Einleitung und Ziel: Diabetes Mellitus Typ 1 kann die Hodenfunktion und Reproduktivität negativ beeinflussen, allerdings sind die Pathomechanismen auf testikulärer Ebene bisher unklar. Daher war das Ziel der Studie, die Auswirkungen einer Diabetes mellitus Typ 1-Erkrankung auf die Hodenfunktion zu untersuchen.

Testicular samples obtained for fertility preservation often need to be transported between clinics. This study aimed to mimic this short-term hypothermic storage (4–8°C) and explore the impact of these conditions and the transport medium composition on prepubertal rat testicular tissue samples. Testicular tissue samples obtained from 7 days post-partum rats were transferred to six compositionally different basal culture media and a balanced salt solution, which had been kept at 4–8°C prior to transfer. The samples were preserved for either 12 or 24 h in these hypothermic conditions. The potential effects of the short-term storage were evaluated by assessing the morphology, measuring the testosterone levels by radioimmunoassay and analysing 96 genes with TaqMan Low-Density Arrays. Levels of gene expression related to energy, apoptosis, and angiogenesis pathways were altered after hypothermic storage for 12 and especially 24 h. We observed only minor differences in gene expression profiles for germ and testicular somatic cells and no differences in tissue morphology and testosterone production levels. Short-term hypothermic storage of testicular tissue with a maximum duration of 24 h does not affect the overall expression profile of testicular cell-specific genes; however, in a minor way, it affects the expression of specific cellular genes. Lay summary Male fertility depends on the proper functioning of cells which develop into reproductive cells. Due to an increasing number of childhood cancer survivors suffering from treatment-related fertility problems, as well as recent reports showing a dramatic decrease in sperm counts over the last decades, male fertility preservation has become an important research topic. To date, there is no method to restore fertility for men who are not able to produce sperm. One promising method to preserve the potential fertility of these patients is freezing tissue or cells from the testicles for future fertility treatments. A critical phase in freezing testicular tissue or cells is the time between removing the tissue from the testicles and freezing it. To better understand the impact of this phase on the quality of the testicular tissue, we used the testes of rats as a model for our research. We found that cooling testis tissue has only minor effects on the expression of genes that are important for testis function.

Brief summary: In this study the authors reported the adult height and timing of puberty of a cohort of Danish children born small for gestational age (SGA) and treated with rhGH in comparison with national growth standards. rhGH treatment significantly increased height SDS in adulthood as compared to the height SDS at treatment start. Age at peak height velocity did not differ compared with the reference cohort, although peak height velocity was reduced in SGA subjects. SGA boys had an earlier onset of puberty compared with the reference cohort.

Journal Article Letter to the Editor From Arroyo et al: “Timing of Puberty, Pubertal Growth, and Adult Height in Short Children Born Small for Gestational Age Treated With Growth Hormone” Get access Ramón Arroyo Ruiz, Ramón Arroyo Ruiz Department of Pediatrics, Complejo asistencial Universitario de Salamanca, Salamanca 37007, SpainResearch Unit for Infectious, Inflammatory and Metabolic, Clinical Pediatrics, Instituto de Investigación Biomédica de Salamanca, Salamanca 37007, Spain https://orcid.org/0000-0003-1424-4236 Search for other works by this author on: Oxford Academic Google Scholar Isabel Leiva-Gea, Isabel Leiva-Gea Department of Pediatric Endocrinology, Hospital Regional Universitario de Málaga, Málaga 29010, SpainGroup on Paediatric Research, Instituto de Investigación Biomédica de Málaga, Málaga 29590, Spain Correspondence: Isabel Leiva-Gea, MD, Department of Pediatric Endocrinology, Hospital Regional Universitario de Málaga, Maternal and Child Hospital, Av. del Arroyo de los Ángeles, Málaga 29010, Spain. Email: isabeleivag@gmail.com. https://orcid.org/0000-0002-2259-7253 Search for other works by this author on: Oxford Academic Google Scholar Juan Pedro López-Siguero Juan Pedro López-Siguero Department of Pediatric Endocrinology, Hospital Regional Universitario de Málaga, Málaga 29010, SpainGroup on Paediatric Research, Instituto de Investigación Biomédica de Málaga, Málaga 29590, Spain https://orcid.org/0000-0001-9282-9139 Search for other works by this author on: Oxford Academic Google Scholar The Journal of Clinical Endocrinology & Metabolism, Volume 108, Issue 10, October 2023, Pages e1157–e1158, https://doi.org/10.1210/clinem/dgad197 Published: 25 April 2023 Article history Received: 17 March 2023 Editorial decision: 03 April 2023 Published: 25 April 2023 Corrected and typeset: 28 April 2023

