Benign Bladder Research Articles

Role of Diffusion MRI and Apparent Diffusion Coefficient Measurement in the Diagnosis, Staging and Pathological Classification of Bladder Tumors

Objectives: Our aim was to investigate the role of simultaneous apparent diffusion coefficient (ADC) measurement and diffusion-weighted MRI (DWMRI) in the diagnosis and differentiation of bladder masses. Methods: Weexamined 45 patients presenting with hematuria by dynamic and diffusion MRI (at b100, b600, b1000 gradients) and cystoscopy. Thirty healthy volunteers without hematuria were also included in the study. Results from imaging techniques were compared to cystoscopic and histopathologic findings. The ADC values of benign and malignant bladder masses, and bladder tissue in healthy subjects were measured and compared with each other. Results: The mean ADC values of benign (n = 10) and malignant lesions (n = 35) were significantly lower than the mean ADC values of adjacent bladder walls and bladder walls of healthy subjects at all 3 gradients (p < 0.05). However, no significant difference was detected between benign and malignant lesions (p > 0.05). Mean ADC values of invasive tumors were significantly lower than superficial malignancies at b600 and b1000 gradients. The mean ADC values of transitional cell tumors were significantly higher than squamous cell tumors at b100 and b600 gradients. Conclusion: Simultaneous measurement ofADC and DWMRI provided useful information in the diagnosis, staging and histopathological classification of bladder tumors.

UP-02.028 The Incidence of Erectile Dysfunction and Retrograde Ejaculation Following Thulium Laser Prostate Vaporesection for Benign Bladder Outflow Obstruction: A Retrospective Study

UP-02.028 The Incidence of Erectile Dysfunction and Retrograde Ejaculation Following Thulium Laser Prostate Vaporesection for Benign Bladder Outflow Obstruction: A Retrospective Study

ERG–TMPRSS2 rearrangement is shared by concurrent prostatic adenocarcinoma and prostatic small cell carcinoma and absent in small cell carcinoma of the urinary bladder: evidence supporting monoclonal origin

Prostatic carcinoma is a heterogeneous disease with frequent multifocality and variability in morphology. Particularly, prostatic small cell carcinoma is a rare variant with aggressive behavior. Distinction between small cell carcinoma of the prostate and urinary bladder may be challenging, especially in small biopsy specimens without associated prostatic adenocarcinoma or urothelial carcinoma. Recently, gene fusions between ETS genes, particularly ETS-related gene (ERG), and transmembrane protease, serine 2 (TMPRSS2) have been identified as a frequent event in prostate cancer. Thus, molecular methods may be helpful in determining the primary site of small cell carcinoma. Thirty cases of prostatic small cell carcinoma from the authors' archives were studied, among which 13 had concurrent prostatic adenocarcinoma. Tricolor fluorescence in situ hybridization (FISH) was performed on formalin-fixed paraffin-embedded tissue sections with a probe cocktail for 3′/5′ ERG and TMPRSS2. Cases of small cell carcinoma of the bladder and conventional prostatic adenocarcinoma (25 each) were also tested as controls. ERG gene alterations were found only in prostate malignancies and not in benign prostatic tissue or bladder small cell carcinoma. TMPRSS2–ERG gene fusion was found in 47% (14/30) of prostatic small cell carcinoma. Of cases with concurrent prostatic adenocarcinoma, 85% (11/13) had identical findings in both components. In 20% of rearranged cases, the ERG abnormality was associated with 5′ ERG deletion. In 17% (5/30) of cases, gain of the 21q22 locus was present. Two cases showed discordant aberrations in the small cell carcinoma and adenocarcinoma, one with deletion of 5′ ERG and one with gain of chromosome 21q, both in only the adenocarcinoma component. Small cell carcinoma of the prostate demonstrates TMPRSS2–ERG rearrangement with comparable frequency to prostatic adenocarcinoma. In cases with concurrent adenocarcinoma and small cell carcinoma, the majority showed identical abnormalities in both components, indicating a likely common clonal origin. Discordant alterations were present in rare cases, suggesting that acquisition of additional genetic changes in multifocal tumors may be responsible for disease progression to a more aggressive phenotype. TMPRSS2–ERG fusion is absent in bladder small cell carcinoma, supporting the utility of FISH in distinguishing prostate from bladder primary tumors and identifying metastatic small cell carcinoma of unknown origin.

