Voided Urine Samples Research Articles

Detection of bladder cancer in urine sediments by a hypermethylation panel of selected tumor suppressor genes.

Promoter hypermethylation can be a useful biomarker for early detection and prognosis of bladder cancer, monitoring response to treatment and complement classical diagnostic procedures. The molecular test was performed on DNA from bladder cancer cells in voided urine samples, tumor tissue DNA and normal control DNAs. We aimed to assess the diagnostic potential of epigenetic changes in urine DNA from bladder cancer cases at various clinico-pathological stages of the disease. The methylation status of 5 genes (p14ARF, p16INK4A, RASSF1A, DAPK, APC) in 113 tumor samples paired with voided urine specimens was analyzed by MSP. We compared the results of methylation analysis with UroVysion test. The methylation profile in tumor/urine DNA was significantly correlated (p ≤ 0,05) with tumor grade in p14ARF, RASSF1a, APC/p14ARF, APC genes, respectively and with stage in p14ARF, RASSF1a/p14ARF genes, respectively. The results of UroVysion test were in correlation with hypermethylation both in tumor and urine DNA in p14ARF, RASSF1a and APC genes (p = 0,008; 0,02 and 0,04, respectively). Promoter hypermethylation of tumor suppressor genes is a frequent mechanism in bladder cancer. We found promoter hypermethylation in all grades and stages of all cases examined. Methylation profile of selected suppressor genes may be a potential useful biomarker and enhance early detection of bladder cancer using a noninvasive urine test.

Detection of Chlamydia trachomatis infections by polymerase chain reaction in asymptomatic pregnant women with special reference to the utility of the pooling of urine specimens.

Background & objectives:Genital Chlamydia trachomatis (CT) infections are one of the most prevalent sexually transmitted infections across the world. In pregnant women, if not detected and treated early, these may result in poor pregnancy outcomes and complications. The present study was aimed to screen CT infections from first void urine (FVU) samples of asymptomatic pregnant women using molecular methods. The secondary objective was to evaluate cost-effectiveness in pooling FVU samples for their diagnostic application.Methods:FVU samples were collected from 1000 asymptomatic pregnant women over a period of three years. Pooling was done by including five specimens in one pool in the amount of 10 µl and subjected to polymerase chain reaction (PCR) and further confirmed by direct fluorescent antibody assay (DFA).Results:The age of study participants ranged from 18 to 43 yr with the median±standard deviation of 26±3.84 yr. Majority of positive participants were younger than 25 years. A total of 200 pools were prepared and 20 of these were PCR positive. When individual specimen in 20 positive pools was tested, 20 PCR-positive specimens were identified from 19 pools, of which 16 were positive by DFA. Thus, CT was detected in 1.6 per cent asymptomatic pregnant women in India and pooling strategy resulted in 70 per cent reduction in a number of tests performed.Interpretation & conclusions:Our study detected C. trachomatis infection in 1.6 per cent asymptomatic pregnant women, and pooling of FVU specimens for PCR testing was found to be a cost-saving strategy in comparison to testing individual samples. Further evaluation and studies on the bigger sample size are warranted to validate these results.

A microRNA biomarker panel for the non-invasive detection of bladder cancer

The development of accurate, non-invasive urinary assays for bladder cancer would greatly facilitate the detection and management of a disease that has a high rate of recurrence and progression. In this study, we employed a discovery and validation strategy to identify microRNA signatures that can perform as a non-invasive bladder cancer diagnostic assay. Expression profiling of 754 human microRNAs (TaqMan low density arrays) was performed on naturally voided urine samples from a cohort of 85 subjects of known bladder disease status (27 with active BCa). A panel of 46 microRNAs significantly associated with bladder cancer were subsequently monitored in an independent cohort of 121 subjects (61 with active BCa) using quantitative real-time PCR (RT-PCR). Multivariable modeling identified a 25-target diagnostic signature that predicted the presence of BCa with an estimated sensitivity of 87% at a specificity of 100% (AUC 0.982). With additional validation, the monitoring of a urinary microRNA biomarker panel could facilitate the non-invasive evaluation of patients under investigation for BCa.

Prenatal Exposure to Glycol Ethers and Neurocognitive Abilities in 6-Year-Old Children: The PELAGIE Cohort Study.

