Prepuberal Patients Research Articles

P-803 DNA-FISH analysis in testicular tissue cells of prepuberal patients with Klinefelter Syndrome

Abstract Study question Can we accurately assess sex-chromosomes number in testicular cells (germ and somatic) of Klinefelter Syndrome (KS) prepuberal patients by DNA-FISH in paraffin embedded sections? Summary answer We have evaluated the sex-chromosomes content of testicular cells in KS prepuberal patients and the presence of XY lines evidenced a gonadal mosaicism status. What is known already Prepuberal boys with Klinefelter Syndrome usually have fertility problems in adulthood, mainly azoospermia. Due to the symptoms cause by the disease, Spermatogonial Stem Cells (SSCs) are less abundant and spermatogenesis does not occur. These patients do not have any alternative to restore the fertility in the future. When a boy has both XY and XXY cell lines, this patient is mosaic (46,XY/47,XXY). Meaning some cells could undergo spermatogenesis and generate gametes with normal sex-chromosomes number. The aim of this work is to test if KS patients diagnosed as pure can have testicular cell lines with XY chromosome number (mosaic). Study design, size, duration We obtained samples from human prepuberal patients diagnosed with pure Klinefelter Syndrome (47,XXY) for the last 5 years. Testicular biopsy fragments are fixed for histological studies and other fragments are cryopreserved. Some fragments are used for immunofluorescence and subsequently processed by DNA-FISH to determine the sex-chromosomes content of testicular cells. Participants/materials, setting, methods In this study, we used 10 prepuberal patients with KS and 5 prepuberal patients with other fertility problems with normal chromosome set as controls. We performed immunofluorescence to determine expression of germ cells (VASA) and SSCs (MAGEA4) markers and somatic cells markers such as Leydig cells (StAR) and Sertoli cells (SOX9). Afterwards, we perform DNA-FISH, with probes specific for chromosomes X and Y and chromosome 18 as a control. Main results and the role of chance The methodology used allows cytogenetic characterization of testicular tissue in paraffin embedded sections. Testicular mosaicism has been observed in all patients diagnosed as pure KS. We have observed a degree of mosaicism of 66-80% in SSCs, of 20-50% in Sertoli cells and of 30-50% in Leydig cells. We pursued a protocol with a good FISH efficiency that allows colocalization of previous immunocharacterized testicular cells. Limitations, reasons for caution The DNA-FISH technique is made just in a single paraffin section of each KS patient testicular sample. Due to the limited and difficulty to obtain KS testicular tissue, we use a limited number of samples. Wider implications of the findings We succeed to demonstrate the mosaicism of testicular cells in prepuberal patients that are diagnosed with KS. Thus, the better understanding of the SSCs with normal chromosome set (XY) could be useful for future in vitro expansion and stem cell therapies. Trial registration number not applicable

Impact of Puberty in Pediatric Migraine: A Pilot Prospective Study.

Background and PurposeThe short-term evolution of pediatric migraine remains unclear. We aimed to describe the evolution of migraine before and after puberty and its relationship with lifestyle habits.MethodsWe prospectively selected prepuberal patients from a neuropediatric unit who had a migraine diagnosis. Their medical history, migraine characteristics and impact, and lifestyle habits were recorded at the baseline visit. After 2 years we performed a telephone follow-up assessment.ResultsNineteen patients were recruited (age 10.2±2.9 years, mean±SD; 57.9% female), of whom 27.5% had migraine with aura. The accompanying symptoms had changed at the follow-up, with significantly higher prevalence rates of dizziness (44.4% vs. 88.9%), vertigo (11.1% vs. 66.7%), mood changes (38.9% vs 83.3%), confusion (5.6% vs. 77.8%), and allodynia (27.8% vs. 61.1%). Sleep disturbances (5.6% vs. 38.9%) and schedule changes (0% vs. 38.9%) increased significantly as triggers. Prodromal symptoms became more prevalent (16.7% vs. 50%), with a higher proportion of sleep disturbances reported (50.0% vs. 87.5%).ConclusionsProdromal symptoms increase in pediatric migraine after 2 years, and some trigger factors for migraine become more prevalent, including sleep disturbances. New accompanying symptoms are also identified. These changes provide information about how migraine changes during puberty along with physical and lifestyle changes, and represent a dynamic physiopathological process that deserves more research.

Neurofibromatosis de tipo 2 con inicio en la edad pediátrica: identificación de los primeros signos y síntomas

Neurofibromatosis type 2 (NF2) is a dominantly inherited neuroectodermal syndrome that predispose to the development of tumors of the central and peripheral nervous system. Additional features include eye and skin abnormalities. A 12-year old male with diagnosis of MF2 according to Baser et al and presentation in childhood was included. A comprehensive bibliographic review of evolution of the diagnostic criteria for NF2 in children was performed. The pattern of presentation of NF2 in childhood differs from adulthood in many aspects. Ophthalmologic and skin manifestations, and not an auditory dysfunction, are the most common initial symptoms in prepuberal-onset NF2. The most frequent symptoms and signs at presentation are posterior subcapsular cataract, skin manifestations as NF2 plaques and/or peripheral nerve tumors, and neurological dysfunction related to isolated or multiple cranial nerve deficits (other than nerve VIII), brainstem masses or spinal masses. As sensitivity of diagnostic criteria in children is low, those prepuberal patients with congenital or early-onset cataracts and typical skin manifestations of NF2 should be systematically assessed.

