The effect of partial outlet obstruction on calpain and phospholipase-2 activities: analyzed by severity and duration.

In an attempt to better understand the two pathways that lead to bladder decompensation following partial obstruction in rabbits one of which is caused by calcium-activated enzymes and the other by oxidative stress, calpain and phospholipase A2 (PLA2) biochemical assays were conducted to see how bladder decompensation is mediated by these two calcium-activated enzymes. Partial outlet obstructions of varying durations (4, 8, and 12 weeks plus controls) were performed on 32 New Zealand white rabbits. The rabbits were also grouped by severity: control, mild, intermediate, and severe. The activities of Calpain and PLA2 on the muscle tissue of the bladders were analyzed. A stronger correlation was seen between activities and severities as opposed to between activities and durations for both PLA2 and calpain. The activity for PLA2 increased dramatically from control to mild and then stayed constant for both intermediate and severe obstructions. Calpain activity increased steadily from control to mild to intermediate to severe. Based on the increase in levels of the calcium-dependent enzymes, it was clearly shown that calcium levels increased in all stages of bladder decompensation most notably with the mild obstructions. Based on previous studies in which nitrotyrosine and dinitrophenol levels did not increase in mildly obstructed rabbits, the calcium overload pathway may predominate in mild decompensation because cells in mildly obstructed bladders are better able to cope with oxidative stress than increased calcium levels.

Effect of severity and duration of bladder outlet obstruction on catalase and superoxide dismutase activity.

OBJECTIVES: To study the effects of partial bladder outlet obstruction on the cell's anti-oxidant defense mechanisms, superoxide dismutase and catalase, in order to elucidate how the bladder responds to oxidative stress. METHODS: Four groups of eight rabbits were subjected to partial bladder outlet obstruction for 4, 8 and 12 weeks. Eight sham rabbits were used as the control group. The bladders were removed under anesthesia, and the muscle and mucosa were separated, frozen and stored at -80°C for analysis. Superoxide dismutase and catalase assays were carried out on these tissues. The groups were also categorized by severity (mild, intermediate and severe) of decompensation, as well as duration. RESULTS: When separated by duration, catalase activity of the mucosa was significantly higher in the control and the 12-weeks obstructed rabbits. This activity was lower than the control in the 4- and 8-weeks obstructed rabbits. When separated by severity, catalase activity of the mucosa was significantly higher and severely decompensated than the muscle in the controls. When separated by duration or severity, superoxide dismutase activity of the muscle was significantly lower than controls for all obstructed rabbits. The activities of both superoxide dismutase and catalase were significantly reduced in the severely decompensated bladder smooth muscle, but not in the 12-weeks obstructed bladder smooth muscle. CONCLUSIONS: Partial bladder outlet obstruction has significant effects on the activity of both superoxide dismutase and catalase in the bladder, with variations that are dependent on the severity and duration of the obstruction.

A Comparison of Total Antioxidant Capacities of Concord, Purple, Red, and Green Grapes Using the CUPRAC Assay.

Considering how popular grapes are in terms of their antioxidant benefits, we compared concord, purple, red, and green grapes for total antioxidant capacity (TAC) and carbohydrate concentration. All grapes were acquired from commercial sources and samples of each were separated into skinned and not skinned groups. Each whole grape and the skins were individually homogenized and then separated into pulp and supernatant fractions. Each fraction was analyzed for total TAC and carbohydrates. The concord grapes and purple grapes had significantly higher TAC in the homogenates than did the red or green grapes. The concord grapes and green grapes had significantly higher TAC in the pulp than in the cytosol whereas the red and purple grapes had approximately the same amount. The majority of the TAC of the purple and red grapes was in the skin whereas the concord and green grapes had approximately the same TAC in the skin and pulp. The concord and purple grapes had the highest TAC when compared to the red and green grapes, whereas the red and green grapes had approximately the same total TAC.