Background:Glycol ethers (GE) are widely used organic solvents. Despite the potential neurotoxicity of several families of organic solvents, little is known about the impact of GE on the neurodevelopment of infants and children.Objectives:We investigated the relation between urinary concentrations of GE metabolites in pregnant women and neurocognitive abilities in their 6-year-old children in the PELAGIE mother–child cohort.Methods:Five GE metabolites were measured in first morning void urine samples of 204 French pregnant women in early pregnancy (< 19 weeks of gestation). Psychologists assessed the neurocognitive abilities of their 6-year-old children with the Wechsler Intelligence Scale for Children IV (WISC) and the Developmental Neuropsychological Assessment (NEPSY). We analyzed the results with linear (WISC) and Poisson regression models (NEPSY), adjusted for potential confounders, including child’s stimulation at home.Results:GE metabolites were detected in 90–100% of maternal urine samples. The WISC Verbal Comprehension score was significantly lower for children with the highest tertile of urinary phenoxyacetic acid (PhAA) [β (third vs. first tertile) = –6.53; 95% CI: –11.44, –1.62]. Similarly, the NEPSY Design Copying subtest score was lower in those with the highest tertile of urinary ethoxyacetic acid (EAA) [β (third vs. first tertile) = –0.11; 95% CI: –0.21, 0.00]. The other GE metabolites we studied were not significantly associated with WISC or NEPSY scores.Conclusions:Prenatal urine concentrations of two GE metabolites were associated with lower WISC Verbal Comprehension Index scores and NEPSY Design Copying subscale scores, respectively, at age 6 years. PhAA is the primary metabolite of 2-phenoxyethanol (EGPhE), which is commonly found in cosmetics, and precursors of EAA are frequently used in cleaning agents. Additional research is needed to confirm our findings and further explore potential effects of prenatal GE exposures on neurocognitive performance in children.Citation:Béranger R, Garlantézec R, Le Maner-Idrissi G, Lacroix A, Rouget F, Trowbridge J, Warembourg C, Monfort C, Le Gléau F, Jourdin M, Multigner L, Cordier S, Chevrier C. 2017. Prenatal exposure to glycol ethers and neurocognitive abilities in 6-year-old children: the PELAGIE cohort study. Environ Health Perspect 125:684–690; http://dx.doi.org/10.1289/EHP39

Validation of a DNA Methylation-Mutation Urine Assay to Select Patients with Hematuria for Cystoscopy

Only 3% to 28% of patients referred to the urology clinic for hematuria are diagnosed with bladder cancer. Cystoscopy leads to high diagnostic costs and a high patient burden. Therefore, to improve the selection of patients for cystoscopy and reduce costs and over testing we aimed to validate a recently developed diagnostic urine assay. Included in study were 200 patients from a total of 3 European countries who underwent cystoscopy for hematuria, including 97 with bladder cancer and 103 with nonmalignant findings. Voided urine samples were collected prior to cystoscopy. DNA was extracted and analyzed for mutations in FGFR3, TERT and HRAS, and methylation of OTX1, ONECUT2 and TWIST1. Logistic regression was used to analyze the association between predictor variables and bladder cancer. Combining the methylation and mutation markers with age led to an AUC of 0.96 (95% CI 0.92-0.99) with 93% sensitivity and 86% specificity, and an optimism corrected AUC of 0.95. The AUC was higher for T1 or greater tumors compared to Ta tumors (0.99 vs 0.93). The AUC was also higher for high grade tumors compared to low grade tumors (1.00 vs 0.93). Overall negative predictive value was 99% based on the 5% to 10% prevalence of bladder cancer in patients with hematuria. This would lead to a 77% reduction in diagnostic cystoscopy. Analyzing hematuria patients for the risk of bladder cancer using novel molecular markers may lead to a reduction in diagnostic cystoscopy. Combining methylation analysis (OTX1, ONECUT2 and TWIST1) with mutation analysis (FGFR3, TERT and HRAS) and patient age resulted in a validated accurate prediction model.

A multiplex urinary immunoassay for bladder cancer detection: analysis of a Japanese cohort.

BackgroundBladder cancer (BCa) is among the most commonly diagnosed malignancies worldwide, and due the high rate of post-operative disease recurrence, it is one of the most prevalent in many countries. The development of non-invasive molecular assays that can accurately detect and monitor BCa would be a major advance, benefiting both patients and healthcare systems. We have previously identified a urinary protein biomarker panel that is being developed for application in at-risk patient cohorts. Here, we investigated the potential utility of the multiplex assay in a Japanese cohort.MethodsThe Japanese study cohort collected from urology clinics at two institutions was comprised of a total of 288 subjects. The protein biomarker panel (IL8, MMP9, MMP10, ANG, APOE, SDC1, A1AT, PAI1, CA9, VEGFA) was monitored in voided urine samples collected prior to cystoscopy using a custom multiplex ELISA assay. The diagnostic performance of the biomarker panel was assessed using receiver operator curves, predictive modeling and descriptive statistics.ResultsUrinary biomarker concentrations were significantly elevated in cases versus controls, and in cases with high-grade and muscle-invasive tumors. The AUC for the 10-biomarker assay was 0.892 (95 % confidence interval 0.850–0.934), with an overall diagnostic sensitivity specificity of 0.85 and 0.81, respectively. A predictive model trained on the larger institutional cohort correctly identified 99 % of the cases from the second institution.ConclusionsUrinary levels of a 10-biomarker panel enabled discrimination of patients with BCa. The multiplex urinary diagnostic assay has the potential to be developed for the non-invasive detection of BCa in at-risk Japanese patients.

Clinical significance of α‑ and β‑Klotho in urothelial carcinoma of the bladder.