Usefulness of computed tomography performed immediately after excretory urography in patients with delayed opacification or dilated upper urinary tract of unknown cause

To evaluate the diagnostic value of computed tomography (CT) performed immediately after excretory urography (EU) in patients with delayed renal opacification or dilated upper urinary system with nonconclusive diagnosis after EU. CT was performed immediately after EU in 39 patients with delayed opacification or dilated upper urinary system of unknown cause, without additional intravenous contrast administration for the CT study. We classified EU + CT findings as benign or malignant causes and we compared our results with the final diagnosis. The combination of EU + CT correctly diagnosed 38 out of the 39 cases with a sensitivity of 97%. Correct diagnosis was established in all malignant cases (n = 17) but one benign case consistent with blood clots in the upper urinary tract was incorrectly diagnosed as a multicentric urothelial carcinoma. Sensitivity, specificity, and accuracy for the diagnosis of the underlying cause with EU + CT was 100%, 95%, and 97%, respectively. The final diagnoses were: urothelial carcinoma (n = 10), stone disease (n = 10), bladder tumor (n = 4), benign post-treatment ureteral stenosis (n = 4), ureteral invasion (n = 3), benign bladder disease (n = 2), urinary tract infections (n = 2), crossing vessels (n = 1), ureteropelvic junction obstruction (n = 1), retrocaval ureter (n = 1), and blood clots in the upper urinary tract due to bleeding renal metastasis from lung cancer (n = 1). Combined EU and CT study allowed correct diagnosis of the underlying cause of delayed excretion or upper urinary tract dilatation in 97% of cases. The combination of EU and CT provides diagnosis reducing time and radiation.

Overexpression of HER-2\neu receptor protein in Urinary Bladder Carcinoma, An Immunohistochemical Study.

Background: Bladder carcinoma is one of the most common cancer worldwide, it accounts for 6.5% of all cancers, with highest incidence in industrialized countries .It represents the fourth most common cancer in men and the eighth in women. Bladder carcinoma depends in its pathogenesis on a combination of genetic and environmental factors, these factors produce phenotypic changes that allow normal transitional cells to become cancerous and finally acquire the “malignant phenotype". Many attempts had been tried to explore the role of some genetic abnormalities encountered in bladder carcinoma. It has indicated that many genetic abnormalities may underline the pathogenesis of cancer evolution of urinary bladder like VEGF , P53, Bcl2 and RB, but nowadays a scientific efforts have raised the possible role of Her-2/neu in bladder carcinoma, and the rate of its overexpression in bladder carcinoma is ranging from 2% - 74%. Objective: To estimate immunohistochemical expression of HER-2\neu receptor protein in Urinary Bladder Carcinoma in relation to other parameters; sex grade, pattern of tumor and stage of tumor .Methods: The present work is performed in the Department of Pathology and Forensic medicine, College of Medicine, Kufa University. Formalin fixed, Paraffin-embedded blocks from 60 (43 males and 17 females) patients with urinary bladder carcinoma were included in this study. A group of 12 patients with chronic cystitis were included as a control group. Avidin-Biotin Complex method was employed for immunohistochemical detection of HER-2\neu.Results: HER-2\neu overexpression was positive in 41.6% of urinary bladder carcinoma, while there was no expression in benign bladder tissue (P&lt;0.05). HER-2\neu immunohistochemical staining was positively correlated with, grade and stage of urinary bladder carcinoma (P&lt;0.05).Conclusion: These findings support the role of HER-2\neu in the carcinogenesis of urinary bladder carcinoma regarding behavior, and aggressiveness, and thus HER-2\neu could be considered as a poor prognostic parameter in urinary bladder carcinoma.

