Various staining procedures in exfoliative urine cytology. Use in routine urology

Exfoliative urinary cytology should be included in the diagnostics and follow up of bladder carcinomas. Staining according to Papanicolaou is time consuming and expensive. Therefore the accuracy of cytology after methylene blue staining, use of prestained slides and Papanicolaou staining was investigated. Urine and bladder washouts were taken from 77 patients with histologically proven bladder tumors, 152 patients after resection of the tumor and 100 patients without tumor of the urinary tract. The results show comparable accuracy of the different staining techniques. Methylene-blue stained and prestained slides cannot be used for documentation; therefore all specimens, which show tumor or are suspicious should also be stained according to Papanicolaou.     

Flow cytometry versus urinary cytology in the evaluation of patients with bladder cancer

We compared the roles of urinary cytology and flow cytometry in the evaluation of patients with bladder cancer in clinical practice situations at a large general hospital. Specimens included 105 bladder washings from patients being followed for urothelial carcinomas and 28 control washings from individuals undergoing cytoscopy for other reasons. Flow cytometry and cytology were performed on aliquots of the same specimen in all bladder cancer samples. When carcinoma was present at the time of specimen collection it was detected by positive cytology in 75 per cent and deoxyribonucleic acid aneuploidy in 78 per cent of the cases. Combination of flow cytometry and urinary cytology increased the diagnostic yield to 95 per cent. Flow cytometry was slightly more sensitive than urinary cytology for detection of abnormalities in specimens from noninvasive and untreated tumors but the only statistically significant difference between the 2 procedures occurred among specimens from treated invasive cancers in which flow cytometry was a less sensitive method than cytology. Abnormal deoxyribonucleic acid ploidy was documented in a few specimens from noncancer-bearing patients having diseases associated with high urothelial cell turnover rates but the concomitant urinary cytology was negative for neoplasia. When used in conjunction with urinary cytology, flow cytometry was a valuable procedure in the followup of patients with bladder cancer. The diagnostic yield with this combination was such that flow cytometry and cytology may be used to reduce the frequency of cystoscopy and biopsy during clinical management in selected situations.     

New molecular markers for bladder cancer detection

PURPOSE OF REVIEW: Bladder cancer continues to be one of the most common genitourinary malignancies. The mainstay of diagnosis remains cystoscopic visualization with transurethral biopsy or resection. As over two-thirds of bladder tumors recur, vigilant surveillance is required. Due to the invasiveness and expense of frequent cystoscopies and the lack of sensitivity of urinary cytology, especially for low-grade superficial lesions, novel molecular markers have been investigated as a means to detect bladder cancer noninvasively. RECENT FINDINGS: As our understanding of the pathogenesis of urothelial neoplasia improves, coupled with recent advances in molecular biological techniques, an array of new approaches to the diagnosis of bladder cancer has emerged. Several urine-based markers have been tested against the standard of urinary cytology with promising results. However, lack of standardization of technique and heterogeneity of bladder cancer itself may hinder the widespread dissemination of these diagnostic aids. SUMMARY: A host of new molecular markers based on the pathogenesis of bladder cancer have been investigated, such as telomerase, survivin, and multitarget fluorescence in situ hybridization, which may eventually improve detection and management of urothelial malignancies. By improving the sensitivity of urinary cytology for low-grade superficial lesions and detecting recurrent disease noninvasively early in its course, these new molecular markers might someday allow changes in the way bladder cancer is diagnosed and followed. At the present time, however, no single molecular marker provides 100% accuracy. Perhaps panels utilizing the most promising of these markers may alter bladder cancer detection and management policy.     