Non-muscle invasive bladder cancer(NMIBC) accounts for ~70% of all bladder cancers. One of the serious clinical issues related to the management of NMIBC is that it has significant potential to progress to muscle invasive bladder cancer(MIBC) after initial treatments. α‑Klotho(KLα), originally identified as an anti‑aging gene, has recently been reported to have antitumor effects in various malignancies. In contrast, β‑Klotho(KLβ) has been reported to have protumoral functions. However, the associations between KLα/KLβ and the biological behavior of urothelial carcinoma remain unclear. In the present study, we evaluated the association between clinicopathological background factors of NMIBC and the expression levels of KLαorKLβ. A high expression level of KLβ, but notKLα, was an independent predictive factor of short progression‑free survival for NMIBC. An elevated level of KLβ correlated with a higher incidence of lymphovascular invasion(LVI). We addedinvitro assays using human bladder cancer cell lines to investigate the role of KLβ. Treatment with exogenous KLβ protein increased the proliferation, migration, transendothelial migration abilities and anchorage‑independent growth of the cell lines. In addition, the KLβ concentration in voided urine samples obtained before initial transurethral surgery was quantitated with enzyme‑linked immunosorbent assay(ELISA). The urine KLβ concentration was found to be higher in patients with bladder cancer than that in healthy volunteers. Our results suggest that KLβ plays important roles in tumor invasion and progression, and its concentration may be a valuable urine‑based marker for the detection of bladder cancer.

Validation of the diagnostic utility of urinary midkine for the detection of bladder cancer.

As it has been demonstrated previously that midkine (also known as neurite growth-promoting factor 2) protein levels in urine of bladder cancer (BCa) patients are increased compared to healthy controls, the present study validated the diagnostic utility of midkine in an independent patient cohort and compared the observed values with voided urine cytology (VUC), which is the current reference standard for non-invasive diagnosis of BCa. Voided urine samples were prospectively collected from 92 BCa patients and 70 control subjects. Protein levels of midkine were assessed using a commercially available enzyme-linked immunosorbent assay and normalized to urinary creatinine. The diagnostic performance of urinary midkine was evaluated by receiver operating characteristic curves. The best combinations of sensitivities and specificities were determined by Youden's Index. Midkine concentrations were significantly elevated in urine samples from BCa patients compared to controls (P<0.001; Mann-Whitney U Test). The level of midkine was associated with disease progression, with the highest concentrations in urine specimens of patients with pT1 and ≥pT2a, as well as high-grade tumors (P<0.001; Mann-Whitney U test). Sensitivities of urinary midkine and VUC were 69.7 and 87.6%, respectively. The corresponding specificities for midkine and VUC were 77.9 and 87.7%, respectively. The combined use of VUC and midkine improved the sensitivity to 93.3%, but reduced the specificity to 66.2%. Despite its reduced discriminatory power for low-grade and low-stage BCa, urinary midkine can be utilized for the identification of high-grade pT1 and ≥pT2a tumors. This means that midkine may potentially be suitable for the identification of patients with high risk BCa.

Correlation between the presence of degenerated inclusion-bearing cells in voided urine samples and the occurrence of polyomavirus infection.

The purpose of the present, prospective, cohort study was to monitor urine cytology samples from recipients of renal transplants to search for the occurrence of decoy cells and degenerated inclusion-bearingcells with an aim to correlate the existence of these cells with molecular detection of polyomavirus BK (BKV) DNA in urine. This study included patients who underwent renal transplantation. Patients had their urine tested quarterly, during the first year post-transplantation, for the presence of decoy cells and degenerated cells, as well as by quantitative determination of BKV load in the urine and plasma. Three hundred and sixty-one examinations were performed on 101 patients within 12months of attendance. Urine cytology results were: 198 (54.9%) negative and 60 (16.6%) positive for the presence of viral cytopathic effects depending on the presence of BKV infection, 72 (19.9%) positive for the manifestation of degenerated cells and 31 (8.6%) unsatisfactory for analysis. There was a subtle tendency towards the presence of degenerated inclusion-bearing cells in cases in which the virus was detected in voided urine. However, the presence of degenerated cells exhibited a tendency to BKV positivity in months 3, 6 and 9 and, exclusively in month 12, this trend was statistically significant. There were not enough strong morphological and staining elements to state the origin of the degenerated cells or to describe the nature of the infection (viral or bacterial), given that these cells were undergoing an apoptotic process in post renal transplant patients.

O032 Implementing a test and treat pathway forMycoplasma genitaliumin men with urethritis attending a GUM clinic

