Voided Urine Samples Research Articles

The Prevalence and Clinical Significance of Urethritis and Cervicitis in Asymptomatic People by Use of Multiplex Polymerase Chain Reaction

PurposeOur purpose was to conduct a screening test for urethritis or cervicitis as a sexually transmitted disease (STD) by using multiplex polymerase chain reaction(PCR) and to determine the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Mycoplasma hominis, and Trichomonas vaginalis in asymptomatic people.Materials and MethodsFrom July 2010 to December 2010, 709 persons who came to the hospital for a general checkup were tested. Multiplex PCR assays were done with first voided urine samples or endocervical swabs by use of the Seeplex® STD6 ACE Detection kit.ResultsThe mean age in this study was 45.4±8.1 years. Among the 709 persons, 229 (32.3%) had a positive result for at least one microorganism, 48 (6.8%) had two different species, 6 (0.8%) had three different species, and 1 person had four different species. The overall prevalence of asymptomatic STDs such as urethritis or cervicitis was 7.1% (50/709). The prevalence rates of chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Mycoplasma hominis, and Trichomonas vaginalis infection in asymptomatic persons were 5.6% (40/709), 0.4% (3/709), 0.3% (2/709), 22.1% (157/709), 11.6% (82/709), and 1.1% (8/709), respectively.ConclusionsWith only a single sample, we could identify the prevalence rates of six microorganisms and the overall proportion of urethritis or cervicitis in asymptomatic people. This proportion cannot be neglected; therefore, screening tests for sexually transmitted diseases such as urethritis or ervicitis should be recommended to asymptomatic people.

Urinary schistosomiasis transmission in Epe, an urban community of Southwest Nigeria

Background:A survey of Schistosoma haematobium infection in Epe, an urban community in Lagos State, Southwest Nigeria, was carried out to ascertain the possibility that schistosomiasis, otherwise considered a rural disease, could reach urban populations.Materials and Methods:About 100 ml of voided urine samples from 200 pupils aged 6–13 years [109 (54.5%) males and 91 (45.5%) females], attending an Anglican primary school, Ebute Afuye, and a community primary school, Erepoto, were examined parasitologically for hematuria and S. haematobium ova following informed consent obtained from their parents/guardians. All samples were screened using polymerase chain reaction (PCR) amplification of the schistosome Dra1 gene. Fourteen Bulinus snails collected from the two sites, Ebute Afuye (6) and Erepoto (8), were screened for schistosome infection by the PCR amplification of the schistosome Dra1 gene. PCR-RFLP of the snails’ its region was analyzed for species identification and a subregion of the cox1 gene from four infected snails (two from each site) was amplified and sequenced.Results:In the Anglican primary school, Ebute Afuye, and community primary school, Erepoto, 16% and 29% were positive for hematuria, and 16% and 17% had schistosome ova, respectively. PCR analysis showed that 57% and 40% were positive for the infection in Anglican primary school, Ebute Afuye, and community primary school, Erepoto, respectively. PCR screening of the snails confirmed that four from Ebute Afuye and three from Erepoto were infected with schistosomes. PCR-RFLP showed that all the 14 snails were Bulinus truncatus while phylogenetic analysis of the sequenced partial cox1 gene corroborated the PCR-RFLP results.Conclusions:There was a high prevalence of S. haematobium infection among the participants detected by PCR, which was able to detect infection in cases otherwise shown to be negative by hematuria. We also observed that B. truncatus is one of the snail species responsible for the transmission of urinary schistosomiasis in the Epe community. For national control programs, it is very important that trends in the prevalence and intensity of schistosomiasis in urban cities be monitored.