Defining the role of NMP22 in bladder cancer surveillance

Despite advances in treatment and knowledge of its pathogenesis, urothelial carcinoma of the bladder remains a significant cause of morbidity and mortality. Experience with the natural course of bladder cancer has revealed that early diagnosis of primary and recurrent disease improves patient prognosis. In this regard, cystoscopy (usually in combination with urinary cytology) has long been regarded as the gold standard for the diagnosis and surveillance of bladder cancer. However, the disadvantages inherent to cystoscopy, including invasiveness and cost, have stimulated a search for alternative methods for detecting urothelial malignancy. The ideal alternative test would duplicate the high accuracy of cystoscopy for detecting bladder tumors while eschewing its invasiveness, attendant morbidity, and high cost. The vast majority of bladder cancers arise from the urothelium, which continually sheds cells as well as intracellular contents into the urine, thereby providing a potential source of cancer-specific markers. Voided cytology and urinalysis are established tests that have been the standard tools for detection of such substances. The last decade has seen the rise of a myriad of novel urine-based bladder tumor markers, including bladder tumor antigen, urinary bladder cancer antigen, fibronectin, telomerase, and nuclear matrix proteins (e.g., NMP22). The NMP22 assay in particular has been the subject of considerable study and has demonstrated some promise as a potential adjunct to cystoscopy and cytology. Through a critical review of the literature, we seek to define the role, if any, of NMP22 in the follow-up of patients with a previous history of urothelial carcinoma of the bladder.     

Value of bladder washing cytology in the diagnosis and follow-up of bladder tumors

It is reported on the accuracy of the cytological examination of freshly voided urine of 247 patients with 318 urothelial bladder tumors, and the results are compared with the evaluation of bladder washing cytology in 82 patients with 105 bladder tumors. If voided urine is used for examination, there is no evidence in tumors with 0 and 1 grade of malignancy, whereas the bladder washing cytology of grade 1 tumors shows a correlation for the histological findings in 26.3%. In tumors with malignant grades GII and GIII, the bladder washing cytology shows a significantly higher accuracy (81.4%) than in examination of voided urine (67.2%). In both methods the accuracy of cytology rises with infiltration and differentiation grades of the tumor.     

Significance of atypical urinary cytology in the evaluation of patients with end‐stage renal disease for kidney transplantation – a retrospective study

To determine what percentage of renal transplant candidates have atypical urinary cytology, what proportion have urothelial carcinoma and whether cystoscopy is necessary with atypical cytology. All end‐stage renal disease (ESRD) patients (703) presenting for renal transplantation at our institution were retrospectively reviewed. Individuals producing sufficient urine were screened with urine cytology and those with atypical cytology or risk factors for bladder cancer underwent cystoscopy. Four hundred and thirty patients had available urinary cytology and, of these, 151 (35%) had atypical cytology. Of patients with atypical cytology, three were identified to have urothelial carcinoma. However, three additional patients with urothelial carcinoma did not present with atypical cytology. In total, 6 of 703 (0.85%) patients had bladder cancer. All were treated with transurethral resection and eventually underwent renal transplant. One patient has had disease progression post‐transplant to distant metastases. This is the largest study to date evaluating the incidence of urothelial carcinoma in ESRD patients presenting for transplant workup. We found the incidence of bladder cancer to be higher than in the general Canadian population, however, most lesions were low grade. We found atypical cytology in transplant candidates to be a poor predictor for these low‐grade lesions and do not recommend routine cystoscopy for atypical cytology.     

Clinical application of NMP22 and urinary cytology in patients with hematuria or a history of urothelial carcinoma

For evaluation of the clinical application of immunoassay for nuclear matrix protein 22 (NMP22 immunoassay) and urinary cytology for early diagnosis and detection of bladder cancer in patients with hematuria and/or a previous history of bladder cancer, 209 urine samples obtained from 137 patients presenting episodes of hematuria or a history of bladder cancer were assayed for NMP22 levels and/or prepared for cytology examination. Biopsy was performed when any visible tumor was identified during cystoscopy examination. The median NMP22 concentrations measured in samples taken from patients with active bladder cancer, from patients with a history of bladder cancer but no active disease, from patients with hematuria, and from healthy volunteers were 18.95, 5.45, 6.39, and 3.75 U/ml, respectively. The urinary NMP22 level recorded for patients with urothelial carcinoma was significantly higher than that noted for individuals without active disease. The sensitivity of the NMP22 assay and of urinary cytology in diagnosing bladder cancer was 69% and 67%, respectively. In contrast, the specificity of these two diagnostic modalities reached 72% and 93%, respectively. The NMP22 assay is slightly more sensitive but less specific than urinary cytology in detecting bladder cancer. This study indicates that determination of urinary NMP22 levels is a useful and noninvasive tool for the detection of bladder cancer because of its high sensitivity. The urinary NMP22 assay may be used as a first-line routine screening method; however, it cannot replace the use of urinary cytology because of its lower specificity.     