Claudin-11 decreases the invasiveness of bladder cancer cells

The expression of claudin-11 in benign and malignant bladder tissue and the effect of forced expression of claudin-11 on tight junction function and invasiveness of bladder cancer cells were studied. Claudin-11 expression was tested in bladder cancer cell lines (T24/83, RT 112/84 and EJ138) using reverse transcription-polymerase chain reaction (RT-PCR) and in benign and malignant bladder tissue by quantitative RT-PCR and immunohistochemistry. T24/83 cells were transfected with the pcDNA.1/NT-GFP-TOPO vector containing full-length human claudin-11 sequence. Stable-transfected cells overexpressing claudin-11 (T24Cl-11Ex), wild-type cells (T24WT) and the empty plasmid control clone (T24GFP) were compared using transurothelial resistance (TUR), in vitro adhesion, invasion and growth assays. Claudin-11 was strongly expressed in the non-invasive RT112/84 cell line compared to the invasive T24/83 and EJ138 TCC cell lines. Benign bladder tissue demonstrated equal expression of claudin-11 mRNA as carcinoma, but displayed more intense staining than malignant tissue on immunohistochemistry. Forced-expression of claudin-11 in T24/83 cells was confirmed by PCR, immunoprecipitation and by immunofluorescence, which demonstrated increased perinuclear claudin-11 staining. Forced expression of claudin-11 did not affect TUR (p = 0.243), but significantly reduced invasion (p = 0.001) while increasing cell matrix adhesion (p = 0.001) and growth rates (p = 0.001). The greater expression of claudin-11 in benign vs. malignant tissue and non-invasive vs. invasive cell lines, and its effect in reducing bladder cancer cell invasiveness suggests that claudin-11 may have a role in preventing cancer progression and may serve as a therapeutic target in reducing metastasis.

Utilizing mRNA extracted from small, archival formalin-fixed paraffin-embedded prostate samples for translational research: assessment of the effect of increasing sample age and storage temperature

Translational prostate cancer research is hampered by long intervals from diagnosis to patient progression and difficulty in obtaining cancer tissue for investigation. As such, it is imperative to utilise aging formalin-fixed paraffin-embedded (FFPE) tissue samples from the pathology archive with linked patient outcome data to allow current day research. This study aimed to assess the adequacy and quantity of mRNA extracted from formalin fixed paraffin embedded (FFPE) prostate tissue, including prostate biopsies, up to 15 years old. The decay of mRNA over time and under differing storage conditions was also assessed. Archived FFPE benign prostatic tissue up to 15 years old from transurethral resection of the prostate (TURP) and transrectal ultrasound guided (TRUS) biopsies as well as fresh tissue obtained from patients undergoing TURP for benign bladder outlet obstruction were used. Following mRNA extraction beta-actin real-time PCR was carried out using a set of 4 different primer/probes to assess mRNA quality and quantity. There was no difference in mRNA quantity/quality extracted from "fresh" FFPE tissue from the same patient over a 4-month period following surgery. The temperature of block storage did not alter quality/quantity of the mRNA (P > 0.05, unpaired t test). Fresh tissue had a higher quality/quantity, indicated by a lower C ( T ) value, than FFPE samples from the same patient (P ≤ 0.03, one-way ANOVA). Despite being up to 15 years old, all archived FFPE TURP and TRUS biopsy samples had "high" or "very high" levels of expression making them suitable for further analysis. However, the quality of the mRNA in archived FFPE samples did significantly decline with increasing sample age. It is possible to extract mRNA of sufficient standard for further transcriptomic analysis from minute FFPE samples up to 15 years old. This work adds to the literature suggesting that exploitation of retrospective prostate tissue collections with robust associated clinical data is possible.

