Ju Hyun Shin

APE1/Ref-1 as a Serological Biomarker for the Detection of Bladder Cancer

Purpose Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein that shows elevated expression in a number of cancers. We attempted to determine whether serum APE1/Ref-1 is elevated in patients with bladder cancer. Materials and Methods Serum APE1/Ref-1 levels were determined using enzyme-linked immunosorbent assay in serum from patients with bladder cancer who had not received chemotherapy or radiotherapy (n=51) and non-tumor controls (n=55). The area under the receiver operating characteristic area under the curve was applied to determine the correlation between clinical factors and the serum levels of APE1/Ref-1. Results Serum levels of APE1/Ref-1 in bladder cancer patients were significantly elevated compared to those of the control group (3.548±0.333 ng/100 μL [n=51] for bladder cancer vs. 1.547±0.319 ng/100 μL [n=55] for the control group), with a sensitivity and specificity of 93% and 59%, respectively. Serum APE1/Ref-1 levels are associated with tumor stage, grade, muscle invasion, and recurrence. Conclusion Serum APE1/Ref-1 might be useful as a potential serologic biomarker for bladder cancer.

Expression of Nitric Oxide Synthase and Aquaporin-3 in Cyclophosphamide Treated Rat Bladder

PURPOSE The expression of Nitric oxide Synthase (NOS) and aquaporin (AQP) water channels in rat bladder is recently reported. The aim of this study is to evaluate the expression of inducible NOS (iNOS), aquaporin-3 (AQP-3) in cyclophosphamide (CYP) induced rat bladder. MATERIALS AND METHODS The 32 Sprague-Dawley rats were divided into cystitis group (n=20) and control group (n=12). In cystitis group, 100mg/kg CYP was injected every second day for 1 week whereas in control group, normal saline was injected. After extracting of the bladder and dividing dome, body and trigone of the bladder, independently H&E staining and immunohistochemical staining for iNOS and AQP-3 were performed. Expressions of iNOS and AQP-3 were analyzed with a confocal laser scanning microscope and an image analyzer. RESULTS The expression of iNOS significantly increased in the mucosa, submucosa layer of dome in cystitis group (p<0.05). The expression of AQP-3 significantly increased in the mucosa, submucosa, vessel layer of dome in cystitis group (p<0.05). CONCLUSIONS These results suggest that inflammatory change activates NOS and AQP-3 expression in the bladder tissue of rats. These may imply that NOS and AQP-3 have a pathophyiological role in the cyclophophamide induced interstitial cystitis. Further study on the NOS and AQP-3 in bladder is needed for clinical application.

Comparison of TOA and TOT for Treating Female Stress Urinary Incontinence: Short-Term Outcomes

Purpose The transobturator adjustable tape (TOA) sling operation is a new procedure that allows for the adjustment of tension after surgical intervention, thus permitting correction of postoperative incontinence or obstruction. The aim of this study was to compare the efficacy and safety of TOA with that of the transobturator tape (TOT) procedure. Materials and Methods Between 2008 and 2009, women with stress urinary incontinence (SUI) underwent TOT (n=63) or TOA (n=40). The preoperative evaluation included history taking, physical examination, voiding diary, stress and 1-hour pad tests, and a comprehensive urodynamic examination. Postoperative evaluation was performed at the 1-week and 3-month postoperative follow-up visits. Results The overall cure rate was 90.0% for the TOA group and 85.7% for the TOT group. The rate of satisfaction was higher in the TOA group than in the TOT group (95.0% vs. 85.6%). Four patients in the TOA group needed reduced tension as the result of urinary obstruction. The tension of the mesh was tightened in 1 patient because of a certain degree of continuing incontinence. The residual urine volume was significantly lower in the TOA group than in the TOT group (7.8 ml vs. 43 ml, p=0.01). Conclusions TOA allowed postoperative readjustment for a number of days after surgical intervention, which allowed for good short-term treatment outcomes. These data suggest that better subjective and objective results and residual urine volume can be obtained in the TOA group than those achieved with the traditional non-adjustable mesh and without significant postoperative complications.

