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BACKGROUND: Bone age determination is usually employed to evaluate growth disorders and their treatment. The Greulich-Pyle method is the simplest and most frequently used type of evaluation, but it presents huge interobserver variability. The BoneXpert is a computer-automated method developed to avoid significant bone age variability among distinct observers. OBJECTIVE: To compare the BoneXpert and Greulich-Pyle methods of bone age determination in eutrophic children and adolescents, as well as in overweight and obese pediatric patients. MATERIALS AND METHODS: The sample comprised 515 participants, 253 boys (159 eutrophic, 53 overweight and 41 obese) and 262 girls (146 eutrophic, 76 overweight and 40 obese). Left hand and wrist radiographs were acquired for bone age determination using both methods. RESULTS: There was a positive correlation between chronological age and Greulich-Pyle, chronological age and BoneXpert, and Greulich-Pyle and BoneXpert. There was a significant increase (P<0.05) in bone age in both the Greulich-Pyle and BoneXpert methods in obese boys when compared to eutrophic or overweight boys of the same age. In girls, there was an increase in bone age in both obese and overweight individuals when compared to eutrophic girls (P<0.05). The Greulich-Pyle bone age was advanced in comparison to that of BoneXpert in all groups, except in obese boys, in which bone age was similarly advanced in both methods. CONCLUSION: The BoneXpert computer-automated bone age determination method showed a significant positive correlation with chronological age and Greulich-Pyle. Furthermore, the impact of being overweight or obese on bone age could be identified by both methods.
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Determinação da Idade pelo Esqueleto/métodos , Obesidade , Sobrepeso , Magreza , Adolescente , Brasil , Criança , Pré-Escolar , Estudos Transversais , Feminino , Mãos/diagnóstico por imagem , Humanos , Masculino , Estudos Prospectivos , Punho/diagnóstico por imagemRESUMO
Aim: X-linked hypophosphatemia (XLH) is the most common inherited form of rickets, and it is caused by pathogenic inactivating variants of the phosphate-regulating endopeptidase homolog X-linked (PHEX) gene. The main purpose of this study is to identify the presence of a genotype-phenotype correlation in a cohort of XLH patients. Methods: This is a retrospective study including patients diagnosed with hypophosphatemic rickets, confirmed by clinical, radiological, and laboratory findings. Medical records were reviewed for phenotypic analyses. Genomic DNA was extracted from the peripheral blood lymphocytes, and PHEX sequencing was performed by exomic NGS sequencing. The Wilcoxon rank-sum test and the two-tailed Fisher's exact test were employed for the statistical analyses of this study. Results: A total of 41 patients were included in this study, and 63.41% (26/41) of the patients were female. The mutation analyses identified 29.27% missense variants and 29.72% nonsense variants, most of them were considered deleterious (66.41%). Six novel deleterious variants in the PHEX gene were detected in seven patients. The median concentrations of pretreatment serum calcium, phosphorus, and parathyroid hormone (PTH) were not significantly different among patients with different genotypes. An orthopedic surgery due to bone deformity was required in 57.69%. Conclusions: Our analysis did not identify any specific genotype as a predictor. No significant genotype-phenotype correlation was found, suggesting that the recognition of subjacent pathogenic mutation in the PHEX gene may have limited prognostic value. Despite this finding, genetic testing may be useful for identifying affected individuals early and providing appropriate treatment.
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The human chorionic gonadotropin (hCG) stimulation test that evaluates gonadal steroidogenesis is crucial in the assessment of patients with 46,XY disorders of sex development (DSD). This study aimed to determine a testosterone (T) cutoff level that indicates an adequate testicular function using LC-MS/MS after stimulation with recombinant human chorionic gonadotropin (rhCG) in a single dose. Nineteen prepubertal children with 46,XY DSD and normal T secretion were evaluated. T and dihydrotestosterone (DHT) levels were measured by liquid chromatography technique with tandem mass spectrometry (LC-MS/MS) before and 7 days after rhCG application at 250 µg. We suggest 0.89 ng/mL as the cutoff point for T after rhCG stimulation analyzed by LC-MS/MS.
