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      Gonadal response after a single-dose stimulation test with recombinant human chorionic gonadotropin (rhCG) in patients with isolated prepubertal cryptorchidism

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          Abstract

          Background

          The evaluation of prepubertal gonadal Leydig cells secretion requires gonadotropin stimulation. Urinary hCG (human chorionic gonadotropin) is currently unavailable in many countries, however, recombinant hCG (rhCG) can be used. Our aim was to evaluate rhCG-stimulated testicular hormones in a group of patients with cryptorchidism.

          Methods

          We evaluated 31 prepubertal boys (age range, 0.75–9.0 years) presenting with unilateral ( n = 24) or bilateral ( n = 7) cryptorchidism. Patients with other genital abnormalities, previous use of hCG or testosterone or previous surgeries were excluded. Blood samples were obtained at baseline and 7 days after a single subcutaneous dose of rhCG (Ovidrel® 250 mcg) to measure the testosterone, DHT (dihydrotestosterone), AMH (anti-Mullerian hormone), and inhibin B levels.

          Results

          rhCG stimulation significantly increased testosterone levels from 10 ng/dl to 247.8 ± 135.8 ng/dl, increased DHT levels from 4.6 ± 0.8 to 32.3 ± 18.0 ng/dl, and increased the T/DHT ratio from 2.2 ± 0.4 to 8.0 ± 3.5. There was also a significant increase in inhibin B (from 105.8 ± 65.2 to 132.4 ± 56.1 pg/ml; p < 0.05) and AMH levels (from 109.4 ± 52.6 to 152.9 ± 65.2 ng/ml; p < 0.01) after the rhCG stimulation.

          Conclusions

          In this cohort, hormonal responses can be elicited after the rhCG stimulation test, suggesting that rhCG is a promising stimulation test to replace the urinary hCG test during the evaluation of gonadal Leydig cells function. The clinical applicability and adequate performance of rhCG testing must be investigated in future studies.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12610-016-0039-2) contains supplementary material, which is available to authorized users.

          Résumé

          Contexte

          L’évaluation de la sécrétion des cellules gonadiques de Leydig prépubères nécessite une stimulation par les gonadotrophines. La gonadotrophine chorionique humaine (hCG) urinaire est actuellement indisponible dans de nombreux pays; toutefois, l’hCG recombinante (rhCG) peut être utilisée. Notre objectif était d’évaluer les hormones testiculaires sous stimulation par rhCG dans un groupe de patients qui présentaient une cryptorchidie.

          Méthodes

          Nous avons évalué 31 garçons prépubères (âgés de 0,75 à 9 ans) qui présentaient une cryptorchidie uni ( n=24) ou bilatérale ( n=7). Ont été exclus les patients avec d’autres anomalies génitales, ceux qui avaient été traités auparavant par hCG ou testostérone ou par chirurgie. Des échantillons sanguins ont été prélevés au départ et à 7 jours après une unique dose sous cutanée de rhCG (Ovidrel® 250 mcg) pour les mesures des taux de testostérone, dihydrotestostérone (DHT), hormone antimüllérienne (AMH) et inhibine B.

          Résultats

          La stimulation par rhCG augmente de façon significative les taux de testostérone de 10 ng/dl à 247.8±135.8 ng/dl, de DHT de 4.6±0.8 à 32.3±18.0 ng/dl, et le ratio T/DHT de 2.2±0.4 à 8.0±3.5. Les taux d’inhibine B (de 105.8±65.2 à 132.4±56.1 pg/ml; p< 0.05) et d’AMH (de 109.4±52.6 to 152.9±65.2 ng/ml; p< 0.01) ont aussi été significativement augmentés après stimulation par rhCG.

          Conclusion

          Dans cette cohorte, des réponses hormonales ont été obtenues après le test de stimulation par rhCG; ceci suggère que rhCG est un test de stimulation prometteur pour remplacer le test par l’hCG urinaire lors de l’évaluation de la fonction gonadique des cellules de Leydig. L’applicabilité clinique et l’évaluation adéquate de la performance du test par rhCG doivent être étudiées dans de futures études.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12610-016-0039-2) contains supplementary material, which is available to authorized users.

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          Most cited references18

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          Total testosterone assays in women with polycystic ovary syndrome: precision and correlation with hirsutism.

          There is no standardized assay of testosterone in women. Liquid chromatography mass spectrometry (LC/MS) has been proposed as the preferable assay by an Endocrine Society Position Statement. The aim was to compare assay results from a direct RIA with two LC/MS. We conducted a blinded laboratory study including masked duplicate samples at three laboratories--two academic (University of Virginia, RIA; and Mayo Clinic, LC/MS) and one commercial (Quest, LC/MS). Baseline testosterone levels from 596 women with PCOS who participated in a large, multicenter, randomized controlled infertility trial performed at academic health centers in the United States were run by varying assays, and results were compared. We measured assay precision and correlation and baseline Ferriman-Gallwey hirsutism scores. Median testosterone levels were highest with RIA. The correlations between the blinded samples that were run in duplicate were comparable. The correlation coefficient (CC) between LC/MS at Quest and Mayo was 0.83 [95% confidence interval (CI), 0.80-0.85], between RIA and LC/MS at Mayo was 0.79 (95% CI, 0.76-0.82), and between RIA and LC/MS at Quest was 0.67 (95% CI, 0.63-0.72). Interassay variation was highest at the lower levels of total testosterone (≤50 ng/dl). The CC for Quest LC/MS was significantly different from those derived from the other assays. We found similar correlations between total testosterone levels and hirsutism score with the RIA (CC=0.24), LC/MS at Mayo (CC=0.15), or Quest (CC=0.17). A testosterone RIA is comparable to LC/MS assays. There is significant variability between LC/MS assays and poor precision with all assays at low testosterone levels.
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            Biochemical and pharmacological control of the multiplicity of coupling at G-protein-coupled receptors.

