A. Lenore Ackerman

Glt1 glutamate receptor mediates the establishment and perpetuation of chronic visceral pain in an animal model of stress-induced bladder hyperalgesia

Psychological stress exacerbates interstitial cystitis/bladder pain syndrome (IC/BPS), a lower urinary tract pain disorder characterized by increased urinary frequency and bladder pain. Glutamate (Glu) is the primary excitatory neurotransmitter modulating nociceptive networks. Glt1, an astrocytic transporter responsible for Glu clearance, is critical in pain signaling termination. We sought to examine the role of Glt1 in stress-induced bladder hyperalgesia and urinary frequency. In a model of stress-induced bladder hyperalgesia with high construct validity to human IC/BPS, female Wistar-Kyoto (WKY) rats were subjected to 10-day water avoidance stress (WAS). Referred hyperalgesia and tactile allodynia were assessed after WAS with von Frey filaments. After behavioral testing, we assessed Glt1 expression in the spinal cord by immunoblotting. We also examined the influence of dihydrokainate (DHK) and ceftriaxone (CTX), which downregulate and upregulate Glt1, respectively, on pain development. Rats exposed to WAS demonstrated increased voiding frequency, increased colonic motility, anxiety-like behaviors, and enhanced visceral hyperalgesia and tactile allodynia. This behavioral phenotype correlated with decreases in spinal Glt1 expression. Exogenous Glt1 downregulation by DHK resulted in hyperalgesia similar to that following WAS. Exogenous Glt1 upregulation via intraperitoneal CTX injection inhibited the development of and reversed preexisting pain and voiding dysfunction induced by WAS. Repeated psychological stress results in voiding dysfunction and hyperalgesia that correlate with altered central nervous system glutamate processing. Manipulation of Glu handling altered the allodynia developing after psychological stress, implicating Glu neurotransmission in the pathophysiology of bladder hyperalgesia in the WAS model of IC/BPS.

Comparison of Times to Ureteral Efflux after Administration of Sodium Fluorescein and Phenazopyridine

Purpose: There is currently a national shortage of indigo carmine. In efforts to identify the most efficient aid for visualizing ureteral efflux intraoperatively we investigated the time to excretion of phenazopyridine vs a newly identified alternative, sodium fluorescein. Materials and Methods: We analyzed prospectively collected data on a cohort of women who underwent pelvic reconstructive surgery in 2015. Per provider preference patterns a number of patients were administered 200 mg phenazopyridine orally with a sip of water 1 hour prior to the start of operative time. Other patients were given 0.5 ml 10% sodium fluorescein intravenously in the operating room. In all cases time was measured between the administration of the agent and the visualization of color changes consistent with agent efflux in an indwelling catheter, which was placed at the start of the operation. Differences in excretion times between the groups were compared with the Wilcoxon rank sum test. Results: Seven women received phenazopyridine and 5 received sodium fluorescein. Mean excretion time was significantly longer in the phenazopyridine group compared to the sodium fluorescein group (81.9 vs 5.1 minutes, p = 0.0057). Median excretion time for phenazopyridine was 70 minutes (range 59 to 127) and for sodium fluorescein it was 5 minutes (range 3 to 9). Conclusions: Sodium fluorescein is excreted significantly faster in the operating room compared to phenazopyridine. Depending on the cost of these agents at an institution, in addition to the desire to decrease operative time, this may impact practice patterns and agent selection.

PD19-02 CHARACTERIZATION OF BACTERIAL IDENTIFIED ON EXPLANTED MESH SLINGS USING NEXT-GENERATION SEQUENCING TECHNIQUES

vaginal swab specimens. Using a custom, manually curated fungal database focused on mammalian-associated genera (THFv1.5), we were able to increase the number of annotated sequences from 62% (using the UNITE database) to 86%. K-means clustering was performed using MatLab. RESULTS: The fungal communities present in vaginal samples from 251 asymptomatic women aged 15-44 were predominantly of low fungal diversity, with a median Shannon diversity index of 0.61 0.52. Using k-means clustering analysis, we identified 3 major community patterns: Candida-predominant, Wallemia-predominant and a third mixed community containing a Pichia-predominant subset. These patterns did not correlate with exogenous hormonal medication use. CONCLUSIONS: This is the first large dataset of vaginal specimens analyzed to define the composition of vaginal mycobiome in asymptomatic, reproductive-age women. While we reconfirmed Candida as the most common predominant species present in the vagina, this reanalysis identifies a large minority of subjects with the halophilic fungus, Wallemia. The dramatically different growth requirements of these two genera suggests that the fungal mycobiome may reflect fundamentally different microenvironments within the vagina of these subject subsets, despite both being asymptomatic. Future studies will aim to examine whether these differences impact the risks for other genitourinary tract pathology.

