Bladder Cancer Screening Research Articles

Bladder cancer: ESMO Practice Guidelines for diagnosis, treatment and follow-up

Bladder cancer: ESMO Practice Guidelines for diagnosis, treatment and follow-up

Cell-free urinary microRNA-99a and microRNA-125b are diagnostic markers for the non-invasive screening of bladder cancer.

BackgroundEvidence implicated the diagnostic significance of microRNAs in whole urine/urine sediments in urothelial carcinoma of the bladder (UCB). However, the contaminated blood cells in patients with haematouria significantly altered the expression profiles of urinary microRNA, influencing the test accuracy.MethodsMicroRNA profiles of the urine supernatants of UCB patients and controls without any malignancy and profiles of malignant and corresponding normal mucosa tissues from the patients were determined by microRNA microarray and compared to identify differentially expressed microRNAs. The differential expression was verified in the tissues of an independent patient cohort by RT-qPCR. The diagnostic significance of selected microRNAs as biomarkers in the urine supernatant was investigated in the expanded cohorts.ResultsMicroRNA-99a and microRNA-125b were down-regulated in the urine supernatants of UCB patients. The degree of down-regulation was associated with the tumor grade. A diagnostic model was developed using a combined index of the levels of microRNA-99a and microRNA-125b in the urine supernatant with a sensitivity of 86.7%, a specificity of 81.1% and a positive predicted value (PPV) of 91.8%. Discriminating between high- and low-grade UCB, the model using the level of microRNA-125b alone exhibited a sensitivity of 81.4%, a specificity of 87.0% and a PPV of 93.4%.ConclusionsThe results revealed a unique microRNA expression signature in the urine supernatants of UCB patients for the development of molecular diagnostic tests. An effective cell-free urinary microRNA-based model was developed using a combined index of the levels of microRNA-99a and microRNA-125b to detect UCB with good discriminating power, high sensitivity and high specificity.

Low- and high-grade bladder cancer appraisal via serum-based proteomics approach

ObjectiveTo address the shortcomings of urine cytology and cystoscopy for screening and grading of urinary bladder cancer (BC) we applied a serum-based proteomics approach as a surrogate tactic for rapid BC probing. MethodsThis study was performed on 90 sera samples comprising of low-grade (LG, n=33) and high-grade (HG, n=32) BC, and healthy controls (HC, n=25). Two-dimensional gel electrophoresis (2DE) tactic was executed to describe serum proteome. MALDI-TOF-MS (MS) was used to identify the characteristics of aberrantly expressed proteins in 2DE and validated using Western blot (WB) and ELISA approach. Receiver operating characteristics (ROC) curve analysis was also performed to determine the clinical usefulness of these proteins to discriminate among LG, HG and HC cohorts. ResultsThis comprehensive approach of 2DE, MS, WB and ELISA reveals five differentially expressed proteins. Among them two biomarkers (S100A8 and S100A9) were able to accurately (ROC, 0.946) distinguish 81% of BC (LG+HG) cases compared to HC with highest sensitivity and specificity. With a comparable tactic, two biomarkers (S100A8 and S100A4) were able to precisely (ROC, 0.941) discriminate 92% of LG cases from HG with utmost sensitivity and specificity. ConclusionsSerum proteomics probing appears to be an encouraging and least-invasive tactic for screening and grading of BC.

Bladder cancer risk: Use of PLCO and NLST to identify a suitable screening cohort.

4546 Background: Bladder cancer (BC) screening is not accepted in part due to low overall incidence. We utilized the PLCO and NLST to identify optimal high-risk populations most likely benefit from screening. Methods: Data were extracted from PLCO and NLST to stratify risk of BC by overall population, gender, race, age at inclusion and smoking status. Incidence rates between groups were compared using chi-square test. Results: BC was identified in 1430/154,898 in PLCO and 439/53,173 in NLST. BCs were grades III/IV in 36.8% and 41.3%, respectively. Incidence rates were significantly higher in men than women (PLCO: 1.4 vs. 0.31/1000 person-years, NLST: 1.84 vs. 0.6/1000 person-years, both p 70 with smoking exposure of 30 pack years (PY) and more, incidence rates were as high as 11.92 (PLCO) and 5.23 (NLST) (per 1000 person-years). In current high int...

