Cause Of Hypospadias Research Articles

A Potential Pathogenic SRD5A2 Mutation and rs632148, rs523349 and rs522638 Polymorphisms in Increasing the Risk of Syndromic Hypospadias in Indonesian Population

BACKGROUND: Hypospadias, a congenital birth defect in male, is the opening of the urethra located on the ventral side of the penis. Several mutations in SRD5A2 encoding steroid 5 alpha-reductase type 2 protein have been identified in hypospadias and polymorphisms in this gene have been known to be associated with an increased risk of hypospadias. In this study, several crucial molecular analyses of the SRD5A2 gene and the association of the identified variants to the risk of syndromic hypospadias in Indonesian population were conducted.METHODS: Thirty-two isolated and 29 syndromic hypospadias patients were enrolled in this study. DNA was isolated from whole blood for the amplification of all exons and exon-intron boundaries of SRD5A2 by polymerase chain reaction (PCR), followed by Sanger sequencing. In silico analysis was performed using PolyPhen-2, Sorting Intolerant from Tolerant (SIFT) and Align GVGD. Statistical analysis was performed using Chi-squared test.RESULTS: A novel missense mutation c.32T>C/p.Leu11Pro was identified in one isolated hypospadias patient and the in silico analysis predicted the mutation to be pathogenic. Three polymorphisms were identified, two in the non-coding region (c.-62G>C/rs632148 and c.281+15T>C/rs522638) and one in exon-1 (c.265C>G/p.Val89Leu/rs523349). Mutant alleles of these polymorphisms were significantly associated with syndromic hypospadias with odds ratios (OR) of 3.4, 3.13 and 2.54 respectively.CONCLUSION: This study suggests that SRD5A2 mutation is one of the causes of hypospadias in Indonesian population and rs632148, rs523349 and rs522638 polymorphisms are significantly associated with an increased risk of syndromic hypospadias.KEYWORDS: mutation, polymorphism, SRD5A2, syndromic hypospadias

Identifying infrequent genetic changes in monozygotic twins afflicted with hypospadias via targeted panel sequencing.

We aimed to identify the genetic causes of hypospadias in children using targeted gene panel sequencing for disorders of sex development (DSD). This study included 18 twin boys with hypospadias: seven and two pairs were monozygotic and dizygotic twins, respectively, and six were discordant and three were concordant twins. Targeted gene panel sequencing for 67 known DSD genes was performed. Sequence variants were classified into five different categories, pathogenic, likely pathogenic, variants of uncertain significance, likely benign, and benign, following the American College of Medical Genetics and Genomics Standards and Guidelines. The mean gestational age and birth weight were 35.3±2.0 weeks and 1.96±0.61 kg, respectively, with seven patients being small for gestational age. Hypospadias was present in 12 patients, with posterior type in 33.3% and anterior type in 66.7%. In three families with twins, both siblings had hypospadias. In addition, cryptorchidism was observed in one subject. Surgical correction of hypospadias was performed at a mean age of 22.1 months. Molecular analysis identified 12 different genetic variants, including two pathogenic mutations in the AMH (p.E389*) and SRD5A2 (p.R246Q) genes, found in subjects with hypospadias, respectively. However, only heterozygous mutations were detected. This study did not identify a definitive genetic component contributing to the development of hypospadias; however, the findings suggest that intrauterine growth retardation may play a significant role.

OR07-6 Disrupting DNA-binding Domain Dimerization In The Androgen Receptor In Mice Results In a Partial Androgen Insensitivity Model