The clinical relevance of urine-based markers for diagnosis of bladder cancer

The aim of the present study was to evaluate the diagnostic relevance of urinary fibronectin (FN), telomerase (RTA), and cytokeratin 20 (CK20) mRNA in comparison with voided urine cytology (VUC). The study included 132 patients with bladder cancer, 60 patients with benign bladder lesions, and 48 healthy individuals. All were subjected to urine cytology, estimation of fibronectin by ELISA, RTA by TRAP, and CK20 mRNA by conventional RT-PCR in urothelial cells from voided urine. The best cutoff point for FN was determined by receiver operating characteristic curve (41.7 ng/mg protein) revealed the highest sensitivity for malignant (80%) followed by the benign (70%) than the healthy individuals (4.1%) at P < 0.001. Also, RTA and VUC showed significant difference among the three investigated groups (P < 0.001). The overall sensitivity (89.3%) and specificity (98.4%) were the highest for CK20 mRNA. Combined sensitivity of VUC with FN, RTA, and CK20 mRNA together (98.4%) was higher than either the combined sensitivity of VUC with any of them or than that of the biomarker alone. Accordingly, when the diagnostic efficacy was considered, CK20 mRNA had the highest sensitivity and specificity compared to all investigated markers.

GALLBLADDER CARCINOMA;

Objective: Chronic Cholecystitis is one of the commonest diseases presenting in surgical department and is subjected to cholecystectomy each time. A suspicious gall bladder on ultrasound initiates further investigations to rule out carcinoma of gall bladder yet some times a benign looking gall bladder on ultrasound turns out to be carcinoma of gall bladder on histopathology. Design: Descriptive study. Place &amp; Duration of study: District Head Quarters Hospital, Sargodha, from September 2007 to March 2009. Patients &amp; Methods: 200 patients who underwent both open and Laparoscopic cholecystectomy for cholelithiasis were subjected to this study. All relevant data was documented on a standardized data form. Patients were between the ages of 28-74. Patients already diagnosed as Gallbladder Carcinoma, empyema gall bladder, mucocele and gall bladder polyp were excluded from the study. Gall bladders removed after each surgery ware sent to laboratory for histopathological evaluation. Results: A total of 200 cases were studied (161 females, 39 males; M: F ratio 1:4). The mean age was 45 years (range 28-74 years). The most common presenting complaint was pain right hypochondrium with nausea and vomiting (85%).The average operating time was 50 minutes in case of open cholecystectomy and 1 hour in laparoscopic cholecystectomy. Complications included biliary leak in 2 patients (1%), Wound Infection in 3 patients (1.5%) and death in one case (0.5%). The overall rate of complications was 3%. There were a total of 5 patients of laparoscopic cholecystectomy who required extension of the incision, for Carcinoma Gallbladder. Conclusions: Carcinoma of gall bladder is a very aggressive malignancy and usually presents at a very advance stage as its symptoms mostly are marked by symptoms of cholecystitis. Detection of gall bladder carcinoma is very difficult in early stages on ultrasound. Any findings in ultrasound suggesting malignancy should be confirmed on further investigations like CT scan .Carcinoma of Gall bladder is not very common in cases of cholecystectomy for chronic Cholecystitis but once found should be dealt with extreme precision following established operating protocols.

Urinary Met level as a novel biomarker for urothelial carcinoma of the bladder.

257 Background: To determine whether urinary soluble Met (sMet) can differentiate between benign conditions and bladder cancer (CaB), and in cases of bladder cancer, between different stages of transitional cell carcinoma (TCC). Methods: Urinary samples from patients with (Total: 63, pTa: 12, pTis: 22, pT1: 13, ≥ pT2: 16) and without (Total: 27) CaB from three different institutions were prospectively collected prior to cystosopy, TURBT or cystectomy. sMet levels were determined by electrochemiluminescence immunoassay and normalized to urinary creatinine values. Normalized sMet values were compared to final pathologic stage. AUC values were obtained comparing patients with and without TCC. Results: Urinary sMet levels accurately differentiated between patients with and without CaB (AUC: 78%, sensitivity, specificity and negative predictive value were: 68%, 78% and 95%, respectively), patients with no CaB and those with lamina propria invasion (AUC: 79%, sensitivity, specificity and negative predictive value were: 65%, 81% and 95%, respectively) and patients with no CaB and those with muscle invasive CaB (AUC: 85%, sensitivity, specificity and negative predictive value were: 75%, 83% and 97%, respectively). Conclusions: Urinary sMet levels accurately distinguish patients with CaB from those without, and between patients with different CaB stages. These results suggest that urinary sMet may have utility as a bladder cancer marker for screening, treatment follow-up and clinical trial design. No significant financial relationships to disclose.