PCA3 molecular urine assay for prostate cancer: association with pathologic features and impact of collection protocols

IntroductionPCA3 is a non-coding mRNA molecule that is overexpressed in prostate cancer. The purpose of this study is to evaluate the utility of the PCA3 molecular urine test scores to predict adverse pathologic features and catheterized specimen collection.MethodsHundred men with clinically localized prostate cancer scheduled to undergo robotic prostatectomy were enrolled in the study following a standard consent process. The study protocol consisted of providing four urine samples. Voided urine obtained following digital rectal examination (DRE) pre-operatively (Vl), catheterized urine without DRE (V2), and l0-day and 6-week postoperative voided (V3 and V4) urine samples were collected and analyzed. These four urine specimens underwent target capture, transcription-mediated amplification, and hybridization in order to quantify both PCA3 and PSA mRNA. The PCA3 score was calculated as the ratio of PCA3 to PSA.ResultsInformative rates (sufficient mRNA for analysis) for VI, V2, V3 and V4 were 91, 85, 0 and 2%, respectively. There was no significant associations with pathological stage, Gleason score >6. Higher PCA3 scores at V1 correlated with increased risk for perineural invasion (P = 0.0479).ConclusionsInformative PCA3 scores can be obtained from post-DRE voided urine as well as catheterized urine without a DRE. The PCA3 test does not seem to predict adverse pathologic features, though, may have an association with perineural invasion. The ability of PCA3 score to predict clinical outcome remains to be determined.

Evaluation of upper urinary tract tumors by FISH in Chinese patients

Upper urinary tract tumor (UUTT) usually presents a high grade and stage, and recurs frequently. The aim of this study was to evaluate the utility of a fluorescence in situ hybridization (FISH) assay on chromosomes 3, 7, 9, and 17 as a reliable and noninvasive method for the diagnosis of Chinese patients with UUTT. Urine specimens from 50 patients with UUTT and 25 donors without evidence of urothelial tumors were analyzed by cytology and FISH. Voided urine samples from 20 normal individuals were used to establish the cut-off values for FISH assay. The McNemar test was applied for sensitivity and specificity. The overall sensitivity of FISH was statistically significantly greater than that of cytology (84.0 vs. 40.0%, P=0.000). The overall specificities of FISH and urine cytology were all 96.0% ( P=1.000). Polysomy in chromosomes 3, 7, and 17 were 38, 42, and 30%, respectively. Heterozygous and homozygous loss of the p16 locus was found in 36 and 32%, respectively. FISH analysis performed on cells collected from voided urine is feasible, and FISH could prove to be a reliable and less invasive ancillary test and improve the sensitivity of urine cytology in the diagnosis of UUTT.

Comparison of the sensitivity and specificity of urine cytology, urinary nuclear matrix protein-22 and multitarget fluorescence in situ hybridization assay in the detection of bladder cancer

This study aimed to compare the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of urine cytology, BladderChek® nuclear matrix protein-22 (NMP22) and UroVysion™ fluorescence in situ hybridization (FISH) tests in patients with newly diagnosed bladder cancer, those with recurrent bladder cancer, and those with bladder cancer but in remission during surveillance. Voided urine samples obtained from 178 patients with suspected or known bladder cancer about to undergo diagnostic or surveillance cystoscopy and 25 control subjects without the disease were divided into four and used for urine culture and cytology, NMP22 BladderChek and UroVysion FISH tests. The sensitivity, specificity, PPV and NPV for each test were calculated. Comparison was made between the ability of each test to detect bladder cancer in the three category of patients listed. Of the 178 patients with bladder cancer, 43 were newly diagnosed, 58 had recurrent disease and 77 were in remission. The sensitivity of each test in newly diagnosed patients was: urine cytology 28%, NMP22 88% and FISH 80%; and in patients with recurrent disease: urine cytology 33%, NMP22 57% and FISH 85%. The mean specificity for urine cytology, NMP22 and FISH was 95%, 67% and 48%, respectively. Of the tests used in the study for detection of bladder cancer, NMP22 appeared to be most cost-effective and rapid, with relatively high sensitivity and specificity in all categories of patients. The NMP22 test may be considered a new gold standard for the assessment of patients with known or suspected bladder cancer.

Human urine-derived stem cells seeded in a modified 3D porous small intestinal submucosa scaffold for urethral tissue engineering