Abstract The androgen receptor (AR) plays a crucial role in the development and maintenance of the male phenotype, as shown in patients with androgen insensitivity syndrome (AIS). The AR is a nuclear receptor that needs to homodimerize to execute its role as transcription factor. Dimerization can occur through three different modes: via the DNA-binding domain (DBD), via the ligand-binding domain and via an interaction between the LBD and the aminoterminal domain. Dimerization via the DBD is very well known and occurs through the D-box located in the second zinc finger. DBD dimerization is generally accepted for all steroid receptors, but some of them show that D-box mutations do not inactivate them completely. However, for the AR, no monomeric transactivation capacity was described yet. To study the role of D-box independent activity of the AR, we introduced a double point mutation in the D-box that is predicted to disrupt DBD dimerization (ARDmon). These two point mutations were found in separate patients suffering from partial AIS. In vitro studies with this mutant suggest that the remaining activity depends on the androgen response element under investigation. We show that ligand binding was not affected by the D-box mutation. Furthermore, BioID assays indicated no changes in coregulatory interactions. When introducing the corresponding mutation in mice, this led to a phenotype that is intermediate between global ARKO and wild type (WT) males. Based on their anogenital distance (AGD) and the absence of nipples at early age, the ARDmon/Y mice were identified as males. Follow-up of their AGD and body weight, however, revealed clear differences with both WT males and females. Furthermore, hypospadias was observed in the ARDmon/Y mice. Testes were smaller in size compared to the WT males and first analysis showed almost no expression of well-known AR regulated genes. Reminiscent structures of the epididymis were found. In summary, the ARDmon mouse model shows an intermediate AIS phenotype. Via orchidectomy in combination with androgen replacement therapy, we will now investigate remaining androgen responses in the ARDmon/Y mice and elucidate response elements on which the AR can act in a D-box independent way. Additionally, we generated the first rodent PAIS model with a genetic cause of hypospadias. Presentation: Saturday, June 11, 2022 12:45 p.m. - 1:00 p.m.

Hypospadias and the Use of the Ages and Stages Questionnaire to Evaluate Neurodevelopmental Status of Boys with Hypospadias.

Placental insufficiency is one of the reasons for the reduction of hormone production. Thus, if one of the suspected causes of hypospadias is placental insufficiency, then the neurodevelopmental status of boys with hypospadias may be impaired. The aim of this study was to evaluate the neurodevelopmental status of boys with hypospadias and guide the parents of those who need support to related departments for early intervention. Boys were divided into two groups, those with hypospadias (group H) and healthy children undergoing traditional circumcision (group C). The parents of the boys completed the Ages and Stages Questionnaire (ASQ) and ASQ-Social Emotional (SE), both of which are screening instruments for the early identification of developmental and social-emotional problems, respectively. Seventy-eight boys had hypospadias and 59 were admitted for traditional circumcision. The group H had statistically significant more impaired scores than group C in communication, gross motor, and personal-social skill sections. The multivariate logistic regression analysis revealed that hypospadias was the independent predictive factor for communication and personal-social skills. Hypospadias and neurologic impairment may share common etiologic factors. Accordingly, physicians should keep in mind that if a boy presents with hypospadias, the possibility of having neurologic impairment is higher than normal population and early intervention has crucial importance. Every boy with hypospadias should be evaluated for neurodevelopmental status.

Case Report: Disorder of Sex Development 46, XY with Proximal Hypospadias in a 10-Year-Old Twin Boy: Disorder of Sex Development 46, XY with Proximal Hypospadias

Hypospadias is a congenital defect in which the opening of the urethra is located on the ventral side of the penis proximal to the tip of the glans penis. The cause of hypospadias is still not clear and believed to have several factors. Hypospadias occurs due to disruption of urethral fold fusion at the 8 th – 15 th week in the uterus which is influenced by dihydrotestosterone. Hypospadias can cause a disorder of sex development according to the conditions. A case of proximal hypospadias was reported to occur in a 10-year-old twin boy accompanied by undescended testis causing a disorder of sex development. For this case, various examinations were carried out along with evaluations to establish the diagnosis, the cause, the treatment plan, and the necessary evaluation. Keywords : hypospadias, disorder of sex development, undescended testis DOI: 10.7176/JHMN/80-01 Publication date: September 30 th 2020

Molecular genetic analysis of AKR1C2-4 and HSD17B6 genes in subjects 46,XY with hypospadias

