Our data suggest that in this group of premature infants the circadian maturation of the hypothalamic-pituitary-adrenal axis occurred at the same postnatal age as reported for term infants and that there was a parallelism between the appearance of such rhythm and the onset of sleep rhythm.

Objective: To determine differences in dietary calcium and vitamin D intakes between female never, former and current smokers. Design: Population-based survey using a validated food frequency questionnaire. Setting: The Bus Sante  2000, epidemiologic observatory of Geneva, Switzerland. Subjects: 2319 women resident in Geneva, Switzerland between 1993 and 1997. Results: Daily calcium intake was 798 mg among current heavy smokers ( 20 cigarettesaday), 882 mg among current moderate smokers (1 ± 19 cigarettesaday) and 945 mg among never smokers (P 0.0001). There was a difference of almost 50 mgaday in median calcium intake from cheese (P 0.04), which corresponded to about one-third of the total difference in calcium intake between heavy smokers and never smokers. Of the current heavy smokers, 21% did not eat yogurt compared to 10% of never smokers (P 0.001). Among yogurt eaters, heavy smokers consumed 90 mgaday of calcium from yogurt vs 115 mgaday in never smokers (P 0.003). Smokers ate more butter and cream (P 0.02) or milk (P 0.06) than never smokers, but these were minor sources of calcium. Fish was the main cause of higher intake of vitamin D in never smokers (0.81 mgaday) compared with heavy smokers (0.53 mgaday) and moderate smokers (0.70 mgaday). The diet of ex-smokers after 5 y or more of smoking cessation tended to resemble that of never smokers for calcium (about 900 mgaday) and vitamin D (about 2.55 mgaday). Conclusion: Female current smokers have lower dietary intakes of calcium and vitamin D than never smokers. Sponsorship: This study was funded by the Swiss National Fund for Scienti®c Research (grants 32.31.326.91,).

Using the proposed algorithm, all alleles were elucidated. False-positive results in MLPA occurred when mutations or polymorphisms were located close to the probe-binding regions. These difficulties were overcome by the association of MLPA with ASO-PCR and paternal segregation. Using these approaches, we can successfully use MLPA in a cost-effective laboratory routine for the molecular diagnosis of CAH-21OHD.

OBJECTIVES:Idiopathic central precocious puberty and its postponement with a (gonadotropin-releasing hormone) GnRH agonist are complex conditions, the final effects of which on bone mass are difficult to define. We evaluated bone mass, body composition, and bone remodeling in two groups of girls with idiopathic central precocious puberty, namely one group that was assessed at diagnosis and a second group that was assessed three years after GnRH agonist treatment.METHODS:The precocious puberty diagnosis and precocious puberty treatment groups consisted of 12 girls matched for age and weight to corresponding control groups of 12 (CD) and 14 (CT) girls, respectively. Bone mineral density and body composition were assessed by dual X-ray absorptiometry. Lumbar spine bone mineral density was estimated after correction for bone age and the mathematical calculation of volumetric bone mineral density. CONEP: CAAE-0311.0.004.000-06.RESULTS:Lumbar spine bone mineral density was slightly increased in individuals diagnosed with precocious puberty compared with controls; however, after correction for bone age, this tendency disappeared (CD = -0.74±0.9 vs. precocious puberty diagnosis = -1.73±1.2). The bone mineral density values of girls in the precocious puberty treatment group did not differ from those observed in the CT group.CONCLUSION:There is an increase in bone mineral density in girls diagnosed with idiopathic central precocious puberty. Our data indicate that the increase in bone mineral density in girls with idiopathic central precocious puberty is insufficient to compensate for the marked advancement in bone age observed at diagnosis. GnRH agonist treatment seems to have no detrimental effect on bone mineral density.

Deregulation of the IGF system observed in human tumors indicates a role in malignant cell transformation and in tumor cell proliferation. Although overexpression of the IGF2 and IGF1R genes was described in adrenocortical tumors (ACTs), few studies reported their profiles in pediatric ACTs. In this study, the IGF2 and IGF1R expression was evaluated by RT-qPCR according to the patient's clinical/pathological features in 60 pediatric ACT samples, and IGF1R protein was investigated in 45 samples by immunohistochemistry (IHC). Whole transcriptome and functional assays were conducted after IGF1R inhibition with OSI-906 in NCI-H295A cell line. Significant IGF2 overexpression was found in tumor samples when compared with non-neoplastic samples (P < 0.001), significantly higher levels of IGF1R in patients with relapse/metastasis (P = 0.031) and moderate/strong IGF1R immunostaining in 62.2% of ACTs, but no other relationship with patient survival and clinical/pathological features was observed. OSI-906 treatment downregulated genes associated with MAPK activity, induced limited reduction of cell viability and increased the apoptosis rate. After 24 h, the treatment also decreased the expression of genes related to the steroid biosynthetic process, the protein levels of the steroidogenic acute regulatory protein (STAR), and androgen secretion in cell medium, supporting the role of IGF1R in steroidogenesis of adrenocortical carcinoma cells. Our data showed that the IGF1R overexpression could be indicative of aggressive ACTs in children. However, in vitro treatments with high concentrations of OSI-906 (>1 μM) showed limited reduction of cell viability, suggesting that OSI-906 alone could not be a suitable therapy to abolish carcinoma cell growth. 23:6Research R C P Lira et al.IGF1R in pediatric adrenocortical tumor IGF2 and IGF1R in pediatric adrenocortical tumors: roles in metastasis and steroidogenesis

