1370 URINE ANALYSIS OF CONFIRMED UTI PATIENTS REVEAL HIGHER LEVELS OF CXC CHEMOKINES COMPARED TO PATIENTS WITH LUTS WITHOUT UTI Pradeep Tyagi*, Royal Oak, MI; Vikas Tyagi, Royal Oak, MI; Harvey Qu, Rochester, MI; Kenneth Peters, Royal Oak, MI; Yao-Chi Chuang, Kaohsiung, Taiwan; Hsin-Tzu Liu, Hann-Chorng Kuo, Hualien, Taiwan; Michael Chancellor, Royal Oak, MI INTRODUCTION AND OBJECTIVES: Reliable prediction of UTI is hindered by the lack of non-invasive predictive urine biomarkers. Past studies have focused on IL-8 (CXCL-8) with the recruitment of neutrophils after bacterial invasion of uroepithelial cells. In the present study, we investigated whether other members of IL-8 family of chemo- kines i.e. CXC chemokines are also elevated following UTI and whether they can together form a specific multi-marker panel that is more useful as a surrogate marker for rapid laboratory diagnosis of UTI and pre- vention of urosepsis. METHODS: Single time point urine specimens were collected from 42 patients with confirmed UTI (+ve bacterial culture report) prior to initiation of antibiotic treatment and compared them with 4 other cohorts of either sex without any infection ( ruled out by urine dipstick test) in the age range of (25–90 yrs). There were Overactive bladder (n= 115), interstitial cystitis/painful bladder syndrome ( IC/PBS); n=59 and asymptomatic controls (n=44). Differentiation between IC/ PBS, OAB-wet and OAB-dry was based on absence or presence of pain symptoms and a 3-day voiding diary. Ten chemokines in urine were measured by MILLIPLEX™ MAP Human Cytokine/Chemokine multi- plex immunoassay and ELISA. Multivariate logistic regression was used to determine association and Kruskal-wallis ANOVA was used to check for significance. RESULTS: The multivariate regression analysis highlighted that existence of UTI was associated with elevated CXCL-10 levels (odds ratio [OR]:1.05; 95% CI, (1.0187, 1.0930), p=0.002). Levels of CXCL-10, CXCL1 and IL-8 in confirmed UTI urine were also higher than in OAB; (1203537.2 vs 39.3117.51pg/ml), (96.1628.19 vs 11.157.50pg/ml) and (11329.45 vs 29.113.54pg/ml), respectively. Urine levels of non-CXC chemokines like MCP-1, IL-1ra and PDGF were higher in population without underlying urinary tract infection such as IC/PBS and control subjects, and higher urinary NGF were noted in OAB wet patients. CONCLUSIONS: Discriminatory multivariate modeling of 10 urinary chemokines identified UTI associated CXC chemokines in urine and thereby demonstrated the utility of mathematical modeling for identifying disease-associated urine biomarkers. Although a single marker such as urinary IL-8 may be a sensitive for UTI, but it is poorly specific as it is also elevated in a variety of other diseases. A muti- marker panel that also includes CXCL-10 and CXCL-1 can be used as a urinary finger print for UTI with increased specificity. Source of Funding: OAB-LUTS grant from Pfizer 1371 NF-/INOS/CAMP SIGNALING CASCADE MEDIATES INFLAMMATORY CYTOKINES-INDUCED UPREGULATION OF CONNEXIN43 EXPRESSION AND FUNCTION IN URINARY TRACT INFECTION Kai Li*, Jian Yao, Norifumi Sawada, Hiroshi Nakagomi, Chuo City, Japan; Tatsuya Miyamoto, Chuo City, Japan; Satoru Kira, Hideki Kobayashi, Takayuki Tsuchida, Masanori Kitamura, Masayuki Takeda, Chuo City, Japan INTRODUCTION AND OBJECTIVES: Urinary tract infection (UTI) is one of the most common infectious diseases in the clinic, which can produce symptoms similar to those experienced with overactive bladder. At present little information is available regarding the mecha- nisms involved in the urinary inflammation-related symptoms and pathogenesis. Given that intercellular communication via gap junctions (GJ) plays an important role in inflammatory responses and that the enhanced GJ protein connexin43 (Cx43) contributes to myogenic blad- der