The formation of the male urethra depends to enzyme-mediated testosterone (T) conversion into 5α-dihydrotestosterone (DHT). Two metabolic pathways could be operating in the fetal testis to synthesize androgens: 1) the "classic" route (T→DHT) mediated by SRD5A2 and 2) a "backdoor" pathway in which DHT is synthesized by aldo-keto reductase family 1, member C2 (AKR1C2), AKR1C3, and AKR1C4 enzymes without formation of a T intermediate. We studied four genes of the "backdoor" pathway in karyotypic males with hypospadias to ascertain whether gene defects in AKRs impair urethral DHT formation that result in hypospadias. The coding regions of the AKR1C2-4 and HSD17B6 genes were analyzed by PCR-SSCP and sequencing in a cohort of 25 Mexican patients (0.3-9 year-old-children) with 46,XY-hypospadias. Chi-squared tests was performed to evaluate the distribution of genotypes, alleles, and the Hardy-Weinberg (H-W) equilibrium. The effect of the genetic variants was investigated by in silico studies. Screening studies revealed distinct genotypic patterns at different exons of AKR1C2-4 whereas HSD17B6 presented a wild-type sequence. The DNA analyses detected two synonymous variants (c.327C>T, c.666T>C/unreported) in AKR1C2. The AKR1C3 had two variants (c.15C>G, c.230A>G), two unreported variants (c.538T>C, c.596G>A), and one silent variant (c.312G>A). Two variants (c.434C>G, c.931C>G) were identified in AKR1C4. All variants were in H-W equilibrium without structural changes. Hypospadias have been associated with defects that alter androgen biosynthesis in the human fetal testis, specifically 5α-DHT. We selected four candidate genes involved in the "backdoor" pathway for the formation of 5α-DHT. Molecular assays of the AKR1C2, AKR1C3, and AKR1C4 genes revealed a total of nine genetic single nucleotide variants. Several variants in the AKR1C genes have been associated with a variety of human pathologies. However, our studies suggest that active steroid biosynthesis via AKR1C might not be involved in hypospadias. Additionally, genetic research suggests a low involvement in the "backdoor" 5α-DHT pathway during human sexual development, specifically, the differentiation of male external genitalia. These results indicate that substitutions in AKR1C2-4 are polymorphisms and all genetic variants lacks deleterious significant association with hypospadias. The data suggest that inactivating mutations in the AKR1C2-4 and HSD17B6 genes are an infrequent cause of hypospadias, which might weaken the contribution of the "backdoor" pathway to embryonic urethral masculinization.

Identification of gene variants in 130 Han Chinese patients with hypospadias by targeted next-generation sequencing.

BackgroundHypospadias is a common congenital malformation of male external genitalia, which mainly manifests as an abnormal urethral opening on the ventral side of the penis. The etiology and clinical phenotype of hypospadias is highly heterogeneous, and its clinical diagnosis is challenging. Currently, over 70% of patients have an unknown etiology. Here, we performed a targeted analysis of gene mutations in 130 patients with hypospadias of unknown etiology to find the precise genetic cause.MethodsWe developed a targeted next‐generation sequencing (NGS) panel, encompassing the exon coding regions of 105 genes involved in external genitalia and urogenital tract development and performed sequencing analysis on 130 children with hypospadias of unknown etiology.ResultsIn total, 25 patients with hypospadias (19.2%) were found to have 20 mutations among the nine genes involved in external genitalia and urogenital tract development, including 16 reported and four novel mutation sites. Twenty‐two patients (16.9%) had diagnostic variants. Multiple genetic mutations were identified in three of the 25 patients. Hypospadias combined with micropenis was the most common phenotype (68%) in 25 patients.ConclusionsHigher frequency mutations were identified in SRD5A2 (52%) and AR (24%) in our patient cohort. Middle or posterior hypospadias with micropenis may be significant indicators of genetic variations. Polygenic inheritance may be a rare genetic cause of hypospadias.

Эпидемиология и теории происхождения гипоспадии

The increase in the prevalence of hypospadias, which has lasted for several decades, signals the need to find ways to prevent the disease. Development of measures aimed to reduce morbidity is impossible without a unified concept of the etiology and pathogenesis of hypospadias. The article analyzes the existing theories of the origin of hypospadias. According to the literature, there are 5 types of causes of hypospadias, some of which are currently established. Among the causes under investigation, the emphasis is on placental factors.