Urinary APE1/Ref-1: A Potential Bladder Cancer Biomarker

Bladder cancer (BCa) is one of the most common urothelial cancers with still noticeable incidence rate. Early detection of BCa is highly correlated with successful therapeutic outcomes. We previously showed that apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) was expressed at an increased level in the serum of BCa patients when compared to the level in healthy controls. In this study, we investigated whether urinary APE1/Ref-1 was also elevated in patients with BCa. In this case-control study, voided urine was collected from 277 subjects including 169 BCa patients and 108 non-BCa controls. Urinary APE1/Ref-1 level was assessed by enzyme-linked immunosorbent assay (ELISA). APE1/Ref-1 levels were significantly elevated in BCa patients relative to levels in non-BCa controls and were correlated with tumor grade and stage. Urinary APE1/Ref-1 levels were also higher in patients with recurrence history of BCa. The receiver operating characteristics (ROC) curve of APE1/Ref-1 showed an area under the curve of 0.83, indicating the reliability and validity of this biomarker. The optimal combination of sensitivity and specificity was determined to be 82% and 80% at a cut-off value of 0.376 ng/100 μL for detection of APE1/Ref-1 in urine. In conclusion, urinary APE1/Ref-1 levels measured from noninvasively obtained body fluids would be clinically applicable for diagnosis of BCa.

A Comparison of the Clinical Efficacy of the Transobturator Adjustable Tape (TOA) and Transobturator Tape (TOT) for Treating Female Stress Urinary Incontinence with Intrinsic Sphincter Deficiency: Short-term Results

Purpose The transobturator adjustable tape (TOA) allows adjustment of tension after surgical intervention, thus permitting correction of postoperative incontinence or obstruction. The aim of this study was to compare the efficacy and safety of TOA versus transobturator tape (TOT) for the treatment of stress urinary incontinence with intrinsic sphincter deficiency (ISD). Materials and Methods Patients underwent TOA (n=33 with ISD) or TOT (n=47 with ISD) insertion by one experienced surgeon. The patients were considered to have ISD on the basis of a Valsalva leak point pressure <60 cmH2O or a maximum urethral closure pressure <20 cmH2O. The preoperative evaluation included history taking, physical examination, voiding diary, stress and 1-hour pad tests, and a comprehensive urodynamic examination. Postoperative evaluation included a stress test, 1-hour pad test, questionnaire, and uroflowmetry with postvoid residuals. Results After 6 months of follow-up, the rate of cure (TOA, 75.6% vs. TOT, 72.3%) was similar between the two groups. The rate of satisfaction was not significantly higher in the TOA group than in the TOT group (84.8% vs. 78.7%; p=0.05). Four patients in the TOA group (12.1%) needed a reduction in tension because of urinary obstruction (flow <10 ml/sec and/or residual urine >50 ml). The tension of the mesh was tightened in 5 patients (15.2%) because of the persistence of a certain degree of incontinence. The residual urine volume at postoperative 7 days was significantly lower in the TOA group than in the TOT group (19.5 ml vs. 41 ml; p=0.016, repeated-measures analysis of variance test). Conclusions The TOA allows postoperative readjustment of the suburethral sling pressure for a number of days after surgical intervention, which allows for the achievement of good short-term results. These data suggest that better lower obstructive voiding symptoms than those achieved with the traditional nonadjustable mesh can be obtained with the TOA.

Expression of Aquaporin-3 in Ipsilateral Rat Kidney With Unilateral Partial Ureteral Obstruction

Purpose Aquaporin (AQP), a protein located in the cellular membrane, allows rapid passage of water across the cell membrane. Various AQP subtypes have been associated with ureteral obstruction. In particular, AQP3 has two functions: water and glycerol transport. The aim of this study was to investigate the expression of AQP3 in the ipsilateral rat kidney in unilateral partial ureteral obstruction (UPUO). Materials and Methods Sprague-Dawley rats (n=30, 200-250 g) were divided into two groups. A sham operation was performed in the control group (n=10) and UPUO of the left upper ureter with a silicone tube was induced in the UPUO group (n=20). The left kidney was obtained from both groups 7 days after the operations. The kidney specimens underwent immunofluorescent staining with AQP3 monoclonal antibody, and the density of AQP3 in the tissue was measured with an image analyzer. Results In the UPUO group, thinning of the epithelial layer and infiltration of inflammatory cells was seen along with the localized expression of AQP3 in the basolateral aspect of the principal collecting duct cells. The mean optical density of AQP3 was significantly lower in the UPUO group than in the control group (100.9±17.5 compared with 131.7±16.9; p<0.001). Conclusions These results suggest that a decrease in the expression of AQP3 may be the result of a urinary stasis reaction caused by UPUO in response to local and intrarenal factors. These changes suggest that AQP3 may have a pathophysiological role in UPUO.

