Voided Urine Samples Research Articles

Comparison of two liquid preservatives for SurePath™ slides prepared from voided urine

Blood-rich gynecologic specimens can be problematic in the processing of liquid-based cytology. However, little is known about the influence of erythrocytes and protein on urine specimens. In addition, the SurePath™ system has two preservatives for non-gynecologic specimens. In this study, we compared the epithelial cell counts and cytomorphology obtained from CytoRich™ (CR) Red and CR Blue. A total of 98 voided urine samples were processed using both CR Red and CR Blue. We made an assessment of the epithelial cell counts, fixation, and staining quality, and backgrounds of both slides. Urine protein and urine erythrocyte counts were analyzed, and those data were compared with the epithelial cell counts in CR Red and CR Blue slides. Overall, epithelial cell counts were equivalent for both CR Red and CR Blue slides. However, in high-level proteinuria cases, the CR Red slides showed higher epithelial cell counts than the CR Blue slides. On the other hand, in microscopic hematuria cases, the CR Blue slides showed higher epithelial cell counts than the CR Red slides. We have found both CR Red and CR Blue to be available for urine cytology. However, it is important to note that CR Blue is inferior to CR Red in epithelial cell recovery rates in cases of high-level proteinuria.

Significance of atypical category in voided urine specimens prepared by liquid-based technology: Experience of a single institution

The goal of this study was to evaluate the clinical significance of atypical cytology in voided urine samples. We also studied any differences in outcome that may exist between the patients being surveyed versus high risk for urothelial carcinoma (UC). This was a retrospective study of all voided urine specimens with "atypical" cytology over a 10-year period. The patients were categorized into those with and without a prior diagnosis of UC as the "surveillance" and "de novo" (DG) groups. Follow-up was obtained. Clinical impact and outcomes of the 2 groups were compared. In this study, 5.7% of voided urine specimens were atypical. Mean age of patients in years, male/female ratio, and time to diagnosis in days was 59 versus 71, 23:15 versus 22:1, and 95 versus 43 in the DG and surveillance group, respectively. Rate of progression to UC was similar in both groups. High-grade UC was significantly higher in the DG. Approximately 20% of patients in the DG were subsequently diagnosed with UC. The common causes for the atypical diagnosis that did not progress to UC were stones and benign prostatic hyperplasia. In the absence of an etiology for the atypia, further investigations are warranted.

S99 Concurrent comparisons between results of antimicrobial susceptibility tests performed on percutaneous nephrostomy tube drainage fluids, and voided urine samples

S99 Concurrent comparisons between results of antimicrobial susceptibility tests performed on percutaneous nephrostomy tube drainage fluids, and voided urine samples

Urinary Expression of Novel Tissue Markers of Kidney Injury After Ureteroscopy, Shockwave Lithotripsy, and in Normal Healthy Controls

Shockwave lithotripsy (SWL) and ureteroscopy (URS) are minimally invasive treatment alternatives for kidney stones. Although less invasive, SWL subjects the renal parenchyma to a high level of energy and the potential to cause renal injury. The ability to detect renal injury post-SWL in a reliable and noninvasive way would be clinically beneficial. Kidney injury molecule 1 (KIM-1) and N-acetyl-β-D-glucosaminidase (NAG) are two proteins secreted by the kidney into the urine and have been found to be sensitive markers of acute kidney injury in transplant patients. The aim of this work was to measure urinary levels of KIM-1 and NAG in patients with kidney stone who were treated by SWL or URS and in nonstone volunteers. Patients with kidney stones who were treated by SWL (n = 50) or URS (n = 10) were recruited. Voided urine samples were collected before and 2 to 3 hours after URS and SWL. In addition, further urinary specimens were collected 2 days and 2 weeks post-SWL treatment. Voided urine samples from healthy volunteers were also collected. Mean KIM-1 values were increased in patients with kidney stones when compared with volunteers. KIM-1 and NAG levels significantly increased post-SWL and returned to baseline within 2 weeks post-SWL. Poor kidney function was significantly associated with increased biomarker activity both in baseline and post-SWL measurements. There was no significant change in urinary KIM-1 and NAG concentrations before and after URS. Kim-1 and NAG levels significantly increased post-SWL treatment suggesting a potential role for these urinary markers in identifying patients at higher risk of tissue injury.