Description of mutation spectrum and polymorphism of Wilms' tumor 1 (WT1) gene in hypospadias patients in the Indonesian population

Hypospadias is one of the most common congenital anomalies of the penis. Previous studies reported mutation of the Wilms' tumor 1 (WT1) gene as a cause of hypospadias. The aim of this study is to describe the WT1 mutation spectrum and polymorphism in hypospadias patients in Indonesia. DNA was isolated from 74 hypospadias patients at the Division of Pediatric Surgery, Department of Surgery Hasan Sadikin Hospital. All exons in the WT1 gene were amplified by a PCR method, followed by Sanger sequencing. Mutation analysis was performed using BioEdit software and in silico analysis using Mutation Taster, Polymorphism Phenotyping-2 (PolyPhen-2), and Sorting Intolerant from Tolerant (SIFT). DNA analysis results showed two types of heterozygous mutations in five subjects (Table), hence the frequency of WT1 mutations was 6.7% (10/148 allele). The first mutation was a missense mutation identified in twin boys. The second was a novel heterozygous alteration in the non-coding region nine bp upstream of exon 6 (c.366-9T>C), which was identified in three patients. One heterozygous polymorphism in the coding region of exon 7 (c.471A>G/rs16754) was identified in 10 subjects. This variant did not cause any change in amino acid products (silence polymorphism). Allele frequency for the G allele (mutant allele) and A allele (wild type) was 13.5% and 86.5%, respectively. WT1 is one of the best known hypospadias genes. The WT1 gene is involved in male genital development in the early and late periods of sex determination, and hence is known as a long-term expression gene in genitalia development. Mutation analysis of WT1 in a Chinese population identified that the WT1 mutation frequency was 4.4%. The WT1 mutation frequency identified in the present study was higher, at 6.7%. Coincidentally, research subjects with p.R158H variants were monozygotic twin siblings with midshaft hypospadias accompanied by undescended testis in one and penoscrotal hypospadia with micropenis in the other. The incidence of familial hypospadias in male siblings suffering from hypospadias was reported to be 9.6% in a study conducted by Sorensen etal. Moreover, in the present study polymorphism c.471A>G(rs16754) at exon 7 was identified heterozygously in 10 research subjects (minor allele frequency 13.5%). WT1 mutations were identified in only a few cases of hypospadias and most of these were syndromic. This result implies that mutation of WT1 is not a common cause of hypospadias in the Indonesian population.

PHENOTYPE-GENOTYPE AND PEDIGREE ANALYSIS OF ISOLATED HYPOSPADIAS PATIENTS

Objective: Hypospadias is a malformation in urethra which has many range of severity. A patient with Isolated hypospadias (IH), a mild disorder of sex development (DSD) has a hypospadias phenotype only. Hypospadias is considered as multifactorial disease in which genetic factors contribute to its development. Chromosome analysis in DSD including hypospadias is conducted for gender assignment and other possible genetic contributions. This analysis solely could not elucidate all genetic causes of hypospadias. Polymorphism of V89L in SRD5A2 is suggested as one of genetic risk factors of hypospadias. To determine the genetic risk factor and pattern of inheritance, a good pedigree construction is required. Material & methods: 35 eligible subjects with IH admitted to Center for Biomedical Research (CEBIOR) during 2012-2015 were randomly selected. 35 normal male as control were included in this study. Data on three generation pedigrees were collected from medical records in 35 affected subjects. Chromosome analyses were done by using G-banding technique. Polymorphism analysis of V89L in SRD5A2 gene was done using PCR-RFLP technique in all samples. Results: From the 35 affected subjects, the most frequent phenotype was penile hypospadias (47%), a pair of twins were monozygotic and one had a cousin diagnosed with urogenital abnormalities (i.e micropenis and chordae). All subjects had 46,XY chromosome. No chromosomal aberration was found. No positive correlation between polymorphism of V89L in SRD5A2 and risk of hypospadias (PR of CC+CG vs GG=1.0, 95% CI: 0.342−2.921, p value=1.0). Conclusion: The pedigree data from our study implies tendency of genetic involvement in hypospadias cases. There were no chromosomal aberrations in hypospadias cases. The finding on polymorphism of V89L in SRD5A2 gene does not support that of previous studies.

