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2, coated with ∼150 Å of gold, and examined with a Hitachi S-4500 FEG Scanning Electron Microscope. Two or more bladders were examined for each type of sample.
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4, 10 mM glucose, and 5 mM Hepes (pH 7.4)] and gently rinsed. One-half of each bladder was incubated with, and one half without, Ringer solution supplemented with gentamicin (100 μg/ml) for 90 min at room temperature. This incubation with gentamicin was sufficient to kill any external bacteria. Bladder halves were washed with PBS, weighed, and homogenized in 1 ml of 0.025% Triton X-100/PBS, and surviving bacteria were plated (2).
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We thank M. Veith for his help with SEM and E. M. Johnson and M. Deshmukh for their helpful suggestions and reagents. This work was supported by NIH grants R01AI29549 and R01DK51406. M.A.M. was supported by a Lucille P. Markey Special Emphasis Pathway in Human Pathobiology postdoctoral fellowship and by NIH fellowship AI09787. All animal experiments were performed under accredited conditions after approval of protocols by the local Animal Studies Committee.
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