Should Antibiotics Be Given Prior to Outpatient Cystoscopy? A Plea to Urologists to Practice Antibiotic Stewardship

BackgroundAntimicrobial resistance is a major health problem, caused primarily by overuse of antibiotics in clinical situations in which they are not necessary. Practice guidelines recommend that antibiotics be given before outpatient cystoscopy to prevent symptomatic urinary tract infection (UTI). ObjectiveTo determine the frequency of febrile UTI after outpatient flexible cystoscopy in antibiotic-naive bladder tumor patients. Design, setting, and participantsA total of 2010 consecutive outpatients with bladder tumors were entered into a prospective registry study. All patients underwent cystoscopy after they submitted a voided urine sample for culture. Significant bacteriuria was defined as >104 colony-forming units per milliliter with a single organism. Patients were stratified for known risk factors for UTI. InterventionPatients underwent flexible cystoscopy and received no antibiotics immediately before or after cystoscopy. They were followed for 30 d for onset of febrile UTI. Outcome measurements and statistical analysisThe end point was incidence of febrile UTI within 30 d of cystoscopy. Febrile UTI was defined as temperature >38°C and dysuria, or having received antibiotics from an outside physician for urinary symptoms. Results and limitationsOf the 2010 patient cystoscopies, 489 (24%) had asymptomatic bacteriuria, and 1521 (76%) had sterile urine. Thirty-nine patients (1.9%) developed febrile UTI ≤30 d after cystoscopy—4.5% in colonized patients and 1.1% in uninfected patients (p=0.02). All UTIs resolved in ≤12–24h with oral antibiotics. None of the patients was admitted for bacterial sepsis. Limitations of the study are that it is a single-surgeon experience in one institution, and results may not apply to other patient populations. ConclusionsAntibacterial therapy before outpatient flexible cystoscopy does not appear necessary in bladder tumor patients who have no clinical signs or symptoms of acute UTI, including asymptomatic bacteriuria. Antibiotic stewardship is the responsibility of all urologists.

Urinary analysis reveals high deoxynivalenol exposure in pregnant women from Croatia

In this pilot survey the levels of various mycotoxin biomarkers were determined in third trimester pregnant women from eastern Croatia. First void urine samples were collected and analysed using a "dilute and shoot" LC-ESI-MS/MS multi biomarker method. Deoxynivalenol (DON) and its metabolites: deoxynivalenol-15-glucuronide and deoxynivalenol-3-glucuronide were detected in 97.5% of the studied samples, partly at exceptionally high levels, while ochratoxin A was found in 10% of the samples. DON exposure was primarily reflected by the presence of deoxynivalenol-15-glucuronide with a mean concentration of 120 μg L(-1), while free DON was detected with a mean concentration of 18.3 μg L(-1). Several highly contaminated urine samples contained a third DON conjugate, tentatively identified as deoxynivalenol-7-glucuronide by MS/MS scans. The levels of urinary DON and its metabolites measured in this study are the highest ever reported, and 48% of subjects were estimated to exceed the provisional maximum tolerable daily intake (1 μg kg(-1) b.w.).