Karyotyping and analysis of 5α -reductase-2 gene mutation in 25 patients with hypospadias

To analyze the karyotypes and SRD5A2 gene mutations in 25 patients with sporadic or familial hypospadias. The patients included 10 adults and 15 children, whose chromosomes were analyzed by G-banded karyotyping, and the SRD5A2 genes were sequenced. Two patients were found to have an abnormal karyotype, while eight have carried compound heterozygous mutations of the SRD5A2 gene, which included 5 genotypes formed by 6 types of mutations, i.e., p.G203S/p.R227Q, p.R227Q/p.R246Q, p.Q6X/p.Q71X, p.L20P/p.G203S, and p.Q71X/p.R227Q. Mutations of the SRD5A2 gene were present in 32% (8/25) of all patients, 35% (8/23) in those with a normal karyotype, and 44.4% (8/18) in those with proximal type hypospadia. Bioinformatic analysis, literature review and pedigree analysis confirmed that all such mutations are pathogenic. Chromosomal anomalies and mutations of the SRD5A2 gene are the main cause of hypospadias. Sequencing of the SRD5A2 gene may explain the etiology of nearly half of the patients with proximal type of hypospadas but a normal karyotype, which can facilitate genetic consulting.

Fetal growth restriction but not preterm birth is a risk factor for severe hypospadias.

Hypospadias has multifactorial causes and occurs at a high frequency among very low-birthweight infants. Placental insufficiency is hypothesized to be one cause of hypospadias; that is, decreased human chorionic gonadotropin (hCG) secretion caused by placental insufficiency is suspected to result in abnormal male external genitalia, but there is little direct evidence to support this. The aim of this study was therefore to identify the features of hypospadias and to clarify the male genital abnormalities caused by fetal growth restriction (FGR). We reviewed the clinical data of boys who underwent hypospadias repair between 2005 and 2011 at Kyoto University Hospital. Twenty boys were included in this study. Fifteen (75%) of the subjects were preterm or low-birthweight infants. Thirteen (65%) had FGR, 60% of whom had severe hypospadias regardless of gestational age. In addition, 92% of the FGR infants also had other genital anomalies, such as cryptorchidism, bifid scrotum, or micropenis. In contrast, only 14% and 43% of the non-FGR infants had severe hypospadias or genital anomalies other than hypospadias, respectively. Placental histopathology was available in eight FGR infants, in seven of whom it was suggestive of blood flow deficiency such as infarction and single umbilical artery. Infants with FGR have a high incidence of hypospadias. FGR caused by placental dysfunction, but not low birthweight, is a risk factor for severe hypospadias associated with multiple genital anomalies.

Hypospadias: One Defect, Multiple Causes, Acting Through Shared Pathways

Hypospadias is one of the most common birth defects among males. Although the birth prevalence of hypospadias has been reported as increasing in some geographic regions, inconsistencies in ascertainment of mild cases and variability in reporting standards are likely contributing to these apparent trends. While hypospadias is highly heritable, there is limited knowledge about the specific genetic and epigenetic factors that play a role in its etiology. Risk factors for hypospadias include family history, older maternal age, nulliparity, high maternal prepregnancy body mass index, hypertension or preeclampsia, multiple gestations, reduced birth weight, and small for gestational age. Of the various prenatal exposures to medications that have been studied, the strongest evidence supports valproic acid as a contributor to hypospadias. Studies evaluating the impact of assisted reproductive technologies (ART) on hypospadias are inconclusive because of potential confounding by subfertility. Many causes of hypospadias may act through a few shared pathways, such as placental dysfunction.

Prevalence and possible causes of hypospadias

Hypospadias is the most common congenital malformation of the male external genitalia. After the heart and circulatory system, it is the second most common developmental disorder in males. It is due to a midline fusion defect of the male urethra, which results in a misplaced urethral meatus. Hypospadias may be distal, medial and proximal. It may occur as an isolated defect or it may develop together with other genital disorders (retention of testes in one or both sides, microphallus, bifid scrotum) or with malformation of other organs. In some cases syndromic forms may also occur. Genetic factors play a crucial role in the occurrence of early developmental defect, but endocrine and environmental factors may also be important in the aetiology of hypospadias. It may be associated with various sex and autosomal chromosomal abnormalities. Monogenic and chromosomal causes of hypospadias accounts for about in 30% of all cases, while genetic factors remain unknown in 70% of cases. The authors summarize the development of the male external genitalia, the prevalence and possible causes of hypospadias. They propose that better understanding of the pathogenesis of the disease may contribute to the prevention and decreased prevalence of the disease.