The goal of this study was to determine whether urothelial cells (UC) and smooth muscle cells (SMC) derived from the differentiation of urine-derived stem cells (USC) could be used to form engineered urethral tissue when seeded on a modified 3-D porous small intestinal submucosa (SIS) scaffold. Cells were obtained from 12 voided urine samples from 4 healthy individuals. USC were isolated, characterized and induced to differentiate into UC and SMC. Fresh SIS derived from pigs was decellularized with 5% peracetic acid (PAA). Differentiated UC and SMC derived from USC were seeded onto SIS scaffolds with highly porous microstructure in a layered co-culture fashion and cultured under dynamic conditions for one week. The seeded cells formed multiple uniform layers on the SIS and penetrated deeper into the porous matrix during dynamic culture. USC that were induced to differentiate also expressed UC markers (Uroplakin-III and AE1/AE3) or SMC markers (α-SM actin, desmin, and myosin) after implantation into athymic mice for one month, and the resulting tissues were similar to those formed when UC and SMC derived from native ureter were used. In conclusion, UC and SMC derived from USC could be maintained on 3-D porous SIS scaffold. The dynamic culture system promoted 3-D cell–matrix ingrowth and development of a multilayer mucosal structure similar to that of native urinary tract tissue. USC may serve as an alternative cell source in cell-based tissue engineering for urethral reconstruction or other urological tissue repair.

Polyomavirus infection and urothelial carcinoma

Polyomavirus infections are common in the general (adult) population with a reported prevalence of more than 80%. Polyomavirus can infect urothelial carcinoma and change the morphology of these malignant cells, as is shown in this paper. An eighty year old Hispanic male was referred to the urology clinic for diagnosis and treatment. The submitted voided urine sample was concentrated, processed, and a SurePath preparation was made. The Papanicolaou-stained slides contained single cells with very large hyperchromatic nuclei with a glassy appearance in addition to atypical neoplastic cells. The single cells with enlarged nuclei proved to stain positive for Simian virus 40 (SV40) antigen. The nuclei of these cells were 2 to 4 times larger than that of the surrounding noninfected atypical cells. These findings were confirmed on histologic sections prepared from tissue biopsy. In conclusion we report that it is difficult to distinguish benign polyomavirus infected cells from their malignant counterparts in cytology but not in histology.

Dual effect of adipose tissue on bone health during growth

Recent studies suggest association between body fat and childhood bone health, although conflicting findings have also been reported. The aim of the present study was to examine the association between body fat content and areal BMD, volumetric BMD, bone turnover markers, and calcitropic factors in apparently healthy children and adolescents. The study was carried out as a school based cross-sectional cohort study with 186 subjects (61% girls) aged from 7 to 19 years. Background characteristics, including medical history and lifestyle factors, were collected. Anthropometry, BMD and body composition measurements with DXA and pQCT, a fasting blood sample and a second morning void urine sample were obtained. The subjects were divided into three groups, representing low, intermediate and high adiposity, based on age and gender specific fat percentage Z-scores. Multivariate analysis was performed to all bone variables using lean body mass, pubertal development, dietary intake of calcium and intensity of physical activity as covariates. In pQCT, a significant difference between the groups was observed in cortical BMD (MANOVA; p = 0.02) in proximal radius, the intermediate group having the highest values. Similarly, DXA-derived LS and WB BMD Z-scores differed significantly between the groups (MANOVA; p = 0.026 and p = 0.036, respectively), with highest values in the intermediate group. High body fat content associated also with low bone turnover markers, low PTH, high S-Ca and high urinary calcium excretion. The differences in biochemical markers explain our and others' findings of bone health in children and adolescents with different body fat content. These findings suggest that normal body fat content is beneficial for bone health in growing children and adolescents while both low and high body fat content have adverse skeletal effects.

Cigarette smoking and second-hand smoking exposure in adolescents with chronic kidney disease: a study from the Midwest Pediatric Nephrology Consortium

Smoking and second-hand smoking [SHS] cause significant cardiovascular mortality and morbidity. In healthy individuals and adults with chronic kidney disease [CKD], cigarette smoking is associated with albuminuria, increased risk for CKD, increased graft loss and progression of renal insufficiency. In children, SHS has been associated with higher blood pressure variability, blood pressure load, elevated C-reactive protein and decreased cognitive function. Using a survey document and urine cotinine, we sought to investigate prevalence of cigarette use and SHS in adolescents with CKD. A cross-sectional study was conducted in which adolescents aged 13 to 18 years with CKD were asked to complete a single anonymous self-administered survey. In addition, a single freshly voided urine sample for cotinine measurement was obtained from eligible subjects. Of 182 subjects, 60 (34%), 25 (14%) and 93 (52%) were transplant recipients, were dialysis dependent and had a glomerulopathy, respectively. Renal status was lacking in four. Twenty-four per cent (24%) had smoked at some point in their lives, and 13% had smoked within the last 30 days of taking the survey. Fifty-two per cent (52%) of all respondents reported living with an adult who smoked, and 54% reported having friends that smoked. Forty-seven per cent (47%) and 44% of those who had never smoked lived with an adult and had friends that smoked, respectively. There was a discrepancy rate of 7% between self-reported non-smokers and urine cotinine, suggesting smoking rates were higher. The highest cotinine/creatinine levels among the non-smokers were observed in those who lived with a smoker and had friends that smoked. Among adolescents with CKD, cigarette smoking and SHS exposure are prevalent and may be important variables to consider when evaluating renal and cardiovascular risk factors and outcomes in children with CKD.

