Scatter-plot of the neighborhood SES and alpha-diversity.

<p>Figs depict associations between neighborhood SES and alpha-diversity in biopsies excised from sigmoid mucosa (upper plots) and feces collected from sigmoid lumen (lower plots). Shannon and Chao1 indices are different metrics for calculating alpha-diversity.</p

Map depicting the neighborhood SES and alpha-diversity.

<p>Map of greater Chicago region, illustrating associations between neighborhood SES and alpha-diversity in biopsies excised from sigmoid mucosa. Depicted are neighborhood SES (left panel), and two metrics of diversity, the Shannon (middle panel) and Chao1 (right panel) indices. For each variable, the sample was divided into quartiles, and neighborhoods were colored as follows: Red = lower 25% of sample; Yellow = 26–50%; Green = 51–75%; Blue = upper 25%. Geospatial data sources: Urban place area from TIGER/Line US Census 2010; Illinois and Wisconsin zip codes from TIGER/Line US Census 2010; Water area from TIGER/Line US Census 2010; Interstate lines from TIGER/Line US Census 2010.</p

Methamphetamine self-administration in non-Tg and Tg rats.

<p><b>(A)</b> Non-Tg and Tg rats similarly performed the methamphetamine self-administration task. Shown is the average number of nose-pokes per 2h session across 21 days by non-Tg and Tg rats. There were a greater number of active nose-pokes <i>vs</i> inactive nose-pokes for both genotypes. Non-Tg and Tg rats did not differ in the <b>(B)</b> average number of methamphetamine infusions per session, or <b>(C)</b> total methamphetamine intake (Student’s <i>t</i>-test, * <i>p</i><0.05; ** <i>p</i><0.01).</p

Scatter-plot of neighborhood SES and bacterial genera.

<p>Figs depict associations between neighborhood SES and <i>Prevotella</i> to <i>Bacteroides</i> ratio (upper panel), as well as relative abundance of <i>Prevotella</i> (middle panel) and <i>Bacteroides</i> (lower panel). Specimens are biopsies excised from sigmoid mucosa.</p

ZO-1 immunofluorescence.

<p><b>(A)</b> Representative photomicrographs of ZO-1 immunofluorescence (red) in the distal colon of saline-yoked <b>(top)</b> and methamphetamine self-administering <b>(bottom)</b> non-Tg <b>(left)</b> and Tg <b>(right)</b> rats. Sections were counterstained with DAPI (blue) to identify crypts. Saline-yoked non-Tg exhibited continuous ZO-1 immunofluorescence around the epithelial cells within the crypts <b>(top, left)</b>; whereas non-Tg rats that self-administered methamphetamine had discontinuous ZO-1 immunofluorescence <b>(bottom, left).</b> Saline-yoked non-Tg and Tg rats that self-administered methamphetamine also had discontinuous ZO-1 immunofluorescence <b>(right)</b>. White bar = 25μm. There was no group difference in intensity of ZO-1 immunofluorescence. <b>(B)</b> A scale from 0–4 was used to score ZO-1 immunofluorescence (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0190078#sec002" target="_blank">Methods</a>). As indicated by horizontal lines above the bars, scores differed significantly between saline-yoked (open bars) non-Tg and saline-yoked Tg rats, and saline-yoked non-Tg and methamphetamine self-administering (filled bars) non-Tg rats. Mann-Whitney <i>U</i>-tests corrected for multiple comparisons (* <i>p</i><0.025).</p

Colon permeability.

<p>Tg rats, regardless of methamphetamine self-administration, exhibited a significantly more permeable colon, as assessed the by the sucralose/lactulose ratio, compared to non-Tg saline yoked-rats (<i>p</i> = 0.001). Methamphetamine self-administration did not result in a significant difference (<i>p</i> = 0.25), and there was no interaction between genotype and meth (<i>p</i> = 0.96). Horizontal lines indicate planned contrasts that showed differences with a Newman-Keuls <i>post hoc</i> test (* <i>p</i><0.05).</p

Claudin-1 protein expression.

<p><b>(A) I</b>mmunoblot illustrating claudin-1 (23kDa) in the distal colon from 8 different samples (s, saline; m, meth). Arrow indicates molecular weight at 25kDa. <b>(B)</b> Overall, Tg rats had a significantly less claudin-1 compared to non-Tg rats (<i>p</i> = 0.01). Methamphetamine (meth) self-administration did not result in a significant difference (<i>p</i> = 0.34), and there was no interaction between genotype and meth (<i>p</i> = 0.70). Horizontal lines indicate an overall effect of genotype from the two-way ANOVA (* <i>p</i><0.05).</p

Environmental disruption of circadian organization increases intestinal permeability.

<p>Young adult (7–9 week) male C57BL/6J mice underwent three months of weekly 12 hour phase shifts of the LD cycle (S) or maintained on a constant 12∶12 LD cycle (NS). Shifted animals had significantly greater intestinal permeability. ***, p<0.001, Student's <i>t</i>-test.</p

Cytoplasmic tight junction protein occludin levels in the proximal colon are significantly elevated in <i>Clock^Δ19/Δ19</i> mutant mice.

<p>(<b>A</b>) <i>Clock^Δ19/Δ19</i> mutant mice on the control diet (CD) had significantly elevated levels of cytoplasmic occludin in the proximal colon compared to wild-type littermates on the control diet (WTD), with an overall significant effect of genotype (p<0.05). *p<0.05, two-way ANOVA followed by <i>post-hoc</i> Fisher's LSD Multiple-Comparison test. (<b>B</b>) <i>Occludin</i> mRNA was not significantly altered.</p

Genetic disruption of circadian organization promotes alcohol-induced hepatic steatosis.

<p>(<b>A</b>) Alcohol-fed <i>Clock^Δ19/Δ19</i> mutant mice (CA) had a significantly greater liver/body weight ratio than control-fed mutants (CD), with an overall significant effect of diet (p<0.01). **p<0.01, two-way ANOVA followed by <i>post-hoc</i> Tukey-Kramer Multiple-Comparison test. (<b>B</b>) Alcohol-fed <i>Clock^Δ19/Δ19</i> mutant mice (CA) exhibited significant steatosis compared to all other experimental groups, with a significant overall effect of genotype (p<0.001), diet (p<0.001), as well as a significant interaction (p<0.05). *p<0.05, **p<0.01; two-way ANOVA followed by <i>post-hoc</i> Tukey-Kramer Multiple-Comparison test. No between group differences were observed for histological assessment of lobular inflammation (<b>C</b>) or ballooning degeneration (<b>D</b>). Histological assessments were performed by a blinded gastrointestinal pathologist. Steatosis score was based on % hepatocyte involvement: 0 = <5%, 1 = 5–33%, 2 = 34–66%, 3 = >67%. Lobular inflammation score was based on the number of foci/200× field: 0 = none, 1 = 1, 2 = 2–4, 3 = >4. Ballooning degeneration score was based on the presence and frequency of ballooned cells: 0 = none, 1 = few, 2 = prominent/many.</p