Defective splicing of the RB1 transcript is the dominant cause of retinoblastomas.

Defective splicing is a common cause of genetic diseases. On average, 13.4% of all hereditary disease alleles are classified as splicing mutations with most mapping to the critical GT or AG nucleotides within the 5' and 3' splice sites. However, splicing mutations are underreported and the fraction of splicing mutations that compose all disease alleles varies greatly across disease gene. For example, there is a great excess (46%; ~threefold) of hereditary disease alleles that map to splice sites in RB1 that cause retinoblastoma. Furthermore, mutations in the exons and deeper intronic position may also affect splicing. We recently developed a high-throughput method that assays reported disease mutations for their ability to disrupt pre-mRNA splicing. Surprisingly, 27% of RB1-coding mutations tested also disrupt splicing. High-throughput in vitro spliceosomal assembly assay reveals heterogeneity in which stage of spliceosomal assembly is affected by splicing mutations. 58% of exonic splicing mutations were primarily blocked at the A complex in transition to the B complex and 33% were blocked at the B complex. Several mutants appear to reduce more than one step in the assembly. As RB1 splicing mutants are enriched in retinoblastoma disease alleles, additional priority should be allocated to this class of allele while interpreting clinical sequencing experiments. Analysis of the spectrum of RB1 variants observed in 60,706 exomes identifies 197 variants that have enough potential to disrupt splicing to warrant further consideration.

Lack of compensation of energy intake explains the success of alternate day feeding to produce weight loss.

BACKGROUND AND AIMS: This study was designed to obtain daily weighed food intake of participants engaged in Alternate Day Feeding (ADF). Prior ADF studies have used self-reported food intake, a method that has received criticism for its limited accuracy. SUBJECTS AND METHODS: Forty-nine university students received academic credit for participating in the study. Following a 10-day baseline period, participants underwent ADF for the next 8 days. Restricted daily intake to ∼ 75% of baseline food intake levels was followed by ad libitum intake on alternate days. Food intake was weighed before and after each meal. Daily body weight was also tracked. INTERVENTION: After the baseline period, participants underwent 8 days of ADF during which they consumed ∼ 75% of baseline energy intake by weight followed by ad libitum intake on alternate days. The trial concluded with 2 additional days of ad libitum feeding, for a total study duration of 10 days. RESULTS: Daily food intake was constant during the baseline period (slope = -0.93 g/d, p = 0.56), and did not differ significantly (995 g (95% CI [752, 1198]) from the total consumed on ad libitum ADF days (951 g (95% CI [777, 1227]). Intake on ad libitum days did not show a trend to increase during the intervention. Body weight declined significantly during ADF. CONCLUSIONS: ADF produces significant weight loss because food intake does not increase on ad libitum feeding days to compensate for reduced intake on restricted energy days. Data are consistent with prior work that suggests humans do not fully compensate for imposed deficits in energy intake.