Bone mass, vitamin D levels and immunosuppressive treatment in patients with liver transplant and risedronate treatment

As bone matures type I collagen undegoes β isomerisation at the DG motif of the CTX epitope. Measurements of αCTX and βCTX allow us to distinguish between bone turnover in newly formed and mature bone, respectively. In states of pathologically high bone turnover such as Paget's and malignant bone diseases, there is a relatively greater increase in the resorption of newly formed bone. Treatment of these with bisphosphonates results in a relatively greater decrease in the resorption of newly formed bone. The aims were to investigate 1) the relative increase in αCTX in postmenopausal osteoporosis and 2) the relative decrease in αCTX with bisphosphonate treatment. We recruited 60 postmenopausal women (mean age 68: range 55-85) with a BMD T-score of <2.5 or <1 and a fracture. They were randomised into 3 treatment groups: ibandronate (n=19), alendronate (n=23) and risedronate (n=18). 75 healthy premenopausal women (mean age 38: range 30-45) were recruited. We measured α and βCTX (Nordic Biosciences Diagnostics, Copenhagen, Denmark) in second morning void urine samples and serum βCTX (Roche Diagnostics, Pensberg, Germany). The median urinary α and βCTX and serum βCTX were significantly lower in healthy premenopausal compared to postmenopausal osteoporotic women at baseline 62%, 110% and 65% respectively (P<0.0001). The median urinary α and βCTX and serum βCTX showed a significant decrease by 68%, 88%, and 79% from baseline respectively (P<0.0001) after 13 weeks of treatment. The α/β CTX ratio increased by 62% (P<0.05). We conclude that resorption of type I collagen in postmenopausal osteoporosis is greater compared to healthy premenopausal women. There is no relative greater increase in the resorption of newly formed bone. With bisphosphonate treatment, the resorption of type I collagen is decreased after 13 weeks of treatment, but there is no greater decrease in the relative resorption of newly formed bone. In fact, there is a preferential decrease in the resorption of mature bone.

Fibroblast Growth Factor Receptor 3 Mutation Analysis on Voided Urine for Surveillance of Patients with Low-Grade Non-Muscle–Invasive Bladder Cancer

Mutations in the fibroblast growth factor receptor 3 (FGFR3) have been found in 70% of the low-grade non-muscle-invasive bladder cancer (NMI-BC) tumors. We aim to determine the potential of FGFR3 mutation analysis on voided urine to detect recurrences during surveillance of patients with low-grade NMI-BC. FGFR3 mutation status of the study inclusion tumor was determined from 200 low-grade NMI-BC patients. Patients with an FGFR3-mutant inclusion tumor were selected for analysis and monitored by cystoscopy, and voided urine samples were collected. FGFR3 mutation analysis was done on 463 prospectively collected urines. Sensitivity and predictive value of the assay were determined for detection of concomitant recurrences. Longitudinal and Cox time-to-event analyses were done to determine the predictive value for detection of future recurrences. Median follow-up was 3.5 years. The sensitivity of the assay for detection of concomitant recurrences was 26 of 45 (58%). Of the 105 positive urine samples, 85 (81%) were associated with a concomitant or a future recurrence. An FGFR3-positive urine was associated with a 3.8-fold (P < 0.0001) higher risk of having a recurrence in the Cox analysis. In contrast, only 41 of 358 (11%) FGFR3-negative urine samples were associated with a recurrence. Positive predictive value increased from 25% to 90% in patients having consecutive FGFR3-positive urine tests. FGFR3 mutation analysis on voided urine is a simple and noninvasive diagnostic method for detection of recurrences during surveillance of patients presenting with a low-grade FGFR3-mutant NMI-BC tumor.