Is Urinary Soluble Fas an Independent Predictor of Non-Muscle-Invasive Bladder Cancer? A Prospective Chart Study

Objectives: To evaluate whether soluble Fas (sFas) in urine is an independent predictor of non-muscle-invasive bladder cancer (NMIBC). Methods: We performed a prospective chart review which included 128 subjects with NMIBC and 88 controls. The first morning voided urine sample (10-20 ml) was obtained from preoperative patients and controls. Expression levels of sFas in urine were analyzed using enzyme-linked immunosorbent assay. Clinical and pathological data, European Organisation for Research and Treatment of Cancer (EORTC) risk group category, follow-up data and urinary sFas values were gathered from each patient, and each prognostic outcome was evaluated. Results: sFas levels were significantly higher in the urine of patients with NMIBC than of those without NMIBC (p = 0.000). The level was significantly higher in cases with a higher stage or grade or high-risk and recurrent disease than in those with a lower stage or grade or low-risk and nonrecurrent disease (each p < 0.05). Kaplan-Meier estimates revealed a significant difference in time to recurrence based on sFas levels in the urine of the NMIBC patients (log-rank test; p = 0.000). On multivariate Cox regression analysis, EORTC risk group category (hazards ratio [HR] = 3.250, p = 0.000) and urinary sFas level (HR = 1.403, p = 0.015) were the independent predictors of NMIBC recurrence. Conclusions: Our study indicated that urinary sFas assay results may help identify NMIBC patients at risk of tumor recurrence. These data can be used to design a future follow-up schedule and treatment strategy for NMIBC patients.

P3.020 Prevalence of Genital Chlamydia Trachomatis and Neisseria Gonorrhoeae Infections Among Adolescents in Northern Italy

BackgroundSexually transmitted infections are widespread globally, especially among people aged 15–25. Chlamydia trachomatis is the most common sexually-acquired bacterial disease. This infection is not notified in Italy; prevalence data on...

Mycoplasmas infection in male HIV/AIDS patients in Jiangsu, China

Mycoplasmas are widely distributed among animals, plants, and human. The four species namely, Mycoplasmas genitalium(Mg), Mycoplasmas fermentans(Mf), Mycoplasmas pentrans(Mpe), Mycoplasmas pirum(Mpi) are also called AIDS-associated mycoplasmas due to their involvement in the development and outcome of AIDS. To investigate the infection prevalence of Mg, Mf, Mpe and Mpi among male HIV/AIDS patients in Jiangsu Province and to analyze the relationship between pathogenic mycoplasmas and cellular immune function of them. First void urine and venous blood samples were collected and epidemiology questionnaires were administered after informed consent. Nested PCR was performed to determine the infection of Mg, Mf, Mpe and Mpi while ELISA assay was applied to detect interleukin-2(IL-2), interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α). SAS 9.0 software was applied to analyze the data. A total of 713 HIV/AIDS patients were recruited in this study. The overall infection rates of Mg, Mf, Mpe and Mpi are 27.9%, 9.7%, 1.0% and 18.4% respectively. Generally, the infection rates of Mg(χ2 = 10.311, P = 0.006) and Mpi were declined as the CD4+ cell counts increased, while Mf infection was higher in CD4+ T cell>350/μl group. The levels of cytokines are different with the variance of mycoplasmas infection. Mycoplasma infection among male HIV/AIDS patients is associated with changes in cellular immune response (cytokines). However, the affect of mycoplasmas on the immune function is complex, further studies are still required to elucidate whether mycoplasmas interact with HIV by interfering host immune system.

Multiplex Protein Signature for the Detection of Bladder Cancer in Voided Urine Samples

Accurate urine assays for bladder cancer detection would benefit patients and health care systems. Through extensive genomic and proteomic profiling of urine components we previously identified a panel of 8 biomarkers that can facilitate the detection of bladder cancer in voided urine samples. In this study we confirmed this diagnostic molecular signature in a diverse multicenter cohort. We performed a case-control, phase II study in which we analyzed voided urine from 102 subjects with bladder cancer and 206 with varying urological disorders. The urinary concentration of 8 biomarkers (IL-8, MMP-9 and 10, PAI-1, VEGF, ANG, CA9 and APOE) was assessed by enzyme-linked immunosorbent assay. Diagnostic performance of the panel of tested biomarkers was evaluated using ROCs and descriptive statistical values, eg sensitivity and specificity. Seven of the 8 urine biomarkers were increased in subjects with bladder cancer relative to those without bladder cancer. The 7 biomarkers were assessed in a new model, which had an AUROC of 0.88 (95% CI 0.84-0.93), and 74% sensitivity and 90% specificity. In contrast, the sensitivity of voided urine cytology and the UroVysion® cytogenetic test in this cohort was 39% and 54%, respectively. Study limitations include analysis performed on banked urine samples and the lack of voided urine cytology and cytogenetic test data on controls. The study provides further evidence that the reported panel of diagnostic biomarkers can reliably achieve the noninvasive detection of bladder cancer with higher sensitivity than currently available urine based assays.