Nest-Like Features in Bladder, Simulating the Nested Variant of Urothelial Carcinoma

Nest-like structures are the cardinal lesion of different benign and malignant bladder entities. These entities emphasize the biological relevance of the subtle morphological nest plasticity. The authors present 25 benign and malignant neoplasms, sharing the presence of florid nest-like growth and the scarcity or absence of superficial papillary components. Differential diagnostic clues are discussed.

The value of diffusion-weighted MRI in the diagnosis of malignant and benign urinary bladder lesions

To investigate the role of diffusion-weighted MRI (DWI) in the diagnosis of urinary bladder (UB) tumours by means of measuring apparent diffusion coefficient (ADC) values. A total of 83 people aged between 18 and 86 years were included in the study: 63 patients with UB pathology (46 malignant, 17 benign) constituted the case group; 20 individuals without any UB pathology constituted the control group. DWI was applied to all individuals. The ADC values were measured based on the tissue of the UB mass entities and normal UB wall in the control group. The mean ADC value in the UB carcinoma group was significantly lower than that in the control group: 1.0684 ± 0.26 × 10(-3) mm(2) s(-1) and 2.010 ± 0.11 × 10(-3) mm(2) s(-1), respectively (p<0.01). There was a significant difference among the mean ADC values of different grades of malignant tumours, corresponding to 0.9185 ± 0.20 mm(2) s(-1) and 1.281 ± 0.18 mm(2) s(-1) in high-grade and low-grade malignant UB carcinomas, respectively (p<0.01). The ADC value in the carcinoma group was significantly lower than that in the benign lesion group: 1.0684 ± 0.26 × 10(-3) mm(2) s(-1) and 1.803 ± 0.19 × 10(-3) mm(2) s(-1), respectively (p<0.01). All 46 malignant lesions displayed a restriction in diffusion; 4 of the 17 benign lesions displayed a mild restriction in diffusion. The sensitivity, specificity and accuracy of DWI in the diagnosis of malignant UB lesions was 100%, 76.5% and 93.65%, respectively. DWI can be beneficial in the differentiation of benign and malignant UB lesions, as well as of high-grade and low-grade UB carcinomas, using quantitative ADC measurements.

Gross Haematuria among Adult Nigerians: Current Trend

To determine if there is any change in the common causes of gross haematuria among adults Nigerians and to determine what proportion of these aetiologies is due to urological malignancies. Consecutive adults with presenting with gross haematuria seen in our centre were studied prospectively between February 2006 and January 2007. All patients had detailed clinical and diagnostic evaluation including urine cytology, flexible/rigid urethrocystoscopy, ultrasonography and intravenous urography to determine the causes of their haematuria. Other special investigations including CT scan and tissue biopsies were only used in selected patients as indicated. The cause of haematuria was determined in 74 patients (93.7%) while haematuria was of undetermined origin in 5 patients (6.3%). Fifteen different specific diagnoses were made as the causes of haematuria in this study. The three commonest individual causes of haematuria were benign prostatic hyperplasia (30.4%), bladder carcinoma (12.6 %) and carcinoma of the prostate (10.1%). Urological malignancies were diagnosed in 29.1 % of the patients. The Urological malignancies detected were bladder cancers (12.6 %), prostate cancers (10.1%), renal Cell Carcinoma (5.1%) and urethral Carcinoma (1.3%). The incidence of the cancers increased with age; 82.6% of the patients with malignancies were aged 50years and above while only 17.4% were below 50 years of age (P= 0.045). Urological malignancies were also more common in men (73.9 %) than in women (26.1 %) (P < 0.05). We conclude that there is changing trend in the aetiologies of gross haematuria among adult Nigerian patients with urological malignancies being now more prevalent than previously reported in our environment.