Gene Expression Signature in Urine for Diagnosing and Assessing Aggressiveness of Bladder Urothelial Carcinoma

To develop an accurate and noninvasive method for bladder cancer diagnosis and prediction of disease aggressiveness based on the gene expression patterns of urine samples. Gene expression patterns of 341 urine samples from bladder urothelial cell carcinoma (UCC) patients and 235 controls were analyzed via TaqMan Arrays. In a first phase of the study, three consecutive gene selection steps were done to identify a gene set expression signature to detect and stratify UCC in urine. Subsequently, those genes more informative for UCC diagnosis and prediction of tumor aggressiveness were combined to obtain a classification system of bladder cancer samples. In a second phase, the obtained gene set signature was evaluated in a routine clinical scenario analyzing only voided urine samples. We have identified a 12+2 gene expression signature for UCC diagnosis and prediction of tumor aggressiveness on urine samples. Overall, this gene set panel had 98% sensitivity (SN) and 99% specificity (SP) in discriminating between UCC and control samples and 79% SN and 92% SP in predicting tumor aggressiveness. The translation of the model to the clinically applicable format corroborates that the 12+2 gene set panel described maintains a high accuracy for UCC diagnosis (SN = 89% and SP = 95%) and tumor aggressiveness prediction (SN = 79% and SP = 91%) in voided urine samples. The 12+2 gene expression signature described in urine is able to identify patients suffering from UCC and predict tumor aggressiveness. We show that a panel of molecular markers may improve the schedule for diagnosis and follow-up in UCC patients.

Reliability of spot urine samples in assessing arsenic exposure.

Urinary arsenic concentration is often used as a biomarker of arsenic exposure. First morning void (FMV) and spot urine samples from 131 participants in southeastern Michigan were analyzed using an HPLC-IC-PMS system for six different arsenic species: arsenobetaine (AsB), arsenite (As[III]), arsenate (As[V]), methylarsonous acid MMA[III], methylarsenic acid MMA[V], and dimethylarsenic DMA[V]. Bland-Altman plots, intraclass correlation coefficients (ICCs), and Pearson correlation procedures were used to evaluate the relationship between the arsenic species in FMV and spot urine collections after normalizing the samples by specific gravity. DMA[III] and MMA[III] were not detected in any of the samples. The sum of As[III], As[V], MMA[V], and DMA[V] was designated SumAs. The ICC between SumAs in FMV and SumAs in spot samples was 0.90. The ICC showed that 90% of variation comes from between individuals and not within individuals. A significant correlation (r=0.80 p<0.001) was observed between FMV and spot samples for SumAs. The spot sample were a good predictor of the MMA[V] (r=0.83 p<0.0001), and DMA[V] (r=0.77 p<0.0001) in the FMV sample. These associations suggest that either FMV or spot samples can be used as an adequate bioindicator of arsenic metabolites in human urine. The benefit of using spot urine samples, instead of 24-h or FMV urine samples, is the potential reduction in budgetary and logistic requirements in epidemiological studies.

Urinary and Serum Carbohydrate Antigen 19-9 as a Biomarker in Ureteropelvic Junction Obstruction in Children

We evaluated the predictive role of serum and urinary carbohydrate antigen 19-9 in the diagnosis and followup of pediatric ureteropelvic junction obstruction. The study included 27 children with ureteropelvic junction obstruction who underwent pyeloplasty (group 1), and 41 controls consisting of 27 healthy children (group 2) and 14 children with hydrocele/renal cyst (group 3). Serum and voided urine were evaluated for carbohydrate antigen 19-9 in each group. Additionally urine from the affected pelvis and fluid in hydrocele/renal cyst were collected at surgery in groups 1 and 3. Serum and voided urine samples were obtained at 3, 6 and 9 months after pyeloplasty for carbohydrate antigen 19-9 assessment, and were correlated with clinical factors. Preoperative carbohydrate antigen 19-9 level was significantly greater in group 1 than in controls. The best cutoff values for serum and urinary carbohydrate antigen 19-9 were 13.21 U/ml and 30.6 U/ml, respectively, with significantly higher sensitivity and specificity for urinary values. Obstruction release was followed by improvement of renal function together with significant reduction in urinary and serum carbohydrate antigen 19-9 at 3 months. Initial pelvis diameter and renographic function significantly correlated with urinary carbohydrate antigen 19-9. No significant correlation was found regarding serum carbohydrate antigen 19-9. Voided urine carbohydrate antigen 19-9 is a noninvasive, clinically applicable marker in congenital obstructive nephropathy. The practical implications of these data for diagnosis and long-term followup in ureteropelvic junction obstruction are significant. Our findings suggest that proper decrease in urinary carbohydrate antigen 19-9 after pyeloplasty is predictive of excellent surgical outcomes and resolution of renal damage.