MP86-15 EFFECTS OF GROUP REHABILITATION UPON WOMEN UNDERGOING SURGERY FOR OBSTETRIC FISTULA

RESULTS: 627 men were eligible for the study. Age, ethnicity, primary language, education and Charlson comorbidity did not differ across PSA strata. Compared to the referent group PSA <10, those with PSA 50 were more likely to receive androgen deprivation therapy as their primary form of treatment (p <0.01). Patients with PSAs 10-19.9 were more likely to have sexual bother (b1⁄411.1, p<0.03) compared to the referent group. (See Table) There were no other differences in other HRQOL domains across PSA strata. CONCLUSIONS: In this population, we found no statistically significant difference in HRQOL outcomes by PSA level. The finding that patients with very elevated PSA levels having outcomes that were no worse than patients with less aggressive disease is important clinically because most quality of life detriments tend to be from treatment of localized disease. Further, these findings will inform physicians on patient symptomatology despite PSA level.

PD44-12 FEMALE SEXUAL DYSFUNCTION TREATMENT: A META-ANALYSIS OF THE PLACEBO EFFECT ACROSS RANDOMIZED CONTROLLED TRIALS

INTRODUCTION AND OBJECTIVES: Sexual dysfunction has a significant impact on quality of life. The use of pornography among females and its impact on sexual dysfunction is poorly described. As an exploratory outcome of a study primarily investigating the relationship between pornography and erectile dysfunction, we attempt to better define pornography use and any contribution to sexual dysfunction in women. METHODS: After IRB approval, all patients presenting to a urology clinic of ages 20-40 years between February and August, 2016 were offered an anonymous survey consisting of self-reported medical history and demographic questions, validated questionnaires and novel questions addressing sexual function, pornography use and addictive behavior. Accrual continues, and we report a planned interim analysis. Descriptive data was compiled, and strength of correlation between subdomains of female sexual function, obsessive or craving behaviors and pornography use were examined. All variables were analyzed with linear regression. RESULTS: Of the first 48 females who agreed to take the survey included in the analysis, the mean age was 28 years. The subjects reported minimal medical comorbidities or risk factors with the most common being depression (16%), PTSD (12%) and smoking (31%). The sample was primarily white (62%), married (60%), heterosexual (81%), and active duty military (58%). The majority of respondents denied pornography use (61%) and 25% used less than weekly. Of those that used pornography, 72% reported duration of 15 minutes or less. The primary access was internet (68%) and phone (55%). The mean Female Sexual Function Inventory total score was 64. There was no observed correlation between female sexual function and pornography use. CONCLUSIONS: Interim results better describe pornography use among females. In a sample of women ages 20-40, pornography use is not uncommon with the main access being through internet or phone. There does not appear to be any correlation between its use and sexual dysfunction as determined by self-reported questionnaire. Further study may better elucidate any relationship between pornography and female sexual dysfunction.

