1158 TREATMENT EFFECTS OF RECOMBINANT HUMAN SOLUBLE THROMBOMODULIN IN PATIENTS WITH SEPTIC URINARY TRACT INFECTION-INDUCED DISSEMINATED INTRAVASCULAR COAGULATION

Vol. 189, No. 4S, Supplement, Monday, May 6, 2013

1157 RAPID DRUG SUSCEPTIBILITY TESTING AND GROWTH QUANTIFICATION OF UROPATHOGENIC CANDIDA ALBICANS BY ISOTHERMAL MICROCALORIMETRY Gernot Bonkat*, Lukas Wernli, Olivier Braissant, Malte Rieken, Georg Mu¨ller, Stephen Wyler, Thomas C. Gasser, Alexander Bachmann, Basel, Switzerland INTRODUCTION AND OBJECTIVES: Urinary tract infections (UTI) caused by Candida albicans are challenging. The frequency of these infections is increasing in parallel with the rapid advances of medical progress. However, much remains unknown and has still to be learned regarding the pathogenesis, diagnosis, and management of candidal infections. The aim of the present study was to (i) investigate the influence of glucose and (ii) different antifungal drugs on the growth rate (GR) of Candida albicans in artificial urine using isothermal microcalorimetry (IMC). METHODS: A microcalorimeter equipped with 48 channels (TAM48, TA Instruments, New Castle, DE) was used. Heat produced during the growth of Candida albicans was monitored in artificial urine added with increasing concentration of glucose or antifungal drugs (Amphotericin B, Fluconazole, Tioconazole and Flucytosine). RESULTS: The GR of Candida albicans increased logarithmically with rising concentrations of glucose. At a concentration of 20 mgo`L-1 glucose the GR was 0.024 h-1, increasing to 0.203 h-1 at 1640 mgo`L-1 glucose. Noteworthy at a concentration of about 1g of glucose per litre GR levelled off. With increasing concentrations of the various antifungal drugs the GR clearly decreased. A gradual decrease was observed using Amphotericin B and Flucytosine for which the minimal inhibitory concentrations (MICs) were 0.5␮g䡠mL-1 (0.54␮M) and 5␮g䡠mL-1 (38.7␮M) respectively. Using Fluconazole and Tioconazole, a steeper decrease was observed. The MICs of these two azole compounds, were 0.8␮g䡠mL-1 (2.61␮M) and 0.5␮g䡠mL-1 (1.29␮M), respectively. CONCLUSIONS: IMC is an innovative approach to (i) measure the GR of Candida albicans in artificial urine at various glucose concentrations, and to (ii) determine the MIC of different antifungal drugs. The method is technically simple and results are provided in a short time. Given the limitations of current laboratory techniques, IMC warrants further study in the clinical setting of candidal UTI. Source of Funding: None

1158 TREATMENT EFFECTS OF RECOMBINANT HUMAN SOLUBLE THROMBOMODULIN IN PATIENTS WITH SEPTIC URINARY TRACT INFECTION-INDUCED DISSEMINATED INTRAVASCULAR COAGULATION Yoshihiro Hashimoto*, Toyota, Japan; Noriyasu Kawai, Keiichi Tozawa, Kenjiro Kohri, Nagoya, Japan INTRODUCTION AND OBJECTIVES: Urinary tract infection (UTI) complicates sepsis and/or disseminated intravascular coagulation (DIC) when the disease becomes severe; mortalities of septic DIC is reported to reach to 37% despite current therapy. Cross-talk between the coagulation system and inflammatory reactions during sepsis causes organ damage followed by multiple organ dysfunction syndrome or even death. Therefore, anticoagulant therapies have been expected to be beneficial in the treatment of severe sepsis. Recombinant human soluble thrombomodulin (rhTM) binds to thrombin to inactivate coagulation, and the thrombin-rhTM complex activates protein C to produce activated protein C. The purpose of this study was to examine the efficacy of rhTM for treating patients with septic UTIinduced DIC. METHODS: This study comprised 45 patients with septic UTIinduced DIC. All patients fulfilled the criteria of severe sepsis and the International Society on Thrombosis and Haemostasis criteria for overt DIC. The initial 25 patients were treated without rhTM (control group), and the following 20 consecutive patients were treated with rhTM (0.06

