No abstract

Hirschsprung disease (HSCR) is a major cause of chronic constipation in children. HSCR can be caused by germline mutations in RET and EDNRB. Defining causality of the mutations identified is difficult and almost exclusively based on in silico predictions. Therefore, the reported frequency of pathogenic mutations might be overestimated. We combined mutation analysis with functional assays to determine the frequencies of proven pathogenic RET and EDNRB mutations in HSCR. We sequenced RET and EDNRB in 57 HSCR patients. The identified RET-coding variants were introduced into RET constructs and these were transfected into HEK293 cells to determine RET phosphorylation and activation via ERK. An exon trap experiment was performed to check a possible splice-site mutation. We identified eight rare RET-coding variants, one possible splice-site variant, but no rare EDNRB variants. Western blotting showed that three coding variants p. (Pr270Leu) (Tyr1062Cys)) resulted in reduced ERK activation. Splice-site assays on c.1880-11A4G could not confirm its pathogenicity. Our data suggest that indeed almost half of the identified rare variants are proven pathogenic and that, hence, functional studies are essential for proper genetic counseling.

We report a Japanese Becker muscular dystrophy (BMD) patient with occasional myalgia and cramps during normal activity that developed at the age of 28 months. His family history was negative for neuromuscular diseases. Muscle biopsy analyses, including dystrophin immunostaining, disclosed no clinically relevant findings. The diagnosis of BMD was initially made at the age of 10 years, when indications of persistent high serum levels of CK prompted us to screen deletions in the dystrophin gene by amplification of 19 deletion-prone exons from the genomic DNA by the polymerase chain reaction (PCR). Among the exons examined, exons 13 and 17 were deleted. To clarify the size of the deletion, the dystrophin transcript was analyzed by reverse transcription PCR. The determined nucleotide sequence of the amplified product encompassing exons 10 to 20 disclosed that the entire segment corresponding to exons 13 to 18 (810 bp) was absent, a deletion that would be expected to cause the production of a dystrophin protein lacking 270 amino acids from the rod domain. This result indicates that occasional myalgia and cramps could be early clinical manifestations of mild BMD, especially in patients who have a deletion in the rod domain, and that deletion screening of the dystrophin gene might be the only reliable method to diagnose such cases.

The first cases of SMEI and SMEB are reported in South-East Asian populations. Two novel SCN1A mutations are also identified in these patients, p.V1612I and p.C1756G, which may lead to neuronal excitability or convulsions.

Risk Factors of Hypospadias among Children in Dr. Sardjito General Hospital F. Jerry Tangkudung, S. Yudha Patria, Eggi ArguniBackground. It has been reported increased incidence of hypospadias. There are many risk factors for hypospadias, one of which is exposure to pesticides. Other risk factors are obstetric and neonatal period histories. Objective. To determine the risk factors for hypospadias in children. Method. We conducted a case-control study in Dr. Sardjito General Hospital using hospital medical records from 2008 through 2011. The research sample consisted of 120 subjects.The data were analyzed using chi-square and logistic regression. Results. A total of 120 children were enrolled, of whom 50% (60) had hypospadias and 50% (60) had no hypospadias. Children with mother's age of more than 35 years had a greater risk for having hypospadia compared to those whose mother's age of less than 35 years (adjusted odds ratio 4.17 (CI 95% 1.12 -15.49; p=0.033). Low birth weight, maternal vegetarian diet during pregnancy, maternal hypertension during pregnancy and rice field environment had no association with the development of hypospadias. Conclusion. Mother age of more than 35 years old during pregnancy is a risk factor for the development of hypospadia in children. Sari Pediatri 2016;17(5):396-400.