Clinical utility of urinary soluble Fas in screening for bladder cancer

Early diagnosis of carcinoma of urinary bladder remains a challenge. Urine cytology, as an adjunct to cystoscopy, is less sensitive for low-grade tumors. Soluble Fas (sFas), a cell-surface receptor and member of the tumor necrosis factor superfamily, is frequently expressed in urinary bladder carcinoma. The objective of this study was to investigate the urinary sFas for diagnosis of transitional cell carcinoma (TCC) of urinary bladder. We examined urinary sFas concentration in 74 controls and 117 cases of TCC, both primary and recurrent disease, by using enzyme-linked immunosorbent assay and compared it with urinary cytology. Urinary sFas concentration was found to be significantly higher in the patient as compared to control group (P < 0.05). An optimal cutoff value of 174.0 pg/mL was proposed. The urinary sFas level was found to have an approximate sensitivity and specificity of 88.03% and 89.19% (P < 0.001), whereas urine cytology had sensitivity of 66.67% and specificity of 95.95%. sFas had better sensitivity in higher grade and both primary and recurrent cases of urinary bladder cancer in comparison with cytology. Out of 15 node positive bladder cancer cases, 13 had high urinary sFas levels, whereas 12 were urinary cytology positive for malignancy. Urinary sFas can be used as a non-invasive diagnostic biomarker for TCC of urinary bladder, both for primary and recurrent disease.

Prospective evaluation of fluorescence-guided cystoscopy to detect bladder cancer in a high-risk population: results from the UroScreen-Study

ObjectiveTo prospectively evaluate the role of fluorescence-guided cystoscopy in a high-risk bladder cancer population undergoing screening based on a multi-marker panel of urine-tests (UroScreen-study).Patients and methodsUroScreen was conducted as a validation study for tumor markers within the frame of a health surveillance program of workers with occupational exposure to aromatic amines. Voluntary annual screens were done in 1,609 men. Cytology, quantitative NMP22® assay, and UroVysion (FISH) were applied to 7091 urine samples. Subjects with at least one positive urine-based tumor marker and/or persisting microscopic hematuria were offered fluorescence-guided (PDD) instead of white light cystoscopy. In case of suspicious findings histopathological evaluation by transurethral biopsy was performed. Data were statistically summarized and compared to tumors found by the standard algorithm of the screening study.ResultsTwenty-two subjects with a mean age of 58 years (39–72) underwent PDD cystoscopy. Of those 3 had positive NMP22 tests, 14 positive FISH tests and 9 suspicious cytologies. Two had persisting microscopic hematuria only. PDD cystoscopy revealed enhanced unifocal fluorescence in 14. All had subsequent transurethral biopsy or resection. In total, 1 urothelial carcinoma (pTaG1, low grade) was diagnosed. In the other participants urothelial cancer of the bladder was ruled out. Chronic cystitis was revealed in 8 of 14 biopsies. No higher detection rate was found using PDD than with the standard algorithm of the UroScreen study in which 17 tumors were detected by white light cystoscopy.ConclusionThe use of PDD does not lead to a higher cancer detection rate in a high-risk screening population. Larger sample sizes may be needed to ultimately asses the value of PDD for bladder cancer screening.

Shortcomings in bladder cancer etiology research and a model for its prevention.