Abstract 4705: Distinct gene promoter hypermethylation profile in bladder cancer patients from different Chinese populations and its implication in cancer detection using voided urine samples

Abstract Bladder cancer is the 6th most common cancer in the world and the incidence is particularly high in southwestern Taiwan. Bladder cancer patients would need to have long term follow-up and repeated urine cytology, however the sensitivity for urine cytology is known to be low. Several studies including ours had identified several tumor-related genes that are hypermethylated in bladder cancer patients and can be detected in voided urine samples (Chan et al., Clin Cancer Res 8: 464-470, 2002). However, comprehensive methylation profile of bladder cancer in Taiwan is currently unknown. On the other hand, studies also suggested that the non-random methylation profile in cancer may be related to the exposure of different environmental carcinogen in different locality. In this regard, we aim to investigate the DNA methylation profile of multiple tumor suppressor genes in bladder cancer patients from different Chinese populations including Taiwan (104 cases), Hong Kong (98 cases) and Beijing (24 cases) by methylation specific PCR (MSP). Genes showing distinct methylation in Taiwanese population will be selected as biomarkers for cancer detection in voided urine samples. Our result showed that frequent methylation was detected in p14 (84.4%), IRF8 (67.4%), sFRP1 (62.8%), and DAPK (52.2%) from cancer patients of Taiwan. While methylation was also detected in RASSF1A (37.0%), p15 (32.6%), APC (28.3%), hMLH1 (21.7%), SOCS-1 (19.6%), and CDH1 (15.2%). The methylation profile demonstrated bimodal distribution which is a characteristic of CpG island methylator phentotype (CIMP). Besides, the number of methylated genes in each patient were significantly correlated with pathological grade (P &amp;lt;0.001) and stage (P&amp;lt;0.05). Interestingly, methylation of CDH1, hMLH1, p14 and p15 showed obvious diversity among different Chinese populations. Importantly, the sensitivity and specificity of methylation detection of any one of the methylated gene (IRF8, p14, sFRP1 and RASSF1A) in voided urine of patients using qMSP is 89.5% and 94.4% respectively. To further identify novel targets that are epigenetically silenced in bladder cancer from Taiwan, we performed differential methylation analysis (DMH) using Agilent 244K CpG island microarray coupled with expression microarray in BFTC905 and TSGH8301 bladder cancer cell lines. ARHGEF17 and CDKN1C were identified to be concurrently methylated in these two bladder cancer cell lines as compared with SV-HUC-1 normal bladder cell line. In conclusion, our result indicated that there are distinct DNA methylation profiles among different Chinese populations. These profiles demonstrate gradual increases with cancer progression. Finally, detection of gene methylation in voided urine with these distinct DNA methylation markers is more sensitive than urine cytology. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4705.

1158 IDENTIFICATION OF A GENE SET PANEL ON URINE SAMPLES FOR THE NONINVASIVE DIAGNOSIS AND AGGRESSIVENESS ASSESSMENT OF BLADDER UROTHELIAL CARCINOMA