Molecular Detection of Prostate Cancer by Methylation-Specific Polymerase Chain Reaction from Urine Specimens

Summary Background: Prostate cancer (PCa) represents the second most prevalent malignancy among males, which is characterized by a high mortality rate. The aim of our study was to evaluate the methylation status of glutathione S-transferase P1 (GSTP1) in urine specimens from males with PCa and benign prostatic hyperplasia (BPH) and its usefulness in distinguishing between males with PCa and BPH by noninvasive methods. Methods: Voided urine specimens were collected from 65 patients with PCa and 45 patients with BPH. Genomic DNA was isolated and subjected to bisulfite modification. Methylation status of the GSTP1 gene was determined by conventional methylation-specific polymerase chain reaction (MSP) analysis. Results: Promoter hypermethylation of the GSTP1 gene in voided urine samples was found in 63 of 65 (97%) males with PCa and in 5 of 45 (11%) males with BPH. The sensitivity and specificity of GSTP1 in discriminating between PCa and BPH males were 98% and 89%, respectively. Conclusions: Gene analysis of GSTP1 using conventional MSP in urine specimens can be used as a noninvasive biomarker to distinguish between men with malignant and benign prostatic diseases.

Screening and identification of specific markers for bladder transitional cell carcinoma from urine urothelial cells with suppressive subtractive hybridization and cDNA microarray

Objective: The objective of this study was to screen and identifydifferentially expressed genes in invasive bladder transitional cellcarcinoma (BTCC).Methods: Voided urine samples were collected from consecutivepatients with BTCC and patients under surveillance for bladdercancer recurrence; voided urine samples from patients with nonmalignantdiseases served as control. We identified the differentiallyexpressed genes by comparing urine samples of bladdercarcinoma to that of the control group with suppressive subtractivehybridization (SSH) and cDNA microarray. The differentiallyexpressed genes were verified by quantitative real-time polymerasechain reaction (QPCR).Results: From the 762 white colonies, a total of 449 positive cloneswere obtained in which 112 were found to be upregulated in BTCC.Sequencing and homology analysis were performed for these 112clonies. The detection rates of some known genes (including IGF-1, human telomerase reverse transcriptase [hTERT], bladder cancerspecific nuclear matrix protein 4 [BLCA-4] and homeobox A13[HOXA13]) for BTCC at the Ta, T1 and &gt;T1 stages were 48%, 90%and 100%, respectively, with a specificity of 85%. The test specificitywas 80% for the 30 control patients with urinary tract infections. Thecombination of BLCA-4 and HOXA13 could distinguish betweenlow- and high-grade tumours, with specificity and sensitivity of 80%.Conclusion: We successfully constructed a reliable SSH library ofBTCC and found that combination detection insulin-like growthfactor 1 (IGF-1), hTERT, BLCA-4 and HOXA13 genes could helpto evaluate BTCC at different stages.

Abstract 1922: Plasminogen activator inhibitor type-1 (PAI-1) enhances bladder cancer tumor progression.

