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Androgenic and estrogenic indices in human newborns and infants: the MIREC-ID study

  • T-V. Nguyen (a1) (a2) (a3), P. Monnier (a2) (a3), G. Muckle (a4), S. Sathyanarayana (a5) (a6) (a7), E. Ouellet (a4), M. P. Velez (a8), L. Dodds (a9) and T. E. Arbuckle (a10)...


Prenatal sex steroid exposure plays an important role in determining child development. Yet, measurement of prenatal hormonal exposure has been limited by the paucity of newborn/infant data and the invasiveness of fetal hormonal sampling. Here we provide descriptive data from the MIREC-ID study (n=173 girls; 162 boys) on a range of minimally invasive physical indices thought to reflect prenatal exposure to androgens [anogenital distances (AGDs); penile length/width, scrotal/vulvar pigmentation], to estrogens [vaginal maturation index (VMI) – the degree of maturation of vaginal wall cells] or to both androgens/estrogens [2nd-to-4th digit ratio (2D:4D); areolar pigmentation, triceps/sub-scapular skinfold thickness, arm circumference]. VMI was found to be associated with triceps skinfold thickness (β=0.265, P=0.005), suggesting that this marker may be sensitive to estrogen levels produced by adipose tissue in girls. Both estrogenic and androgenic markers (VMI: β=0.338, P=0.031; 2D:4D – right: β=−0.207, P=0.040; left: β=−0.276, P=0.006; AGD-fourchette − β=0.253, P=0.036) were associated with areolar pigmentation in girls, supporting a role for the latter as an index of both androgen and estrogen exposure. We also found AGD-penis (distance from the anus to the penis) to be associated with scrotal pigmentation (β=0.290, P=0.048), as well as right arm circumference (β=0.462, P<0.0001), supporting the notion that these indices may be used together as markers of androgen exposure in boys. In sum, these findings support the use of several physical indices at birth to convey a more comprehensive picture of prenatal exposure to sex hormones.


Corresponding author

Author for correspondence: T-V. Nguyen, Department of Psychiatry, McGill University, Royal Victoria Hospital, 1001 Decarie, C6.1188 Suite, Montreal, QC, H4A 3J1, Canada. E-mail:


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1.Abbott, DH, Padmanabhan, V, Dumesic, DA. Contributions of androgen and estrogen to fetal programming of ovarian dysfunction. Reprod Biol Endocrinol. 2006; 4, 17.
2.Hines, M. Prenatal endocrine influences on sexual orientation and on sexually differentiated childhood behavior. Front Neuroendocrinol. 2011; 32, 170182.
3.Hollier, LP, Keelan, JA, Hickey, M, Maybery, MT, Whitehouse, AJ. Measurement of androgen and estrogen concentrations in cord blood: accuracy, biological interpretation, and applications to understanding human behavioral development. Front Endocrinol (Lausanne). 2014; 5, 64.
4.Keelan, JA, Mattes, E, Tan, HW, et al. Androgen concentrations in umbilical cord blood and their association with maternal, fetal and obstetric factors. PLoS One. 2012; 7, e42827.
5.Foradori, CD, Weiser, MJ, Handa, RJ. Non-genomic actions of androgens. Front Neuroendocrinol. 2008; 29, 169181.
6.Luconi, M, Forti, G, Baldi, E. Genomic and nongenomic effects of estrogens: molecular mechanisms of action and clinical implications for male reproduction. J Steroid Biochem Mol Biol. 2002; 80, 369381.
7.Reyes, FI, Winter, JS, Faiman, C. Studies on human sexual development. I. Fetal gonadal and adrenal sex steroids. J Clin Endocrinol Metab. 1973; 37, 7478.
8.Reyes, FI, Boroditsky, RS, Winter, JS, Faiman, C. Studies on human sexual development. II. Fetal and maternal serum gonadotropin and sex steroid concentrations. J Clin Endocrinol Metab. 1974; 38, 612617.
9.Eisenberg, ML, Hsieh, MH, Walters, RC, Krasnow, R, Lipshultz, LI. The relationship between anogenital distance, fatherhood, and fertility in adult men. PLoS One. 2011; 6, e18973.
10.Hsieh, MH, Breyer, BN, Eisenberg, ML, Baskin, LS. Associations among hypospadias, cryptorchidism, anogenital distance, and endocrine disruption. Curr Urol Rep. 2008; 9, 137142.
11.Mitchell, RT, Mungall, W, McKinnell, C, et al. Anogenital distance plasticity in adulthood: implications for its use as a biomarker of fetal androgen action. Endocrinology. 2015; 156, 2431.
12.Dean, A, Sharpe, RM. Anogenital distance or digit length ratio as measures of fetal androgen exposure: relationship to male reproductive development and its disorders. J Clin Endocrinol Metab. 2013; 98, 22302238.
13.Thankamony, A, Pasterski, V, Ong, KK, Acerini, CL, Hughes, IA. Anogenital distance as a marker of androgen exposure in humans. Andrology. 2016; 4, 616625.
14.Harlid, S, Adgent, M, Jefferson, WN, et al. Soy formula and epigenetic modifications: analysis of vaginal epithelial cells from infant girls in the IFED study. Environ Health Perspect. 2017; 125, 447452.
15.van der Laak, JAWM, de Bie, LMT, de Leeuw, H, de Wilde, PCM, Hanselaar, AGJM. The effect of Replens (R) on vaginal cytology in the treatment of postmenopausal atrophy: cytomorphology versus computerised cytometry. J Clin Pathol. 2002; 55, 446451.
16.van der Laak, JAWM, Schijf, CP, Kerstens, HMJ, et al. Development and validation of a computerized cytomorphometric method to assess the maturation of vaginal epithelial cells. Cytometry. 1999; 35, 196202.
17.Wilson, MJ, Spaziani, E. Testosterone regulation of pigmentation in scrotal epidermis of the rat. Z Zellforsch Mikrosk Anat. 1973; 140, 451458.
18.Natale, CA, Duperret, EK, Zhang, JQ, et al. Sex steroids regulate skin pigmentation through nonclassical membrane-bound receptors. Elife. 2016; 5, e15104.
19.Ali, A, Berens, P, Siddiqui, G, Ali, V. Nipple and areolar hyperpigmentation secondary to the use of estradiol spray on the ipsilateral forearm skin: a report of two cases. J Womens Health. 2012; 21, 363365.
20.Diven, DG, Crawford, JM. Ipsilateral areolar hyperpigmentation following unilateral application of estradiol spray. Am J Obstetr Gynecol. 2010; 203, E8.
21.Tadokoro, T, Itami, S, Hosokawa, K, Terashi, H, Takayasu, S. Human genital melanocytes as androgen target cells. J Invest Dermatol. 1997; 109, 513517.
22.Fullerton, A, Fischer, T, Lahti, A, et al. Guidelines for measurement of skin colour and erythema – a report from the standardization group of the European society of contact dermatitis. Contact Dermatitis. 1996; 35, 110.
23.Takiwaki, H, Overgaard, L, Serup, J. Comparison of narrow-band reflectance spectrophotometric and tristimulus colorimetric measurements of skin color - 23 anatomical sites evaluated by the dermaspectrometer(R) and the chroma-meter-Cr-200(R). Skin Pharmacol. 1994; 7, 217225.
24.Garn, SM, Hertzog, KP, Poznanski, AK, Nagy, JM. Metacarpophalangeal length in the evaluation of skeletal malformation. Radiology. 1972; 105, 375381.
25.Galis, F, Ten Broek, CM, Van Dongen, S, Wijnaendts, LC. Sexual dimorphism in the prenatal digit ratio (2D:4D). Arch Sex Behav. 2010; 39, 5762.