Bladder cancer (BC) is the most expensive cancer to treat. Its incidence and mortality have not decreased in the last three decades. Numerous uncertainties are still surrounding the etiology of BC. There is a need for a low-cost screening test for BC that would be applicable for early detection in asymptomatic persons, a test that would preferably be noninvasive and have satisfactory sensitivity and specificity. The first part of this paper addresses critical issues in the research into BC etiology, which we classified as entrances, toxicity and metabolism, amounts, and duration of exposure to carcinogens in the bladder. In the second part, based on the proven risk factors for BC, we present a simple scoring system as part of a new BC screening method. The heterogeneous results of studies on BC etiology are largely due to a lack of research into the compounds (and their mutual interactions) present in the urinary bladder, carcinogens absorbed through the skin and/or inhaled, and the daily dynamics of exposure to exogenous risk factors. We have calculated a score for BC screening which is an integral component of a new, four-level system of BC prevention. Interactions of carcinogens and their daily dynamics deserve more attention in further clarifying BC etiology. New attempts in BC screening should be focused on urine content analyses (carcinogens, antioxidants, vitamins, minerals) and not only on hematuria and currently used biomarkers. We propose a score for BC pre-evaluation and recruitment for screening and a new model of BC prevention.

Screening for urinary bladder cancer with the use of nuclear matrix protein (NMP) 22: is it feasible?

Background Tobacco use accounts for the majority of urinary bladder cancer (UBC) cases both in men and women. It is widely accepted that smokers have increased risk of UBC [1-4]. An early diagnosis of UBC is crucial because patients with superficial UBC have much better prognosis than those with invasive UBC [1,2,4,5]. During the past years a variety of urine based markers have been introduced for screening of UBC. Nuclear matrix protein 22 (NMP22) is a nuclear protein that is accountable for chromatid regulation and cell separation during replication. The detection of NMP22 has been used from many clinical studies to evaluate the possibility of UBC screening [2,6-8]. The purpose of this review is to access the current literature in order to determine the usefulness of NMP22 in screening of UBC.

Predicting Risk of Bladder Cancer Using Clinical and Demographic Information from Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial Participants

Effective screening and prevention strategies for bladder cancer require accurate risk stratification models. We developed models to predict the risk of bladder cancer based on clinical and sociodemographic data on participants in the Prostate, Lung, Colorectal, and Ovarian Cancer (PLCO) screening trial. Baseline clinical and sociodemographic data were obtained from 149,542 PLCO participants, ages 55 to 74 years, without a prior history of bladder cancer. Cox proportional hazards models were used to predict the risk of all bladder cancers (ABC) and of high-grade bladder cancers (HGBC) from baseline information. We used the HGBC risk model to design a hypothetical bladder cancer mortality prevention trial. Over a median follow-up of 12 years, 1,124 men and 259 women developed bladder cancer (including 392 and 72 with HGBC, respectively). The incidence in men and in women was 133.6 and 29.6 cases per 100,000 person-years, respectively. Nomograms constructed for predicting the risk of ABC and HGBC had c-indices of 0.746 and 0.759, respectively. Age, race, education, smoking (intensity and duration), comorbidity, prostatitis, syphilis, and hormone replacement therapy use were statistically significant predictors in the models. We show that our risk model can be used to design a bladder cancer mortality prevention trial half the size of a trial designed without risk stratification. Models to predict the risk of ABC and HGBC have been developed and validated. Using the upper 40th percentile from the HGBC model, a suitable cohort for a screening or chemoprevention trial could be identified, although the size and follow-up of such a trial would be costly.

Screening for bladder cancer with urinary tumor markers in chemical workers with exposure to aromatic amines

To validate urinary markers for the early detection of bladder cancer (BC) in chemical workers. UroScreen was conducted as a validation study for tumor markers within the frame of a health surveillance program of the German Social Accident Insurance for active or retired workers with former exposure to aromatic amines. From 2003 to 2010, 1,609 men took part in voluntary annual screens. Cytology, the quantitative NMP22(®) assay, and UroVysion™ were applied to 7,091 urine samples. Fifteen out of 21 tumors were detected following test positivity. The UroVysion/NMP22 panel detected 14 out of 21 tumors versus 8 tumors with cytology alone (sensitivity 66.7 vs. 44.4%, specificity 94.5 vs. 98.5%). The sensitivity of the panel increased to 85.7% in samples collected ≤12months before diagnosis and when papillomas were excluded, compared to 58.3% with cytology. About 3% of NMP22 tests were false-positive. UroVysion results overlapped with cytology due to the preselection of atypical cells. NMP22 was less and UroVysion more frequently positive in diluted urine samples. Leukocytes confounded NMP22 but not UroVysion. The low incidence of BC in this study population yielded low positive predictive values of the markers and high costs per tumor detected with screening. UroVysion in combination with NMP22 detected more cases than cytology alone, at the expense of a lower specificity. High costs per detected case resulted from a lower BC incidence than in the past when levels of occupational exposure to aromatic amines were higher. Currently, it cannot be recommended to apply these markers for screening in asymptomatic workers. The increase in sensitivity is not balanced by the high costs of UroVysion and the false-positive tests of NMP22.

