Longitudinal hormonal evaluation in a patient with disorder of sexual development, 46,XY karyotype and one NR5A1 mutation.

Steroidogenic factor 1 (encoded by the NR5A1 gene) is a critical regulator of reproduction, controlling transcription of key genes involved in sexual dimorphism. To date, NR5A1 variants have been found in individuals with a 46,XY karyotype and gonadal dysgenesis, as well as with a wide spectrum of genital anomalies and, in some patients, with adrenal insufficiency. We describe evolution of gonadal function, from the neonatal period to puberty, in a patient with a 46,XY karyotype, a disorder of sexual development, and a mutation (c.691_699dupCTGCAGCTG) in the NR5A1 gene. The patient, ascertained at birth due to ambiguous genitalia, showed normal values of plasma testosterone in the late neonatal period. Evaluation of the hormonal profile over time indicated severe tubular testicular hypofunction suggestive for a 46,XY disorder of gonadal development. A comprehensive review of published reports of 46,XY and disordered sexual development related to the NR5A1 gene confirmed the clinical and hormonal variability in patients with NR5A1 mutations. Analysis of multiple data allowed us to define the most common features associated with NR5A1 mutations. We further confirmed the indication to perform NR5A1 screening in patients with 46,XY karyotype and disordered sexual development even when Müllerian structures appear to be absent and plasma testosterone levels are within the normal range for age.

Vaginoplasty for disorders of sex development.

One of the most common problem found in patients with Disorders of Sexual Developments is the absence or extreme hypoplasia of the vagina. The type of patients presenting this anomaly may belong to completely different groups: (1) Patients with a urogenital sinus with urethra and vagina fusing together to form a common channel. (2) Patients with absent Müllerian structures and different degrees of external virilization. (3) Complex malformations. Treatment options: treatment of these patients is under discussion and may consist, basically, in non-operative dilation methods or surgical creation of a neovagina. Consensus is far to be reached among the various surgical subspecialties regarding the optimal method of vaginal replacement. Adequate number of long-term follow up patients are still non-available so that most conclusions are based on small number series. The authors describe the different treatment options in detail.

A new suture material for hypospadias surgery: a comparative study.

PURPOSE: We compared the results of hypospadias repair using polyglytone versus polydioxanone to evaluate the potential benefit of using a suture with a rapid absorption time. MATERIALS AND METHODS: A total of 100 patients 8 to 24 months old affected by distal isolated penile hypospadias were considered for this study. Patients were randomized and assigned to 2 different groups according to the suture material used during the surgical procedure (tubularized incised plate repair with or without preputial reconstruction). Polyglytone was used in group A and polydioxanone was used in group B. All patients were evaluated at 4 intervals (1 week, 1 month, 6 months and 2 years postoperatively). Persistence of sutures on penile skin, urethral fistulas, skin dehiscence, infection and skin tracks were recorded. Statistical analysis was performed using chi-square test. RESULTS: Followup data documented the absence of significant differences in terms of urethral fistula rate, skin dehiscence and acute skin infection. Persistence of sutures and multiple skin tracks at long-term followup were significantly greater in patients in group B. CONCLUSIONS: Both sutures are adequate for hypospadias surgery in small children. The use of a rapid absorption monofilament may allow much more rapid disappearance of the skin sutures. In the long term this outcome means almost complete absence of suture tracks. No statistically significant difference in terms of urethrocutaneous fistula was observed, suggesting that the tensile strength of polyglytone is adequate.

Retroperitoneoscopic renal surgery in children: our experience.

BACKGROUND: Laparoscopic renal surgery has become an accepted approach for benign disease in adult and pediatric urology. We present our experience in renal laparoscopy in infants during the past 5 years and evaluate our series to establish the safety and efficacy of such procedures in children. MATERIALS AND METHODS: From August 1999 to December 2004, we performed 32 renal laparoscopic procedures on 31 children aged 12 months to 16 years (mean, 42 months). Twenty-seven patients underwent unilateral retroperitoneal nephrectomy; 1 child with Denis Drash syndrome underwent transperitoneal bilateral nephrectomy; 2 children underwent renal cyst unroofing; and 1 child with lymphoma underwent retroperitoneal laparoscopic renal biopsy. Indications for surgery were: renal function <9% in cases of unilateral nephrectomy; the prevention of renal neoplastic changes in the patient with Denys-Drash syndrome; symptomatic large renal cysts; and suspected lymphoma not diagnosed with a previous percutaneous biopsy. RESULTS: All procedures were completed laparoscopically. In 6 cases, the accidental opening of the peritoneum did not require conversion to open surgery. Intraoperative blood loss was minimal. One patient who underwent a retroperitoneal nephrectomy required a blood transfusion for postoperative bleeding into the retroperitoneal space. Twenty-four of 27 unilateral retroperitoneal nephrectomy patients were discharged on postoperative day 2. Mean follow-up was 30 months (range, 6-64 months). Cosmetic results were excellent in all patients and no long-term complications have been encountered so far. CONCLUSION: Laparoscopic urologic surgery may be performed in children with minimal morbidity, minimal postoperative discomfort, improved cosmetic results, and a short hospital stay.