Gestational obesity and risk of hypospadias in male newborns. A population based case-control study

Aim: Hypospadias is a relatively common male genital anomaly that may be caused by anomalies in maternal hormone levels and/or blood glucose levels as well as nutritional deficiencies. Maternal obesity, which increases the risk of diabetes, may alter hormone levels, and may thereby be associated with risk of hypospadias. This study was designed to determine the impact of these risk factors on hypospadias in boys born at the largest tertiary hospital in Qatar. Methods: This population-based, case-control study used linked birth-hospital discharge data from Hamad General Hospital in Doha, Qatar, from January 2007 to December 2012. Boys with hypospadias were identified, and risk factors were determined. Adjusted odds ratios (aORs) and 95% confidence intervals (CIs) for hypospadias were estimated by multivariable logistic regression models adjusted for maternal age, parity and body mass index (BMI). Results: Seventy-three newborn male infants with hypospadias and 100 without the condition, with birth weight above 2500 g, were compared. Maternal obesity (BMI ≥ 30 kg/m2) was not associated with risk of hypospadias (aOR 1.07; 95% 0.95 - 1.21), and no trend in risk with increasing maternal BMI was observed. Conclusion: Although the causes of male genital malformation are multifactorial, the present data do not support the hypothesis that gestational maternal obesity is a cause of hypospadias in male infants.

Individual Variation of the Genetic Response to Bisphenol A in Human Foreskin Fibroblast Cells Derived from Cryptorchidism and Hypospadias Patients

Background/PurposeWe hypothesized that polymorphic differences among individuals might cause variations in the effect that environmental endocrine disruptors (EEDs) have on male genital malformations (MGMs). In this study, individual variation in the genetic response to low-dose bisphenol A (BPA) was investigated in human foreskin fibroblast cells (hFFCs) derived from child cryptorchidism (CO) and hypospadias (HS) patients.Methodology/Principal FindingshFFCs were collected from control children without MGMs (n = 5) and child CO and HS patients (n = 8 and 21, respectively). BPA exposure (10 nM) was found to inhibit matrix metalloproteinase-11 (MMP11) expression in the HS group (0.74-fold, P = 0.0034) but not in the control group (0.93-fold, P = 0.84) and CO group (0.94-fold, P = 0.70). Significantly lower levels of MMP11 expression were observed in the HS group compared with the control group (0.80-fold, P = 0.0088) and CO group (0.79-fold, P = 0.039) in response to 10 nM BPA. The effect of single-nucleotide polymorphism rs5000770 (G>A), located within the aryl hydrocarbon receptor nuclear translocator 2 (ARNT2) locus, on individual sensitivity to low-dose BPA was investigated in the HS group. A significant difference in neurotensin receptor 1 (NTSR1) expression in response to 10 nM BPA was observed between AA and AG/GG groups (n = 6 and 15, respectively. P = 0.031). However, no significant difference in ARNT2 expression was observed (P = 0.18).Conclusions/SignificanceThis study advances our understanding of the specificity of low-dose BPA effects on human reproductive health. Our results suggest that genetic variability among individuals affects susceptibility to the effects of EEDs exposure as a potential cause of HS.

X-linked congenital adrenal hypoplasia associated with hypospadias in an Egyptian baby: a case report