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Espectrometria de Massas em Tandem , Testosterona , Criança , Humanos , Testosterona/farmacologia , Cromatografia Líquida , Gonadotropina Coriônica/farmacologiaRESUMO
Context: Ectopic posterior pituitary (EPP), a condition in which the posterior pituitary gland is displaced due to defective neuronal migration, is frequently associated with hypopituitarism. Genetic variants play a role, but many cases remain unexplained. Objective: A large EPP cohort was studied to explore the importance of genetic variants and how they correlate with clinical findings. Methods: Whole exome sequencing was performed on a discovery sample of 27 cases to identify rare variants. The variants that met the criteria for rarity and biological relevance, or that were previously associated with EPP (ROBO1 and HESX1), were then resequenced in the 27 cases plus a replication sample of 51 cases. Results: We identified 16 different variants in 12 genes in 15 of the 78 cases (19.2%). Complete anterior pituitary deficiency was twice as common in cases with variants of interest compared to cases without variants (9/15 [60%] vs 19/63 [30.1%], respectively; Z test, Pâ =â 0.06). Breech presentation was more frequent in the variant positive group (5/15 vs 1/63; Z test, Pâ =â 0.003). Four cases had variants in ROBO1 and 1 in HESX1, genes previously associated with EPP. The ROBO1 p.S18* variant has not been reported previously; ROBO1 p.Q1227H has not been associated with EPP previously. Conclusion: EPP cases with variants of interest identified in this study were more likely to present with severe clinical disease. Several variants were identified in genes not previously associated with EPP. Our findings confirm that EPP is a multigenic disorder. Future studies are needed to identify additional genes.
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The spectrum and incidence of gene fusions in papillary thyroid carcinoma (PTC) can differ significantly depending on the age of onset, histological subtype or radiation exposure history. In sporadic pediatric PTC, RET/PTC1-3 and AGK-BRAF fusions are common genetic alterations. The role of RET/PTC as a prognostic marker in pediatric PTC is still under investigation. We recently showed that AGK-BRAF fusion is prevalent in young patients (mean 10 years) and associated with specific and aggressive pathological features such as multifocality and lung metastasis. In this pilot study, we report a unique patient harboring three different foci: the first was positive for AGK-BRAF fusion, the second was positive for just RET/PTC3 fusion and the third was negative for both rearrangements. To investigate whether AGK-BRAF and RET/PTC3 are associated with genomic instability and chromatin modifications, we performed quantitative fluorescence in situ hybridization (Q-FISH) of telomere repeats followed by 3D imaging analysis and 3D super-resolution Structured Illumination Microscopy (3D-SIM) to analyze the DNA structure from the foci. We demonstrated in this preliminary study that AGK-BRAF is likely associated with higher levels of telomere-related genomic instability and chromatin remodeling in comparison with RET/PTC3 foci. Our results suggest a progressive disruption in chromatin structure in AGK-BRAF-positive cells, which might explain a more aggressive disease outcome in patients harboring this rearrangement.
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Background The intima-media thickness of the carotid artery (cIMT) and endothelial dysfunction are associated with cardiovascular (CV) disease. Objectives To evaluate the correlation between cIMT, brachial intraluminal diameter and flow-mediated vasodilation on the reactive hyperemia phase in adolescents with obesity with predictors of CV risk. Methods Seventy-three pubertal patients with overweight or obesity were evaluated (45 girls) with a mean (standard deviation [SD]) age of 12.9 (2.5) years. Patients underwent anthropometric measurements and had the lipid profile, oral glucose tolerance test (oGTT) and serum intercellular adhesion molecule-1 (sICAM-1) levels analyzed. The ratios of the waist circumference (WC)/height (WHtR) and triglycerides (TG)/high-density lipoprotein cholesterol (HDL-C), homeostatic model assessment of insulin resistance (HOMA-IR), the Matsuda index and insulin area under the curve (AUC) were calculated. All patients were evaluated for cIMT and arterial blood flow velocity of the brachial artery. Results 75.3% of the patients had high cIMT values. We found a positive correlation between WHtR and cIMT (r = 0.233; p = 0.050). There was a positive correlation between sICAM-1 and insulin AUC (r = 0.323; p = 0.012) and WHtR (r = 0.258; p = 0.047). Patients with abnormal arterial dilation had higher sICAM-1 values (p = 0.02) despite having smaller WHtR (p = 0.046). Conclusions These adolescents with obesity had high cIMT values. Insulin resistance was associated with sICAM-1. Endothelial dysfunction was positively correlated with sICAM-1. There is no consensus about what the best laboratorial approach to evaluate insulin resistance in adolescents is, and the cutoff values of each method are arbitrary. So, as we saw earlier, the association between anthropometric data (WHtR) and ultrasound findings could be useful to evaluate the CV risk of these adolescents with obesity, because of its practical, direct and low-cost value.
