Colon-specific reduction/reversal of polyps.

<p>Initiation of treatment with ATB-346 (14.5 mg/kg once daily for 14 days) at week 12 had no effect on the small intestinal polyp score (assessed at week 14), but markedly reduced polyp formation in the colon (**p<0.01).</p

Dose-related preventative effects of oral treatment with ATB-346 or naproxen on intestinal polyp formation in APC^Min/+ mice.

<p>Groups of at least 6 mice each were treated daily with vehicle, naproxen (1 or 10 mg/kg), or equimolar doses of ATB-346 or of the hydrogen sulfide-releasing moiety of ATB-346 (TBZ; 4-hydroxythiobenzamide). Treatments were started at week 6 of life, and the numbers and areas (in mm²) of polyps in the small intestine and colon were blindly assessed at week 14 of life (panels A, B and C show examples; arrows indicate polyps). Panel D shows the ‘total polyp score’ data (the sum of the areas of all polyps in each mouse). Data are shown as the mean ± SEM. **p<0.01, ***p<0.001 versus the vehicle-treated group (one-way ANOVA and Dunnett’s test). ^Ψp<0.05 versus the group treated with naproxen at the same dose (Student’s t-test).</p

Summary of transcriptomics analysis.

<p>Genes altered by >1.5-fold in the colon of APC^Min/+ mice as compared to wild-type mice, and the effects of daily treatment for one week with naproxen or ATB-346. Fold-change values represent direction of change from the first listed condition to the second. During the 6^th week of age, mice were treated with vehicle, naproxen at a dose of 10 mg/kg, or ATB-346 at an equimolar dose. Tissue samples were harvested at the start of the 7^th week of age. *p<0.05 by one-way ANOVA with Student-Neuman-Keuls post-hoc test and Storey’s bootstrapping multiple comparison correction. n = 6-8/group.</p

Treatment with ATB-346 reduced intestinal β-catenin and cMyc levels in APC^Min/+ mice.

<p>Expression of both β-catenin (panel A) and cMyc (panel B) were significantly elevated in colonic tissue from APC^Min/+ mice as compared to wild-type mice. Treatment of APC^Min/+ mice with ATB-346 (14.5 mg/kg) significantly reduced β-catenin and cMyc expression to levels comparable to those in wild-type mice, while naproxen treatment (10 mg/kg) reduced cMyc but not β-catenin. Tissue samples were collected from the mice at 14 weeks of age, while drug treatment occurred in the 6^th and 7^th weeks of age. Each bar represents the mean ± SEM for at least 4 mice (p<0.05 vs. wild-type mice; ^ψp<0.05 vs. wild type. *p<0.05 vs. naproxen-treated).</p

Polyp formation in the small intestine and colon of APC^Min+ mice at 12 weeks of age.

<p>Panel A shows ileum from a wild-type (C57Bl/6) mouse, while panel B shows ileum from an APC^Min+ mouse (with numerous polyps visible in the latter). Panel C shows the Polyp Score for 12 APC^Min+ mice, illustrating the regional variation through the small intestine and colon. Each line represents the results from one mouse. The ileum exhibited the greatest polyp score, while the colon exhibited the lowest.</p

Targeted metabolic profiling of amines and methylarginines in human plasma from healthy donors and patients with sepsis.

<p>Amine and methylarginine levels were measured using <b>(A)</b> UHPLC-MS/MS, LC-MS/MS and/or ELISA in the plasma of healthy donors and patients with sepsis at diagnosis (0 hours), 24 hours and 72 hours post diagnosis. Comparisons between levels of L-arginine from <b>(B)</b> UHPLC-MS/MS, LC-MS/MS and ELISA and <b>(C)</b> ADMA from LC-MS/MS and ELISA are shown at diagnosis (0 hours), 24 hours and 72 hours, post diagnosis. <b>(D)</b> Canonical pathways extracted from IPA software were based on an input of read outs comparing plasma levels of amines in healthy donors and patients with sepsis as a ratio. Data are ± SEM for n = 21 healthy donors and n = 38 patients with sepsis. Data was analysed by one-way ANOVA with Dunnett’s post-hoc test and Benjamini-Hochberg test with a false discovery rate of 0.05; *p<0.05.</p

iNOS activity in LPS-activated mouse macrophages treated in plasma from healthy donors and patients with sepsis.

<p><b>(A)</b> iNOS activity in plasma from healthy vs sepsis patients at 0, 24 and 72 hours. Correlations in ADMA:L-arginine ratio with iNOS activity at <b>(B)</b> 0–24 hours and <b>(C)</b> 24–72 hours post diagnosis. Data are mean ± SEM for n = 21 healthy donors and n = 38 patients with sepsis. Data was analysed by <b>(A)</b> Kruskal-Wallis one-way ANOVA with Dunn’s post-hoc test and <b>(B-C)</b> linear regression. *p<0.05 when compared to healthy.</p

iNOS activity in LPS-activated mouse macrophages treated in culture media.

<p>iNOS activity in macrophages was measured in cells cultured in the presence of <b>(A,B)</b> 50μM or <b>(C,D)</b> 80μM L-arginine. Data are ± SEM for n = 4–6. Data was analysed by <b>(A,C)</b> paired t-test and <b>(B,D)</b> one-way ANOVA with Dunnett’s post-hoc test. *p<0.05.</p

ADMA:L-arginine ratio in healthy donors vs patients with sepsis.

<p>ADMA and L-arginine were measured from plasma of healthy donors and patients with sepsis at diagnosis (0 hours), 24 and 72 hours post diagnosis using ELISA. Data are ± SEM for n = 21 healthy donors and n = 38 patients with sepsis. Data was analysed by one-way ANOVA with Dunnett’s post-hoc test. *p<0.05.</p