IntroductionX-linked congenital adrenal hypoplasia is a rare developmental disorder of the human adrenal cortex and is caused by deletion or mutation of the dosage-sensitive sex reversal adrenal hypoplasia congenita critical region of the X chromosome, gene 1 (DAX-1) gene. Most affected children present with failure to thrive, salt wasting and hypoglycemic convulsions in the first months of life. Hypospadias affects approximately one in 250 live male births. Mutations in the mastermind-like domain-containing 1 (MAMLD1) gene have been implicated as one of the causes of hypospadias in children. To the best of our knowledge, an association between congenital adrenal hypoplasia due to a DAX-1 mutation and hypospadias due to mutation of the MAMLD1 gene has not previously been reported in the literature.Case presentationA 35-day-old male Egyptian baby was referred to our institution for the evaluation of a two-week history of recurrent vomiting associated with electrolyte imbalance. On examination, our patient was found to have hypotension and dehydration. A genital examination showed distal penile hypospadias with chordee and normal testes. He had hyponatremia, hyperkalemia, hypoglycemia and metabolic acidosis. Endocrinological investigations revealed low levels of cortisol, 17-hydroxyprogesterone and aldosterone, with a high level of adrenocorticotrophic hormone. A provisional diagnosis of congenital adrenal hypoplasia associated with hypospadias was made. A molecular genetics study confirmed the diagnosis of X-linked congenital adrenal hypoplasia due to DAX-1 mutations and hypospadias due to MAMLD1 mutation. He was started on hydrocortisone and fludrocortisone treatment. After three weeks of treatment, his symptoms improved and his blood sugar, sodium, potassium and cortisol levels normalized.ConclusionsWe report the case of an Egyptian baby with an association of congenital adrenal hypoplasia due to DAX-1 mutation and hypospadias due to MAMLD1 mutation. Early diagnosis of this association and determining its optimal treatment are vital in helping to avoid its fatal course.

The association between some endocrine disruptors and hypospadias in biological samples

Hypospadias is a birth defect found in boys in which the urinary tract opening is not at the tip of the penis. The etiology of hypospadias is still unidentified, but endocrine disruptors are considered as one possible cause of hypospadias. In this study, target endocrine disruptor compounds were established for an assay. The target compounds included 5 phthalates (di-(2-ethylhexyl)phthalate (DEHP), di-n-butyl-phthalate (DBP), mono-(2- ethylhexyl) phthalate (MEHP), mono-n-butyl-phthalate (MBP) and phthalic acid (PA)), 2 alkylphenols (n-nonylphenol (n-NP) and t-octylphenol (t-OP)) and bisphenol A. The association between these 8 endocrine disruptors and hypospadias was studied. The levels of endocrine disruptors in the urine and plasma of a control group were compared with those of a patient group. DEHP (P = 0.006) and n-NP (P = 7.26e-6) in the urine samples and PA (P = 0.009) and BPA (P = 7.22e-10) in the plasma samples showed a significant association with hypospadias. The levels of endocrine disruptors in the urine and plasma of the mothers were also compared to those of the patients to investigate the metastasis of the endocrine disruptors from the mother. These levels did not, however, show a relationship with hypospadias (R2 = 0.001–0.563).