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Artéria Braquial/diagnóstico por imagem , Doenças Cardiovasculares/epidemiologia , Artérias Carótidas/diagnóstico por imagem , Espessura Intima-Media Carotídea , Obesidade/diagnóstico por imagem , Sobrepeso/diagnóstico por imagem , Adolescente , Brasil , Estudos Transversais , Endotélio/fisiopatologia , Feminino , Teste de Tolerância a Glucose , Humanos , Resistência à Insulina , Molécula 1 de Adesão Intercelular/sangue , Lipídeos/sangue , Masculino , Fatores de Risco , Circunferência da CinturaRESUMO
Pituitary developmental defects lead to partial or complete hormone deficiency and significant health problems. The majority of cases are sporadic and of unknown cause. We screened 28 patients with pituitary stalk interruption syndrome (PSIS) for mutations in the FAT/DCHS family of protocadherins that have high functional redundancy. We identified seven variants, four of which putatively damaging, in FAT2 and DCHS2 in six patients with pituitary developmental defects recruited through a cohort of patients with mostly ectopic posterior pituitary gland and/or pituitary stalk interruption. All patients had growth hormone deficiency and two presented with multiple hormone deficiencies and small glands. FAT2 and DCHS2 were strongly expressed in the mesenchyme surrounding the normal developing human pituitary. We analyzed Dchs2-/- mouse mutants and identified anterior pituitary hypoplasia and partially penetrant infundibular defects. Overlapping infundibular abnormalities and distinct anterior pituitary morphogenesis defects were observed in Fat4-/- and Dchs1-/- mouse mutants but all animal models displayed normal commitment to the anterior pituitary cell type. Together our data implicate FAT/DCHS protocadherins in normal hypothalamic-pituitary development and identify FAT2 and DCHS2 as candidates underlying pituitary gland developmental defects such as ectopic pituitary gland and/or pituitary stalk interruption.
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Proteínas Relacionadas a Caderinas/genética , Caderinas/genética , Doenças da Hipófise/genética , Adolescente , Animais , Proteínas Relacionadas a Caderinas/metabolismo , Caderinas/metabolismo , Feminino , Humanos , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mutação , Hipófise/crescimento & desenvolvimento , Hipófise/metabolismo , Adulto JovemRESUMO
BACKGROUND: Women with 21-hydroxylase deficiency present much variability in external genitalia virilization, even among those with similar impairments of 21-hydroxylase (21OH) activity. OBJECTIVE: To evaluate if the number of CAG (nCAG) repeats of the androgen receptor gene influences the degree of external genitalia virilization in women with CYP21A2 mutations, grouped according to impairment of 21OH activity. PATIENTS: The nCAG was determined in 106 congenital adrenal hyperplasia (CAH) patients and in 302 controls. The patients were divided, according to their CYP21A2 genotypes, into Groups A and B, which confer total and severe impairment of 21OH activity, respectively. METHODS: The inactivation pattern of the X-chromosome was studied through genomic DNA digestion with Hpa II. The CAG repeat region was amplified by polymerase chain reaction (PCR) and analysed by GeneScan. RESULTS: The nCAG and the frequency of severe skewed X-inactivation did not differ between normal women and patients. The nCAG median in genotype A was 20.7 (IQR 2.3) for Prader I + II, 22.5 (3.6) for Prader III and 21 (2.9) for Prader IV + V (P < 0.05 for Prader III and Prader IV + V). The nCAG median in genotype B was 21.3 (1.1) for Prader I + II, 20.5 (2.9) for Prader III and 22 (2.8) for Prader IV + V (P > 0.05). A significant difference was found regarding the nCAG median in patients presenting Prader III from genotypes A and B. CONCLUSIONS: We observed great variability in the degree of external genitalia virilization in both CYP21A2 genotypes, and we showed that the CAG repeats of the androgen receptor gene influences this phenotypic variability.