Drug treatment for lower urinary tract symptoms

Lower urinary tract symptoms (LUTS) are classified into three groups: storage, voiding, and post-micturition symptoms. The most popular causes of LUTS are benign prostatic hyperplasia (BPH) and overactive bladder (OAB). Although BPH is a pathologic term, clinically, we use this when patients have LUTS due to benign prostatic enlargement and obstruction. OAB is defined as urgency, with or without urge incontinence, usually with frequency and nocturia. Currently α1-adrenoceptor antagonists are the most common drug treatment for BPH, and are thought to act by relaxing the prostatic smooth muscle. They are all effective for the treatment of LUTS/BPH. 5α-reductase inhibitors, such as fiansteride and dutasteride, are another treatment option for BPH symptoms, which reduce the prostatic volume by inducing epithelial atrophy. Long-term combination therapy with alpha-1-blockers and 5α-reductase inhibitors reduces the risk of the overall clinical progression of BPH significantly more than does treatment with either drug alone. Antimuscarinics are the mainstay for the treatment of OAB. Antimuscarinics competitively block muscarinic receptors of all subtypes but with variations in selectivity for the different subtypes. When they are used for the treatment of OAB, they are active during the storage phase of the bladder, with little or no effect on voiding contractions. Desmopressin acetate is a synthetic analogue of Arginin vasopressin, which has been proven effective for the treatment of nocturnal polyuria in LUTS.

Imp3 expression in urothelial carcinomas of the urinary bladder

Superficial tumors including Ta, Tis, and T1 make up 75% of urothelial carcinomas of the bladder. While the behavior of these superficial urothelial cancers is relatively benign, invasive tumors have a significant mortality rate. However, Ta and T1 tumors might display different biological behavior. There is therefore a great need for biomarkers that can accurately distinguish the behavior of urothelial carcinomas in addition to tumor grade and stage. Our aim was to determine the immunohistochemical expression profile of insulin like growth factor II mRNA binding Protein 3 (IMP3) and its correlation with tumor stage and grade in benign urothelium and bladder urothelial carcinomas. The expression of IMP3 in 91 patients with benign urothelium (20 cases), low grade invasive (17 cases) / noninvasive (20 cases) urothelial carcinoma and high grade invasive (20 cases) / non-invasive (14 cases) urothelial carcinoma was evaluated by immunohistochemistry in this study. IMP3 was not expressed in benign urothelium, low-grade non-invasive urothelial carcinoma and high grade non-invasive urothelial carcinoma. Expression of IMP3 was found in 11.76% of low-grade invasive urothelial carcinomas and 55% of high grade invasive urothelial carcinomas. Statistical analysis including chi2 tests showed that IMP3 expression of invasive urothelial carcinomas was statistically significant (p < 0.000). The detection of IMP3 only in invasive carcinomas although some of them were low grade showed that the expression of IMP3 may be related to aggressive behavior of urothelial carcinomas.

Determination of Calcium and Iron in Prostate and Bladder Tumors by Atomic Absorption Spectrophotometer

Background and objective: Serum calcium and iron were used as a biochemical markers for investigation of the neoplastic diseases ,since there is a growing concern about the possible links between these minerals and tumors of various organs and tissues ,the objectives of this study were to determine the changes in the serum levels of Ca &amp; Fe ions in patients with benign prostatic hyperplasia and bladder cancer and to identify the role of these markers in diagnosis and prognosis of these patients . Methods: This study was done in Baghdad at Al-Kadhemia Teaching Hospital between 10/9/2002 and 1/9/2003 .Serum samples were obtained for measurement of calcium and iron.,which were determined by flame atomic absorption spectrophotometer . Results: Hypercalcemia affects up to 30% of patients with bladder and prostate cancers. In addition there were a decrease in the iron serum level as a result of tumor growth . Conclusion: Hypercalcemia, is one of the most common metabolic disorders accompanying neoplasia.The cancers most often associated with hypercalcemia, in addition there were a reduction in serum level of iron this biochemical marker may be associated with the pathogenesis of cancer.