Altered Secretory Activity of APE1/Ref-1 D148E Variants Identified in Human Patients With Bladder Cancer

Purpose: Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein involved in DNA repair and redox modulation. Recently, serum and urinary APE1/Ref-1 levels were reported to be increased in patients with bladder cancer. Genetic variations of APE/Ref-1 are associated with the risk of cancer. However, the effect of APE1/Ref-1 variants on its secretory activity is yet unknown. Methods: APE1/Ref-1 variants were evaluated by DNA sequencing analysis of reverse transcription polymerase chain reaction products in coding DNA sequences (CDS) of APE1/Ref-1 in bladder tissue samples from patients with bladder cancer (n=10). Secretory activity of APE1/Ref-1 variants was evaluated with immunoblot and enzyme-linked immunosorbent assay of the culture medium supernatants. Results: Four different substitution mutants (D148E, I64V/D148E, W67R/D148E, and E86G/D148E) of APE1/Ref-1 were identified in bladder cancer specimens. However, deletion mutants of APE1/Ref-1 CDS were not found. The secretory activity of the APE1/Ref-1 variants (D148E, I64V/D148E, and E86G/D148E) was increased compared to that of wild type APE1/Ref-1. Furthermore, the secretory activity in basal or hyperacetylated conditions was much higher than that in APE1/Ref-1 D148E-transfected HEK293 cells. Conclusions: Taken together, our data suggest that the increased secretory activity of D148E might contribute to increased serum levels of APE1/Ref-1 in patients with bladder cancer.

Prospective Study Comparing the Suprapubic Arc (Sparc) Procedure and the Transobturator (Monarc) Procedure for Treating Female Stress Urinary Incontinence

Objectives: In a comparative trial we evaluated the efficacy and safety of the suprapubic arch (Sparc) and transobturator (Monarc) procedures for the treatment of female stress urinary incontinence (SUI).

Proteomic Analysis of Urothelium of Rats with Detrusor Overactivity Induced by Bladder Outlet Obstruction

Overactive bladder (OAB) syndrome is a condition that has four symptoms: urgency, urinary frequency, nocturia, and urge incontinence and negatively affects a patients life. Recently, it is considered that the urinary bladder urothelium is closely linked to pathogenesis of OAB. However, the mechanisms of pathogenesis of OAB at the molecular level remain poorly understood, mainly because of lack of modern molecular analysis. The goal of this study is to identify a potential target protein that could act as a predictive factor for effective diagnosis and aid in the development of therapeutic strategies for the treatment of OAB syndrome. We produced OAB in a rat model and performed the first proteomic analysis on the mucosal layer (urothelium) of the bladders of sham control and OAB rats. The resulting data revealed the differential expression of 355 proteins in the bladder urothelium of OAB rats compared with sham subjects. Signaling pathway analysis revealed that the differentially expressed proteins were mainly involved in the inflammatory response and apoptosis. Our findings suggest a new target for accurate diagnosis of OAB that can provide essential information for the development of drug treatment strategies as well as establish criteria for screening patients in the clinical environment.

Pathophysiology of Bladder Cancer

Abstract Bladder cancer has a complex and multifactorial pathophysiology. Numerous environmental, molecular, or genetic factors are involved in the development of urothelial cell cancers. There is a growing interest in understanding the kinds of genetic alterations or its cellular signaling pathways that contribute to the pathophysiology for progression or recurrence of bladder cancer. Genetic or epigenetic alterations of tumor suppressor genes, growth factor receptors, and DNA repair genes are all thought to be responsible. Established molecular biomarkers and expression profiles of bladder cancer are useful candidates in the early prediction or recurrence of bladder cancers.