Natural history of vesicoureteral reflux associated with kidney anomalies.

OBJECTIVES: To evaluate the cystographic follow-up of patients with multicystic dysplastic kidney (MCDK), renal agenesis, and renal ectopia with associated primary vesicoureteral reflux (VUR). METHODS: Patients with primary associated VUR (grade 2 or more) and with a minimal follow-up of 24 months were included in this study. RESULTS: Of the children with renal agenesis, 24% had VUR. The median grade of VUR was significantly greater in the boys than in the girls (4 versus 2, respectively; P < 0.05). All girls and 34% of the boys experienced spontaneous resolution 1 year after diagnosis; 66% of the boys required operative treatment. Of the patients with MCDK, 16% had VUR. The median grade of VUR in the kidney contralateral to the MCDK was greater in the boys than in the girls (3.5 versus 2, respectively; P = 0.06). All girls and 60% of the boys had spontaneous resolution 1 year after diagnosis; 40% of the boys underwent operative treatment. Of the children with renal ectopia, 30% had VUR. The median grade of VUR for the refluxing unit was significantly greater in the girls than in the boys (3 versus 2, respectively; P < 0.05). Spontaneous resolution of VUR was observed in all boys (66% at 1 year) and in 40% of the girls; 60% of the girls required operative treatment. CONCLUSIONS: Spontaneous resolution of VUR can be anticipated in girls with MCDK or renal agenesis and in boys with renal ectopia. These data can be used in planning the proper follow-up schedule for babies with a kidney anomaly detected by ultrasonography.

The incidence of associated urological abnormalities in children with renal ectopia.

PURPOSE: We evaluate the incidence of associated urological abnormalities in children with renal ectopia (RE). MATERIALS AND METHODS: We retrospectively reviewed database records of 99 consecutive pediatric cases of RE diagnosed at our hospital between 1988 and 2001. RESULTS: Mean patient age at diagnosis was 13 months (range 0 to 9 years, SD 2.2 years). Twenty children were evaluated for symptoms, while the remainder were asymptomatic and evaluated after prenatal or postnatal ultrasound finding of RE. There were 10 cases of crossed renal ectopia and 89 cases of simple renal ectopia (S-RE), of which 82 were unilateral and 7 bilateral. The most frequent associated urological abnormality was vesicoureteral reflux (VUR), which was present in 20% of patients with crossed renal ectopia and 30% with S-RE. In patients with bilateral S-RE the incidence of VUR increased to 71%. In patients with unilateral S-RE VUR involved the orthotopic kidney in more than 85% of the cases. Moreover, technetium dimercapto-succinic acid scans documented primitive reduction of function of the ectopic kidney in 87% of the unilateral S-RE cases. Other associated urological anomalies were present in 14% of patients, the most frequent of which were contralateral renal dysplasia (4%), cryptorchidism (5%) and hypospadias (5%). CONCLUSIONS: A high proportion of children with renal ectopia have associated urological anomalies, and VUR is the most common. The presence of VUR in the normally positioned kidney together with decreased function of the ectopic kidney might predispose these children if not recognized and treated to renal function impairment.

Upregulated expression of EGF and TGF-alpha in the proximal respiratory epithelium in the human hypoplastic lung in congenital diaphragmatic hernia.

Newborn infants with congenital diaphragmatic hernia (CDH) still have a high mortality rate. Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) are peptide growth factors involved in the fetal lung growth and development. The EGF and TGF-alpha have been reported to promote pulmonary branching activity and alveolar type-II pneumocyte proliferation. Epidermal growth factor and TGF-alpha immunoreactivity and mRNA expression in the bronchial and bronchiolar epithelium is maximal during early fetal life and barely detectable in the proximal airways of neonatal lung. The purpose of this study was to determine protein and gene expression of EGF and TGF-alpha in CDH lung in order to elucidate the potential role of these growth factors in the pathogenesis of pulmonary hypoplasia in CDH. Lung tissue specimens were obtained from archival lung tissue from 11 patients with CDH and 5 controls. Indirect immunohistochemistry was performed using ABC method with anti-EGF and anti-TGF-alpha antibodies. In situ hybridization was performed using EGF and TGF-alpha specific digoxigenin-labeled oligonucleotide probes. The most striking difference between hypoplastic CDH lung and control lung was the strong EGF and TGF-alpha mRNA expression and immunoreactivity in the bronchial and bronchiolar epithelium in CDH lung. The upregulated protein and gene expression of EGF and TGF-alpha in the proximal airways in the CDH hypoplastic lung suggests persistence of fetal stage of pulmonary airway development in CDH.