Role of computed tomography urography in the clinical evaluation of upper tract urothelial carcinoma

Intravenous urography has been widely used for the evaluation of upper tract urothelial carcinoma. However, computed tomography urography presently has a higher diagnostic accuracy for upper tract urothelial carcinoma (94.2–99.6%) than intravenous urography (80.8–84.9%), and has replaced intravenous urography as the first‐line imaging test for investigating patients with a high risk of upper tract urothelial carcinoma. Although the detection rate for bladder tumors using standard computed tomography urography is not yet high enough to replace cystoscopy, the addition of a 60‐ to 80‐s delayed scan after the administration of contrast material for the whole pelvis improves the detection rate. A drawback to computed tomography urography is the higher radiation dose of 15–35 mSv, compared with a mean effective dose of 5–10 mSv for intravenous urography. Among several approaches to reducing the radiation dose, the use of an iterative reconstruction algorithm is most likely to become an effective solution because of its simplicity. One advantage of computed tomography urography over intravenous urography is its ability to reliably differentiate between upper tract urothelial carcinoma and calculi or blood clots. Computed tomography urography also shows characteristic findings of other benign conditions. These findings, in combination with negative cytology, are very important diagnostic clues for avoiding an unnecessary nephroureterectomy. For the clinical staging, a recent study has reported the high diagnostic accuracy of computed tomography urography with respect to ≥pT3 tumors. The present review shows the current status of computed tomography urography for the evaluation of upper tract urothelial carcinoma.     

Urinary cytology has a poor performance for predicting invasive or high-grade upper-tract urothelial carcinoma

Study Type – Therapy (case series) Level of Evidence 4 What’s known on the subject? and What does the study add? Accurate preoperative staging for upper‐tract urothelial carcinoma (UTUC) lesions is presently limited. Urinary cytology has shown promise for characterizing pathological features of bladder cancer. The role of cytology for UTUC is at present poorly defined. In this large multi‐institutional cohort of patients, urinary cytology was limited in its ability to accurately predict the grade and stage of upper‐tract lesions. Selective ureteral sampling improved the diagnostic accuracy of cytology when compared to bladder specimens. Improved preoperative surrogate markers for staging UTUC remain necessary.

OBJECTIVE

? To evaluate the diagnostic accuracy of urine cytology for detecting aggressive disease in a multi‐institutional cohort of patients undergoing extirpative surgery for upper‐tract urothelial carcinoma (UTUC).

METHODS

? We reviewed the records of 326 patients with urinary cytology data who underwent a radical nephroureterectomy or distal ureterectomy without concurrent or previous bladder cancer. ? We assessed the association of cytology (positive, negative and atypical) with final pathology. Sensitivity and positive predictive value (PPV) of a positive (± atypical) cytology for high‐grade and muscle‐invasive UTUC was calculated.

RESULTS

? On final pathology, 53% of patients had non‐muscle invasive disease (pTa, pTis, pT1) and 47% had invasive disease (≥pT2). Low‐grade and high‐grade cancers were present in 33% and 67% of patients, respectively. ? Positive, atypical and negative urine cytology was noted in 40%, 40% and 20% of cases. Positive urinary cytology had sensitivity and PPV of 56% and 54% for high‐grade and 62% and 44% for muscle‐invasive UTUC. ? Inclusion of atypical cytology with positive cytology improved the sensitivity and PPV for high‐grade (74% and 63%) and muscle‐invasive (77% and 45%) UTUC. Restricting analysis to patients with selective ureteral cytologies further improved the diagnostic accuracy when compared with bladder specimens (PPV > 85% for high‐grade and muscle‐invasive UTUC).

CONCLUSIONS

? In this cohort of patients with UTUC treated with radical surgery, urine cytology in isolation lacked performance characteristics to accurately predict muscle‐invasive or high‐grade disease. ? Improved surrogate markers for pathological grade and stage are necessary, particularly when considering endoscopic modalities for UTUC.     