26.Manning, JT, Fink, B. Sexual dimorphism in the ontogeny of second (2D) and fourth (4D) digit lengths, and digit ratio (2D:4D). Am J Hum Biol. 2018; 30, e23138.
27.Malas, MA, Dogan, S, Evcil, EH, Desdicioglu, K. Fetal development of the hand, digits and digit ratio (2D:4D). Early Hum Dev. 2006; 82, 469475.
28.Fink, B, Manning, JT. Direct versus indirect measurement of digit ratio: new data from Austria and a critical consideration of clarity of report in 2D:4D studies. Early Hum Dev. 2018; 127, 2832.
29.Ribeiro, E, Neave, N, Morais, RN, Manning, JT. Direct versus indirect measurement of digit ratio (2D:4D): a critical review of the literature and new data. Evol Psychol-Us. 2016; 14, 18.
30.Szwed, A, Kosinska, M, Manning, JT. Digit ratio (2D:4D) and month of birth: a link to the solstitial-melatonin-testosterone effect. Early Hum Dev. 2017; 104, 2326.
31.Velez, MP, Arbuckle, TE, Monnier, P, Fraser, WD. Is maternal periconceptional smoking associated with 2D:4D digit ratio in their children? J Dev Orig Hlth Dis. 2017; 8, 597603.
32.Velez, MP, Arbuckle, TE, Monnier, P, Fraser, WD. Female digit length ratio (2D:4D) and time-to-pregnancy. Hum Reproduct. 2016; 31, 21282134.
33.Welsh, M, MacLeod, DJ, Walker, M, Smith, LB, Sharpe, RM. Critical androgen-sensitive periods of rat penis and clitoris development. Int J Androl. 2010; 33, e144152.
34.Fritsch, H, Hoermann, R, Bitsche, M, Pechriggl, E, Reich, O. Development of epithelial and mesenchymal regionalization of the human fetal utero-vaginal anlagen. J Anat. 2013; 222, 462472.
35.Wilson, MJ. Inhibition of development of both androgen-dependent and androgen-independent pigment-cells in scrotal skin dermis of the rat by anti-androgen treatment during fetal growth. Endocrinology. 1983; 112, 321325.
36.Javed, A, Lteif, A. Development of the human breast. Semin Plast Surg. 2013; 27, 512.
37.Wilson, JD, Leihy, MW, Shaw, G, Renfree, MB. Androgen physiology: unsolved problems at the millennium. Mol Cell Endocrinol. 2002; 198, 15.
38.Fritsch, M, Orfanos, CE, Zouboulis, C. Sebocytes are the key regulators of androgen homeostasis in human skin. J Invest Dermatol. 2001; 117, 785785.
39.Thornton, MJ. The biological actions of estrogens on skin. Exp Dermatol. 2002; 11, 487502.
40.Dusek, A, Bartos, L. Variation in ano-genital distance in spontaneously cycling female mice. Reprod Domest Anim. 2012; 47, 984987.
41.Manning, JT, Scutt, D, Wilson, J, Lewis-Jones, DI. The ratio of 2nd to 4th digit length: a predictor of sperm numbers and concentrations of testosterone, luteinizing hormone and oestrogen. Hum Reprod. 1998; 13, 30003004.
42.Sathyanarayana, S, Beard, L, Zhou, C, Grady, R. Measurement and correlates of ano-genital distance in healthy, newborn infants. Int J Androl. 2010; 33, 317323.
43.Swan, SH, Main, KM, Liu, F, et al. Decrease in anogenital distance among male infants with prenatal phthalate exposure. Environ Health Persp. 2005; 113, 10561061.
44.Santos, S, Gaillard, R, Oliveira, A, et al. Associations of infant subcutaneous fat mass with total and abdominal fat mass at school-age: the generation R study. Paediatr Perinat Ep. 2016; 30, 511520.
45.Gale, C, Logan, KM, Jeffries, S, et al. Sexual dimorphism in relation to adipose tissue and intrahepatocellular lipid deposition in early infancy. Int J Obesity. 2015; 39, 629632.
46.Bluher, M. Importance of estrogen receptors in adipose tissue function. Mol Metab. 2013; 2, 130132.
47.Arbuckle, TE, Fraser, WD, Fisher, M, et al. Cohort profile: the maternal-infant research on environmental chemicals research platform. Paediatr Perinat Ep. 2013; 27, 415425.
48.Bernbaum, JC, Umbach, DM, Ragan, NB, et al. Pilot studies of estrogen-related physical findings in infants. Environ Health Perspect. 2008; 116, 416420.
49.Farage, M, Maibach, H. Lifetime changes in the vulva and vagina. Arch Gynecol Obstet. 2006; 273, 195202.
50.Adgent, MA, Umbach, DM, Zemel, BS, et al. A longitudinal study of estrogen-responsive tissues and hormone concentrations in infants fed soy formula. J Clin Endocrinol Metab. 2018; 103, 18991909.
51.Pedersen, SB, Bruun, JM, Hube, F, Kristensen, K, Hauner, H, et al. Demonstration of estrogen receptor subtypes alpha and beta in human adipose tissue: influences of adipose cell differentiation and fat depot localization. Mol Cell Endocrinol. 2001; 182, 2737.
52.Bhasin, S. Effects of testosterone administration on fat distribution, insulin sensitivity, and atherosclerosis progression. Clin Infect Dis. 2003; 37, S142S149.
53.Bhasin, S, Taylor, WE, Singh, R, et al. The mechanisms of androgen effects on body composition: Mesenchymal pluripotent cell as the target of androgen action. J Gerontol a-Biol. 2003; 58, 11031110.


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