Validation of RiskCheck Bladder Cancer ©, Version 5.0 for Risk-Adapted Screening of Bladder Cancer

Introduction: The aim of the study was to assess the strength of the online tool RiskCheck Bladder Cancer©, version 5.0 (RCBC) for early detection of bladder cancer (BC). Materials and Methods: RCBC was evaluated retrospectively based on the data of 241 patients, of which 141 were suffering from BC. Statistical analysis was performed by descriptive statistics, nonparametric group comparison, classification tree analysis and ROC analysis. Results: ROC analysis of the risk classification showed a sensitivity of 71.6%, a specificity of 56.5%, a positive predictive value of 67.8%, a negative predictive value of 52% and an accuracy of 63.5%. BC risk factors ranked by importance are time of smoking (p < 0.0001), gender (within the nonsmoking group: p < 0.009), occupational toxin exposure (within the group <35 years of smoking: p < 0.048) and amount of consumed cigarettes resulting in a 95% association with BC (within the group >35 years of smoking: p < 0.0001). Conclusions: The high predictive power of RCBC for the identification of asymptomatic patients living under risk could be demonstrated.

Urinary markers for bladder cancer

Bladder cancer has the fifth highest incidence of all malignancies in the United States, with a propensity to recur, requiring lifelong surveillance after diagnosis. Urinary markers of disease have been of extreme interest in this field in an effort to simplify surveillance schedules and improve early detection of tumors. Many markers have been described, but most remain investigational. However, some markers have undergone clinical trials and are approved for clinical use. In this review, urinary markers and their application for screening and surveillance of bladder cancer are discussed.

Evaluation of urovysion and cytology for bladder cancer detection

Urine cytology has been used for screening of bladder cancer but has been limited by its low sensitivity. UroVysion is a multiprobe fluorescence in situ hybridization (FISH) assay that detects common chromosome abnormalities in bladder cancers. For this study, the authors evaluated the effectiveness of multiprobe FISH and urine cytology in detecting urothelial cell carcinoma (UCC) in the same urine sample. In total, 1835 cases with the following criteria were selected: valid results from both the multiprobe FISH assay and urine cytology in the same urine sample, histologic and/or cystoscopic follow-up within 4 months of the original tests, or at least 3 years of clinical follow-up information. The results of FISH and cytology were correlated with clinical outcomes derived from a combination of histologic, cystoscopic, and clinical follow-up information. Of 1835 cases, 1045 cases were from patients undergoing surveillance of recurrent UCC, and 790 were for hematuria. The overall sensitivity, specificity, positive predictive value, and negative predictive value in detecting UCC were 61.9%, 89.7%, 53.9%, and 92.4%, respectively, for FISH and 29.1%, 96.9%, 64.4%, and 87.5%, respectively, for cytology. The performance of both FISH and cytology generally was better in the surveillance population and in samples with high-grade UCC. In 95 of 296 cases with atypical cytology that were proven to have UCC, 61 cases, mostly high-grade UCC, were positive using the multiprobe FISH assay. The UroVysion multiprobe FISH assay was more sensitive than urine cytology in detecting UCC, but it produced more false-positive results. The current data suggest that the use of FISH as a reflex test after an equivocal cytologic diagnosis may play an effective role in detecting UCC.