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Hiperplasia Suprarrenal Congênita/genética , Receptores Androgênicos/genética , Repetições de Trinucleotídeos/genética , Virilismo/genética , Feminino , Genótipo , Humanos , Masculino , Esteroide 21-Hidroxilase/genética , Esteroide 21-Hidroxilase/metabolismoRESUMO
Hydrocortisone acetate is usually employed in the treatment of classic congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency. In Brazil, however, oral hydrocortisone acetate is only available from manipulation pharmacies. Prednisolone has stable oral pharmaceutical formulations commercially available, with the advantage of a single daily dose. The aim of this study was to compare the efficacy of oral prednisolone and oral hydrocortisone in the treatment of CAH due to 21-hydroxylase deficiency. Fifteen patients with mean (SD) chronological age of 7.2 (3.6) years, were evaluated in two consecutive 1-year periods. In the first year, hydrocortisone (17.5 mg/m2/day, divided in three doses) was used in the treatment, followed by the use of prednisolone (3 mg/m2/day, once in the morning) in the second year. The comparison between the two treatments was assessed after a one-year treatment period by: variation of height standard deviation score (SDS) (delta Height SDS), variation of height SDS according to bone age (delta BA SDS), variation of body mass SDS (delta BMI SDS) and serum levels of androstenedione. No significant difference was observed in relation to the delta Height SDS, delta BA SDS and delta BMI SDS. No significant difference was observed in the serum levels of androstenedione. We conclude that the efficacy of prednisolone administered once a day orally is comparable to the oral use of hydrocortisone three times a day. Oral prednisolone may be an option for patients with CAH due to 21-hydroxylase deficiency.
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Hiperplasia Suprarrenal Congênita/tratamento farmacológico , Anti-Inflamatórios/uso terapêutico , Glucocorticoides/uso terapêutico , Prednisolona/uso terapêutico , Esteroide 21-Hidroxilase/metabolismo , Adolescente , Anti-Inflamatórios/administração & dosagem , Estatura , Índice de Massa Corporal , Criança , Pré-Escolar , Feminino , Glucocorticoides/administração & dosagem , Humanos , Lactente , Masculino , Prednisolona/administração & dosagemRESUMO
Aromatase is a cytochrome P450 enzyme (CYP19A1 isoform) able to catalyze the conversion of androgens to estrogens. The aromatase gene mutations highlighted the action of estrogen as one of the main regulators of bone maturation and closure of bone plate. The use of aromatase inhibitors (AI) in boys with short stature has showed its capability to improve the predicted final height. Anastrozole (ANZ) and letrozole (LTZ) are nonsteroidal inhibitors able to bind reversibly to the heme group of cytochrome P450. In this review, we describe the pharmacokinetic profile of both drugs, discussing possible drug interactions between ANZ and LTZ with other drugs. AIs are triazolic compounds that can induce or suppress cytochrome P450 enzymes, interfering with metabolism of other compounds. Hydroxilation, N-dealkylation and glucoronidation are involved in the metabolism of AIs. Drug interactions can occur with azole antifungals, such as ketoconazole, by inhibiting CYP3A4 and by reducing the clearance of AIs. Antiepileptic drugs (lamotrigine, phenobarbital, and phenytoin) also inhibit aromatase. Concomitant use of phenobarbital or valproate has a synergistic effect on aromatase inhibition. Therefore, it is important to understand the pharmacokinetics of AIs, recognizing and avoiding possible drug interactions and offering a safer prescription profile of this class of aromatase inhibitors. Arch Endocrinol Metab. 2017;61(3):391-7.