Background/introduction National guidelines recommend testing men with non-gonococcal urethritis(NGU) for Mycoplasma genitalium (MG) where testing is available. Recent studies have shown concerning levels of macrolide resistance and high rates of treatment failure with 1 g azithromycin. In response to this, we changed our standard treatment of NGU to doxycycline and implemented a test and treat pathway for MG in male NGU. Aim(s)/objectives To determine the prevalence of MG and to measure clearance rates of infection post-treatment. Methods From 1 st September 2015 first void urine samples from men with NGU were routinely tested for MG using the Fast Track Diagnostics™ urethritis PCR. Men with confirmed MG were given 5days azithromycin and offered test of cure at 4 weeks. Results 304 men had NGU over 5months. Mean age 33.2 years, 64.5% heterosexual, 77.6% white ethnicity and 8.2% HIV-positive. Tests for MG were performed in 230/304 (76%). 16.5% (38/230) were MG positive compared with 22.5% chlamydia. 6 (15.8%) men with MG were co-infected with chlamydia and 1 (2.6%) had urethral gonorrhoea. 20/38 men with MG (52.6%) were given azithromycin 5days, 8/38 (21.1%) had azithromycin 1g stat. 7/38 (18.4%) re-tested positive at 4 weeks, 6 of whom had initially had azithromycin 5days. All were given moxifloxacin 400mg for either 7, 10 or 14 days. Discussion/conclusion The high rates of MG found support routine testing in men with NGU. Despite appropriate treatment, some men returned with positive TOC suggestive of macrolide failure. This demonstrates a need for more widely available MG diagnostics with resistance testing in the UK and attention to antimicrobial stewardship so that NGU can be better managed.

Urinalysis in chinchillas (Chinchilla lanigera).

To evaluate urine variables in chinchillas (Chinchilla lanigera). Evaluation study. Urine samples from 41 chinchillas. Voided urine samples were collected from clinically normal chinchillas that were exhibited during a breeder exposition. Urinalysis was performed within 1 hour after collection. Urine specific gravity (USG) was measured before and after centrifugation with a handheld veterinary refractometer. Urine dipstick analysis and microscopic sedimentation examination were performed on all samples. Additionally, a urine sulfosalicylic acid (SSA) precipitation test and quantitative protein analysis were performed on samples with sufficient volume. 17 of 41 (41%) samples had a USG ≥ 1.050, and USG ranged from 1.014 to > 1.060. The USG before centrifugation did not differ significantly from that after centrifugation. Protein was detected in all urine samples on dipstick analysis. The SSA precipitation test yielded negative results for all samples tested. Results of the quantitative protein analyses were not correlated with the results of the dipstick analyses or SSA tests. The recorded pH for all samples was 8.5, which was the upper limit of detection for the reagent strip. Glucose and ketones were detected in 5 and 6 samples, respectively. Crystals were observed in 28 of 41 (68%) samples; 27 of those samples contained amorphous crystals. Urinalysis results for clinically normal chinchillas were provided. For chinchilla urine samples, measurement of USG by refractometry prior to centrifugation is acceptable and protein concentration should be determined by quantitative protein analysis rather than dipstick analysis or the SSA test.

Expression Level of Urinary MicroRNA-146a-5p Is Increased in Patients With Bladder Cancer and Decreased in Those After Transurethral Resection

BackgroundBladder cancer is the most prevalent malignancy involving the urinary system and exhibits a markedly high recurrence rate. Therefore, reliable and noninvasive diagnostic and surveillance methods are desperately needed. Patients and MethodsCandidate microRNAs (miRNAs) were selected from the miRNAs that were differentially expressed in bladder cancer cell lines (T24 and RT4) compared to normal ureteral epithelial tissue using miRNA-microarray analysis. The candidate miRNAs were validated by quantitative reverse transcription polymerase chain reaction assay using voided urine samples. ResultsWe identified 3 miRNAs (miR-301b, -563, and -146a-5p) that demonstrated > 2-fold higher expression levels in cancer cell lines than in the normal ureteral epithelial tissue. Of these, only miR-146a-5p was consistently and significantly higher in urine samples from the patients with bladder cancer than in those from the normal individuals (P = .0014). The patients with high-grade tumors exhibited significantly higher urinary miR-146a-5p levels than those with low-grade tumors, and the patients with invasive tumors tended to show higher urinary miR-146a-5p levels than those with noninvasive tumors. Elevated urinary miR-146a-5p levels in patients with bladder cancer were decreased to the normal level after transurethral resection of the tumors (P = .0214). ConclusionOur study suggested that urinary miR-146a-5p might be useful as a new noninvasive diagnostic marker, therapeutic target, or anticancer agent for bladder cancer, as well as for increasing our understanding of cancer biology.

Effects of cranberry extract on prevention of urinary tract infection in dogs and on adhesion of Escherichia coli to Madin-Darby canine kidney cells

To determine effects of cranberry extract on development of urinary tract infection (UTI) in dogs and on adherence of Escherichia coli to Madin-Darby canine kidney (MDCK) cells. 12 client-owned dogs (in vivo experiment) and 6 client-owned dogs (in vitro experiment). 12 dogs with a history of recurrent UTI received an antimicrobial (n = 6) or cranberry extract (6) orally for 6 months. Dogs were monitored for a UTI. For the in vitro experiment, cranberry extract was orally administered to 6 dogs for 60 days. Voided urine samples were collected from each dog before and 30 and 60 days after onset of extract administration. Urine was evaluated by use of a bacteriostasis assay. An antiadhesion assay and microscopic examination were used to determine inhibition of bacterial adherence to MDCK cells. None of the 12 dogs developed a UTI. The bacteriostasis assay revealed no zone of inhibition for any urine samples. Bacterial adhesion was significantly reduced after culture with urine samples obtained at 30 and 60 days, compared with results for urine samples obtained before extract administration. Microscopic examination revealed that bacterial adherence to MDCK cells was significantly reduced after culture with urine samples obtained at 30 and 60 days, compared with results after culture with urine samples obtained before extract administration. Oral administration of cranberry extract prevented development of a UTI and prevented E coli adherence to MDCK cells, which may indicate it has benefit for preventing UTIs in dogs.