            For decades, it has been generally proposed that a given receptor always interacts with a particular GTP-binding protein (G-protein) or with multiple G-proteins within one family. However, for several G-protein-coupled receptors (GPCR), it now becomes generally accepted that simultaneous functional coupling with distinct unrelated G-proteins can be observed, leading to the activation of multiple intracellular effectors with distinct efficacies and/or potencies. Multiplicity in G-protein coupling is frequently observed in artificial expression systems where high densities of receptors are obtained, raising the question of whether such complex signalling reveals artefactual promiscuous coupling or is a genuine property of GPCRs. Multiple biochemical and pharmacological evidence in favour of an intrinsic property of GPCRs were obtained in recent studies. Thus, there are now many examples showing that the coupling to multiple signalling pathways is dependent on the agonist used (agonist trafficking of receptor signals). In addition, the different couplings were demonstrated to involve distinct molecular determinants of the receptor and to show distinct desensitisation kinetics. Such multiplicity of signalling at the level of G-protein coupling leads to a further complexity in the functional response to agonist stimulation of one of the most elaborate cellular transmission systems. Indeed, the physiological relevance of such versatility in signalling associated with a single receptor requires the existence of critical mechanisms of dynamic regulation of the expression, the compartmentalisation, and the activity of the signalling partners. This review aims at summarising the different studies that support the concept of multiplicity of G-protein coupling. The physiological and pharmacological relevance of this coupling promiscuity will be discussed.
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              The concordance between serum anti-Mullerian hormone and testosterone concentrations depends on duration of hCG stimulation in boys undergoing investigation of gonadal function.

              In boys undergoing investigation of gonadal function, the relationship between a single measurement of serum anti-Mullerian hormone (AMH) and hCG stimulated serum testosterone is unclear. The aim of the study was to assess concordance between serum AMH and testosterone concentrations following hCG stimulation of two different durations. Samples from 284 children (M : F, 154 : 130) with a median age of 8 years (10th, 90th centiles, 0.25, 14) were used to establish an AMH reference range. Clinical data were reviewed in boys undergoing investigation of gonadal function and who had an AMH measurement and a hCG stimulated (3-day or 3-week) (n = 26) testosterone. Of these 26 boys, 11 had combined genital anomalies, whereas the rest had conditions such as isolated hypospadias, undescended testes or microphallus. Normal testosterone response to hCG stimulation was defined as a level greater than 3.5 nmol at day 4 and 9.5 nmol/l at day 22. In the reference group, the 5th centile AMH for boys below 1 year was 215 pmol/l and between 1 and 8 years 180 pmol/l. The 95th centile for girls for these respective age groups was 30 pmol/l and 25 pmol/l. In those cases where serum testosterone concentrations were available at day 1, day 4 and day 22 of the 3 week-hCG test, five cases had a normal serum testosterone at day 4 and three cases only showed such a response by day 22. In those where serum AMH was less than 180 pmol/l, a poor testosterone response of less than 3.5 nmol was observed in approximately seven of eight (88%) cases with a 3-day hCG stimulation test or the 3-week test. An AMH of greater than 180 pmol/l was associated with a normal testosterone response at day 4 in 10 out of 15 (67%) cases and at day 22 in eight of 11 (73%) cases. However, a low serum testosterone concentration of less than 3.5 nmol after the 3-day hCG test was only associated with a likelihood of a low AMH in three of eight (37%) cases. With the 3-week hCG test, a low day 22 testosterone of 9.5 mmol/l or less was associated with a low AMH of 180 pmol/l or less in four of seven (57%) cases. In boys undergoing investigation of gonadal function, the concordance between AMH and testosterone is better at day 22 than day 4. A normal AMH may provide useful information on overall testicular function but does not exclude the need for an hCG stimulation test.
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                Author and article information

                Contributors
                letiribeiro@yahoo.com.br
                thais_kataoka@yahoo.com.br
                renata.wznk@yahoo.com.br
                vinbrito@uol.com.br
                +55-11-33677768 , carloslongui@msn.com
                Journal
                Basic Clin Androl
                Basic Clin Androl
                Basic and Clinical Andrology
                BioMed Central (London )
                2051-4190
                28 October 2016
                28 October 2016
                2016
                : 26
                : 13
                Affiliations
                [1 ]Pediatric Endocrinology Unit, Pediatrics Department, Irmandade da Santa Casa de Misericórdia de São Paulo, and Santa Casa de São Paulo School of Medical Sciences, Rua Dr. Cesário Mota Jr, 112, Vila Buarque, São Paulo, CEP 01221-020 Brazil
                [2 ]Developmental Endocrinology Unit, University of São Paulo Faculty of Medicine Clinics Hospital - USP, São Paulo, Brazil
                Article
                39
                10.1186/s12610-016-0039-2
                5084384
                030534db-283b-46f8-93ac-ad0bdb67a62a
                © The Author(s). 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 24 May 2016
                : 25 August 2016
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2016

                gonadotrophine chorionique humaine,cryptorchidie,testostérone,hormone antimüllérienne,rhcg recombinante,human chorionic gonadotropin,cryptorchidism,testosterone,anti-mullerian hormone,recombinant hcg

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