Cathepsin S is activated during colitis and causes visceral hyperalgesia by a PAR2-dependent mechanism in mice.

BACKGROUND & AIMS: Although proteases control inflammation and pain, the identity, cellular origin, mechanism of action, and causative role of proteases that are activated during disease are not defined. We investigated the activation and function of cysteine cathepsins (Cat) in colitis. METHODS: Because protease activity, rather than expression, is regulated, we treated mice with fluorescent activity-based probes that covalently modify activated cathepsins. Activated proteases were localized by tomographic imaging of intact mice and confocal imaging of tissues, and were identified by electrophoresis and immunoprecipitation. We examined the effects of activated cathepsins on excitability of colonic nociceptors and on colonic pain, and determined their role in colonic inflammatory pain by gene deletion. RESULTS: Tomography and magnetic resonance imaging localized activated cathepsins to the inflamed colon of piroxicam-treated il10(-/-) mice. Confocal imaging detected activated cathepsins in colonic macrophages and spinal neurons and microglial cells of mice with colitis. Gel electrophoresis and immunoprecipitation identified activated Cat-B, Cat-L, and Cat-S in colon and spinal cord, and Cat-S was preferentially secreted into the colonic lumen. Intraluminal Cat-S amplified visceromotor responses to colorectal distension and induced hyperexcitability of colonic nociceptors, which required expression of protease-activated receptor-2. Cat-S deletion attenuated colonic inflammatory pain induced with trinitrobenzene sulfonic acid. CONCLUSIONS: Activity-based probes enable noninvasive detection, cellular localization, and proteomic identification of proteases activated during colitis and are potential diagnostic tools for detection of predictive disease biomarkers. Macrophage cathepsins are activated during colitis, and Cat-S activates nociceptors to induce visceral pain via protease-activated receptor-2. Cat-S mediates colitis pain and is a potential therapeutic target.

Gallstone detection at CT in vitro: effect of peak voltage setting.

This study was a retrospective single-institutional study approved by the Committee on Human Research and was HIPAA compliant. A waiver for informed consent was granted. The purpose of the study was to evaluate the effect of four peak voltage settings on the in vitro conspicuity of gallstones in an anthropomorphic phantom at computed tomography (CT). An anthropomorphic phantom was scanned with (n = 86) or without (n = 85) gallstones at CT by using 80, 100, 120, and 140 kVp. The sensitivity for gallstone detection was significantly higher at 140 kVp (86% [74 of 86] for reader 1 and 81% [70 of 86] for reader 2) than at lower voltage settings (up to 67% [58 of 86] for reader 1 and 63% [54 of 86] for reader 2, P < .05 for each reader), regardless of gallstone size (<1.0 cm vs > or =1.0 cm in diameter, P < .05 for each reader). CT attenuation measurements were not useful for determination of gallstone composition. Abdominal CT performed at 140 kVp may be useful when gallstone disease is of clinical concern.