Research Aim Diabetes type 1 can negatively influence testicular function and fertility but the pathomechanisms on testicular level remain to be elucidated. Therefore, the aim of the study was to evaluate effects of diabetes mellitus type 1 on testicular function.

Convergent evidence associates exposure to endocrine disrupting chemicals (EDCs) with major human diseases, even at regulation-compliant concentrations. This might be because humans are exposed to EDC mixtures, whereas chemical regulation is based on a risk assessment of individual compounds. Here, we developed a mixture-centered risk assessment strategy that integrates epidemiological and experimental evidence. We identified that exposure to an EDC mixture in early pregnancy is associated with language delay in offspring. At human-relevant concentrations, this mixture disrupted hormone-regulated and disease-relevant regulatory networks in human brain organoids and in the model organisms Xenopus leavis and Danio rerio , as well as behavioral responses. Reinterrogating epidemiological data, we found that up to 54% of the children had prenatal exposures above experimentally derived levels of concern, reaching, for the upper decile compared with the lowest decile of exposure, a 3.3 times higher risk of language delay.

Context Growth hormone (GH) is used to treat short children born small for gestational age (SGA); however, the effects of treatment on pubertal timing and adult height are rarely studied. Objective To evaluate adult height and peak height velocity in short GH-treated SGA children. Methods Prospective longitudinal multicenter study. Participants were short children born SGA treated with GH therapy (n = 102). Adult height was reported in 47 children. A reference cohort of Danish children was used. Main outcome measures were adult height, peak height velocity, age at peak height, and pubertal onset. Pubertal onset was converted to SD score (SDS) using Danish reference data. Results Gain in height SDS from start of treatment until adult height was significant in both girls (0.94 [0.75; 1.53] SDS, P = .02) and boys (1.57 [1.13; 2.15] SDS, P < .001). No difference in adult height between GH dosage groups was observed. Peak height velocity was lower than a reference cohort for girls (6.5 [5.9; 7.6] cm/year vs 7.9 [7.4; 8.5] cm/year, P < .001) and boys (9.5 [8.4; 10.7] cm/year vs 10.1 [9.7; 10.7] cm/year, P = .002), but no difference in age at peak height velocity was seen. Puberty onset was earlier in SGA boys than a reference cohort (1.06 [-0.03; 1.96] SDS vs 0 SDS, P = .002) but not in girls (0.38 [-0.19; 1.05] SDS vs 0 SDS, P = .18). Conclusion GH treatment improved adult height. Peak height velocity was reduced, but age at peak height velocity did not differ compared with the reference cohort. SGA boys had an earlier pubertal onset compared with the reference cohort.

Fertility preservation for male childhood cancer survivors not yet capable of producing mature spermatozoa, relies on experimental approaches such as testicular explant culture. Although the first steps in somatic maturation can be observed in human testicular explant cultures, germ cell depletion is a common obstacle. Hence, understanding the spermatogonial stem cell (SSC) niche environment and in particular, specific components such as the seminiferous basement membrane (BM) will allow progression of testicular explant cultures. Here, we revealed that the seminiferous BM is established from 6 weeks post conception with the expression of laminin alpha 1 (LAMA 1) and type IV collagen, which persist as key components throughout development. With prepubertal testicular explant culture we found that seminiferous LAMA 1 expression is disrupted and depleted with culture time correlating with germ cell loss. These findings highlight the importance of LAMA 1 for the human SSC niche and its sensitivity to culture conditions.