Background: Loss-of-function mutations in the imprinted gene MKRN3 represent the most common known genetic defects associated with central precocious puberty (CPP). Methods: We report the first case of a girl carrying an MKRN3 mutation detected in childhood and followed until the development of pubertal signs. Results: The girl was screened at the age of 4 years because of a positive family history; her sister had developed CPP at 6 years of age and was found to harbor the MKRN3 p.Pro161Argfs*16 mutation, inherited from their asymptomatic father. During close follow-up, she initially developed increased growth velocity at 6 years (9 cm/year), followed by a slightly increased basal luteinizing hormone level (0.4 mIU/ml) and, ultimately, clinical thelarche with rapid progression (Tanner stage 1-3) between 6.3 and 6.7 years. In the context of a loss-of-function MKRN3 mutation and a positive family history, these features established the diagnosis of CPP and supported the initiation of treatment with a gonadotropin-releasing hormone analog. The absence of significant bone age advancement, pubic or axillary hair, or behavioral or social problems could be ascribed to the early diagnosis. Conclusion: The identification of carriers of MKRN3 mutations may contribute to early diagnosis of CPP, facilitating treatment decisions and guiding genetic counseling and prompt intervention in familial cases.

Purpose To explore pituitary tumors by methylome and transcriptome signatures in a heterogeneous ethnic population. Design Retrospective cross-sectional study. Patients and Methods Clinicopathological features, methylome and transcriptome were evaluated in pituitary tumors from 77 patients (61% women, age: 12-72 years) followed due to functioning (FPT: GH-secreting n = 18, ACTH-secreting n = 14) and non-functioning pituitary tumors (NFPT, n = 45) at Ribeirao Preto Medical School, University of Sao Paulo. Results Unsupervised hierarchical clustering analysis (UHCA) of methylome (n = 77) and transcriptome (n = 65 out of 77) revealed three clusters each: one enriched by FPT, other by NFPT, and another by ACTH-secreting and NFPT. Comparison between each omics-derived clusters identified 3,568 and 5,994 differentially methylated and expressed genes, respectively, which were associated with each other, with tumor clinical presentation, and with 2017 and 2022 WHO classifications. UHCA considering 11 transcripts related to pituitary development/differentiation also supported three clusters: POU1F1-driven somatotroph, TBX19-driven corticotroph, and NR5A1-driven gonadotroph adenomas, with rare exceptions (NR5A1 expressed in few GH-secreting and corticotroph-silent adenomas; POU1F1 in few ACTH-secreting adenomas; and TBX19 in few NFPTs). Conclusions This large heterogenic ethnic Brazilian cohort confirms that integrated methylome and transcriptome signatures classify FPT and NFPT, which are associated with clinical presentation and tumor invasiveness. Moreover, the cluster NFPT/ACTH-secreting adenomas raises interest regarding tumor heterogeneity, supporting the challenge raised by the 2017 and 2022 WHO definition regarding the discrepancy, in rare cases, between clinical presentation and pituitary lineage markers. Finally, making our data publicly available enables further studies to validate genes/pathways involved in pituitary tumor pathogenesis and prognosis.

Objective Insulin sensitivity evaluation by hyperinsulinemic-euglycaemic clamp in non-classical congenital adrenal hyperplasia (NC-CAH) due to 21-hydroxilase deficiency. Design and Setting Cross-sectional study at University Hospital outpatient clinics. Patients and Methods NC-CAH patients (25F/5M; 24 ± 10 years) subdivided into C/NC (compound heterozygous for one classical and one non-classical allele) and NC/NC (two non-classical alleles) genotypes were compared to controls. Results At diagnosis, C/NC patients presented higher basal and ACTH-stimulated 17OHP and androstenedione levels than NC/NC genotype. Patients and controls presented similar weight, BMI, abdominal circumference, and total fat body mass. NC-CAH patients showed higher waist-to-hip ratio, lower adiponectin and HDL cholesterol levels with no changes in fasting plasma glucose, HbA1c, HOMA-IR, adiponectin, leptin, IL-6, TNF-alfa, CRP, and carotid-IMT. All patients had used glucocorticoid (mean time of 73 months). Among the twenty-two patients with successful clamp, thirteen were still receiving glucocorticoid - three patients using cortisone acetate, nine dexamethasone, and one prednisone (hydrocortisone equivalent dose of 5.5 mg/m²/day), while nine patients were off glucocorticoid but had previously used (hydrocortisone equivalent dose of 5.9 mg/m 2/day). The NC-CAH patients presented lower Mffm than controls (31±20 vs 55±23 µmol.min -1.kg -1, p=0.002). The Mffm values were inversely correlated with the duration of glucocorticoid treatment (r=-0.44, p=0.04). There was association of insulin resistance and glucocorticoid type but not with androgen levels. Conclusion Using the gold standard method, the hyperinsulinemic-euglycemic clamp, insulin resistance was present in NC-CAH patients and related to prolonged use and long-acting glucocorticoid treatment. Glucocorticoid replacement and cardiometabolic risks should be monitored regularly in NC-CAH.