This provided a quantitative basis for comparing each age-matched alcohol/control pair and also for comparing the total mean density counts for all alcohol-exposed brains combined with the total mean density counts for all control brains combined.
To determine if differences between the experimental and control brains were statistically significant, an unpaired students t test with Welch correction, where appropriate, was performed comparing the total mean (± SEM) density counts for the alcohol brains combined (n = 5) versus the total mean (± SEM) density counts for the control brains combined (n = 4).
We compared individual density counts for all regions of interest and, to make our results comparable with the reference range in children (6), we computed cerebro-cerebellar indexes by dividing density counts of particular regions by those of the cerebellum.
Tests of normality were undertaken using the Jarque-Bera test (JB test) on the mean SNP density counts for all regions as described in Table 1 for each chromosome and the null hypothesis of normality could not be rejected[ 13].
RING1B, SUZ12 and EZH2 interval sets were annotated with normalized read density counts for untreated and tamoxifen-treated samples from both Ring1a −/− Ring1b fl/fl and Kdm2b fl/fl experiments, and the untreated to treated fold change was calculated.
Dividing the total volume into the total profile count yielded a density count (profiles per mm) for each alcohol-exposed and each control brain.
Smears were initially graded for the presence/absence of Plasmodium and approximate parasite density, then parasite density counts per 200 leukocytes were made from positive smears.
We used data from Sinden et al. (2007) because density counts are available for individual mosquitoes.
A random subsample was used for density counts (only adult females were taken into consideration).
For cell density counting, digitized images, acquired using a conventional microscope and a charge-coupled device camera, were used.
For RGC density counts, rats were deeply anesthetized and perfused transcardially with 4% paraformaldehyde (PFA) in 0.1 M phosphate buffer following which both eyes were immediately enucleated.