Measuring Personal Exposure to Organophosphate Flame Retardants Using Silicone Wristbands and Hand Wipes.

Organophosphate flame retardants (PFRs) are widely used as replacements for polybrominated diphenyl ethers in consumer products. With high detection in indoor environments and increasing toxicological evidence suggesting a potential for adverse health effects, there is a growing need for reliable exposure metrics to examine individual exposures to PFRs. Silicone wristbands have been used as passive air samplers for quantifying exposure in the general population and occupational exposure to polycyclic aromatic hydrocarbons. Here we investigated the utility of silicone wristbands in measuring exposure and internal dose of PFRs through measurement of urinary metabolite concentrations. Wristbands were also compared to hand wipes as metrics of exposure. Participants wore wristbands for 5 consecutive days and collected first morning void urine samples on 3 alternating days. Urine samples were pooled across 3 days and analyzed for metabolites of the following PFRs: tris(1,3-dichloroisopropyl) phosphate (TDCIPP), tris(1-chloro-2-isopropyl) phosphate (TCIPP), triphenyl phosphate (TPHP), and monosubstituted isopropylated triaryl phosphate (mono-ITP). All four PFRs and their urinary metabolites were ubiquitously detected. Correlations between TDCIPP and TCIPP and their corresponding urinary metabolites were highly significant on the wristbands (rs = 0.5-0.65, p < 0.001), which suggest that wristbands can serve as strong predictors of cumulative, 5-day exposure and may be an improved metric compared to hand wipes.

Multigene Methylation Analysis And The Noninvasive Diagnosis Of Prostate Cancer From Body Fluids

Introduction During prostatic carcinogenesis, DNA hypermethylation occurs, thus representing a promising biomarker for the early detection of this malignancy. In our study, we aim to determine the usefulness of a molecular and multigene test for prostate cancer. However, this is based on the quantitative methylation-specific polymerase chain reaction (qMSP) of three genes from voided urine specimens by noninvasive methods. Materials and Methods In this study, the voided urine specimens were collected from 89 patients with prostate cancer and 69 controls. Genomic DNA was isolated and subjected to bisulfite modification. Consequently, we tested the methylation status of genomic DNA of three genes, namely: GSTP1, APC, and MDR1. This was done using the quantitative methylationspecific PCR method. Therefore, the obtained results were correlated with the clinicopathologic findings. Results Promoter methylation of GSTP1 gene in voided urine samples was found in 87 out of 89 (97.8%) PCa patients and in 13 out of 62 (21 %) BPH men. In APC gene, methylated levels have been found in 61 out of 89 (68.5%) PCa patients and in 8 out of 62 (12.9%) BPH men. MDR1 gene was found to be hypermethylated in 60 out of 89 (67.4%) PCa patients and in 4 out of 62 (6.5%) BPH men. In addition, we obtained a sensitivity of 88.99% and a specificity of 85.5% for the multigene panel. The AUC in this case was 0.927. Conclusion The analysis of a multigene panel of three methylated genes in prostate cancer by qMSP, can be used to distinguish between men with malignant and benign prostatic diseases from voided urine specimens. Also, it can be used for the follow-up of those men who are presenting increased risk of prostate cancer by noninvasive methods.

PI-LBA07 THE DEVELOPMENT AND CLINICAL VALIDATION OF A HIGH SENSITIVITY URINE BIOMARKER TEST FOR THE DETERMINATION OF RECURRENCE IN UROTHELIAL CARCINOMA PATIENTS