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Inibidores da Aromatase/farmacocinética , Estatura/efeitos dos fármacos , Nitrilas/farmacocinética , Triazóis/farmacocinética , Anastrozol , Inibidores da Aromatase/uso terapêutico , Sistema Enzimático do Citocromo P-450/metabolismo , Interações Medicamentosas , Feminino , Humanos , Letrozol , MasculinoRESUMO
Thyroid cancer is the fastest increasing cancer worldwide in all age groups. Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer in both adults and children. PTC genomic landscape has been extensively studied in adults, but information regarding sporadic pediatric patients is lacking. Although BRAF V600E mutation is highly prevalent in adults, this mutation is uncommon in pediatric cases. As adult and pediatric PTC is a mitogen-activated protein kinase-driven cancer, this altered pathway might be activated by different genetic events. The aim of this study was to investigate the occurrence of AGK-BRAF fusion gene, recently described in radiation-exposed pediatric PTC, in a cohort of exclusively sporadic pediatric PTC. The series consisted of 30 pediatric PTC younger than 18 years of age at the time of diagnosis and 15 matched lymph node metastases (LNM). Primary tumors and matched LNM were screened for the presence of the AGK-BRAF fusion transcript by RT-PCR. To confirm the identity of the amplified products, randomly selected samples positive for the presence of the fusion transcripts were sequenced. Moreover, BRAF dual-color, break-apart probes confirmed BRAF rearrangement. Overall, the AGK-BRAF fusion gene was detected in 10% (3/30) of primary tumors. For one of these cases, paired LNM was also available, which also shows the presence of AGK-BRAF fusion gene. This study described, for the first time, the presence of AGK-BRAF in sporadic pediatric PTC. Understanding the molecular events underlying pediatric PTC may improve preoperative diagnosis, allow molecular prognostication and define a therapeutic approach toward sporadic PTC patients.
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Proteínas Semelhantes a Angiopoietina/genética , Proteínas de Fusão Oncogênica/genética , Proteínas Proto-Oncogênicas B-raf/genética , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/patologia , Adolescente , Proteína 6 Semelhante a Angiopoietina , Carcinoma Papilar/genética , Carcinoma Papilar/patologia , Criança , Pré-Escolar , Feminino , Humanos , Hibridização in Situ Fluorescente , Metástase Linfática , Masculino , Mutação , Metástase Neoplásica , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Câncer Papilífero da Tireoide , Translocação Genética , Carga TumoralRESUMO
BACKGROUND: In 21-hydroxylase deficiency (21-OHD), there is an influence of genotype on the severity of external genitalia virilization. However, females carrying mutations predicting a similar impairment of enzymatic activity present a wide variability of genital phenotypes. In such cases, interindividual variability in genes related to the sex steroid hormone pathway could play a role. OBJECTIVE: To evaluate the influence of POR, HSD17B5 and SRD5A2 variants on the severity of external genitalia virilization in 21-OHD females. DESIGN AND PATIENTS: Prader stages were evaluated in 178 females with 21-OHD from a multicenter study. The 21-OHD genotypes were divided into two groups according to their severity: severe and moderate. The influences of the POR p.A503V, HSD17B5 c.-71A>G, HSD17B5 c.-210A>C, and SRD5A2 p.A49T and p.V89L variants on the degree of external genitalia virilization were analyzed. RESULTS: The POR p.A503V, HSD17B5 c.-71A>G, HSD17B5 c.-210A>C, and SRD5A2 p.A49T and p.V89L variants were found in 25, 33, 17, 1, and 31% of the alleles, respectively. In uni- and multilinear regression, HSD17B5 c.-210A>C has a significant influence on the degree of external genitalia virilization. This variant was also identified with a higher frequency in the most severely virilized females. CONCLUSION: We demonstrated that a variant in the promoter region of HSD17B5 related to fetal androgen synthesis influences the genital phenotype in 21-OHD females.