overactivity, we asked whether and how gap junctions in bladder are influenced by inflammatory cytokines. METHODS: Bladder smooth muscle cells (BSMCs) isolated from Sprague-Dawley rats were analyzed for Cx43 expression and function after stimulation with inflammatory cytokines, interleukin 1 beta (IL-1) and tumor necrosis factor alpha (TNF), by Western blot, immunofluorescent staining and scrape-loading dye-transfer assay. The implicated signal mechanisms were identified by using various kinase inhibitors and specific siRNA. In vivo mouse inflammatory model was induced by intraperitoneal injection of LPS. The expressions of Cx43 in bladder were examined by histocytochemical and western blot analysis. RESULTS: (1) Incubation of BSMCs with IL-1 and TNF resulted in a time-dependent elevation in Cx43 protein levels. This was associated with obviously increased membrane localization of Cx43 protein and GJ function. (2) Increment of Cx43 by the inflammatory cytokines was preceded by an activation of PKA, as revealed by VASP phosphorylation. Blockade of PKA signaling pathway with PKA inhibitor H89, cAMP inhibitor Rp-cAMP or CREB siRNA could largely abolish the effects. (3) The Cx43-elevating effects could also be abolished by NFb inhibitor SC514 and by iNOS inhibitor L-NAME. Further studies revealed that SC514 abrogated iNOS expression, and exogenous NO donor SNAP, also induced an elevation in Cx43. (4) Intraperitoneal injection of LPS into mouse resulted in an elevated Cx43 expression and membrane localization in bladder. CONCLUSIONS: In conclusion, we demonstrated, for the first time, that inflammatory cytokines induced Cx43 expression and func- tion in BSMCs through mechanisms involving NFB/iNOS/ cAMP sig- naling cascade. Our finding may open a new window towards our understanding of the molecular mechanisms implicated in inflammatory urinary disorders and provide a possible explanation for urge symp- toms seen in bladder infection. Source of Funding: None 1372 FGF-10/FGFR2B SIGNALING DURING ACUTE CYCLOPHOSPHAMIDE-INDUCED BLADDER UROTHELIAL INJURY IN MICE Jonathan Yamzon, Kyung Hwa Lee, Los Angeles, CA; Koh-ichi Kuremoto, Osaka, Japan; In-Seon Choi, Sara Parsa, Saverio Bellusci, David Warburton, Chester Koh*, Los Angeles, CA INTRODUCTION AND OBJECTIVES: Fibroblast growth fac- tor-10 (FGF-10)/FGFr2b transgenic mice have been used in several organ systems, such as the lung, to investigate the role of FGF-10 in organ wound healing/regeneration. The cyclophosphamide (CYP)-in- duced bladder injury model is a well-established model for the study of acute bladder injury. However, the characterization of bladder urothe- lium during wound healing with regards to FGF- 10/FGFR2b signaling has not been previously described. We described the role of FGF-10/ FGFR2b signaling in the wound healing response during acute CYP- induced bladder injury in FGF-10 transgenic mice. METHODS: Bladders from inducible transgenic FGF-10 atten- uated mice via FGFr2b attenuation (rtTA; tet(o)FGFr2b) as well as from controls were exposed to weight-based intraperitoneal dosages of CYP. Retrieved bladders from various time points after CYP injection were analyzed using histology, immunohistochemistry, and RT-PCR. RESULTS: Histological section of the urinary bladders showed peak bladder injury, with epithelial ulceration, hemorrhage, and sub- mucosal edema, occurring at 4 hours after CYP injection (Figure 1). In the FGF-10 attenuated/FGFr2b attenuated mice, peak bladder injury was noted at 8 hours after CYP injury. The temporal delay in inflam- mation correlated with decreased mRNA expression of FGF-10 gene activity. CONCLUSIONS: The inflammatory response to acute cyclo- phosphamide-induced bladder injury is mediated by FGF-10/FGFr2b Vol. 185, No. 4S, Supplement, Monday, May 16, 2011 THE JOURNAL OF UROLOGY e547