attie ◽  
Edward O. Reiter ◽  
Paul Saenger ◽  
Joyce Baptista the Genentech Inc. Coopera
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Final Height ◽  
Recombinant Human Growth Hormone ◽  
Human Growth ◽  
High Dose ◽  
Gh Treatment

Quantification of urinary insulin-like growth factors (IGFs) and IGF binding protein 3 in healthy volunteers before and after stimulation with recombinant human growth hormone

1995 ◽  
Vol 132 (4) ◽  
pp. 433-437 ◽  
Author(s):  
Burkhard Tönshoff ◽  
Werner F Blum ◽  
Mark Vickers ◽  
Sabine Kurilenko ◽  
Otto Mehls ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Factors ◽  
Human Growth Hormone ◽  
Binding Protein ◽  
Recombinant Human Growth Hormone ◽  
Human Growth ◽  
Igf I ◽  
Before And After ◽  
Igfbp 3 ◽  
Insulin Like Growth Factors

Tönshoff B, Blum WF, Vickers M, Kurilenko S, Mehls 0, Ritz E. Quantification of urinary insulin-like growth factors (IGFs) and IGF binding protein 3 in healthy volunteers before and after stimulation with recombinant human growth hormone. Eur J Endocrinol 1995;132:433–7. ISSN 0804–4643 We examined excretion of urinary insulin-like growth factors I and II (IGF-I and IGF-II) and their major binding protein IGFBP-3 in comparison to their respective serum concentration in nine healthy female volunteers (median age 25 years, range 22–27) under baseline conditions and after stimulation with recombinant human growth hormone (rhGH), 4.5 IU twice daily subcutaneously for a period of 3 days. The IGFs were measured in unconcentrated urine by use of recently developed, highly sensitive radioimmunoassays. The IGFBP-3 was measured by a specific radioimmunoassay. The mean (±sd) urinary concentrations of IGF-I (0.08 ± 0.07 μg/l), IGF-II (1.02 ± 0.47 μg/l) and IGFBP-3 (19.1 ± 6.9 μg/l) were two to three orders of magnitude lower than in serum. The ratio of IGF-II over IGF-I concentration in urine (13:1) was five times higher than in serum (2.5:1), and the ratio of IGFBP-3 over the sum of IGF-I and IGF-II in urine (17:1) was four times higher than in serum (4:1). Urinary excretion was 63.3 ± 46.6 ng·m−2 · 24 h−1 for IGF-I, 1002 ± 598 ng·m−2 · 24 h−1 for IGFII and 18039 ± 4983 ng·m−2·24 h−1 for IGFBP-3. Using fast protein liquid exclusion chromatography, only immunoreactive IGFBP-3 components of less than 60 kD were detected in urine, with a major peak at 20kD. Urinary IGFBP-3 excretion correlated with serum IGFBP-3 (r = 0.61, p < 0.01) and the glomerular filtration rate (r = 0.56, p < 0.05) measured by steady-state inulin infusion clearances. Administration of rhGH stimulated significantly (p < 0.005) the serum IGF-I concentration by 50%, but not the urinary IGF-I excretion. In conclusion: the considerably higher ratio of IGF-II to IGF-I in urine compared to serum indicates that urinary IGF excretion does not represent only filtered IGFs, urinary IGF-I is a less sensitive indicator of GH activity than serum IGF-I, and as urinary IGFBP-3 excretion is in proportion to the glomerular filtration rate and serum IGFBP-3, it presumably reflects renal filtration of small immunoreactive IGFBP-3 fragments from the circulation. Burkhard Tönshoff, University Children's Hospital, Im Neuenheimer Feld 150, 69120 Heidelberg, Germany

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