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3-Hidroxiesteroide Desidrogenases/genética , Hiperplasia Suprarrenal Congênita/genética , Alelos , Hidroxiprostaglandina Desidrogenases/genética , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas , Virilismo/genética , 3-Oxo-5-alfa-Esteroide 4-Desidrogenase/genética , Hiperplasia Suprarrenal Congênita/patologia , Membro C3 da Família 1 de alfa-Ceto Redutase , Feminino , Humanos , Proteínas de Membrana/genética , Estudos Retrospectivos , Virilismo/patologiaRESUMO
Relative antiinflammatory and immunosuppressive potencies of glucocorticoids (GC) were previously well defined. Nonetheless, GC also regulate cell proliferation and programmed death (apoptosis). The aim of this study was to determine the relative potency of different GC on the modulation of cell survival. The GC-sensitive lymphoblast cell line CEM-c7/14 was submitted to 48 h-exposure to GC (dose-response curve from 10(-8) to 10(-5) M). Cell survival was analyzed employing the DimethylTiazol-Tetrazolium (MTT) test. For each GC at least 4 experiments were performed in quadruplicate. Responses to different GC at the same molarity were analyzed by ANOVA on Ranks. Cell responses to the same GC in different concentrations were tested by repeated measures ANOVA. The EC50 for each GC was calculated with the GraphPad Prism 3.0 software. The use of low concentrations (10(-8) and 10(-7) M) of hydrocortisone and methylprednisolone determined a similar effects on cell survival, which was less prominent than that observed with betamethasone, budesonide or momethasone. Momethasone was the most potent GC, inducing the most intense dexamethasone reduction on cell survival at the lowest concentration (10(-8) M). Momethasone and methylprednisolone were the two GC with the strongest impact on cell survival. Our findings suggest that antiproliferative and apoptotic potencies of GC are different from those previously reported antiinflammatory and immunosuppressive actions.
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Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Glucocorticoides/farmacologia , Imunossupressores/farmacologia , Análise de Variância , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica , HumanosRESUMO
OBJECTIVES: To describe population reference values for shoes size, and to identify possible disproportional foot growth during GH therapy. MATERIALS AND METHODS: Construction of percentile chart based on 3,651 controls (male: 1,838; female: 1,813). The GH treated group included 13 children with idiopathic short stature (ISS) and 50 children with normal height, but with height prediction below their target height; male: 26 and female: 37 mean ± SD age 13.3 ± 1.9 and 12.9 ± 1.5 years, respectively. GH (0.05 mg/kg/day) was used for 3.2 ± 1.6 years, ranging from 1.0-10.3 years. Height expressed as SDS, target height (TH) SDS, self-reported shoes size and target shoes size (TSS) SDS were recorded. RESULTS: Reference values were established showed as a foot SDS calculator available online at www.clinicalcaselearning.com/v2. Definitive shoes size was attained in controls at mean age of 13y in girls and 14y in boys (average values 37 and 40, respectively). In the study group, shoes size was -0.15 ± 0.9 and -0.02 ± 1.3 SDS, with target feet of 0.08 ± 0.8 and -0.27 ± 0.7 SDS in males and females, respectively. There was a significant positive correlation between shoes size and familial TSS, between shoes size and height and between TSS and TH. There was no correlation between duration of GH treatment and shoes size. Our data suggest that during long-term treatment with GH, patients maintain proportional growth in shoes size and height, and the expected correlation with the familial target. CONCLUSIONS: We conclude that there is no excessive increase in the size of foot as estimated by the size of shoes in individuals under long term GH therapy.