Abstract Introduction: Bladder cancer (BCa) is among the five most common malignancies worldwide and has the highest recurrence rate of any tumor type. BCa biology is poorly understood and early detection remains one of the most urgent issues in BCa research. Previously we have identified a preliminary molecular signature of BCa in voided urine samples from BCa patients. Our results revealed aberrant overexpression of the serine protease inhibitor (serpin) plasminogen activator inhibitor type-1 (PAI-1), both at the genomic and protein level. Increasing evidence indicate that elevated levels of PAI-1 correlates to a poor prognosis in many tumor types including breast, colorectal, gastric, ovarian, and more recently bladder cancer. PAI-1 has emerged as a potential target for BCa treatment and the aim of this study is to provide a greater understanding of its role in bladder cancer progression. Methods: PAI-1 expression levels in UROtsa (benign urothelial cell), KU7 (low-grade papillary urothelial cancer cell), and UMUC14 and T24 (high-grade invasive urothelial cancer cell lines) was tested by real-time PCR (RTPCR) and Western blot analysis. Secreted PAI-1 levels were measured by ELISA. KU7 stable clones overexpressing PAI-1 was established by transfection of plasmid DNA encoding human PAI-1. T24 and UMUC14 stable clones with shRNA knockdown of PAI-1 were established by infection with retroviral particles. Cell proliferation, migration, invasion, and apoptosis assays were performed in stable clones. Results: RTPCR analysis revealed an increase of PAI-1 in T24 and UMUC14 cells (27.9 and 5.2 times, respectively) while KU7 cells exhibited significantly reduced levels as compared to the UROtsa control. Similar results were observed for PAI-1 protein concentrations in cell culture supernatants and western blot analysis. While overexpression of PAI-1 in KU7 cells promoted an increase in proliferation, migration and invasion, these were significantly reduced upon inactivation of PAI-1 in T24 and UMUC14 cells stably expressing shRNA PAI-1. Loss of PAI-1 function also led to a substantial increase in apoptotic cell death. Conclusions: These results suggest that PAI-1 overexpression in BCa promotes tumor progression and therapeutic strategies targeting PAI-1 may serve clinically beneficial outcomes. Funding provided by research grants from Florida Department of Health James and Esther King Team Science Award 10KT-01 (CJR) and National Cancer Institute RO1 CA116161 (SG). Citation Format: Evan Gomes, Steven Goodison, Makito Miyake, Shanti Ross, Ge Zhang, Charles Rosser. Plasminogen activator inhibitor type-1 (PAI-1) enhances bladder cancer tumor progression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1922. doi:10.1158/1538-7445.AM2013-1922

Abstract 644: Identification of a novel hypermethylated tumor suppressor gene in human bladder cancer and its implication in non-invasive cancer detection using voided urine.

Abstract Bladder cancer is the sixth most common cancer in the world and the incidence is particularly high in southwestern Taiwan. Although bladder cancer patients have a low mortality rate, long term follow-up with repeated cystoscopy is required due to the high recurrence nature of the tumor. Therefore, a non-invasive detection assay is urgently required for bladder cancer patients. Aberrant promoter hypermethylation which is considered as a hallmark of cancer for over a decade, plays an important role in controlling cancer progression. Nevertheless, detection of promoter hypermethylation in bodily fluid has been implicated as a non-invasive and sensitive tool for cancer diagnosis. In this regard, by using illumina 27K CpG island methylation array, we identified a novel gene that are hypermethylated in 7 bladder cancer patient samples but not in primary normal human urothelim. In vitro promoter methylation assay and epigenetic treatment confirmed that the expression of this gene is epigenetically controlled in bladder cancer cell lines. Bisulphite pyrosequencing demonstrated a significant correlation between higher methylation level of this gene and cancer grade (P&amp;lt;0.01) and recurrence free survival (P&amp;lt;0.05) in bladder cancer patient samples (n=100). To investigate the diagnostic potential of this novel epigenetic marker in non-invasive cancer detection, we performed qMSP assay in voided urine samples from bladder cancer patients (n=27) as well as normal healthy control (n=19). The sensitivity and specificity of this gene in cancer detection is 42.3% and 100% respectively. The sensitivity can increase to 93.3% when using combination of this gene and our previously identified epigenetic markers (IRF8, p14, sFRP1). Re-expression of this gene in bladder cancer cell line inhibited cancer growth in soft agar colony formation assay and invasion assay. In conclusion, this gene may be a potential tumor suppressor in bladder cancer and its potential in non-invasive diagnosis of bladder cancer using voided urine warrants further investigation. Citation Format: Chia-Ming Yeh, Pi-Che Chen, Yu-Hsin Chen, Lin-Chien Hung, Yi-Ting Wang, Ming-Hsuan Tsai, Martin MS Huang, Cheng-Huang Shen, Cheng-Da Hsu, Michael WY Chan. Identification of a novel hypermethylated tumor suppressor gene in human bladder cancer and its implication in non-invasive cancer detection using voided urine. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 644. doi:10.1158/1538-7445.AM2013-644