ABSTRACTIntroductionSexual development in females and males are routinely measured according to the Tanner Stages. Sparse data exist on the timing of pubertal milestones in Pakistan. To fill this gap, the age of attainment of pubertal milestones and their relationship with nutritional status was explored among children and adolescents living in the rural district of Matiari, Pakistan.MethodsAnthropometry, nutrition biomarkers and Tanner Stage were assessed among girls aged 9.0–14.9 years (n = 723) and boys aged 10.0–15.9 years (n = 662) who were free from known disease in the rural District of Matiari, Pakistan. Median age was calculated for all Tanner Stages and menarche. Multivariable linear regressions were undertaken to determine covariates associated with the timing (age) of pubertal milestones.ResultsAmong participants living in this rural community, the median age of puberty onset for girls was 11.9 years (95%CI:10.9; 12.5) and boys was 12.3 years (95%CI:11.5; 12.9). Age at first menarche was 12.9 years (95%CI:12.1; 13.3). Undernutrition was widespread among adolescents in this community. Thirty‐seven percent of females and 27.0% of males were stunted while 20.5% of females and 31.3% of males were thin. Only 8% (n = 58) of females and 12% (n = 78) of males were free from any nutrient deficiency with most adolescents having two or three nutrient deficiencies.ConclusionsUndernutrition (stunting or thinness) was associated with relatively older ages for early puberty stages but not puberty completion. This may decrease the duration of the pubertal growth spurt and curtail potential catch‐up growth that may occur during puberty. Efforts to decrease nutrient deficiencies, stunting and thinness beyond childhood should be made in rural Pakistan.

In recent years, there has been an increased incidence in several of the most common reproductive disorders, including hypospadias and cryptorchidism in newborns, and testicular cancer and lower sperm quality in young adult males. In addition, the timing of puberty has also changed over time. Although the cause of these reproductive effects is a matter of intense debate, a link with the presence of ubiquitous compounds in the environment, or the exposure to specific groups of medications during foetal life, has been suggested. Results from epidemiological and experimental studies, as well as clinical observations in humans indicate that endocrine-disrupting chemicals may be associated with those disorders. In this review, we will summarize the results of epidemiological studies and experimental studies utilising human testicular cells or tissue. Due to increasing public interest and the recently published data, the main focus will be on the effects of prenatal exposure to mild analgesics.

Background Advances in three-dimensional culture technologies have led to progression in systems used to model the gonadal microenvironment in vitro. Despite demonstrating basic functionality, tissue organisation is often limited. We have previously detailed a three-dimensional culture model termed the three-layer gradient system to generate rat testicular organoids in vitro. Here we extend the model to human first-trimester embryonic gonadal tissue. Results Testicular cell suspensions reorganised into testis-like organoids with distinct seminiferous-like cords situated within an interstitial environment after 7 days. In contrast, tissue reorganisation failed to occur when mesonephros, which promotes testicular development in vivo, was included in the tissue digest. Organoids generated from dissociated female gonad cell suspensions formed loosely organised cords after 7 days. In addition to displaying testis-specific architecture, testis-like organoids demonstrated evidence of somatic cell differentiation. Within the 3-LGS, we observed the onset of AMH expression in the cytoplasm of SOX9-positive Sertoli cells within reorganised testicular cords. Leydig cell differentiation and onset of steroidogenic capacity was also revealed in the 3-LGS through the expression of key steroidogenic enzymes StAR and CYP17A1 within the interstitial compartment. While the 3-LGS generates a somatic cell environment capable of supporting germ cell survival in ovarian organoids germ cell loss was observed in testicular organoids. Conclusion The 3-LGS can be used to generate organised whole gonadal organoids within 7 days. The 3-LGS brings a new opportunity to explore gonadal organogenesis and contributes to the development of more complex in vitro models in the field of developmental and regenerative medicine.