You have accessJournal of UrologyBladder Cancer: Detection and Screening1 Apr 20101158 IDENTIFICATION OF A GENE SET PANEL ON URINE SAMPLES FOR THE NONINVASIVE DIAGNOSIS AND AGGRESSIVENESS ASSESSMENT OF BLADDER UROTHELIAL CARCINOMA Lourdes Mengual, Moises Burset, Maria J. Ribal, Elisabet Ars, Mercedes Marín-Aguilera, Manuel Fernández, Mercedes Ingelmo-Torres, Humberto Villavicencio, and Antonio Alcaraz Lourdes MengualLourdes Mengual More articles by this author , Moises BursetMoises Burset More articles by this author , Maria J. RibalMaria J. Ribal More articles by this author , Elisabet ArsElisabet Ars More articles by this author , Mercedes Marín-AguileraMercedes Marín-Aguilera More articles by this author , Manuel FernándezManuel Fernández More articles by this author , Mercedes Ingelmo-TorresMercedes Ingelmo-Torres More articles by this author , Humberto VillavicencioHumberto Villavicencio More articles by this author , and Antonio AlcarazAntonio Alcaraz More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.658AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES To develop an accurate and noninvasive method for bladder cancer diagnosis and prediction of disease aggressiveness based on the gene expression patterns of urine samples. METHODS Gene expression patterns of 341 urine samples from bladder urothelial cell carcinoma (UCC) patients and 235 controls were analyzed by means TaqMan Arrays. In a first phase of the study, 3 consecutively gene selection steps were performed to identify a gene set expression signature to detect and stratify UCC in urine. Subsequently, those genes more informative for UCC diagnosis and prediction of tumor aggressiveness were combined to obtain a classification system of bladder cancer samples. In a second phase, the obtained gene set signature was evaluated in a clinical routine scenario analyzing only voided urine samples. RESULTS We have identified a 12+2 gene expression signature for UCC diagnosis and prediction of tumor aggressiveness on urine samples. Overall, this gene set panel had 98% sensitivity (SN) and 99% specificity (SP) in discriminating between UCC and control samples and 79% SN and 92% SP in predicting tumor aggressiveness. The translation of the model to the clinically applicable format corroborate that the 12+2 gene set panel described maintains a high accuracy for UCC diagnosis (SN=89% and SP=95%) and tumor aggressiveness prediction (SN=79% and a SP=91%) in voided urine samples. CONCLUSIONS The 12+2 gene expression signature described in urine is able to identify patients suffering from UCC and predict tumor aggressiveness. We show that a panel of molecular markers can improve the schedule for diagnosis and follow-up in UUC patients. Barcelona, Spain© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e448 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Lourdes Mengual More articles by this author Moises Burset More articles by this author Maria J. Ribal More articles by this author Elisabet Ars More articles by this author Mercedes Marín-Aguilera More articles by this author Manuel Fernández More articles by this author Mercedes Ingelmo-Torres More articles by this author Humberto Villavicencio More articles by this author Antonio Alcaraz More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Role of Urinary Carbohydrate Antigen 19-9 as a Practical Biomarker for Determining Conservative or Surgical Management of Children with Ureteropelvic Junction Obstruction

Purpose Previously, we reported elevated level of urine carbohydrate antigen 19-9 (CA 19-9) in childern with ureteropelvic junction obstruction (UPJO). In present study we aimed to investigate the role of urinary CA 19-9 as a predictive biomarker to serve as a surrogate in differentiating between children with UPJO in need of pyeloplasty and those with dilated nonobstructed kidneys suitable for conservative treatment. Material and Methods The study included 36 children with UPJO treated with pyeloplasty (GI) and 24 children with dilated nonobstructed kidneys who were treated conservatively (GII). Voided urine samples were obtained for CA 19-9 measurement before treatment, and every 3 months during follow-up. Additionally, ultrasonography in three-month intervals and annual renography were performed. Follow-up ranged from 12-27 months (average 16.4 months). pyeloplasty was considered for patients with >10% loss of renal function. Results In GI preoperative CA 19-9 was significantly higher than in GII and a cutoff value of 41.06 U/ml yielded a sensitivity of 96.7% and a specificity of 93%. Compared to initial measurements, CA 19-9 decreased significantly at 3 months after pyeloplasty in GI and in the 12th month in GII with significant correlation with changes in renal pelvis diameter. In three patients due to rising CA 19-9 levels during follow-up along with decline in renal function (mean 12.8%) pyeloplasty was considered. Conclusions Urinary CA 19-9 is a non-invasive clinically applicable marker for definition of management protocol in children with UPJO. The practical clinical implications of the biomarker for long-term follow-up of children with UPJO after pyeloplasty and those receiving conservative treatment are significant.