Cost-Effectiveness of Fluorescence In Situ Hybridization in Patients with Atypical Cytology for the Detection of Urothelial Carcinoma

Patients with atypical cytology and equivocal or negative cystoscopy pose a challenge due to uncertainty about the presence of cancer. We determined the cost-effectiveness of using fluorescence in situ hybridization assays to determine the need for biopsy in patients with atypical cytology and equivocal or negative cystoscopy. Data from 2 large prospective studies evaluating the usefulness of fluorescence in situ hybridization in the setting of atypical cytology to detect urothelial carcinoma were combined. The data were used to calculate sensitivity and specificity for the UroVysion fluorescence in situ hybridization assay in various clinical scenarios. Cost data were obtained from our institution and Medicare reimbursement rates. Evaluations with or without bladder biopsy and with or without upper tract evaluation were considered. The study included 263 patients with atypical cytology and equivocal (62) or negative (201) cystoscopy. In patients with equivocal cystoscopy (assuming biopsy was performed in the operating room) biopsy based on fluorescence in situ hybridization results saved $1,740 per patient ($3,267 vs $1,527 per patient) and avoided 42 biopsies compared to biopsy in all patients. If office based biopsies were used then cost savings using fluorescence in situ hybridization results were $95 per patient. Among patients with negative cystoscopy biopsy based on fluorescence in situ hybridization resulted in costs savings of $2,241 per patient, avoiding 167 biopsies, compared to biopsy in all patients. Assuming office based biopsy, the cost savings were $216 per patient. The decision to perform biopsy based on fluorescence in situ hybridization assay in patients with atypical cytology and equivocal or negative cystoscopy was associated with a significant decrease in bladder cancer associated costs.

Urinary concentrations of type II collagen cleavage following anterior cruciate ligament reconstruction

Urinary concentrations of type II collagen cleavage following anterior cruciate ligament reconstruction