Bone Metabolism and Endocrine Function in Children with Chronic Myeloid Leukemia (CML) Treated with Imatinib during Puberty.

Imatinib, a BCR-ABL tyrosine kinase inhibitor, is an effective treatment for adults and children with CML. However, the in vivo inhibition of the PDGF receptor and c-kit in normal cells may have clinical side effects: altered bone and mineral metabolism, reduction of testosterone and gynecomastia have been reported in adults treated with imatinib. To evaluate bone and mineral metabolism and puberal development, we studied 4 consecutive prepuberal patients (median age 10.6; range 9–12 years) with Ph+ CML treated with imatinib (340mg/sqm/day) in complete cytogenetic response. To evaluate puberal development we used the Tanner staging and serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), free testosterone (fT), inhibin-B, progesterone (P), 17a-hydroxyprogesterone (17a-OH-P) dihydrotestosterone (DHT), prolactin (Prl), growth hormone (GH), insulin-like growth factor-1 (IGF-1); to evaluate mineral metabolism, we tested osteocalcin levels, C-terminal telopeptides of type I collagen (CTX), parathyroid hormone (PTH), serum and urinary calcium, serum and urinary phosphate levels; bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DEXA). Table 1 reports the serum concentration results at different time points during treatment. The bone resorption marker CTX was up to 2.6 times over the highest reference interval for age matched normal boys and was accompanied by a reduction of BMD for chronologic age (mean z-score -1.3). The longitudinal growth, between the 50th and the 10th percentile before imatinib, slowed below the 10th percentile, although TSH, GH and IGF-1 were within the normal range. All males had progesterone and 17a-OH-P above the highest cut-off point and one (case 2) developed gynecomastia, pubertal delay, low BMD for chronologic age (z-score -2.9), serum calcium in the upper normal limits, urinary calcium higher than normal (896 mg/day, n.v. 100–300 mg/day), while serum PTH, 25-OH-D, urinary phosphate, BAP and osteocalcin were within the normal range. This patient during therapy presented an increase of FSH with a simultaneous decrease of inhibin-B; the decrease inhibin-B/FSH ratio is predictive of a spermatogenesis failure. In conclusion, the reduced BMD, the growth swiftness delay, gynecomastia and the inhibin-B/FSH ratio in our adolescents are probably due to imatinib. To better understand the safety profile of imatinib, it is important to investigate larger series of CML children treated with targeted therapies involving the PDGF receptor and c-kit.FSHLHTInhibin-BP17a-OH-PCTXAge(yrs)/Imatinib (mos)mlU/ml(1–14)mlU/ml(1.5–9.2)ng/ml(2.4–8.2)pg/ml(50–250)ng/dl(0.1–0.9)ng/dl(0.4–3.6)ng/ml(0.23–1.7)Case 1(male)13.9/204.141.6118311.92.5514.9/324.84.63.821771.074.63.5816.1/461.76.04.491032613.73.55Case 2(male)13.0/174.41.9<0.1770.590.491.8315.0/413.93.13.15571.572.94.4516.0/5313.41.53.4094.24.63.22Case 3(male)10.5/174.31.90.12800.720.551.3411.5/294.22.40.621591.122.31.4812.6/422.63.72.67691.523.22.35Case 4(female)11.5/1761.1<0.1390.260.421.9412.6/305.91.6<0.1510.50.351.7514.1/467.21.11.07230.750.681.62

Puberdade precoce associada a tumor misto ovariano (células germinativas- estroma-cordão sexual): aspectos clínicos, diagnósticos e manejo de um caso

Granulosa-stromal tumors comprise 5 to 8% of all primary ovarian neoplasms. The first clinical manifestation is precocious puberty in most prepuberal patients. We report a case of mixed germ cell-cord stromal tumor of ovary in a 7.2 years old girl, who presented with isosexual pseudo-precocious puberty of progressive outcome. Serum testosterone, estradiol and 17alphaOH-progesterone levels were increased. Abdominal-pelvic ultrasound revealed a right ovarian mass. Unilateral salpingo-oophorectomy was performed with complete resection of the tumor. The patient is well 7 years after surgery with normal pubertal and growth development and no signs of tumor relapse. We review the clinical manifestations of ovarian tumors, classification and staging of sex cord-stromal tumors, follow-up, tumor markers, treatment and prognosis.