Towards risk stratification in bladder cancer

Bladder cancer risk assessment is currently based on an individual's age and history of exposure to known carcinogens. A new report illustrates how interactions between cigarette smoking and germline genetic variation can identify those at particularly high risk, who could be targeted in future bladder cancer prevention and screening efforts.

Risk-adapted screening in bladder cancer with the open-access tool RiskCheck Bladder Cancer: Proof of principle by the health service research foundation IQUO.

1571 Background: The open-access questionnaire RiskCheck Bladder Cancer (RCBC) was proven in daily routine work from German urologists organized in the health services research foundation IQUO on asymptomatic patients to identify BC risk exposure and its relation to detectable tumors. Methods: The open-access RCBC questionnaire checked asymptomatic patients for their BC risk exposure in relation to personal-, smoking-, occupation- and medical induced risk. This resulted in a risk stratification with low- intermediate- and high risk. The intermediate and high risk subjects were checked for tumor presence by routine diagnostics. IBM-SPSS 20 was used for descriptive statistics and effectiveness was proven by classification tree analysis and cross-table analysis with Chi-square test. Results: Out of 303 checked asymptomatic persons 274 (90.4%) were negative for tumor and 29 (9.6%) had a detectable tumor. In the group of NED 176 (68.1%) persons were classified as low risk, and 98 (16.2%) with tumor risk. Out of the 29 detected tumors 20 were at intermediate or high risk (68.9%). This resulted in an over all detection rate of 6.6% and focused on the risk population of 16.9%. The RCBC risk assessment was significant (p &lt; 0.01). Sensitivity 69.0%, specificity 64.2%, NPV 95.1%, PPV 16.9%, false positive cases 35.8%, false negative cases 31.0%, accuracy 64.7%. Compared to the common incidence (35/100,000) this is an increase in effectiveness of 188.6 in the screened population and 482.8 in relation to the risk population alone. Conclusions: Preventive medical care becomes effective because RCBC is able to condense a population with focusing investigations on people living under risk. A reasonable preventive care by a yearly recall control in urological offices for the risk population (32.3%) can be organized. In consequence the assessment is work effective, aim achieving and in result cost effective. The questionnaire RCBC integrates evidence based bladder cancer inductors, is easy in use and as a open-access tool available in 10 languages via internet to all medical services. www.riskcheck-bladder-cancer.info.

Screening for bladder cancer: the best opportunity to reduce mortality

Bladder cancer kills more women than cervical cancer and is also a significant cause of mortality in men. Little progress has been made in improving survival in patients with advanced bladder cancer. Two pilot studies using microhematuria screening have shown that screening for bladder cancer results in close to 80% downstaging, with high-grade cancers being detected before they have invaded the bladder wall. Results of long-term follow-up even suggest a striking reduction in bladder cancer mortality. The main obstacles to screening for bladder cancer may be overcome if a higher-risk population is identified by designing a risk scale for exposure to cigarette smoke and occupational carcinogens, and through genetic testing for susceptibility to cancer and home hematuria screening, which in itself identifies a population with approximately 3% to 4% risk of bladder cancer. The feasibility and cost effectiveness of screening for bladder cancer can be significantly improved by incorporating a secondary screening strategy using a more sensitive and specific bladder cancer marker that is currently available, and by limiting urological evaluations to patients who show positive results on one or more of these tests. Bladder cancer is the most costly cancer to treat in the United States and pharmacoeconomic studies suggest that screening for bladder cancer could not only save lives but also reduce costs per year-life saved. A pilot study is underway and the urology community should be very supportive of studies to validate this opportunity.