You have accessJournal of UrologyPlenary I - Tuesday1 Apr 2016PI-LBA07 THE DEVELOPMENT AND CLINICAL VALIDATION OF A HIGH SENSITIVITY URINE BIOMARKER TEST FOR THE DETERMINATION OF RECURRENCE IN UROTHELIAL CARCINOMA PATIENTS Yair Lotan, Jay Raman, Shahrokh Shariat, Laimonis Kavalieris, Chris Frampton, Henry Crist, Elsie Jacobson, Jimmie Suttie, David Darling, Scott Asroff, Doug Scherr, George Adams, Evan Goldfischer, Jeffrey Thill, Joseph Williams, Joshua Stein, and Paul O'Sullivan Yair LotanYair Lotan More articles by this author , Jay RamanJay Raman More articles by this author , Shahrokh ShariatShahrokh Shariat More articles by this author , Laimonis KavalierisLaimonis Kavalieris More articles by this author , Chris FramptonChris Frampton More articles by this author , Henry CristHenry Crist More articles by this author , Elsie JacobsonElsie Jacobson More articles by this author , Jimmie SuttieJimmie Suttie More articles by this author , David DarlingDavid Darling More articles by this author , Scott AsroffScott Asroff More articles by this author , Doug ScherrDoug Scherr More articles by this author , George AdamsGeorge Adams More articles by this author , Evan GoldfischerEvan Goldfischer More articles by this author , Jeffrey ThillJeffrey Thill More articles by this author , Joseph WilliamsJoseph Williams More articles by this author , Joshua SteinJoshua Stein More articles by this author , and Paul O'SullivanPaul O'Sullivan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.03.130AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES To develop a high sensitivity, urine based, gene expression test for the determination of recurrence of Urothelial Carcinoma (UC) and to assess the performance of this test for the segregation of patients with a low probability of recurrence from those requiring further investigation. METHODS 1093 samples were collected from 803 patients prospectively recruited at 11 community and tertiary centres the United States. Voided urine samples were collected pre-cystoscopy for urine dipstick, NMP22 Bladderchek, NMP22 ELISA, urine cytology and Cxbladder. Local cytology and FISH were also collected where available. Patients were partitioned into either a training or validation set based on demographic and risk factors. The training set (n=354 patients) was used to develop a novel algorithm (Cxbladder Monitor) encompassing gene expression data and previous tumor occurrence information. Cxbladder Monitor was validated using an independent validation set (n=449 patients). Final model parameters were derived on all data and bootstrap methods used to estimate 95% CIs and performance characteristics. RESULTS Incidence of confirmed UC within the recruited population was 11%. Confirmed tumor stage was as follows : 64% Ta, 14% T1, 22% Tis. Grade was confirmed as 50% low grade and PUMLMP and 50% high grade disease. Final model Cxbladder Monitor exhibited a sensitivity of 93% (95% CI 82, 97), outperforming all direct comparator tests and locally obtained FISH results, across all stages and grades of tumor; see table 1. Cxbladder Monitor demonstrated an NPV of 97% (95% CI: 93, 98). CONCLUSIONS Cxbladder Monitor has been developed in response to the limitations of current urine based tests, and the unmet clinical need for a high sensitivity, high NPV test to enhance the clinical investigation of recurrent UC. Cxbladder Monitor demonstrated potential to rule out those patients with a low risk of having recurrent disease and therefore to reduce cystoscopy burden on patients undergoing routine investigation. Use of Cxbladder Monitor could reduce the number of invasive procedures required for the evaluation of recurrent UC. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1071 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Yair Lotan More articles by this author Jay Raman More articles by this author Shahrokh Shariat More articles by this author Laimonis Kavalieris More articles by this author Chris Frampton More articles by this author Henry Crist More articles by this author Elsie Jacobson More articles by this author Jimmie Suttie More articles by this author David Darling More articles by this author Scott Asroff More articles by this author Doug Scherr More articles by this author George Adams More articles by this author Evan Goldfischer More articles by this author Jeffrey Thill More articles by this author Joseph Williams More articles by this author Joshua Stein More articles by this author Paul O'Sullivan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

S&T-10 PROSTATE CANCER DIAGNOSIS BY MULTIPLE SECRETED PROTEIN BIOMARKERS IN VOIDED URINE

INTRODUCTION AND OBJECTIVES: Prostate cancer is currently screened by an imperfect serum PSA test. Since PSA is not specific to cancer, this leads to overdiagnosis where men undergo unnecessary biopsies to find no cancer, and overtreatment for tumors that may not be life threatening. Better markers are needed. Using cell type-specific transcriptomes, genes with increased expression in cancer cells (of Gleason patterns 3 and 4) and cancer-associated stromal cells (of different Gleason scores) were identified, and those encoding secreted proteins were selected for marker development: AGR2, AGR3, CCL3, CEACAM5, CEACAM6, CRISP3, CXCL14, IL24, MMP9, POSTN, SFRP4, CD90. METHODS: Voided urine samples were collected from consented patient donors at pre-op, and healthy controls with no diagnosis of cancer. The urine was spin concentrated to 1 ml and the protein concentration was estimated by BCA Protein Assay. PRISM-SRM (high pressure high-resolution separations coupled with intelligent selection and multiplexing-selective reaction monitoring) targeted proteomics was used to measure simultaneously these protein biomarkers in individual samples. The marker amounts were presented as ratios of PSA, which was used for 0normalization0. In data analysis, cancer severity ranking was generated from pathology parameters of the tumors including Gleason score, tumor volume and stage. Statistical analysis and graphical display of the data were carried out with OriginPro 2015. RESULTS: The data from a cohort of 14 cases (UW) and 25 controls (UTSA + UW) was analyzed to produce AUC (area under the curve) values ranging from 0.774 for AGR2, 0.820 for CXCL14, 0.820 for CEACAM5, 0.837 for CD90, 0.871 for MMP9, 0.889 for SFRP4, 0.889 for IL24, 0.894 for CRISP3, to 0.906 for CEACAM6, and 0.914 for the marker panel of nine proteins (P1⁄42.20E-05) in distinguishing cancer from non-cancer. For correlation with tumor pathology, the CEACAM5/ CRISP3 concentration ratios showed increase with severity ranking (R1⁄40.5219) while the CD90/CRISP3 ratios showed decrease with ranking (R1⁄40.445). High CEACAM5/CRISP3 and low CD90/CRISP3 ratios were indicative of high risk disease. Other ratios had lower R values. CONCLUSIONS: For prostate cancer detection, voided urine provides the least invasive means for cancer detection. The AUC value obtained from the marker panel is superior to that of PHI (0.708) and MiPS (Michigan Prostate Score, 0.762). Urine collection is simple (at patients0 convenience) and does not require a vigorous DRE, which makes the test suitable for population screening. The marker ratios could also provide information on high risk vs. low risk regarding the cancer detected. A useful multimarker test could eventually be applied to replace biopsy.

