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All antibiotics were purchased from Sigma Chemical Co. Accumulation of ciprofloxacin Ciprofloxacin accumulation was measured spectrofluorometrically using the method described by Mortimer and Piddock 20 with some modifications. Bacteria were harvested by centrifugation 9000 g at room temperature, washed and concentrated 10-fold in PBS, pH 7.
After the addition of ciprofloxacin, 1 mL samples were removed at different time intervals. Three minutes after the addition of ciprofloxacin, the efflux pump inhibitor CCCP final concentration 100 mM was added when necessary.
The samples were immediately diluted in 1 mL of ice-cold PBS and centrifuged for 1 min at 11 000 rpm in a microcentrifuge.
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It was a good day, it seemed; five out of six contained a small trophy, from field mice to voles, which snuffled around a bit so we could get a look at them before scuttling off into the undergrowth. Today, in total, I will be involved in four separate shows, and this is not unusual.
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The bouncing camera feels a bit too jarring during the opening land-based scenes, but proves to be a great asset in conveying the tension and disorientation during the hijacking, And the danger. Like the captured crew, we never know when a gun barrel will appear as we sneak along the deck.
This vessel is almost an above-the-water submarine with very small windows. In the interior scenes we can almost smell the sweat and breathe the stale air in this big fiberglass coffin.
Although this story dominated headlines for many days, the film still has a sense of urgency and discovery.
The news reports just hinted at the drama.