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Pé/crescimento & desenvolvimento , Transtornos do Crescimento/tratamento farmacológico , Hormônio do Crescimento Humano/uso terapêutico , Sapatos/estatística & dados numéricos , Adolescente , Adulto , Estatura/efeitos dos fármacos , Criança , Pré-Escolar , Estudos Transversais , Feminino , Pé/anatomia & histologia , Hormônio do Crescimento Humano/farmacologia , Humanos , Masculino , Tamanho do Órgão/efeitos dos fármacos , Valores de Referência , Adulto JovemRESUMO
P450c17 deficiency is an autosomal recessive disorder and a rare cause of congenital adrenal hyperplasia characterized by hypertension, hypokalemia, and impaired production of sex hormones. We performed a clinical, hormonal, and molecular study of 11 patients from 6 Brazilian families with the combined 17alpha-hydroxylase/17,20-lyase deficiency phenotype. All patients had elevated basal serum levels of progesterone (1.8-38 ng/ml; 0.57-12 pmol/liter) and suppressed plasma renin activity. CYP17 genotyping identified 5 missense mutations. The compound heterozygous mutation R362C/W406R was found in 1 family, whereas the homozygous mutations R96W, Y329D, and P428L were seen in the other 5 families. The R96W mutation has been described as the cause of p450c17 deficiency in Caucasian patients. The other mutations were not found in 50 normal subjects screened by allele-specific oligonucleotide hybridization (Y329D, R362C, and W406R) or digestion with HphI (P428L) and were recently found in other Brazilian patients. Therefore, we elucidated the genotype of 11 individuals with p450c17 deficiency and concluded that basal progesterone measurement is a useful marker of p450c17 deficiency and that its use should reduce the misdiagnosis of this deficiency in patients presenting with male pseudohermaphroditism, primary or secondary amenorrhea, and mineralocorticoid excess syndrome.
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Hiperplasia Suprarrenal Congênita/diagnóstico , Hiperplasia Suprarrenal Congênita/genética , Progesterona/sangue , Esteroide 17-alfa-Hidroxilase/genética , Adolescente , Hiperplasia Suprarrenal Congênita/sangue , Hiperplasia Suprarrenal Congênita/complicações , Adulto , Sequência de Bases/genética , Brasil , Estudos de Casos e Controles , Criança , Pré-Escolar , Diagnóstico Diferencial , Transtornos do Desenvolvimento Sexual/etiologia , Feminino , Genótipo , Heterozigoto , Homozigoto , Hormônios/sangue , Humanos , Masculino , Mutação de Sentido IncorretoRESUMO
BACKGROUND: The expression of glucocorticoid-receptor (GR) seems to be a key mechanism in the regulation of glucocorticoid (GC) sensitivity and is potentially involved in cases of GC resistance or hypersensitivity. The aim of this study is to describe a method for quantitation of GR alpha isoform (GRalpha) expression using real-time PCR (qrt-PCR) with analytical capabilities to monitor patients, offering standard-curve reproducibility as well as intra- and inter-assay precision. RESULTS: Standard-curves were constructed by employing standardized Jurkat cell culture procedures, both for GRalpha and BCR (breakpoint cluster region), as a normalizing gene. We evaluated standard-curves using five different sets of cell culture passages, RNA extraction, reverse transcription, and qrt-PCR quantification. Intra-assay precision was evaluated using 12 replicates of each gene, for 2 patients, in a single experiment. Inter-assay precision was evaluated on 8 experiments, using duplicate tests of each gene for two patients. Standard-curves were reproducible, with CV (coefficient of variation) of less than 11%, and Pearson correlation coefficients above 0,990 for most comparisons. Intra-assay and inter-assay were 2% and 7%, respectively. CONCLUSION: This is the first method for quantitation of GRalpha expression with technical characteristics that permit patient monitoring, in a fast, simple and robust way.
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Reação em Cadeia da Polimerase/métodos , Receptores de Glucocorticoides/genética , Adulto , Criança , Feminino , Humanos , Células Jurkat , Masculino , Reação em Cadeia da Polimerase/normas , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Receptores de Glucocorticoides/biossíntese , Padrões de Referência , Reprodutibilidade dos TestesRESUMO
Routine magnetic resonance imaging (MRI) is an established standard method to investigate the etiology of pituitary insufficiency. Among the anatomic abnormalities usually observed, ectopic hyperintense signal on T1 sequence is the most frequently associated with pituitary dysfunction. We developed a new protocol (FAST1-MRI) which is able to detect anatomic hypothalamic-pituitary abnormalities with 100% concordance when compared to the routine MRI protocol. FAST1-MRI takes only 3.25 minutes, and is performed without contrast, sedation or anesthesia. We studied 17 controls and 31 patients with growth hormone (GH) deficiency (18/31 with abnormal MRI). Patients with ectopic hyperintense signal were shorter in height, had lower IGF-I and IGFBP-3 levels, and reduced GH response after clonidine. In conclusion, we describe a new simplified MRI protocol that we propose should be used in the diagnosis of GH deficiency.
