Growth Hormone Possible to Detect?
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1 Growth Hormone Possible to Detect?
2
3 GH The most expensive, most fashionable and least understood of the new athletic drugs. (Underground Steroid Handbook 1983) Could be but does it work?
4 GH increases muscle mass in animals
5 4 months GH replacement in an adult GH-deficient patient
6 GH substitution reduces Abdominal fat
7 Clin Endocrinol1996;45:681-8
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13 Mata Analysis of Placebo-Controlled Trials Isometric muscle strength Clinical Endocrinology (2009) 71,
14 Impact of GH substitution on exercise capacity and muscle strength in GH-deficient adults: a meta-analysis of blinded, placebo controlled trials Aerobic exercise capacity Clinical Endocrinology (2009) 71,
15 Ann Intern Med. 2010;152(9): Placebo-controlled, double blind GH trial (2 mg/d) in 96 recreationally trained athletes (63 men and 33 women) for 8 weeks + 6 weeks washout Measurements included body composition, muscle strength, VO2- max, sprint capacity (wingate value) Supported by WADA
16 Sprint Capacity (Wingate Value) 8.3% (P < 0.005) 3.9% (P = 0.05)
17 Challenges in GH abuse detection rhgh is almost identical to pituitary GH GH is secreted in a pulsatile manner Exercise & stress increase GH secretion Urine testing is not feasible The presence of a prohibited substance or its metabolites or markers in an athlete s bodily specimen constitutes an offence (WADA). 17
18 GH secretion is pulsatile and episodic! 24 h profile of serum GH concentration and deconvolution-resolved GH secretory rate Hartman et al, J Clin Endocrinol Metab 1992; 74: 757
19 GH secretion is stimulated by exercise In an intensity-based manner
20 Endogenous GH comes in many forms! Endocrine Review 1991;12:
21 Isoform method Narrow window of opportunity (< 24 h) Detects only biosynthetic 22k doping Does not detect GH secretagogs
22 GH 2000 UK, Sweden, Italy, Denmark Joint Study supported by EU-IOC ( ) Protocols: High dose GH in healthy subjects GH and related markers in athletes Cross-sectional Prospective Before, during and after competition Results: 20 peer-review publications A method
23 Females Males
24 Discriminant analysis: The combination of IGF-I and P-III-P Provided the best model Corrections for age and gender A pre-defined specificity of 1:10000 Sensitivity: Males: 86% Females: 60% GH & IGF Res 2007;17:
25 Detection of dopers with GH formula Non-doped Doped Visit day of 7 study GH treatment Treatment 0.06 IU/kg/day GH Placebo Overall sensitivity 90% GH-2000 formula score Erotokritou-Mulligan GH& IGF Research (5): Epub 2007 Jun 20 25
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27 IOC Rome Workshop 1999 It is possible to detect exogenous GH administration for up to 2 weeks after the last dose Principle was scientifically sound Sensitivity and specificity reasonable Blood testing and immunoassays will be necessary Results obtained were in a predominantly white European population More data on the effect of injury needed and so the GH-2004 project was established
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29 Ethnicity 1200 IGF-I (ng/ml) * Caucasian (n=493) Oriental (n=25) Afro-Caribbean (n=191) Indo-Asian (n=19) Other (n=29) Age (years)
30 Injury Men (n=127) GH score ±3 28±3 84±7 7±3 21±3 42±7 Number of Days since Injury
31 Further work: Adolescents GH-2000 score increases during puberty No false positives
32 Current state according to the inventor The Biomarker is formally approved by WADA/IOC at the London Olympic Games and caught two paraolympians who were missed by the iso-form test. Both admitted doping and went home. The test is currently in limbo as there are commercial assay problems Maybe measurement of IGF-I and PIIINP with MS may solve the problem?
33 Jorgensen Excuse me I know the game is over, but just for the record, I don t think my buzzer was working properly
34 Validation in other populations/ labs Test worked in other studies Germany Australia Rome 34
35 Determining Decision limits to be used in antidoping New limits with currently available assays that should allow introduction of test for anti-doping purposes 35
36 as well as in man
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39 Change in IGF-I and P-III-P with GH IGF-I P-III-P IGF-I (ng/ml) P-III-P (U/ml) Pre GH On GH 0 Pre GH On GH High dose GH Low dose GH Placebo 39 Eur J Endocrinol Jul;163(1):45-54
40 Effect of Exercise on IGF-I IGF-I Exercise Time (minutes) GH Placebo Wallace et al JCEM :
41 Institut für Dopinganalytik und Sportbiochemie Kreischa GH Study 15 healthy male, non-competitive athletes Age yrs, mean 24 years BMI kg/m 2 Two treatment groups placebo (n=5) rhgh 0.06 IU/kg/day (n=10) 14 days therapy by injection Blood samples taken prior to, during and until 4 weeks after treatment. Kniess et al Anal Bioanal Chem Jul;376(5):
42 Response of GH-dependent markers in an ethnically diverse group 31 male & 13 female healthy volunteers recruited from gyms & University of Southampton sports clubs 19 Afro-Caribbean, 21 Indo-Asian, 4 Oriental Randomly assigned to three treatment groups: Placebo 42
43 Effect of Injury No significant change in IGF-I 40-70% rise in P-III-P Dependent on type and severity of injury Men (n=127) Women (n=40) GH score ±3 28±3 84±7 7±3 21±3 42± ±3 28±3 84±7 7±3 21±3 42±7 Number of Days since Injury Erotokritou-Mulligan et al (2008) J Clin Endocrinol & Metab 93(7):
44 STAT5b is the major signaling protein GH p Subject # Muscle t Fat p t 0,25 0,20 0,15 P=0.001 P=0.07 AU 0,10 0,05 Am J Physiol 2006;291:E899-E905 0,00 Saline GH Saline GH Muscle Fat
45 GH signalling pathways documented in vivo in muscle and fat in human subjects GHR-GH-GHR Src JAK2 STAT5b PI3-kinase MAPK PKB/Akt ERK1/2 JNK SOCS IGF-I AJP 2006;291:E JCEM 2008;93:
46 GH (µg/l) ,4 FFA 1,2 (mmol/l) P = ,0 0,8 0,6 0,4 0,2 0, GH Saline Time (minutes) Am J Physiol 2006;291:E899-E905
47 Effect of GH substitution on exercise capacity and muscle strength: a meta-analysis Excluded n = 88 Identified n = 137 Excluded n = 23 Retrieved n = 49 Fulfilled Inclusion n = 26 Non- Extractable data n = 13 Meta-analysis n = 17 New Articles n = 4 Clinical Endocrinology (2009) 71,
48 V02-max (n=148) Weighted mean±se: 0,17 ± 0.02 l/min (p < 0,0001)
49 Muscle Strength n = 113 Total Jørgensen 1989 Whitehead Wallymahmed weighted mean ± SE: 2,8 ± 2,2 % (p =0,20) Vahl Jorgensen 1996
50
51 IGF-I is an important mediator of the anabolic GH effects (but serum IGF-I mainly reflects hepatic production) Juul et al. JCEM 1994, 78:
52 Ethnicity IGF-I (μg/l) GH treatment High dose GH Low dose GH Placebo Day
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