Clinical usefulness of fluorescence in situ hybridization for diagnosis and surveillance of bladder cancer

We evaluated the performance of the fluorescence in situ hybridization (FISH) assay in comparison with that of urinary cytology for the detection of bladder urothelial carcinoma in routine clinical practice. Voided urine samples from 602 patients with hematuria were analyzed. The bladder cancer group consisted of 95 patients who had biopsy-proven bladder cancer, and the control group consisted of 507 patients without bladder cancer. We found a significant difference between the overall sensitivities of FISH and cytology (60% vs. 28.4%, respectively; P < 0.0001). The overall specificity was 99% with cytology and 94.9% with FISH, although this difference was not statistically significant. The mean values for all four probes in the true-positive group were higher than those in the false-positive group. The difference in the mean values between the two groups was significant only for the CEP3 and CEP17 probes. Furthermore, the severity of the genetic alterations detected by FISH showed a positive correlation with both tumor invasiveness (stage Ta → T1, T2) and histological grade (G1, G2 → G3). Together, these findings suggest that FISH can be a useful diagnostic and surveillance tool for patients who are suspected of having new or recurrent bladder cancer.

Comparison of the yield of different screening approaches to detect chronic kidney disease

Screening for chronic kidney disease (CKD) has been advised in high-risk populations. The present study aims to compare the yields of four approaches to select high-risk subjects for CKD screening, which are defined as follows: Approach 1, history of cardiovascular (CV) disease, diabetes mellitus or hypertension (=high CV risk); Approach 2, high CV risk or age >55 years; Approach 3, urinary albumin concentration (UAC) ≥20 mg/L; or Approach 4, UAC ≥10 mg/L at pre-screening. The study population is a sample of the general population of Groningen, the Netherlands (n = 3398). UAC was measured (nephelometry) in a first morning void urine sample collected at home and sent to a laboratory by post. Information on demographics and the presence of CV risk factors was obtained by a questionnaire. The presence of CKD was determined during examination at an outpatient clinic. At baseline, 12% of the subjects met the criteria of Approach 1, 33% of Approach 2, 8% of Approach 3 and 25% of Approach 4. CKD was diagnosed in 370 subjects (11%). Approach 2 detected the most CKD patients (sensitivity 65%), while Approach 3 resulted in the lowest number needed to screen (1.9). During a follow-up of 7 years, only the UAC pre-screening approaches detected CKD patients who had both significantly accelerated renal function loss and increased CV risk compared to subjects without CKD. Only 28% of CKD patients detected by the UAC approaches used antihypertensive/angiotensin-converting enzyme inhibitor treatment prior to screening. This study suggests that pre-screening based on UAC should be favoured in comparison to screening based on CKD risk factors to detect CKD patients at high renal and CV risk.

Clinical experience with survivin as a biomarker for urothelial bladder cancer

This study was carried out as a prospective pilot study to evaluate the potential of survivin mRNA measurement in patients suspicious for urothelial bladder cancer (BC). Data were also analyzed for possible influences of secondary urological findings on survivin measurements. Survivin was measured by an mRNA assay in voided urine samples of 50 patients with suspicion of new or recurrent BC prior to transurethral resection. Sample evaluation was possible in 49 cases. Histopathology revealed no malignancy in 17 (35%) and BC in 32 (65%) patients. Survivin mRNA was quantitated by real-time PCR from frozen cell pellets of centrifuged urine samples. A ROC analysis of the survivin data was performed. ROC analysis identified the best cut-off level at 10,000 mRNA copies, resulting in a sensitivity of 53% and a specificity of 88%. Seven of the 20 pTa tumors (35%), all four pT1 (100%) and all four muscle-invasive tumors (100%) were detected. Of four patients with carcinoma in situ (Cis), 50% could be identified. Only two patients (4%) were assessed as false positive. Histologically confirmed cystitis and concomitant urological findings (inflammatory cells in urine, microhematuria and others) had no detectable influence on survivin measurements. In present group of patients, survivin was a reliable biomarker for high-grade urothelial BC (sensitivity 83%), but not for low grade (sensitivity 35%) urothelial BC with a high specificity (88%). No confounders influencing the results of survivin measurements could be identified.