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Transtornos do Crescimento/etiologia , Hormônio do Crescimento Humano/deficiência , Sistema Hipotálamo-Hipofisário/patologia , Imageamento por Ressonância Magnética/métodos , Doenças da Hipófise/diagnóstico , Adolescente , Criança , Feminino , Hormônio do Crescimento Humano/sangue , Humanos , Masculino , Doenças da Hipófise/complicações , Valores de ReferênciaRESUMO
The effects of glucocorticoids (GC) are mediated by the activation of specific receptors that can be quantified in vitro by several laboratory tests. In vivo, other tests to determine GC sensitivity have been described, but only employing pharmacological doses. In this study, we used a very low dose of dexamethasone, an in vivo model to assess individual GC sensitivity. Fifty-five obese children and adolescents and 17 controls were studied. The patients were submitted to four 12-h urine collections, starting at 22:00 h; dexamethasone was administered orally at the end of the second urine sample. Patients were divided in the following groups: group Ob75 (n = 29) and the control group (n = 17) received dexamethasone 75 microg/m2, and group Ob150 (n = 26) received dexamethasone 150 microg/m2. Urinary cortisol was determined by RIA and expressed as microg/m2/12 h. All patients and controls showed a circadian rhythm before GC, which was maintained after dexamethasone only in controls. In the obese patients the circadian rhythm was abolished following both doses of dexamethasone, but more prominently with the dose of 150 microg/m2. In the obese group given 75 microg/m2, urinary cortisol inhibition was only observed in the first 12 h after dexamethasone, suggesting a partial and shorter suppression of the hypothalamic-pituitary axis. In both control and obese patients, the very low dose of dexamethasone was able to create a gradient of cortisol suppression that could be useful to identify an individual's sensitivity to glucocorticoids.
Assuntos
Dexametasona , Relação Dose-Resposta a Droga , Resistência a Medicamentos/efeitos dos fármacos , Glucocorticoides/antagonistas & inibidores , Glucocorticoides/farmacologia , Obesidade/fisiopatologia , Administração Oral , Adolescente , Criança , Ritmo Circadiano/efeitos dos fármacos , Ritmo Circadiano/fisiologia , Dexametasona/administração & dosagem , Técnicas de Diagnóstico Endócrino , Resistência a Medicamentos/fisiologia , Retroalimentação Fisiológica/efeitos dos fármacos , Feminino , Glucocorticoides/metabolismo , Humanos , Hidrocortisona/antagonistas & inibidores , Hidrocortisona/urina , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/fisiologia , Masculino , Métodos , Obesidade/urinaRESUMO
Southwick's angles measured in the anteroposterior and Lauenstein frog-leg views have been used for planning surgical treatment of deformities caused by slipped capital femoral epiphysis (SCFE). Clinically established epiphysiolysis is associated with a decrease in anteroposterior angles and an increase in Lauenstein angles. Obesity is one of the factors involved in the development of SCFE, but its influence in Southwick's angles has not been previously described. Additionally, few reports describe normal values adjusted for sex and bone age. In this study, we determined Southwick's angles in normal control participants (n=70), in patients with obesity (n=31) and in patients with epiphysiolysis (n=33). The intra-observer variations were 1.7 and 23.3% for the anteroposterior and Lauenstein angles, respectively. No significant differences were detected in the control group when these individuals were compared according to sex or chronological age. Obese patients showed a significant increase in the anterposterior angle when compared with control individuals. The hip with epiphysiolysis presented a significant reduction in the anteroposterior angle and an increase in the Lauenstein angle, when the same was compared with the contralateral hip or with the control values. In patients with unilateral SCFE the Lauenstein angle of the normal hip was detected as increased, which was possibly related to the obesity associated with decreased insulin resistance presented by these patients. We conclude that the detection of risk for developing SCFE should include an expanded clinical and radiological profile characterized by pubertal males who present obesity associated with insulin resistance and an increase in Southwick's anteroposterior angle.