Key issues in bladder cancer management

It is with great enthusiasm that I introduce you to the first bladder cancer supplement of the CUAJ. As Editor of this supplement, I am excited to present you with up-to-date issues in bladder cancer written by Canadian and international physicians and scientists. The supplement highlights key aspects of bladder cancer from the management of T1G3 tumours and BCG failures to the management of advanced cancers, including the role of lymphadenectomy, urethrectomy, perioperative chemotherapy and metastasectomy. The supplement also addresses the future prospects towards the pathways of personalized medicine. Authors review the impact of nonurothelial histology, as well as risk-adapted and treatment adapted surveillance strategies. Controversial issues are debated: Is there a niche for prostate-sparing cystectomy? Is bladder cancer screening ready for prime time? Does chemoprevention have a place in bladder cancer? These are just a few of the questions that the supplement attempts to answer. I congratulate all the authors who have contributed toward this supplement. I know you have worked hard on this endeavour and I hope this final product complements the reader’s needs and your expectations. On a global note, the actual prevalence of bladder cancer is 10 times its incidence, creating a major economic burden on health-care systems.1 As measured on the basis of cumulative per-patient cost from diagnosis until death, bladder cancer is the most expensive cancer to treat, accounting for almost US$3.7 billion (2001 values) of direct costs in the United States. Unfortunately, the lack of public awareness and patient advocacy, disproportionate research funding and decreasing research activity have limited the progress in bladder cancer management.2 We need to be more proactive with this disease. As one of the founding members of the Canadian Bladder Cancer Network (CBCAN), I would like to take this opportunity to introduce this one-of-a-kind organization. The CBCAN is a nonprofit national organization whose mission is to (1) help bladder cancer patients and their support teams in the day-to-day issues with the disease; (2) raise awareness of bladder cancer among the general public and medical community; and (3) advocate for allocation of government and private funds directed to diagnosis, treatment and medical research to eliminate bladder cancer. For more information, I encourage you to visit our website at www.cbcan.org. This site is open to physicians and patients.

1282 RISK-ADAPTED SCREENING PERFORMED WITH THE INTERNET TOOL RISKCHECK BLADDER CANCER© (RCBC) WAS EVALUATED BY THE UROLOGICAL HEALTH SERVICE RESEARCH FOUNDATION IQUO-GERMANY

You have accessJournal of UrologyBladder Cancer: Detection & Screening1 Apr 20131282 RISK-ADAPTED SCREENING PERFORMED WITH THE INTERNET TOOL RISKCHECK BLADDER CANCER© (RCBC) WAS EVALUATED BY THE UROLOGICAL HEALTH SERVICE RESEARCH FOUNDATION IQUO-GERMANY Gerson Lüdecke, Götz Geiges, and Frank König Gerson LüdeckeGerson Lüdecke Giessen, Germany More articles by this author , Götz GeigesGötz Geiges Berlin, Germany More articles by this author , and Frank KönigFrank König Berlin, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.2636AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The open-access questionnaire RCBC was proven in daily routine work from German urologists organized in the health services research foundation IQUO on asymptomatic patients. In consequence we could characterize our patient population in respect to BC risk exposure and the presence of BC in this risk population. METHODS The open-access RCBC questionnaire was used in urological offices to check asymptomatic patients for their BC risk exposure. All people with intermediate and high risk were checked for tumor presence by urine diagnostics, optional by ultrasound and in case of suspect results controlled by cystoscopy. IBM-SPSS 20 was used for statistical analysis; incidence, distribution and correlation between risk stratification and tumor detection. The effectiveness was proven by classification tree analysis and cross-table analysis with Chi-square test. Significance was defined with p < 0.05. RESULTS Out of 303 checked asymptomatic persons 274 (90.4%) were negative for tumor and 29 (9.6%) had a detectable tumor. In the group of NED 176 (68.1%) persons were classified as low risk, and 98 (16.2%) with tumor risk. Out of the 29 detected tumors 20 were at intermediate or high risk (68.9%). This resulted in an over all detection rate of 6.6% and focused on the risk population of 16.9%. The association of tumor presence and increased risk identified by RCBC assessment was significant (p < 0.01). This described a sensitivity of 69.0%, a specificity of 64.2%, NPV of 95.1%, PPV of 16.9%, false positive cases of 35.8%, false negative cases of 31.0% and a accuracy of 64.7%. Compared to the documented incidence rates for Europe and the industrialized countries (35/100,000) this is an increase in effectiveness of 188.6 in the screened study population and 482.8 in relation to the risk population alone. CONCLUSIONS Risk-adapted screening in bladder cancer delivers a reasonable approach to diagnose bladder cancer before emerging symptoms. Preventive medical care becomes effective because RCBC is able to condense the screened population with focusing investigations on people living under risk. With a NPV of 95.1% unnecessary investigations can be minimized effectively. In consequence the assessment is work effective and aim achieving. The quote of over locked tumors (2.9%) was low and all NMIBC with low-risk grading. The questionnaire RCBC integrates evidence based bladder cancer inductors, is easy in use and as an open-access tool available in 10 languages via internet. www.riskcheck-bladder-cancer.info. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e524-e525 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Gerson Lüdecke Giessen, Germany More articles by this author Götz Geiges Berlin, Germany More articles by this author Frank König Berlin, Germany More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...