Increased expression of ICAM-1 and VCAM-1 in the lung of nitrofen-induced congenital diaphragmatic hernia in rats.

Recently, increased expression of inflammatory cytokine, tumor necrosis factor (TNF)-alpha, has been reported in both humans and animal models with CDH and the decreased TNF-alpha expression in CDH lung after antenatal dexamethasone (Dex) treatment. Intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 are induced by several inflammatory cytokines such as TNF-alpha. The aim of this study was to investigate pulmonary ICAM-1 and VCAM-1 expression in CDH lung in rats and to determine the effect of antenatal glucocorticoid. CDH model was induced in pregnant rats following administration of nitrofen on day 9.5 of gestation. In control animals, the same dose of olive oil was given without nitrofen. Dex (0.25 mg/kg) was given on day 18.5 and 19.5 of gestation. RT-PCR was performed to evaluate the relative amount of ICAM-1 and VCAM-1 mRNA expression. Fluorescein immunohistochemistry using anti-ICAM-1 and anti-VCAM-1 antibody was performed using light and confocal microscopy. ICAM-1 and VCAM-1 mRNA expression and ICAM-1 and VCAM-1 immunoreactivity were markedly increased in CDH lung compared to controls. Dex downregulated the expression of both adhesion molecules in the hypoplastic lung. Increased ICAM-1 and VCAM-1 mRNA expression in hypoplastic lungs would suggest that the increased local synthesis of pulmonary adhesion molecules may induce respiratory distress in CDH. Decreased expression of adhesion molecules in CDH lungs after Dex treatment suggests that antenatal glucocorticoids therapy may improve pulmonary immaturity and associated respiratory distress in nitrofen-induced CDH lung.

The adolescent varicocele: the crucial role of hormonal tests in selecting patients with testicular dysfunction.

BACKGROUND/PURPOSE: Early identification and treatment of varicocele during adolescence may reduce the risk of infertility. Prophylactic surgery on all adolescents with varicocele would be unnecessary surgery on a high percentage of them. Suggested guidelines for early intervention have included testicular volume loss and abnormality in the hormonal integrity of the hypothalamic-pituitary-testicular axes. Previous studies have documented no correlation between testicular volume loss and abnormal response to GnRH test. The role of these 2 prognostic factors in predicting semen abnormalities in this age group never has been investigated, so far. In the current study we correlated hormonal tests and orchidometry to semen analysis in a selected group of adolescents. METHODS: Adolescents with varicocele and Tanner stage V underwent semen analysis, Prader orchidometry, and hormonal tests (basal LH and FSH [bLH and bFSH], testosterone [T], LH and FSH after GnRH stimulation test [sLH and sFSH]). According to semen analysis (in relation to Oval Motility Index) patients were divided into 2 groups: group A, adolescents with pathologic semen and group B, adolescents with normal semen. Hormonal tests and testicular volumes (in those with left unilateral varicocele) were compared between the 2 groups. Statistical analysis was performed using Mann-Whitney U test and Student's t test. RESULTS: Of 76 patients, 20 (Group A) had a pathologic semen analysis. Levels of bLH, bFSH, sFSH were significantly higher in group A compared with group B (56 with normal semen). sLH was higher in group A, but the difference was not statistically significant. T levels were not different between the 2 groups. Of 68 patients who had left unilateral varicocele and no previous inguinal--scrotal surgery 14 had pathologic semen analysis. There was no statistically significant difference between the 2 groups for any of the orchidometric parameters investigated. CONCLUSIONS: The evaluation of testicular volumes during physical examination for varicocele in Tanner stage V adolescents is not predictive for testicular dysfunction. LH and FSH dosages with or without GnRH stimulation test can identify those subjects at risk for infertility, and their use should be encouraged for selecting patients who need surgical correction.

Antenatal dexamethasone enhances endothelin-1 synthesis and gene expression in the heart in congenital diaphragmatic hernia in rats.