Type 1 diabetes mellitus (T1DM) is associated with impaired spermatogenesis, lower testosterone levels and epididymal weight. However, the underlying processes in the testis are unknown and remain to be elucidated. Therefore, the present study focused on the effects of T1DM on testicular function in a spontaneously diabetic rat model.BB/OKL rats after diabetes manifestation were divided into three groups: without insulin treatment, insulin treatment for a duration of 2 and of 6 weeks. Anthropometrical data, circulating levels of gonadotrophins, testosterone and inhibin B were measured. Intratesticular testosterone, oxidative stress, inflammation, and apoptosis were analyzed. Key enzymes of steroidogenesis were evaluated in the testis.Untreated diabetic rats had significantly lower serum FSH and LH levels. Serum and intratesticular testosterone levels significantly decreased in the untreated diabetic rat compared to healthy controls. Key markers of Leydig cell function were significantly downregulated on RNA level: Insl3 by 53% (p=0.006), Star by 51% (p=0.004), Cyp11A1 by 80% (p=0.003), 3Beta-Hsd2 by 61% (p=0.005) and Pbr by 52% (p=0.002). In the insulin-treated group only Cyp11A1 and 3Beta-Hsd2 transcripts were significantly lower. Interestingly the long-term insulin treated group showed significant upregulation of most steroidogenic enzymes without affecting testosterone levels. TNF-alpha and apoptosis were significantly increased in the long-term insulin treated rats. In conclusion T1DM, with a severe lack of insulin, has an adverse action on Leydig cell function. This is partially reversible with well compensated blood glucose control. Long-term T1DM adversely affects Leydig cell function due to the process of inflammation and apoptosis.

BACKGROUND:Potentially harmful effects of persistent organic pollutants (POPs) such as polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT) on prenatal development and the endocrine system have been controversially discussed.METHODS:Working with a German cohort of 324 pregnant women, we assessed POP levels and used robust linear regression models to determine potential associations between maternal POP concentrations and pre- and postnatal development in the children, as well as the thyroid hormone status of the mother and child.RESULTS:Maternal p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) and most measured PCBs positively correlated with postnatal weight gain. We detected no correlation between newborn birth weight and head circumference, respectively, and maternal PCB and p,p'-DDE serum levels, while body length at birth was negatively associated with the maternal serum concentration of PCB 183. Maternal p,p'-DDE and nearly all PCB serum levels showed a negative correlation with maternal free triiodothyronine (FT3). p,p'-DDE and PCB 74 and 118 were negatively associated with maternal thyroid-stimulating hormone levels. In addition, we identified significant associations between maternal POP levels and thyroid hormone parameters of the child.CONCLUSIONS:These results indicate that POP exposure likely affects different aspects of pre- and postnatal development and impacts the thyroid hormone status of both mother and child.IMPACT:Pregnant women in a German cohort display a substantial accumulation of POPs. Body mass index and age influence maternal serum POP levels. Maternal POP levels show correlations with the child's length at birth and weight gain, and FT3 levels in the mother and child. Our data provide additional evidence for the potentially harmful influence of POPs. Our data indicate that POPs influence pre- and postnatal development.