PD13-05 FLUORESCENCE IN SITU HYBRIDIZATION (FISH) IN THE DIAGNOSIS OF BLADDER AND UPPER TRACT UROTHELIAL CARCINOMA: A SINGLE-INSTITUTION EXPERIENCE

You have accessJournal of UrologyBladder Cancer: Upper Tract Transitional Cell Carcinoma I1 Apr 2016PD13-05 FLUORESCENCE IN SITU HYBRIDIZATION (FISH) IN THE DIAGNOSIS OF BLADDER AND UPPER TRACT UROTHELIAL CARCINOMA: A SINGLE-INSTITUTION EXPERIENCE Mark Mann, Ryan Cleary, Leonard Gomella, Scott Hubosky, Peter McCue, Demetrius Bagley, Costas Lallas, and Edouard Trabulsi Mark MannMark Mann More articles by this author , Ryan ClearyRyan Cleary More articles by this author , Leonard GomellaLeonard Gomella More articles by this author , Scott HuboskyScott Hubosky More articles by this author , Peter McCuePeter McCue More articles by this author , Demetrius BagleyDemetrius Bagley More articles by this author , Costas LallasCostas Lallas More articles by this author , and Edouard TrabulsiEdouard Trabulsi More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.976AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Bladder cancer (BC) is the second most common urological cancer while upper tract urothelial carcinoma (UTUC) is far rarer with approximately 7000 new cases per year. Urine cytology is the only widespread urinary marker is use but has many limitations including low sensitivity and wide interpreter variability. We evaluated the multi target UroVysion (Abbott Molecular, IL) fluorescence in situ hybridization (FISH) assay for the diagnosis of urothelial cancer (UC) in patients with BC only, UTUC only, and combined BC and UTUC. METHODS A single institution retrospective analysis was conducted comparing sensitivity, specificity, positive predictive value, and negative predictive values. FISH within 6 months of endoscopic evaluation were obtained from outpatient voided urine samples. Our institutional pathology department confirmed disease stage from specimens obtained during endoscopic evaluations for lower tract disease. For upper tract disease, disease was confirmed by retrograde ureteroscopy, biopsies of visual lesions, and site specific upper tract cytology. RESULTS A total of 415 patients submitted voided FISH specimens. Seventy-seven patients after evaluation for UC had no evidence of malignancy. One hundred and seventy-six patients had BC, 65 had UTUC, and 32 had both BC and UTUC. Overall FISH was more sensitive than cytology 54.9% in comparison with cytology 42.2% (p=0.01). However, specificity favored cytology 92.9% compared to 73.5% with FISH (p<0.01). After analyzing BC only patients, the same significant finding of increased sensitivity and decreased specificity was identified. For UTUC, FISH had a trend toward being more sensitive, but it was not significant. For both upper and lower tract disease there was no difference in the sensitivities. Cytology had improved positive predictive value (PPV) over FISH, 76.9% in comparison to 64.6% (p=0.02). Negative predictive value (NPV) also favored cytology 74.2% vs. 64.9% (p = 0.02). When analyzing individual cohorts, cytology had improved PPV for BC alone patients. UTUC showed no difference for PPV and NPV. For both UTUC and BC, NPV was slightly favored for FISH over cytology 93.2% vs. 91.2% (p= 0.03). CONCLUSIONS Voided urine FISH testing does offer a higher detection of UC for BC compared to voided cytology; however specificity was worse. In analysis of the separate cohorts, FISH does not appear to improve detection of UC in patients with either UTUC only or both BC and UTUC. FISH alone should not replace cytology as a standalone urinary tumor assay for detection of UC. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e299 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Mark Mann More articles by this author Ryan Cleary More articles by this author Leonard Gomella More articles by this author Scott Hubosky More articles by this author Peter McCue More articles by this author Demetrius Bagley More articles by this author Costas Lallas More articles by this author Edouard Trabulsi More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP88-12 HIGHLY SENSITIVE AND SPECIFIC NOVEL BIOMARKERS FOR THE DIAGNOSIS OF TRANSITIONAL BLADDER CARCINOMA