1140 EVALUATION OF URINARY EXOSOMES IN BLADDER CANCER PATIENTS

You have accessJournal of UrologyBladder Cancer: Basic Research (III)1 Apr 20131140 EVALUATION OF URINARY EXOSOMES IN BLADDER CANCER PATIENTS Jennifer J. Ahn, Guillermo Salazar, Matthew Seaman, Anna Scott, Piruz Motamedinia, Crystal Castaneda, Wayne Comper, James M. McKiernan, and Leileata Russo Jennifer J. AhnJennifer J. Ahn New York, NY More articles by this author , Guillermo SalazarGuillermo Salazar New York, NY More articles by this author , Matthew SeamanMatthew Seaman New York, NY More articles by this author , Anna ScottAnna Scott New York, NY More articles by this author , Piruz MotamediniaPiruz Motamedinia New York, NY More articles by this author , Crystal CastanedaCrystal Castaneda New York, NY More articles by this author , Wayne ComperWayne Comper New York, NY More articles by this author , James M. McKiernanJames M. McKiernan New York, NY More articles by this author , and Leileata RussoLeileata Russo New York, NY More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.755AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Despite significant strides in understanding the molecular pathogenesis of bladder cancer (BCa), diagnosis and surveillance continue to rely upon cystoscopy and cytology. As a result, there has been substantial interest in the use of biomarkers for diagnosis and risk stratification. However, current assays rely on tumor cells to be shed into the urine, or examine proteins that may not be specific for cancer. We have previously shown that urinary exosomes and microvesicles (collectively referred to as exosomes) are a consistent and reproducible means of sampling high integrity genetic material, but this has yet to be investigated in the context of BCa. Here, we examined the feasibility of evaluating the urinary exosomes of bladder cancer patients. METHODS Following Columbia University Medical Center IRB approval, voided urine samples were collected from 12 patients. BCa status at time of collection, as determined by cystoscopy and biopsy, was high-grade (HG) tumor (n = 5), low-grade (LG) tumor (n = 2), or no evidence of disease (NED) (n = 5). Samples were stored at 4°C and 0.8 μm filtration was used to remove whole cells and debris. Urinary exosomal RNA (exoRNA) was isolated using an in-house technique (Exosome Diagnostics, NY) and analyzed using RT-qPCR for 18 genes previously described as being differentially expressed in BCa. To compare expression levels, genes were standardized to 18S and relative quantitation (RQ) was calculated using DataAssist software (v.3.0). RESULTS Of the 18 genes analyzed, ICAM1 showed significantly higher expression in patients with HG BCa, as compared to NED patients (RQ 37.0, p = 0.0084). A similar trend was seen when comparing patients with any BCa (HG or LG) to NED patients (RQ 14.2, p = 0.051). UPK2 expression levels were not significantly different between the groups HG versus NED (RQ 1.6, p = 0.958) or LG versus NED (RQ 1.0, p = 0.975). CONCLUSIONS We successfully isolated exosomal RNA from the voided urine of patients with concurrent bladder cancer, as well as those with no evidence of disease. Additionally, significant upregulation of ICAM1 was seen in patients with HG disease. Ultimately, this technique shows promise as a non-invasive method of assessing gene expression in bladder cancer, independent of examining urothelial cells. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e465-e466 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jennifer J. Ahn New York, NY More articles by this author Guillermo Salazar New York, NY More articles by this author Matthew Seaman New York, NY More articles by this author Anna Scott New York, NY More articles by this author Piruz Motamedinia New York, NY More articles by this author Crystal Castaneda New York, NY More articles by this author Wayne Comper New York, NY More articles by this author James M. McKiernan New York, NY More articles by this author Leileata Russo New York, NY More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

1286 STEPWISE APPLICATION OF IMMUNOCYTOLOGY AND FISH IN THE SURVEILLANCE OF PATIENTS WITH NON-MUSCLE INVASIVE BLADDER CANCER

detection of BCa. Herein, we report the diagnostic utility of various multivariate combinations of these biomarkers. METHODS: We performed a case-controlled validation study in which voided urines from 127 patients (64 tumor bearing subjects) were analyzed. The urinary concentrations of 14 biomarkers (Interleukin-8 (IL-8), Matrix metalloproteinase-9 (MMP-9), Matrix metalloproteinase-10 (MMP-10), Syndecan 1 (SDC1), Chemokine (C-C motif) ligand 18 (CCL18), plasminogen activator inhibitor-1 (PAI-1), CD44, vascular endothelial growth factor (VEGF), Angiogenin (ANG), Carbonic anhydrase 9 (CA9), Alpha 1-Antitrypsin (A1AT), Osteopontin (OPN), Pentraxin3 (PTX3), and Apolipoprotein E (APOE)) were assessed by enzyme-linked immunosorbent assay (ELISA). Diagnostic performance of each biomarker and multivariate models were compared using receiver operating characteristic curves and the chi-square test. RESULTS: An 8-biomarker diagnostic signature achieved the most accurate BCa diagnosis (sensitivity 92%, specificity 97%). Subsequently, the validate diagnostic signature was tested by ELISA in an independent cohort of 308 patients (102 tumor bearing subjects and 206 diverse controls including UTI, stones, voiding symptoms, hematuria and normals). The 8-biomarker diagnostic signature was accurate in diagnosing BCa in voided urine samples (sensitivity 88%, specificity 91%). CONCLUSIONS: These data show that a multivariate urinebased assay can markedly improve the accuracy of non-invasive BCa detection. Further validation studies are under way to investigate the clinical utility of this panel of biomarkers for BCa diagnosis and disease monitoring.