BACKGROUND/PURPOSE: Although high levels of endothelin-1 (ET-1) in plasma may be relevant in certain pathophysiologic states, such as pulmonary hypertension accompanying congenital diaphragmatic hernia (CDH), experimental evidence favors a local, paracrine, or autocrine role for ET-1 in most tissues. Evidence of ET-1 production has been documented in fetal heart tissue where it exerts growth-enhancing and mitogenic effects. ET-1 also has a potent positive inotrope action on cardiac muscle. ET-1 -/- homozygous mice display a wide variety of cardiac anomalies, which also are features of the human and of the experimental CDH. Autopsy reports have shown that total heart weight is reduced significantly in the presence of CDH, and animal models have documented the presence of cardiac hypoplasia associated with CDH. Experimental and clinical studies have shown that prenatal exposure to corticosteroids improves cardiovascular function in the immediate newborn period. The aim of this study was to determine cardiac gene expression of ET-1 and of its receptor ET(A) and the cardiac ET-1 content in the heart of nitrofen-induced CDH in rats and to evaluate the effect of antenatal Dexamethasone (Dex) treatment. METHODS: A CDH model was induced in pregnant rats after administration of 100 mg of nitrofen on day 9.5 of gestation (term, 22 days). Dex (0.25 mg/kg) was given by intraperitoneal injection on days 18.5 and 19.5 of gestation. Cesarean section was performed on day 21 of gestation. The fetuses were divided into 3 groups: group I, control (n = 8); group II, nitrofen-induced CDH (n = 8); group III, nitrofen-induced CDH with antenatal Dex treatment (n = 8). ET-1 protein was measured using ELISA. RT-PCR was performed to evaluate the relative amount of ET-1 and ET(A) mRNA expression. RESULTS: There was a reduction in ET-1 mRNA (P <.05) and in ET(A) mRNA (P <.01) in the heart of CDH group compared with controls. ET-1 protein level also was reduced in heart of CDH compared with controls. Antenatal Dex treatment increased significantly both ET-1 mRNA and protein levels in the heart of CDH animals (P <.05 and P <.01, respectively). CONCLUSIONS: The reduced cardiac ET-1 gene expression and ET-1 synthesis may be responsible for the heart hypoplasia associated with CDH. Prenatal corticosteroids increase the cardiac production of ET-1, and this may enhance heart growth and cardiac inotropism at birth.

Effect of mechanical ventilation on the pulmonary expression and production of elastin in nitrofen-induced diaphragmatic hernia in rats.

BACKGROUND/PURPOSE: Hypolastic lung in congenital diaphragmatic hernia (CDH) shows markedly thickened alveolar walls, increased interstitial tissue, and markedly diminished alveolar air space, showing morphologic immaturity. Decrease in lung compliance and distensibility often is seen in human CDH as well as experimentally produced CDH. Collagen and elastin, critical components of the lung connective tissue, have been suggested to have important influence on lung compliance and maximal expansion. The barotrauma caused by mechanical ventilation is known to produce structural changes in the pulmonary architecture. The aim of this study was to investigate the expression and production of elastin in the lung in newborn rats with CDH during mechanical ventilation. METHODS: CDH was induced in rat embryos after administration of nitrofen to pregnant dams on day 9.5 of gestation. Cesarean section was performed on day 21 of gestation. The newborn rats were intubated using a 24-gauge Teflon catheter. After ligation of the umbilical cord, the intubated animals were transferred immediately to a warm plate and connected to a modified ventilator. Ventilation was continued for a maximum of 6 hours. The relative amount of soluble elastin in the lung was assessed using an enzyme-linked immunosorbent assay (ELISA) technique. Reverse transcription polymerase chain reaction (RT-PCR) was performed to evaluate the relative amount of tropoelastin mRNA expression in the lung. RESULTS: Elastin mRNA in the CDH lung was increased significantly (P <.01) at one hour after ventilation compared with ventilated controls. Elastin protein significantly increased in the CDH lung at one hour (P <.01) and 6 hours (P <.01) after starting ventilation compared with controls. CONCLUSION: The data show that during mechanical ventilation, elastin production is increased significantly in the CDH lung, and this may further affect lung compliance.

Is endoscopic follow-up needed in pediatric patients who undergo surgery for GERD?

BACKGROUND: This study evaluated the role of endoscopy in the postoperative management of pediatric patients who undergo fundoplication for GERD. METHODS: Medical records of 109 otherwise healthy children who underwent operation for GERD from 1979 to 1996 were reviewed. Patients with respiratory symptoms or esophageal stenosis were excluded. All patients underwent endoscopic surveillance with endoscopy being performed in the early (within 1 year) and late (between 1 and 2 years) postoperative periods. Specifically evaluated were the appearance of the wrap and evidence of esophagitis. The risk of a recurrence of esophagitis based on wrap appearance and the presence of clinical symptoms in patients with endoscopic evidence of esophagitis were also evaluated. RESULTS: At early endoscopy 3 patients with an intact wrap and 8 with a defective wrap had esophagitis (not significant). At late endoscopy, 5 patients with an intact wrap and 17 with a defective wrap had esophagitis (p < 0.05). CONCLUSIONS: An intact wrap does not prevent recurrence of GERD. Such an occurrence is even more likely when endoscopy demonstrates a defective wrap. For all patients who have undergone fundoplication, endoscopic evaluation at 1 to 2 years is recommended to detect esophagitis in the absence of symptoms so treatment can be initiated before symptoms occur.