Differentially methylated loci distinguish ovarian carcinoma histological types: evaluation of a DNA methylation assay in FFPE tissue.

Epigenomic markers can identify tumor subtypes, but few platforms can accommodate formalin-fixed paraffin-embedded (FFPE) tumor tissue. We tested different amounts of bisulfite-converted (bs) DNA from six FFPE ovarian carcinomas (OC) of serous, endometrioid, and clear cell histologies and two HapMap constitutional genomes to evaluate the performance of the GoldenGate methylation assay. Methylation status at each 1,505 CpG site was expressed as ß-values. Comparing 400 ng versus 250 ng bsDNA, reproducibility of the assay ranged from Spearman r(2) = 0.41 to 0.90, indicating that ß-values obtained with a lower DNA amount did not always correlate well with the higher amount. Average methylation for the six samples was higher using 250 ng (ß-value = 0.45, SD = 0.29) than with 400 ng (ß-value = 0.36, SD = 0.32). Reproducibility between duplicate HapMap samples (r(2) = 0.76 to 0.92) was also variable. Using 400 ng input bsDNA, THBS2 and ERG were differentially methylated across all histologic types and between endometrioid and clear cell types at <0.1% false discovery rate. Methylation did not always correlate with gene expression (r(2) = -0.70 to 0.15). We found that lower bsDNA overestimates methylation, and, using higher bsDNA amounts, we confirmed a previous report of higher methylation of THBS2 in clear cell OC, which could provide new insight into biological pathways that distinguish OC histological types.

Early childhood outcomes of infants born with gastroschisis.

PURPOSE: To describe neonatal and childhood outcomes of a contemporary cohort of infants with gastroschisis. METHODS: Observational, single center, inception cohort of children born with gastroschisis from January 2005 to December 2008. RESULTS: Of 63 infants, 61 survived to hospital discharge and 39 were seen for follow-up. Complications included sepsis (37%), necrotizing enterocolitis (10%), parenteral nutrition related cholestasis (25%), and short bowel syndrome (13%). Of survivors, 5% had visual impairment and 10% had hearing loss. No child tested had mental delay or cerebral palsy. Early gestational age predicted death or disability (OR 0.60, 95% CI 0.38, 0.96; p=0.033). There was a high incidence of prescription medications for presumed gastroesophageal reflux (90%). Some infants continued to require tube feeds (15%). There were improvements in longitudinal growth reflected in increasing z-scores. CONCLUSIONS: Although children with gastroschisis are at risk for disability, childhood outcomes are encouraging.

Teaching child development to medical students.

PURPOSE: Several published strategies on teaching the screening of normal child development were integrated into a small group learning experience for second-year medical students to address practical and logistical problems of approaches used individually. This study examines the effectiveness of this integrated approach using student evaluations. METHOD: A total of 191 second-year university medical and dental students were invited to participate. Well-described learning objectives, the Ages and Stages Questionnaire (ASQ), live parent-child dyads and video backup were used. Students rotated through three small group stations. Feedback was provided using a Likert scale (from 1, low, to 5, high) and written comments. Consent was obtained. Live parent-child dyads versus video clip groups were analysed by averaging overall scores. Generalised estimating equation (GEE) analysis in stata (Stata Corporation, College Station, Texas) was used for comparing the two groups. RESULTS: A total of 178 students (93%) agreed to participate and filled out the evaluation forms. The overall score on the Likert scale was 4.6 (range 4-5). On two occasions video clips were substituted for live parent-child dyad presentations in one of the three stations. These students (n=43, rating 4.61/5) rated their experience as comparable with those who had three live family stations (n=135, rating 4.56/5). Student comments were grouped into broad themes, with most being positive about their learning experience. CONCLUSIONS: This integrated approach is highly acceptable. Video clip usage, live dyads, clear written objectives and use of a standardised screening tool preserved the interaction and immediacy of a clinical encounter, while maintaining consistency in content.