A high-throughput assay for the detection of Tyr-phosphorylated proteins in urine of bladder cancer patients

BackgroundBladder cancer has the peculiarity of shedding neoplastic cells and their components in urine representing a valuable opportunity to detect diagnostic markers. Using a semi-quantitative method we previously demonstrated that the levels of Tyr-phosphorylated proteins (TPPs) are highly increased in bladder cancer tissues and that soluble TPPs can also be detected in patient's urine samples. Although the preliminary evaluation showed very promising specificity and sensitivity, insufficient accuracy and very low throughput of the method halted the diagnostic evaluation of the new marker. To overcome this problem we developed a quantitative methodology with high sensitivity and accuracy to measure TPPs in urine. MethodsThe Immobilized Metal Affinity Chromatography (IMAC) was miniaturized in a 96 well format. Luminescence, visible and infrared fluorescence antibody-based detection methods were comparatively evaluated. ResultsDue to their low abundance we evidenced that both phosphoprotein enrichment step and very sensitive detection methods are required to detect TPPs in urine samples. To pursue high throughput, reproducibility and cost containment, which are required for bladder cancer screening programs, we coupled the pre-analytical IMAC procedure with high sensitive detection phases (infrared fluorescence or chemiluminescence) in an automated platform. ConclusionsA high throughput method for measuring with high sensitivity TPP levels in urine samples is now available for large clinical trial for the establishment of the diagnostic and predictive power of TPPs as bladder cancer marker. General significanceThe new assay represents the first quantitative and high throughput method for the measurement of TPPs in urine.

A case–control study on the association between bladder cancer and prior bladder calculus

BackgroundBladder calculus is associated with chronic irritation and inflammation. As there is substantial documentation that inflammation can play a direct role in carcinogenesis, to date the relationship between stone formation and bladder cancer (BC) remains unclear. This study aimed to examine the association between BC and prior bladder calculus using a population-based dataset.MethodsThis case–control study included 2,086 cases who had received their first-time diagnosis of BC between 2001 and 2009 and 10,430 randomly selected controls without BC. Conditional logistic regressions were employed to explore the association between BC and having been previously diagnosed with bladder calculus.ResultsOf the sampled subjects, bladder calculus was found in 71 (3.4%) cases and 105 (1.1%) controls. Conditional logistic regression analysis revealed that the odds ratio (OR) of having been diagnosed with bladder calculus before the index date for cases was 3.42 (95% CI = 2.48-4.72) when compared with controls after adjusting for monthly income, geographic region, hypertension, diabetes, coronary heart disease, and renal disease, tobacco use disorder, obesity, alcohol abuse, and schistosomiasis, bladder outlet obstruction, and urinary tract infection. We further analyzed according to sex and found that among males, the OR of having been previously diagnosed with bladder calculus for cases was 3.45 (95% CI = 2.39-4.99) that of controls. Among females, the OR was 3.05 (95% CI = 1.53-6.08) that of controls.ConclusionsThese results add to the evidence surrounding the conflicting reports regarding the association between BC and prior bladder calculus and highlight a potential target population for bladder cancer screening.