PD50-12 HIGH CATASTROPHIZING IN PATIENTS WITH SELF-REPORTED PAINFUL MESH COMPLICATIONS HAVE POORER OUTCOMES

INTRODUCTION AND OBJECTIVES: Scarring secondary to mesh and prosthetic materials is a serious clinical problem within the GU tract. Fibrotic matrices contain fibronectin; and alpha-smooth muscle actin contributes contraction. Metalloproteinases (MMPs) such as MMP1 and -3 can modulate matrix protein accumulation through degradation. Results from our institution have shown that silica materials can directly induce scarring through the interaction with tissue fibroblasts in vitro. By extension, we hypothesized that other materials may induce fibrotic changes through cellular matrix gene expression. Objectives: 1) to establish a 3D model of human fibroblasts to study patterns of fibroblast response to materials and 2) measure gene expression in human fibroblasts exposed to prosthetic and mesh materials compared to a control. METHODS: Collagen gel was prepared by using 3 mg/ml in final concentration with 0.5% of polyvinyl alcohol (PVA). Mesh or catheter materials and human dermal fibroblasts (70,000 /ml) were added to a collagen gel and seeded in a 24-well plate (0.5 ml of gel in each well) to create a 3D environment for fibroblast response. After polymerization of collagen, another 250,000 cells in 0.5 ml medium were added on the top of gel. Cells were cultured at 37 0C, 5% CO2 for indicated time point. Images of cells were taken under reverse microscopy to determine the pattern of the scarring contraction. Gel cell matrix was harvested and digested with 1 mg/ml of collagenase for 15 minutes, pelleted by centrifugation and RNA was extracted. RT-PCR was performed for 32 cycles to analyze gene expression. RESULTS: After 5 days, fibroblast contractions were identified surrounding prosthetic materials but not within the control. There were increases in type 1 collagen, a-smooth muscle actin and fibronectin expression in fibroblasts exposed to prosthetic materials compared to fibroblasts grown in collagen gel alone. MMP-1 and MMP-3 were also detected. CONCLUSIONS: Fibroblasts exposed to mesh and catheter materials responded with an increase in fibronectin, alpha smooth muscle actin and type 1 collagen that is increased compared to controls. This may indicate why in vivo these materials induce fibrosis. Because fibronectin, type 1 collagen and alpha smooth muscle actin are main components of scarring and their gene expression is elevated, future directions include development of medical devices that could induce downregulation of these genes.

MP31-19 SYMPTOMATIC OVERLAP IN OVERACTIVE BLADDER AND INTERSTITIAL CYSTITIS/PAINFUL BLADDER SYNDROME

INTRODUCTION AND OBJECTIVES: Interstitial cystitis/ bladder pain syndrome (IC/BPS) is a chronic inflammation that results in recurring pain in the bladder and the surrounding pelvic region caused by abnormal excitability of micturition reflexes. Spinal cord stimulation (SCS) is currently in clinical use for the attenuation of neuropathic pain, and has been identified for amelioration of pain in a rodent model of colorectal distention. This study aimed to investigate the potential effect of SCS on attenuation of visceral pain-related visceromotor reflexes (VMR) in rodents with cyclophosphamide-induced cystitis. METHODS: Female Sprague-Dawley rats underwent intraperitoneal injection of cyclophosphamide (CYP, n1⁄48) or saline (controls, n1⁄44). 48 hours after CYP or saline injections, all rats underwent surgical preparations under urethane anesthesia. Stimulation wires were placed on the dorsal surface of the spinal segments of L2 and L3 for SCS. A PE-50 tubing was inserted into the bladder to obtain the intravesical pressure (IVP). Two wires are placed in the left external abdominal oblique muscle to record VMR. Before and after SCS (40 Hz, 0.2 ms for 25 min), VMR and maximum intravesical pressure (IVPmax) were obtained during continuous bladder infusion and isotonic bladder distention (IBD). RESULTS: During continuous bladder infusion (Fig 1A), the ratio of VMR threshold/IVPmax was significantly decreased in CYP rats indicating early VMR appearance compared to controls. SCS significantly increased the ratio indicating the delayed VMR appearance. During IBD with urethral occlusion, SCS delayed the latency of VMR appearance in CYP rats below the voiding thresholds at 10 and 20 cmH2O. SCS also significantly decreased VMR area under the curve (AUC) in CYP rats (Figs 1B-C). CONCLUSIONS: SCS attenuated visceral pain-related VMR in rats with CYP-induced cystitis suggesting a role for SCS in modulating visceral nociception and hyperalgesia. SCS appears to be a potential treatment of IC/PBS.