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mg/kg/day) for six days (rhTM group). The primary outcome measure was 28-day mortality. Stepwise multivariate Cox regression analysis was used to assess which independent variables were associated with mortality. Comparisons of Sequential Organ Failure Assessment (SOFA) score on sequential days between the two groups were analyzed by repeated measures analysis of variance. RESULTS: Cox regression analysis showed 28-day mortality to be significantly lower in the rhTM group than in the control group (adjusted hazard ratio, 0.301; 95% confidence interval, 0.104 to 0.867; P ⫽ 0.028). SOFA score in the rhTM group decreased significantly in comparison with that in the control group (P ⫽ 0.029). In the post hoc test, SOFA score decreased rapidly in the rhTM group compared with that in the control group on day 1 (P ⬍ 0.05). CONCLUSIONS: We found that rhTM administration may improve organ dysfunction in patients with septic UTI-induced DIC. Further clinical investigations are necessary to evaluate the effect of rhTM on the pathophysiology of septic UTI-induced DIC. Source of Funding: None

1159 IDENTIFICATION OF THE IN VIVO BINDING TARGET OF PROTEUS MIRABILIS MR/P FIMBRIAE Jessica Schaffer, Melanie Pearson*, New York, NY INTRODUCTION AND OBJECTIVES: Proteus mirabilis is a common causative agent of urinary tract infections in patients who have undergone long-term catheterization. As part of the infection process, P. mirabilis must attach to the surfaces of the urinary tract. In other uropathogenic bacteria, this is carried out by adhesive organelles called fimbriae. These fimbriae bind to carbohydrates on surfaceexpressed glycoproteins on epithelial cells of the urinary tract. P. mirabilis encodes 17 chaperone-usher fimbriae, including the mannose-resistant Proteus-like (MR/P) fimbria. While the MR/P fimbria is the best characterized, and is essential for causing maximal infection of the urinary tract, the in vivo target of MR/P fimbriae remains unidentified. METHODS: To identify the sugars bound by MR/P fimbriae, we employed two complimentary approaches. First we used a carbohydrate array to identify sugars bound by whole P. mirabilis maximally expressing MR/P fimbriae. Second, we took advantage of the previously characterized MR/P fimbriae-mediated hemagglutination of chicken red blood cells. We used an in vitro overlay assay to identify which proteins P. mirabilis binds in the red blood cell membrane. RESULTS: Of the 30 sugars present on the carbohydrate array, whole P. mirabilis bacteria bound most consistently to fucose, which is present as a terminal sugar on UPIb, a major surface protein in the bladder. Using the in vitro overlay assay, we have determined that P. mirabilis expressing MR/P fimbriae bind two proteins from the membranes of red blood cells. Efforts are currently underway to identify the proteins and their relevant glycosylation. CONCLUSIONS: These methods combined will allow us to identify which sugar residues are bound by MR/P fimbriae, which will contribute to the identification of the in vivo target of MR/P fimbriae. This is the first step in the development of fimbrial binding inhibitors as a treatment for P. mirabilis urinary tract infections. Source of Funding: K22 AI083743 (MMP)

1160 HUMAN BETA DEFENSIN-1 INHIBITS RETROVIRAL TRANSDUCTION Carrie Q Sun*, Rebecca S Arnold, John A Petros, Atlanta, GA INTRODUCTION AND OBJECTIVES: Viral infections are associated with multiple genitourinary (GU) tract malignancies. Human beta defensin-1 (hBD-1) protein, a small cysteine-rich cationic antimicrobial peptide secreted by GU epithelia, plays a role in viral host defense and is a tumor suppressor with cancer specific loss in both