Aim To assess factors associated with the timing of puberty onset (Tanner Stage Breast 2/Genital 2) among adolescents living in an urban slum in Karachi, Pakistan. Methods Girls enrolled at 8-10 years (n = 1009) and boys 9-11 years (n = 863) were followed every 6 months from 2006 to 2010. Parametric survival analysis for interval-censored data was used to estimate median age at puberty onset and assess the association between exposures and timing of puberty onset. Results The overall median age at Tanner Stage Breast 2 (AAB2) was 10.1 years (95% CI: 10.1; 10.5), and the median age at Tanner Stage Genital 2 (AAG2) was 10.1 years (95% CI: 10.1; 10.6). Stunting delayed AAB2 1 year and AAG2 about 6 months when compared to non-stunted peers. In the multivariable model, after adjusting for age at enrolment, stunting, thinness and vitamin A deficiency (VAD) were significantly associated with delayed AAB2, while stunting and anaemia were significantly associated with delayed AAG2. Conclusion Among adolescents living in the Karachi slum, stunting and highly prevalent anaemia delayed AAG2, while stunting, thinness and VAD delayed AAG2. Parental and household factors were not significantly associated with the timing of puberty onset.

To assess factors associated with the timing of puberty onset (Tanner Stage Breast 2/Genital 2) among adolescents living in an urban slum in Karachi, Pakistan. Girls enrolled at 8-10 years (n = 1009) and boys 9-11 years (n = 863) were followed every 6 months from 2006 to 2010. Parametric survival analysis for interval-censored data was used to estimate median age at puberty onset and assess the association between exposures and timing of puberty onset. The overall median age at Tanner Stage Breast 2 (AAB2) was 10.1 years (95% CI: 10.1; 10.5), and the median age at Tanner Stage Genital 2 (AAG2) was 10.1 years (95% CI: 10.1; 10.6). Stunting delayed AAB2 1 year and AAG2 about 6 months when compared to non-stunted peers. In the multivariable model, after adjusting for age at enrolment, stunting, thinness and vitamin A deficiency (VAD) were significantly associated with delayed AAB2, while stunting and anaemia were significantly associated with delayed AAG2. Among adolescents living in the Karachi slum, stunting and highly prevalent anaemia delayed AAG2, while stunting, thinness and VAD delayed AAG2. Parental and household factors were not significantly associated with the timing of puberty onset.

Objectives: Staging sexual maturation is an integral component of adolescent research. The Pubertal Development Scale (PDS) is commonly used as a puberty self-assessment tool because it avoids the use of images. Among the youth living in rural Pakistan, we determined the accuracy of self-reported pubertal assessments using a modified PDS compared to the 'gold standard' of physically assessed Tanner stages by a physician. Methods: The strength of agreement between self-assessed puberty using a modified PDS and the 'gold' standard of physician-assessed Tanner stages was reported using weighted kappa (kappa(W)) for girls (n = 723) of 9.0-14.9 years of age or boys (n = 662) of 10.0-15.9 years of age living in the rural District of Matiari. Results: Agreement between the gold standard and self-assessment for puberty was substantial, with a( )kappa(W) of 0.73 (95% confidence interval [CI]: 0.67; 0.79) for girls and a kappa(W) of 0.61 (95% CI: 0.55; 0.66) for boys. Substantial agreement was observed for both boys and girls classified as thinness but only for girls with a normal body mass index. Those who were classified as severely thin had moderate agreement. The prevalence of overestimation was 18.5% (95% CI: 15.9-21.5) for girls and 2.7% (95% CI: 1.7-4.3) for boys, while the prevalence of underestimation estimation was 8.0% (95% CI: 6.2-10.2) for girls and 29.0% (95% CI: 25.8-32.6) for boys. Conclusions: Most girls and boys assessed their pubertal development with substantial agreement with physician assessment. Girls were better able to assess their puberty, but they were more likely to overestimate. Agreement for boys was also substantial, but they were more likely to underestimate their pubertal development. In this rural Pakistan population, the PDS seems to be a promising tool for self-assessed puberty.

