Prostaglandin E2 Production and Cyclooxygenase-2 Induction in Human Urinary Tract Infections and Bladder Cancer Marcia A. Wheeler, Derek A. Hausladen,

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Prostaglandin E2 Production and Cyclooxygenase-2 Induction in Human Urinary Tract Infections and Bladder Cancer Marcia A. Wheeler, Derek A. Hausladen, Jeong H. Yoon, Robert M. Weiss The Journal of Urology Volume 168, Issue 4, Pages 1568-1573 (October 2002) DOI: 10.1016/S0022-5347(05)64522-3 Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 1 Urinary prostaglandin E2 (PGE2) levels in 18 patients with urinary tract infections (UTI) and 16 age matched controls. A, levels in patients at diagnosis were higher than in controls. B, levels were lower after than before (ACTIVE) antibiotic treatment in 7 patients. Squares indicate mean plus or minus SEM. Asterisk indicates p <0.05. The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 2 Urinary prostaglandin E2 (PGE2) levels in patients with urogenital cancer. Single level in patient with renal cancer of 11,642 pg./ml. urine was included in average but not shown in scattergram. TCC, transitional cell carcinoma. Asterisk indicates p <0.05 versus controls. The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 3 Effect of BCG intravesical instillation on urinary prostaglandin E2 (PGE2) levels in 14 patients with bladder cancer. A, Mean prostaglandin E2 before first instillation (Prior), before highest of second or third instillation (Highest), before sixth instillation (Last) and in month after treatment (Post). B, prostaglandin E2 per patient before first, before second or third BCG and before sixth instillation. Asterisk indicates p <0.05 versus previous values. ‡, p <0.05 versus highest values. The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 4 COX-1 and COX-2 mRNA in human ureter (Ur), bladder urothelium (Ua), bladder muscle (Mus) and in urine from patient with urinary tract infection (Neut) on representative gel showing second round of nested RT-PCR with COX-1 and COX-2 riboprobes. The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 5 Representative immunoblot analysis showing COX-1 protein in ureter (UR) and whole bladder (BL). Monoclonal antibody detected COX-1 after SDS-PAGE in COX-1 standard (Std.) but not in COX-2 standard. The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 6 Representative immunoblot analysis of COX-1 and COX-2 protein in bladder (BL) urothelium (UA) and muscle layers (MU). COX-1 was detected in human bladder urothelial and muscle layers. COX-2 was detected in 1 of 4 bladder muscle preparations but not in bladder urothelium. The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 7 COX-1 and COX-2 protein, and mRNA in urine particulates from patients with urinary tract infection. A, amplification reactions with a β-actin specific primer of 309 bp (lane 2), COX-1 nested primers (lanes 3 and 4) and COX-2 nested primers (lanes 5 and 6). First round amplification of COX-1 (lane 3) and COX-2 (lane 5) showed less product than nested second round amplification of COX-1 (lane 4) and COX-2 (lane 6). COX-1 and COX-2 mRNA was detected in all 3 urine particulates tested. Lane 1, 100 to 1,000 bp DNA ladder. B, immunoblot analysis with COX-1 antibody revealed COX-1 protein in ovine electrophoresis standard (lane 1) and in 1 (lane 3) but not in 2 other (lanes 2 and 4) urine particulate samples. Immunoblot analysis with COX-2 antibody demonstrated COX-2 in different urine particulate sample than COX-1 (lane 5) but not in other 2 samples (lanes 6 and 7). The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions

Fig. 8 COX-2 immunoreactivity was detected in urine particulates from 8 patients with bladder cancer. A, representative immunoblot shows immunoreactivity to COX-2 antibody in urine particulates before (lane 1, Pre), after (lanes 2 and 3, Post) and during (lanes 4 to 7, Treatment) BCG treatment. B, Blots probed for COX-2 protein were scanned digitally. In each patient highest intensity band corresponding to 72 kDa. was considered 100% and each other band was assigned relative value. Total of 4 pretreatment, 30 treatment and 4 posttreatment samples were scanned. Asterisk indicates p <0.05 versus before and after BCG. The Journal of Urology 2002 168, 1568-1573DOI: (10.1016/S0022-5347(05)64522-3) Copyright © 2002 American Urological Association, Inc. Terms and Conditions