The efficacy of transurethral electrocautery resection for small-volume benign prostatic hyperplasia and bladder neck contracture

Objective To explore the efficacy of transurethral electrocautery resection for small-volume benign prostate hyperplasia and bladder neck contracture.Method Transurethral resection of prostate combined with transurethral electrocautery resection of bladder neck was performed on 19 patients with obvious symptoms of urinary tract obstruction due to small-volume benign prostate hyperplasia and bladder neck contracture.Results The symptoms of obstructin was effectively relieved in all the patients.Conclusion Transurethral resection of prostate combined with transurethral electrocautery resection of bladder neck is a safe,reliable surgical procedure for small-volume benign prostate hyperplasia and bladder neck contracture. Key words: Transurethral resection of prostate; Bladder neck contracture; Benign prostate hyperplasia

Congenital bladder diverticulum with benign bladder wall lesion resembling rhabdomyosarcoma.

Congenital bladder diverticula in children are uncommon and rarely present with bladder outlet obstruction. We present a case highlighting an interesting association between a congenital bladder diverticulum and a benign inflammatory bladder wall lesion mimicking a rhabdomyosarcoma. Open surgery was required as different imaging modalities and cystoscopy were insufficient to exclude a malignant process.

Bladder Cancer-associated Protein, a Potential Prognostic Biomarker in Human Bladder Cancer

It is becoming increasingly clear that no single marker will have the sensitivity and specificity necessary to be used on its own for diagnosis/prognosis of tumors. Interpatient and intratumor heterogeneity provides overwhelming odds against the existence of such an ideal marker. With this in mind, our laboratory has been applying a long term systematic approach to identify multiple biomarkers that can be used for clinical purposes. As a result of these studies, we have identified and reported several candidate biomarker proteins that are deregulated in bladder cancer. Following the conceptual biomarker development phases proposed by the Early Detection Research Network, we have taken some of the most promising candidate proteins into postdiscovery validation studies, and here we report on the characterization of one such biomarker, the bladder cancer-associated protein (BLCAP), formerly termed Bc10. To characterize BLCAP protein expression and cellular localization patterns in benign bladder urothelium and urothelial carcinomas (UCs), we used two independent sets of samples from different patient cohorts: a reference set consisting of 120 bladder specimens (formalin-fixed as well as frozen biopsies) and a validation set consisting of 2,108 retrospectively collected UCs with long term clinical follow-up. We could categorize the UCs examined into four groups based on levels of expression and subcellular localization of BLCAP protein and showed that loss of BLCAP expression is associated with tumor progression. The results indicated that increased expression of this protein confers an adverse patient outcome, suggesting that categorization of staining patterns for this protein may have prognostic value. Finally, we applied a combinatorial two-marker discriminator using BLCAP and adipocyte-type fatty acid-binding protein, another UC biomarker previously reported by us, and found that the combination of the two markers correlated more closely with grade and/or stage of disease than the individual markers. The implications of these results in biomarker discovery are discussed.

Quantitative measurement of attenuation coefficients of bladder biopsies using optical coherence tomography for grading urothelial carcinoma of the bladder

Real-time grading of bladder urothelial carcinoma (UC) is clinically important, but the current standard for grading (histopathology) cannot provide this information. Based on optical coherence tomography (OCT)-measured optical attenuation (μ(t)), the grade of bladder UC could potentially be assessed in real time. We evaluate ex vivo whether μ(t) differs between different grades of UC and benign bladder tissue. Human bladder tissue specimens are examined ex vivo by 850-nm OCT using dynamic focusing. Three observers independently determine the μ(t) from the OCT images, and three pathologists independently review the corresponding histology slides. For both methods, a consensus diagnosis is made. We include 76 OCT scans from 54 bladder samples obtained in 20 procedures on 18 patients. The median (interquartile range) μ(t) of benign tissue is 5.75 mm(-1) (4.77 to 6.14) versus 5.52 mm(-1) (3.47 to 5.90), 4.85 mm(-1) (4.25 to 6.50), and 5.62 mm(-1) (5.01 to 6.29) for grade 1, 2, and 3 UC, respectively (p = 0.732). Interobserver agreement of histopathology is "substantial" [Kappa 0.62, 95% confidence interval (IC) 0.54 to 0.70] compared to "almost perfect" [interclass correlation coefficient (ICC) 0.87, 95% CI 0.80 to 0.92] for OCT. Quantitative OCT analysis (by μ(t)) does not detect morphological UC changes. This may be due to factors typical for an ex-vivo experimental setting.