Background Some children born small for gestational age (SGA) experience supra-physiological insulin-like growth factor-I (IGF-I) concentrations during GH treatment. However, measurements of total IGF-I concentrations may not reflect the bioactive fraction of IGF-I which reaches the IGF-I receptor at target organs. We examined endogenous IGF-bioactivity using an IGF-I kinase receptor activation (KIRA) assay that measures the ability of IGF-I to activate the IGF-IR in vitro. Aim To compare responses of bioactive IGF and total IGF-I concentrations in short GH treated SGA children in the North European Small for Gestational Age Study (NESGAS). Material and method In NESGAS, short SGA children (n = 101, 61 males) received GH at 67 µg/kg/day for 1 year. IGF-I concentrations were measured by Immulite immunoassay and bioactive IGF by in-house KIRA assay. Results Bioactive IGF increased with age in healthy pre-pubertal children (n = 94). SGA children had low-normal bioactive IGF levels at baseline (-0.12 (1.8 SD), increasing significantly after one year of high-dose GH treatment to 1.1 (1.4) SD, P < 0.01. Following high-dose GH, 68% (n = 65) of SGA children had a total IGF-I concentration >2SD (mean IGF-I 2.8 SDS), whereas only 15% (n = 15) had levels of bioactive IGF slightly above normal reference values. At baseline, bioactive IGF (SDS) was significantly correlated to height (SDS) (r = 0.29, P = 0.005), in contrast to IGF-I (SDS) (r = 0.17, P = 0.10). IGF-I (SDS) was inversely correlated to delta height (SDS) after one year of high-dose GH treatment (r = -0.22, P = 0.02). Conclusion In contrast to total IGF-I concentrations, bioactive IGF stayed within the normal reference ranges for most SGA children during the first year of GH treatment.

Masculinization of the external genitalia in humans is dependent on formation of 5α-dihydrotestosterone (DHT) through both the canonical androgenic pathway and an alternative (backdoor) pathway. The fetal testes are essential for canonical androgen production but little is known about the synthesis of backdoor androgens despite their known critical role in masculinization. In this study, we have measured plasma and tissue levels of endogenous steroids in second trimester human male fetuses using multi-dimensional and high-resolution mass-spectrometry. Results show that androsterone is the principal backdoor androgen in the fetal circulation and that DHT is undetectable (<1ng/ml). Backdoor pathway intermediates are found primarily in the placenta and fetal liver with significant androsterone levels also in the fetal adrenal. Backdoor intermediates, including androsterone, are mostly undetectable in the fetal testes. This is consistent with transcript levels of enzymes involved in the backdoor pathway (SRD5A1, AKR1C2/4, CYP17A1), as measured by qPCR. These data identify androsterone as the predominant backdoor androgen in the human fetus and show that it is formed primarily in non-gonadal tissue with placental progesterone the likely substrate. Masculinization of the human fetus depends, therefore, on androgen synthesis by both the fetal testes and non-gonadal tissues leading to DHT formation at the genital tubercle. Our findings provide, for the first time, a solid basis to explain why placental insufficiency is associated with disorders of sex development in humans

A low oxygen supply to the fetus causes intrauterine growth restriction and can affect gonadal development of the offspring, having a potential impact on fertility. We investigated histology and gene expression in the postnatal rat ovary after fetal hypoxia induced by uterine artery ligation. Sprague-Dawley rats underwent uterine artery ligation at day 19 of gestation. Offspring were sacrificed at 5, 20 and 40 days post-partum. Follicles were counted and classified in hematoxylin-eosin stained sections. Gene expression of 90 genes was analyzed by TaqMan® Low Density Array. A significantly lower number of total and primordial follicles was detected in 20 days post-partum intrauterine growth restricted animals. Follicle density was not different at 40 days post-partum, suggesting that compensatory mechanisms occurred during the pre-pubertal window. Uterine artery ligation modified the expression of 24 genes involved in different cellular functions, among which proliferation, apoptosis and metabolism. Ovarian follicle pool was affected by fetal hypoxia in early life, but this effect did not persist in puberty. Genes involved in cellular processes were affected at all ages, potentially implying long-term genetic alterations. Further analyses are needed to elucidate later effects of fetal hypoxia on ovarian function and fertility.