Prediction of collection time in ASD children with normal sleep patterns (n = 63).

<p>Prediction of collection time in ASD children with normal sleep patterns (n = 63).</p

List of 11 microbes most related to collection time.

<p>List of 11 microbes most related to collection time.</p

Accurate prediction of collection time from salivary miRNA levels.

<p><b>(A)</b> 11 of the total 61 identified miRNA predictors and their accuracy of prediction for sample set 3. <b>(B)</b> Sine transformed values of the average expression of 1 of the 61 CircaMiRs (miR-199b-3p) for the subjects in sample set 3 (collected at various times across 2 days).</p

Salivary miRNA levels change across time.

<p><b>(A)</b> Heat map clustering of expression data for the 61 miRNAs changed according to collection time in sample set 1. This set consisted of 24 samples from 4 subjects across 3 days of sampling (days 1, 3, 7) at a frequency of 2 times/day (9 am, 9 pm). <b>(B)</b> Heat map clustering of expression data for the 61 miRNAs changed according to collection time in sample set 2. This set consisted of 48 samples from 3 subjects obtained across 4 days of sampling (days 1, 5, 10, 15) at a frequency of 4 times/day (9 am, 1:30 pm, 5:30 pm, 9 pm).</p

Correlations of salivary miRNA and microbial model terms across sample sets.

<p>Correlations of salivary miRNA and microbial model terms across sample sets.</p

Changes in functional microbiome expression across time.

<p><b>(A)</b> The hierarchical heat map displays average abundance values for microbial RNAs representing 22 KEGG/COG metabolic pathways, that displayed nominal differences (p<0.05) in expression across 4 time periods (7–9 AM, 10 AM-2 PM, 3–6 PM, 7–10 PM). The dendrogram (y-axis) represents inter-relatedness of KEGG/COG pathway activity measured by Pearson distance metric across the 120 samples. Red denotes relative increased abundance of KEGG/COG transcripts, while blue denotes relative decrease in related transcripts. Chi-square and raw p-values (Kruskal-Wallis ANOVA) are displayed for each of the 22 pathways. <b>(B)</b> A partial least squares discriminant analysis utilizing mean abundance levels for all 202 KEGG/COG metabolic pathways with microbial RNA mappings is displayed for the four collection time periods. Note that global metabolic activity in these 202 pathways achieves partial separation of the four time periods, while accounting for 20.6% of the variance in the dataset.</p

Relationship between oscillating salivary RNA levels and timing of daily activities.

<p>Relationship between oscillating salivary RNA levels and timing of daily activities.</p

Physiologic pathways over-represented by mRNA targets of the 11 CircaMiRs.

<p>Physiologic pathways over-represented by mRNA targets of the 11 CircaMiRs.</p

Transcripts targeted by CircaMiRs with associated expression levels across time.

<p>Transcripts targeted by CircaMiRs with associated expression levels across time.</p

Quantitative relationships between CircaMiRs and CircaMicrobes.

<p>A Pearson’s correlation analysis was used to compare expression of 11 CircaMiRs and 11 CircaMicrobes. The 22 features are sorted by a complete clustering algorithm, and the hierarchical tree indicates similarity in expression pattern across samples. Blue indicates strong inverse relationships while red indicates strong direct relationships.</p