Early nonpolypoid colorectal cancer: radiographic diagnosis of depth of invasion.

PURPOSE: To diagnose depth of invasion of early flat and depressed nonpolypoid colorectal cancers with double-contrast barium enema examination. MATERIALS AND METHODS: Ninety-seven early nonpolypoid colorectal cancers, initially identified with colonoscopy, were investigated with double-contrast barium enema examination. Depth of invasion in resected specimens obtained with endoscopic mucosal or surgical resection was determined. Cancer confined to the mucosal layer or with focal extension to the submucosal layer was defined as Ca-m; moderate to massive extension of cancer into the submucosal layer was classified Ca-sm. RESULTS: In flat elevated and depressed cancers, converging folds and semilunar deformity were observed significantly more frequently in Ca-sm than in Ca-m tumors (P < .05 and P < .01, respectively). Sensitivities and specificities of these findings for Ca-sm tumor were 42%, 93% and 86%, 85%, respectively. Deep depression, irregular surface of the depression, and tumor size larger than 20 mm were also predictive findings for depressed Ca-sm tumors (P < .05); the specificity of each was 100%. With use of these radiographic predictors, overall accuracy for diagnosing depth of invasion was 85%. CONCLUSION: Findings on double-contrast barium enema study are highly predictive of depth of invasion of early nonpolypoid colorectal cancer. Radiographic findings of converging folds, semilunar deformity, deep depression, irregular surface of the depression, and tumor size are predictors of Ca-sm tumor.

Oxidation process of bovine heart ubiquinol-cytochrome c reductase as studied by stopped-flow rapid-scan spectrophotometry and simulations based on the mechanistic Q cycle model.

Stopped-flow rapid-scan spectrophotometry was employed to study complicated oxidation processes of ubiquinol-cytochrome c reductase (QCR) that was purified from bovine heart mitochondria and maximally contained 0.36 mol of ubiquinone-10/mol of heme c1. When fully reduced QCR was allowed to react with dioxygen in the presence of cytochrome c plus cytochrome c oxidase, the oxidation of b-type hemes accompanied an initial lag, apparently low potential heme bL was oxidized first, followed by high potential heme bH. Antimycin A inhibited the oxidation of both b-type hemes. The oxidation of heme c1 was triphasic and became biphasic in the presence of antimycin A. On the other hand, starting from partially reduced QCR that was poised at a higher redox potential with succinate and succinate-cytochrome c reductase, the b-type hemes were oxidized immediately without a lag. When the ubiquinone content in QCR was as low as 0.1 mol/mol heme c1 the oxidation of the b-type hemes was almost suppressed. As the Q-deficient QCR was supplemented with ubiquinol-2, the rapid oxidation of b-type hemes was restored to some extent. These results indicate that a limited amount of ubiquinone-10 found in purified preparations of QCR is obligatory for electron transfer from the b-type hemes to iron-sulfur protein (ISP) and heme c1. The characteristic oxidation profiles of heme bL, heme bH, and heme c1 were simulated successfully based on a mechanistic Q cycle model. According to the simulations the two-electron oxidation of ubiquinol-10 via the ISP and heme c1 pathway, which is more favorable thermodynamically than the bifurcation of electron flow into both ISP and heme bL, does really occur as long as heme bL is in the reduced state and provides ubiquinone-10 at center i. Mechanistically this process takes time, thus explaining the initial lag in the oxidation of the b-type hemes. With the partially reduced QCR, inherent ubisemiquinone at center i immediately oxidizes reduced heme bH thus eliminating the lag. The mechanistic Q cycle model consists of 56 reaction species, which are interconnected by the reaction paths specified with microscopic rate constants. The simulations further indicate that the rate constants for electron transfer between the redox centers can be from 10(5) to 10(3) s-1 and are rarely rate-limiting. On the other hand, a shuttle of ubiquinone or ubiquinol between center o and center i and the oxidation of heme c1 can be rate-limiting. The interplay of the microscopic rate constants determines the actual reaction pathway that is shown schematically by the "reaction map." Most significantly, the simulations support the consecutive oxidation of ubiquinol in center o as long as both heme bL and heme bH are in the reduced state. Only when heme bL is oxidized and ISP is reduced can SQo donate an electron to heme bL. Thus, we propose that a kinetic control mechanism, or "a kinetic switch," is significant for the bifurcation of electron flow.

Oxidation-reduction behavior of the heme c and heme d moieties of Pseudomonas aeruginosa nitrite reductase and the formation of an oxygenated intermediate at heme d1.

Dithionite reduced the heme c moiety of Pseudomonas nitrite reductase almost instantaneously, whereas the spectral change of heme d proceeded in two steps, requiring at least 15 min for completion. The final spectrum coincided well with that obtained by anaerobic reduction with ascorbate, during which a quasi oxidation-reduction equilibrium was established between the two heme groups. The difference in apparent redox potential was calculated to be 24 mV, heme d being more negative. When the enzyme was supplemented with a reductant and molecular oxygen, an oxygenated intermediate appeared at the heme d moiety.