717 SEXUAL DIFFERENTIATION IN MALE MOUSE FROM DAY 0 TO 10 A DETAILED AND NOVEL MORPHOMETRIC DESCRIPTION

You have accessJournal of UrologyPediatrics: Basic Research1 Apr 2012717 SEXUAL DIFFERENTIATION IN MALE MOUSE FROM DAY 0 TO 10 A DETAILED AND NOVEL MORPHOMETRIC DESCRIPTION Bruce Schlomer, Max Ferretti, Esequiel Rodriguez, Gerald Cunha, and Laurence Baskin Bruce SchlomerBruce Schlomer San Francisco, CA More articles by this author , Max FerrettiMax Ferretti San Francisco, CA More articles by this author , Esequiel RodriguezEsequiel Rodriguez San Francisco, CA More articles by this author , Gerald CunhaGerald Cunha San Francisco, CA More articles by this author , and Laurence BaskinLaurence Baskin San Francisco, CA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.802AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The external genitalia of mice at birth are profoundly undifferentiated and sexual differentiation continues after birth. The time period of sexual differentiation with the most dramatic changes in morphology is day 0 to 10. This period of sexual differentiation in mice corresponds to the embryologic time in humans where an abnormality in sexual differentiation is thought to lead to external genitalia abnormalities such as hypospadias. To study potential genetic and environmental causes of hypospadias in mice, accurate and detailed morphometric measurements and morphological descriptions of this period are needed. METHODS The external genitalia of male wild type CD-1 mice were serially sectioned transversely at 7 μm for histologic staining. Age groups of 0-1 day, 2-3 day, 4-5 day, and 10 day mice were included. Morphometric measurements on well aligned sections were obtained of the male urogenital mating protuberance (MUMP), foreskin, urethra, and other distal penile structures. Anatomical three-dimensional reconstructions (3DR) were created using the serial sections with the BioVis3D software. RESULTS The detailed morphometric measurements obtained are shown in Figure 1. The morphology of the distal penis changes dramatically between 0 and 10 days and is partly characterized by the development of paired extensions of the lateral and ventral erectile structures and a distal paired extension of the MUMP (Figure 2). The anatomical and relational measurements are presented in Table 1. Table 1. Morphometric measurements by age categories Morphometric measurement (μm) Day 0-1 (n=10) Day 2-3 (n=13) Day 4-5 (n=9) Day 10 (n=8) a) Distal extension of MUMP1 3.5(6.2)A 26.4(13.2)B 29.6(13.1)B 72.6(35.4)C b) Lateral erectile structure extension1 2.8(4.2)A 31.8(24.0)B 52.1(20.0)B 169.8(55.8)C c) Ventral erectile structure extension1 17.2(8.6)A 52.5(33.0)B 103.4(18.2)C 172.4(22.0)D d) MUMP tip to ventral erectile structure extension1 207.2(34.0)A 237.7(50.8)A 252.0(23.2)A 378.9(113.8)B e) MUMP tip to lateral erectile structure extension1 126.4(23.0)A 168.8(31.2)AB 194.4(30.9)B 250.7(53.2)C f) MUMP tip to penile urethra 256.2(47.8)A 303.7(49.4)AB 370.2(24.6)B 576.6(123.9)C g) MUMP tip to CCG 180.6(31.3)A 234.8(49.3)B 311.9(33.6)C 454.1(58.3)D h) CCG tip to glans penis without epithelial folds 180.6(55.1)A 209.5(85.5)AB 172.7(45.0)A 308.9(151.3)B i) MUMP tip to glans penis without epithelial folds 361.9(48.7)A 459.3(73.8)B 512.6(25.4)B 791.0(144.9)C MUMP tip to ventral foreskin fusion2 119.7(60.9)A -6.4(71.4)B -92.6(68.5)C -381.5(66.5)D Data presented in μm as mean (standard deviation). Abbreviations: CCG = corpus cavernosum glandis, MUMP = male urogenital mating protuberance. A-D Different superscripts within rows indicate a significant difference between age groups with analysis of variance test with Bonferroni correction (p<0.05). 1 Paired structures used 2 measurements per animal. 2 A negative value signifies ventral foreskin fusion occurs distal to tip of MUMP. CONCLUSIONS Dramatic changes in mouse penile morphology occur from day 0 to 10. We have described these changes using novel, accurate, and relevant morphometric measurements and 3DR. These measurements and morphologic descriptions will be critical in future studies on potential causes of hypospadias. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e294-e295 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Bruce Schlomer San Francisco, CA More articles by this author Max Ferretti San Francisco, CA More articles by this author Esequiel Rodriguez San Francisco, CA More articles by this author Gerald Cunha San Francisco, CA More articles by this author Laurence Baskin San Francisco, CA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

392 Quantitative measurement of the androgen receptor in prepuces of boys with and without hypospadias

Abstract Purpose Identifiable causes of hypospadias, a midline fusion defect of the male ventral urethra, are still infrequently well-known. The aim of this study was to quantify the androgen receptor mRNA and androgen receptor protein in prepuces of boys with and without hypospadias. Material and Methods Forty prepuce specimens of circumcised boys, aged between 12 and 14 months, with (n=20) and without hypospadias (n=20), were investigated. Immediately after surgery all probes were fixed in formaldehyde and small parts were deep frozen in liquid nitrogen at -80° C. The total RNA of the specimens was isolated and cDNA was written. With the aid of real time PCR the amount of present androgen receptor mRNA was measured. For quantification of present androgen receptor protein, Western Blot (Bradford method) and immunohistochemistry for androgen receptor using standardized automated procedures (Discovery XT, Ventana) were performed. Statistical analyses using the the Kolmogorov-Smirnov and Mann-Whitney U-test were done. Results Our observations show that the androgen receptor mRNA is significantly increased in the prepuce of hypospadiac boys compared to specimens of boys with phimosis.(p Conclusions Our results provide evidence that the different expression of androgen receptor mRNA indicates the extent of a defect androgen receptor signalling in boys with hypospadias.