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Hirschsprung disease (HSCR) is a major cause of chronic constipation in children. HSCR can be caused by germline mutations in RETand EDNRB.Defining causality of the mutations identified is difficult and almost exclusively based on in silicopredictions. Therefore, the reported frequency of pathogenic mutations might be overestimated. We combined mutation analysis with functional assays to determine the frequencies of proven pathogenic RETand EDNRBmutations in HSCR. We sequenced RETand EDNRBin 57 HSCR patients. The identified RET-coding variants were introduced into RETconstructs 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 EDNRBvariants. Western blotting showed that three coding variants p.(Pr270Leu), p.(Ala756Val) and p.(Tyr1062Cys) resulted in lower activation of RET. Moreover, only two RETvariants (p.(Ala756Val) and p.(Tyr1062Cys)) resulted in reduced ERK activation. Splice-site assays on c.1880-11A>G 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.

Diarrhea remains a major cause of child morbidity and mortality in low- and middle-income countries. Reliable data on the economic burden of diarrhea is required to support the selection of appropriate health intervention programs. This study aimed to estimate the costs of acute diarrhea in children under five years of age in Indonesia, a large middle-income country with a substantial diarrheal burden.

Rotavirus strains in stool specimens from 111 children aged 3–24 months admitted to hospital in Yogyakarta, Indonesia for treatment of acute diarrhoea were serotyped using VP7serotype specific monoclonal antibodies in a double sandwich enzyme immunoassay. A serotype could be assigned to 59 of 111 specimens (53%). Inability to assign a serotype to 47% of specimens was probably due to loss of the outer capsid during transport of specimens from Indonesia to Australia. All four major human rotavirus serotypes were detected during the 15 month survey from June 1978 to August 1979, including one serotype 1, 5 serotype 2, 31 serotype 3, and 21 serotype 4 strains. One additional strain reacted with serotype 3 and 4 Mabs. Serotype 3 strains showed intratypic variation.

Nine different electropherotypes of rotaviruses occurred among 85 children with rotavirus diarrhea in Yogyakarta, Indonesia. Eighty percent of the electropherotypes had a “long” RNA pattern characteristic of human subgroup 2.