Comparison of fluorescence in situ hybridization, NMP22 bladderchek, and urinary liquid‐based cytology in the detection of bladder urothelial carcinoma

The aim of this study is to evaluate the diagnostic values of the fluorescence in situ hybridization (FISH), NMP22 BladderChek, and liquid-based cytology (LBC) in the detection of bladder urothelial carcinoma (UC). Consecutive voided urine samples were collected from 138 in-house patients with a variety of urologic conditions and 37 healthy individuals as negative controls. FISH, NMP22 BladderChek, and LBC were performed on the specimens. All three tests were evaluated independently in a blinded fashion. In all, 104 out of the 175 patients enrolled in this study had histologically proven UC. LBC, FISH, and NMP22 BladderChek were successfully performed on 175, 149, and 119 cases, respectively. The three tests revealed overall sensitivities of 73.1%, 86.5%, and 67.6%, respectively. FISH was more sensitive than LBC (P=0.022) and NMP22 BladderChek (P=0.004). Combination of all the tests yielded a superior sensitivity of 96.7% compared with LBC (P<0.001), NMP22 BladderChek (P<0.001), and FISH (P=0.016), with the specificity only decreased slightly. Sensitivities of the three tests enhanced significantly with increasing UC grade (P<0.05). The positive rates of FISH and NMP22 BladderChek in equivocal cytologic diagnoses were 85.7% and 61.9% in UC, and 37.5% and 50.0% in non-UC (FISH: P=0.021; NMP22 BladderChek: P=0.683). FISH was more sensitive than LBC and NMP22 BladderChek. FISH had the ability to clarify equivocal cytologic diagnoses. Combination of all three tests showed an improvement in the sensitivity compared to any single test alone in detecting UC with the specificity slightly decreased.

A pilot study of urinary microRNA as a biomarker for urothelial cancer

Objective: MicroRNAs (miRNAs) are part of a class of small ribonucleic acid (RNAs). They are important regulatory molecules, involved in several cell processes, such as developmental timing, stem cell division and apoptosis. Dysregulated miRNAs have been identified in several human malignancies, including bladder cancer tissue samples, and may confer a “tumour signature” that can be exploited for diagnostic purposes. We report on a prospective pilot study investigating the diagnostic capability of miRNAs in the urine of patients with urothelial cancer.Methods: Voided urine samples were collected from patients with urothelial carcinoma just prior to bladder tumour resection, as well as age-matched healthy control patients. Pathology demonstrated both low- and high-grade cancer. Total RNA was isolated and quantitative reverse transcriptase-polymerase chain reaction was performed on the RNA extracts using primers for 4 miRNAs shown previously to be dysregulated in solid urothelial carcinomas with RNU6B as the endogenous control. Standard urine cytology was performed on all samples in a blinded fashion.Results: Two miRNAs of interest were dysregulated in the urine from cancer patients with miR-125b showing an average 10.42-fold decrease (p &lt; 0.01) and miR-126 showing an average 2.70-fold increase (p = 0.30) in the cancer samples compared to the normal controls. The sensitivity and specificity of the cytology on the same urine samples were 50% and 80%, respectively. Using these 2miRNAs only, a decision-tree prediction model was generated for a validation cohort of patients yielding a specificity of 100% and a sensitivity of 80%.Discussion: This preliminary study of candidate urinary miRNA inpatients with low- and high-grade urothelial cancer demonstrated a significantly improved diagnostic accuracy over cytology. These results provide rationale for further studies on discovery and validation of candidate miRNAs in voided urine and may potentially lead to the development of a non-invasive and sensitive test for bladder cancer diagnosis and prognosis.