A widely distributed gene cluster compensates for uricase loss in hominids.

Approximately 15% of US adults have circulating levels of uric acid above its solubility limit, which is causally linked to the disease gout. In most mammals, uric acid elimination is facilitated by the enzyme uricase. However, human uricase is a pseudogene, having been inactivated early in hominid evolution. Though it has long been known that uric acid is eliminated in the gut, the role of the gut microbiota in hyperuricemia has not been studied. Here, we identify a widely distributed bacterial gene cluster that encodes a pathway for uric acid degradation. Stable isotope tracing demonstrates that gut bacteria metabolize uric acid to xanthine or short chain fatty acids. Ablation of the microbiota in uricase-deficient mice causes severe hyperuricemia, and anaerobe-targeted antibiotics increase the risk of gout in humans. These data reveal a role for the gut microbiota in uric acid excretion and highlight the potential for microbiome-targeted therapeutics in hyperuricemia.

Oxidative ornithine metabolism supports non-inflammatory C. difficile colonization.

The enteric pathogen Clostridioides difficile (Cd) is responsible for a toxin-mediated infection that causes more than 200,000 recorded hospitalizations and 13,000 deaths in the United States every year1. However, Cd can colonize the gut in the absence of disease symptoms. Prevalence of asymptomatic colonization by toxigenic Cd in healthy populations is high; asymptomatic carriers are at increased risk of infection compared to noncolonized individuals and may be a reservoir for transmission of Cd infection2,3. Elucidating the molecular mechanisms by which Cd persists in the absence of disease is necessary for understanding pathogenesis and developing refined therapeutic strategies. Here, we show with gut microbiome metatranscriptomic analysis that mice recalcitrant to Cd infection and inflammation exhibit increased community-wide expression of arginine and ornithine metabolic pathways. To query Cd metabolism specifically, we leverage RNA sequencing in gnotobiotic mice infected with two wild-type strains (630 and R20291) and isogenic toxin-deficient mutants of these strains to differentiate inflammation-dependent versus -independent transcriptional states. A single operon encoding oxidative ornithine degradation is consistently upregulated across non-toxigenic Cd strains. Combining untargeted and targeted metabolomics with bacterial and host genetics, we demonstrate that both diet- and host-derived sources of ornithine provide a competitive advantage to Cd, suggesting a mechanism for Cd persistence within a non-inflammatory, healthy gut.

In vitro and in situ techniques yield different estimates of ruminal disappearance of barley.

Objectives were to compare in vitro and in situ disappearance of dry matter (DM), neutral detergent fiber (NDF), and starch of traditional (unprocessed and rolled) and hulless (unprocessed) barley. Experiment 1: three barley sources were compared using in vitro techniques. The sources were: 1) traditional barley that was not processed, 2) traditional barley processed through a roller mill, and 3) hulless barley that was not processed. For in vitro incubation, each barley source was ground through a 1-mm screen. Ground barley sources were weighed into bags (25 micron porosity) and incubated in ruminal fluid from two steers fed 80% rolled corn for 3, 6, 12, 24, 48, or 72 h. Intact bags were assayed for NDF; remaining bags were opened and the residual was removed and analyzed to determine disappearance of DM and starch. Experiment 2: the barley sources used in Exp. 1 were compared using in situ techniques. For in situ analysis, each barley source was ground in a Wiley mill with no screen to mimic mastication. Artificially masticated samples were weighed into Dacron bags (50 ± 10 micron porosity) and incubated in eight ruminally fistulated steers (n = 8) for 3, 6, 12, 24, 48, and 72 h. Residual contents were analyzed to determine in situ disappearance of DM, NDF, and starch. Data were analyzed using the MIXED procedures of SAS (9.4 SAS Institute, Cary, NC) with repeated measures. DM disappearance was greatest (P < 0.05) for hulless barley in vitro and for rolled barley in situ, regardless of time postincubation. For both trials, NDF disappearance was greatest (P < 0.05) for hulless barley, regardless of time postincubation. Starch disappearance at all time points was greatest (P < 0.05) for rolled barley in situ. Starch disappearance was greater (P < 0.05) for hulless barley at 6 h of in vitro incubation compared to rolled and unprocessed barley, whereas starch disappearance in vitro was comparable (P = 0.60) between barley sources. When the grains were compared in vitro, minor differences were noted, presumably because barley sources were finely ground prior to incubation. Compared to in vitro estimates, in situ techniques had greater variation in ruminal degradation estimates. Differences observed between in situ and in vitro techniques are driven largely by differences between the procedures. Although laboratory methods are widely used to estimate ruminal degradation, these techniques did not provide comparable estimates of ruminal degradation of barley.

Precise Targeted Cleavage of a r(CUG) Repeat Expansion in Cells by Using a Small-Molecule-Deglycobleomycin Conjugate.

RNA repeat expansions cause more than 30 neurological and neuromuscular diseases with no known cures. Since repeat expansions operate via diverse pathomechanisms, one potential therapeutic strategy is to rid them from disease-affected cells, using bifunctional small molecules that cleave the aberrant RNA. Such an approach has been previously implemented for the RNA repeat that causes myotonic dystrophy type 1 [DM1, r(CUG)exp] with Cugamycin, which is a small molecule that selectively binds r(CUG)exp conjugated to a bleomycin A5 cleaving module. Herein, we demonstrate that, by replacing bleomycin A5 with deglycobleomycin, an analogue in which the carbohydrate domain of bleomycin A5 is removed, the selectivity of the resulting small-molecule conjugate (DeglycoCugamycin) was enhanced, while maintaining potent and allele-selective cleavage of r(CUG)exp and rescue of DM1-associated defects. In particular, DeglycoCugamycin did not induce the DNA damage that is observed with high concentrations (25 µM) of Cugamycin, while selectively cleaving the disease-causing allele and improving DM1 defects at 1 µM.