MP82-02 DECREASED URINARY FUNGAL BURDEN AND DIVERSITY IN OVERACTIVE BLADDER

INTRODUCTION AND OBJECTIVES: Hedgehog signaling pathway is known to have important role in the urogenital development. Transcription mediators of the pathway, Gli2 and Gli3, have been shown to be heavily involved in proper urogenital sinus formation. Both Gli2 and Gli3 null mice are non-viable, and display severe urogenital ad hindgut malformations. Here, we have generated a compound genetic mutant, Gli2+/-;Gli3D699/+, that is viable well into adulthood, and displaying variable urogenital malformations including urinary incontinence in its females. We aim to characterize the urinary incontinence observed in Gli2+/-; Gli3D699/+ female mice and assess its functional, anatomical, and histological characteristics. METHODS: Gli2+/and Gli3D699/+ mice were crossed to generate the double mutant (Gli2+/-; Gli3D699/+) female mice and wild type female mice were used as comparison controls; which all were verified via Polymerase Chain Reactions. Void measurements, Cystometrogram (CMG) and leak point pressure (LPP) were performed in all genotypes to assess bladder functions. The mice were then sacrificed to harvest the bladders and urethras for gross characterization via ink injection and histological assays. Differences were reported as mean and standard errors of mean (SEM) and analyzed using univariate analysis. Statistical significance set at 0.05. RESULTS: No significant differences between the mutant and wild type mice were detected for 24 hour urinary output [(n1⁄4 13) mean 26.5cc 5 vs ( n1⁄47) mean 22.15cc 6, p1⁄40.13]. CMG studies revealed a decrease in peak micturition pressure values and significantly reduced LPP in Gli2+/-; Gli3D699/+ mice compared to wild type mice [(n1⁄45) 4.28 cmH2O 2.4 vs (n1⁄44) 20.24 cmH2O 6.45, p<0.0001; (n1⁄45) 6.66 cmH2O 1.6 vs (n1⁄45) 26.5cmH2O 5, p<0.05; respectively]. Gross characterization revealed that the ano-genital distance was severely reduced in double mutant mice; however, the urethra, vagina, and anus all remain separate and distinctly identifiable in these mice. Histological analyses revealed Gli2+/-; Gli3D699/+ mice exhibited a widened urethra and a decrease in smooth muscle layer thickness in the bladder outlet and urethra, with increased mucosal folding. CONCLUSIONS: Gli2+/-; Gli3D699/+ female mice display persistent urinary incontinence with evident malformation of the bladder outlet and urethra. This presents a genetic mouse model for female urinary incontinence and alludes to potential genetic factors involved in the human condition.

MP82-18 SHARED ALTERATIONS IN URINARY BACTERIAL COMMUNITIES IN PATIENTS WITH INTERSTITIAL CYSTITIS AND OVERACTIVE BLADDER

INTRODUCTION AND OBJECTIVES: Clinically, mid-urethral sling procedure has become a regular and popular treatment for stress urinary incontinence (SUI). However, it has limited improvement if intrinsic sphincter deficiency dominantly and/or co-existing serious urgency symptom. We applied a new neuro-modulative sling in an acute urinary incontinence rat model to test if it can improve the urinary incontinence and urinary urgency simultaneously. METHODS: The neuro-modulative sling made by sling mesh along with small silver wire electrodes which were connected to electrical stimulator with the free ends in the middle of sling for electrical stimulation. An acute mixed urinary incontinence animal model was made by bilaterally pudendual nerve transaction (PNT, n1⁄421) or sham PNT (n1⁄420) followed by potassium chloride (4M, KCl) or saline (control, 0.9% NaCl) bladder perfusion. Urodynamic testing performed to confirm the changing of the bladder contractions. Leak point pressure (LPP) was also tested during filling cytometry. RESULTS: With current above 10mA and frequency above 50Hz result in significantly increased LPP during electrical stimulation delivering to the neuro-modulative sling (p<0.05). The continuous electrical stimulation with bi-polar square wave at the parameters of frequency to 5 Hz and pulse duration to 150 ms may decrease the increased bladder contractions significantly after PNT and KCl perfusion (p<0.01). CONCLUSIONS: The study demonstrated the neuro-modulative sling with different electrical stimulation parameters may improve mixed urinary incontinence in an acute animal model. Our future direction is studying its effects on a long-term animal model and possible complications if long-term implantation and use.

PD44-05 WRITING IN THE MARGINS OF SEXUAL FUNCTION QUESTIONNAIRES: A QUALITATIVE ANALYSIS FROM WOMEN WITH PELVIC FLOOR DISORDERS

I-PSS score (6.8 vs 6.9, p 1⁄40.69), nor mean FSFI score (22.9 vs 23.2, p1⁄4 0.68) compared to lower intensity cyclists. High intensity cyclists were more likely to develop perineal numbness, OR 1.6 (95% CI 1.3-2), and saddle sores, OR 2.2 (95% CI 1.8-2.8). Bike seat type had no significant effect in any of the above mentioned results. CONCLUSIONS: Contrary to previous literature, we demonstrate that cycling has no appreciable effect on female sexual or urinary function. However; our study suggests that cycling may increase the risk of UTI and perineal numbness.