Pesquiza precoz de nefropatía diabética en niños y adolescentes portadores de diabetes mellitus tipo 1

Introduction: Diabetic nephropathy commences early after the diagnosis of type 1 DM, described risk factors include time after diagnosis, deficient metabolic control, puberty and arterial hypertension. Objective: To evaluate the aforementioned parameters and their association with early renal compromise in children with type 1 DM. Methods: Prospective study measuring microalbuminuria, random morning creatininuria (index microalbuminuria/creatininuria (ACR), plasma creatinine, creatinine clearence, renal ultrasound, actual glycosylated hemoglobin and previous average, age of diagnosis, puberty and blood pressure. Results: 44 patients were studied, 20 males, average age 11.68 ± 3.01 years, with a follow-up since DM diagnosis of 3.8 ± 2.84 years. Average HbA1c was 9.1 ± 1.9%. Hyperfiltration was observed in 7/44 (15.9%) and nephromegalia was noted in 7/28 (25%). 8/44 patients (18,2%) had a raised ACR, and the 25% of prepuberal patients who has increase ACR were diagnosed at an earlier age. There was no correlation between metabolic control, hyperfiltration or nephromegalia with other studied parameters. Conclusions: Microalbuminuria determined by ACR is present in 18.2% of patients, presented more frequent in prepuberal children who debut at youngest age, being a possible risk factor for its future development.

Tumor ovárico de la granulosa: causa infrecuente de telarquia prematura

Precocious thelarche usually results from a physiological process but can sometimes be the first sign of precocious pseudopuberty. Ovarian granulosa cell tumors are highly unusual in childhood, appearing as precocious puberty in most prepuberal patients. During adolescence these tumors may cause menstrual irregularities, virilization and abdominal pain. Their malignancy is low and surgical treatment is usually curative if the tumors are limited to the ovaries. More advanced stages require chemotherapy, are difficult to cure and produce high mortality. We present the case of a 16-month-old girl with a granulosa cell tumor who presented with progressive precocious thelarche over 1 month that was satisfactorily resolved after resective surgery. This case demonstrates that other causes of puberal development should be investigated when precocious thelarche with fast progression is observed, with special attention paid to tumoral disease in the differential diagnosis.

Skeletal maturation in the XY female syndrome.

The assessment of bone age in eleven prepuberal patients with XY-female syndrome shows that skeletal maturation in these patients follow a male pattern. This supports the concept of a factor on the Y chromosome regulating skeletal maturation in the first ten years of life. Tall stature does not seem to be a feature of this syndrome in childhood.

45 TESTICULAR FUNCTION IN LEUKEMIC PATIENTS AFTER CHRONIC TREATMENT WITH CYTOSTATIC DRUGS

Testicular function was studied in 3 adult patients, 1 pubertal stage II, and 6 prepubertal with acute lymphoblastic leukemia, in remission after 4-6 years of treatment with chemotherapy, cranial irradiation and intrathecal methotrexate. Serum levels of: testosterone in adult patients X = 631ng/100ml (570-707), prepuberal, X = 24 (11-32);LH in adult patients X = 39ng/ml LER 907 (18-70), prepuberal X=19 (12-25); FSH in adult patients X = 157ng/ml LER 907 (70-240), prepuberal X = 68(12-120) were normal except plasma FSH of 1 adult patient which was high (240). In the prepuberal patients there was a normal response to HCG and to LH-RH Testicular histology showed sclero-hyalinosis and/or hypoespermatogenesis in 1 puberal and 2 adult patients and leukemic infiltration in 1 azoospermic adult.Sperm count was normal in the other 2. In 3 out of 4 prepuberal patients, unexpectedly early signs of sperm maturation without tisular damage were observed. Our data suggest that the mature testis is more labile to damage by treatment and spermatogenesis in the inmature testis is stimulated by an undetermined mechanism.

Late results after surgical treatment of maldescended testes with special regard to exocrine and endocrine testicular function.

Testicular volumes and exocrine testicular function were determined in 82 men who had undergone orchidopexy (36 bilateral, 46 unilateral) at 5-18 years of age. The testicular volumes in unilateral and bilateral orchdopexied patients correspond to those on normal mature males. Semen analyses in unilateral cryptorchidism were normal in 13, doubtful in 23 and pathological in 10 patients. Out of a total of 36 patients who have undergone bilateral orchidopexy, 3 patients were found to have normal, 7 patients doubtful and 26 patients pathological sperm analyses. Endocrine evaluation in bilateral cryptorchidism (prepuberal, puberal and postpuberal) showed no differences in testosterone levels compared to control groups. Some postpuberal patients with pathological sperm analyses were found to have elevated LH and FSH levels; 22 postpuberal patients with pathological sperm analyses showed hypergonadotropism with markedly elevated LH and FSH levels after GnRH.

Morphologic studies of the thymus in human neoplasia.

Quantitative histologic studies have shown that the thymus in patients with neoplasia differs in several morphologic respects from the thymus in normal controls of the same sex and age. The most consistently altered features were: increased diameter of Hassall's corpuscles in prepuberal patients with neoplasia and decreased diameter in postpuberal patients with neoplasia; decreased total cellularity and decreased number of lymphocytes in postpuberal patients; and increased number of Hassall's corpuscles, decreased number of lymphocytes, increased diameter of the largest Hassall corpusle, and decreased total cellularity in prepuberal patients. Most of these differences were statistically significant. These findings suggest that the thymus is related to human as well as experimental neoplastic disease.