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology V1 Apr 2016MP88-12 HIGHLY SENSITIVE AND SPECIFIC NOVEL BIOMARKERS FOR THE DIAGNOSIS OF TRANSITIONAL BLADDER CARCINOMA Hong Koo Ha, Prashant Kumar, Sayantani Nandi, Tuan Zea Tan, Siok Ghee Ler, Kee Seng Chia, Wei-Yen Lim, Ja Yoon Ku, Chan Ho Lee, Kyung Lee, Kyoung Hwan Kim, Seung Ryong Baek, Hyo Jung Park, Jeong Zoo Lee, and Jean Paul Thiery Hong Koo HaHong Koo Ha More articles by this author , Prashant KumarPrashant Kumar More articles by this author , Sayantani NandiSayantani Nandi More articles by this author , Tuan Zea TanTuan Zea Tan More articles by this author , Siok Ghee LerSiok Ghee Ler More articles by this author , Kee Seng ChiaKee Seng Chia More articles by this author , Wei-Yen LimWei-Yen Lim More articles by this author , Ja Yoon KuJa Yoon Ku More articles by this author , Chan Ho LeeChan Ho Lee More articles by this author , Kyung LeeKyung Lee More articles by this author , Kyoung Hwan KimKyoung Hwan Kim More articles by this author , Seung Ryong BaekSeung Ryong Baek More articles by this author , Hyo Jung ParkHyo Jung Park More articles by this author , Jeong Zoo LeeJeong Zoo Lee More articles by this author , and Jean Paul ThieryJean Paul Thiery More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2430AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Transitional bladder carcinoma (BCa) is prevalent in developed countries, particularly among men. Given that these tumors frequently recur or progress, the early detection and subsequent monitoring of BCa at different stages is critical. Current BCa diagnostic biomarkers are not sufficiently sensitive for substituting or complementing invasive cystoscopy. Here, we sought to identify a robust set of urine biomarkers for BCa detection. METHODS Using a high-resolution, mass spectrometry-based, quantitative proteomics approach, we measured, compared and validated protein variations in 451 voided urine samples from healthy subjects, non-bladder cancer patients and patients with non-invasive and invasive BCa. We identified five robust biomarkers: Coronin-1A, Apolipoprotein A4, Semenogelin-2, Gamma synuclein and DJ-1/PARK7. In diagnosing Ta/T1 BCa, these biomarkers achieved an AUC of 0.92 and 0.98, respectively, using ELISA and western blot data (sensitivity, 79.2% and 93.9%; specificity, 100% and 96.7%, respectively). In diagnosing T2/T3 BCa, an AUC of 0.94 and 1.0 was attained (sensitivity, 86.4% and 100%; specificity, 100%) using the same methods. RESULTS Urine samples from 66 healthy subjects, 110 Ta/T1 BCa patients and 63 T2/T3 BCa patients were analyzed using western blotting. All BCa urine samples exhibited an elevated level in at least three out of the five biomarkers. ROC curve, established using band intensity values, showed an AUC of 0.98 and 1.0 for healthy versus Ta/T1 and healthy versus T2/T3, respectively, using the five biomarkers in combination CONCLUSIONS We established a robust, urinary protein biomarker panel for BCa diagnosis. This panel consists of five biomarkers with significantly higher sensitivity and specificity than that of the currently available, noninvasive diagnostic assays for the detection of BCa. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1133 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Hong Koo Ha More articles by this author Prashant Kumar More articles by this author Sayantani Nandi More articles by this author Tuan Zea Tan More articles by this author Siok Ghee Ler More articles by this author Kee Seng Chia More articles by this author Wei-Yen Lim More articles by this author Ja Yoon Ku More articles by this author Chan Ho Lee More articles by this author Kyung Lee More articles by this author Kyoung Hwan Kim More articles by this author Seung Ryong Baek More articles by this author Hyo Jung Park More articles by this author Jeong Zoo Lee More articles by this author Jean Paul Thiery More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Screening for asymptomatic urogenital Chlamydia trachomatis infection at a large Dublin maternity hospital: results of a pilot study.

There are currently no Irish guidelines on screening for Chlamydia trachomatis infection in pregnancy. Prevalence rates in the antenatal population are not known which has prevented the development of screening recommendations for this group. The objective of this study was to determine the prevalence of asymptomatic urogenital C. trachomatis infection in young women attending for care at a large maternity hospital. All patients aged 25years and under attending the Hospital between December 2011 and December 2013 were offered screening for urogenital C. trachomatis infection. Nucleic acid amplification testing of the C. trachomatis cryptic plasmid was performed on either endocervical swabs or first void urine samples. There were 2687 women tested for C. trachomatis infection, 83.4% (2241/2687) through the antenatal clinics, 7.1% (193/2687) through the gynaecology clinic, and 9.4% (253/2687) through the emergency department. The rate of a positive test result was 5.6% (151/2687) overall. The rates in women ages 16-18, 19-21 and 22-25years were 9.1% (31/340), 6.5% (50/774) and 4.4% (69/1561), respectively. A positive test result was more likely in those who were unemployed (p=0.04), those who were Irish (p=0.03) and those who were unmarried (p<0.01). There were no cases of neonatal C. trachomatis infection in babies born to mothers who were screened in early pregnancy. The prevalence rate of detected C. trachomatis infection was 5.6% in the study population. Screening of antenatal patients may have a role in preventing vertical transmission of infection to the neonate.