Clinical study on urothelial tumors of dye workers in Wakayama City

Between January 1951 and December 31, 1990 urothelial tumors were detected in 112 of 1,085 male dye workers (10.3%) in Wakayama City who were formerly engaged in manufacturing of benzidine and/or beta-naphthylamine. The period from exposure to the chemicals to development of the tumor was a mean of 24.1 +/- 9.4 years. A peak incidence of urothelial tumors was observed also approximately 25 years after the peak period of manufacturing these intermediate products of dyes. The mean period from exposure to such carcinogenic chemicals to the onset of the disease was estimated to be 25 years. Of the 78 patients with primary bladder cancer diagnosed since 1969, 43 (55.1%) had tumors diagnosed mostly as a result of a positive urinary cytology test obtained as part of a screening program and 35 (44.9%) had tumors diagnosed as the result of symptoms. Ten patients (24.4%) in the screened group had been treated with total cystectomy by the last followup examination compared with 17 (50.0%) in the symptomatic group. The 10-year cumulative survival rates were significantly (p less than 0.05) higher in the screened patients (75.1%) than in the symptomatic patients (55.1%). Our results indicate that screening of high risk populations with urinary cytology tests is effective for early diagnosis and treatment of urothelial tumors, and it improves patient prognosis. Furthermore, the biological behavior of occupational urothelial tumors may be different from that of urothelial tumors in the general population.     

Transitional cell carcinoma of the upper urinary tract following total cystectomy for bladder cancer]

From 1975 to 1990, we treated 118 patients with urinary epithelial cancer, including 100 with primary bladder cancer, 13 with primary upper urinary tract cancer, and 5 with both diseases. Thirty-five patients with primary bladder cancer underwent total cystectomy. Upper urinary tract urothelial cancer developed in 4 patients (4.0%) and was detected only after cystectomy. Three patients had multiple bladder tumors before cystectomy and recurrent tumors under long-term bladder-preserving treatment. The other patient had had cystectomy for the primary bladder lesion. Our present policy is to perform urinary cytology once a month and intravenous urography once a year in patients with bladder cancer for early detection of secondary upper urinary tract cancer.     

Bladder treatment. Immunotherapy and chemotherapy.

The urinary bladder is affected by numerous treatments, including surgery, chemotherapy radiotherapy, photodynamic and laser therapy, and immunotherapy. The literature on therapy for bladder cancer is abundant but tends to offer little information concerning the tissue and cellular manifestations related to treatment. A variety of changes in urothelial cells may be seen on urinary cytology or biopsy. Therapy-associated cytologic abnormalities remain the most common cause of false-positive reports in urothelial cytology. Most pathologic changes are characterized by an exuberant inflammatory reaction, blood vessel abnormalities, and degenerative/regenerative urothelial atypia; none of these findings are diagnostic of the outcome of a specific therapy.     

Urinzytologische Diagnostik vor dem Hintergrund der neuen histopathologischen Klassifikation

Urinary cytology is a basic adjunct to cystoscopy and transurethral resection in the diagnosis and characterization of high-grade urothelial carcinomas of the bladder. According to the new WHO classification the former tumor grading G1-3 for non-invasive carcinomas has been replaced by a separation into low-grade and high-grade urothelial carcinomas. An interesting question is where the former non-invasive G2 carcinomas will be positioned in this new classification. In a retrospective analysis we focused on 44 patients with pTaG2 and 17 patients with pT1G2 carcinomas and found that this group of tumors is cytologically heterogeneous but easily differentiated into low-grade and high-grade lesions. A cytometrical analysis significantly underlines the results of the cytological diagnostics. High-grade tumors show a higher recurrence and progression rate. Cytological diagnostics can therefore assist in differentiating low-grade from high-grade urothelial carcinomas.     

Clinical analysis of patients with occupational urothelial tumors

The incidence of occupational urothelial tumor and the accompanied problems were studied on the workers who had manufactured or handled aromatic amines at a certain chemical factory. Twenty-five out of 398 dyestuff workers, who were examined at regular intervals, were found to have urothelial tumors and the incidence rate was 63%. The mean age at onset, the mean period of aromatic exposure and the mean latent period from the initial exposure until tumor development were 61 year-old, 7.2 years and 30 years, respectively. A high incidence rate was found in the long exposed workers and the smoking group. The negative correlation was observed between the age of first exposure to carcinogens and the latent period. The workers who had been exposed to two or three kinds of aromatic amines had the highest incidence followed by those exposed to benzidine and those to alpha-naphthylamine. No urothelial tumor occurred in the workers exposed to beta-naphthylamine. Ninety-four percent of the initial tumors were superficial and transurethral resection of tumors was performed as the initial surgery for the patients with bladder tumors. The recurrence rate in the bladder cavity after the surgery was 39%, which was almost the same rate as that of non-occupational bladder tumors, however, the recurrence rate in the upper urinary tract was high (26%). The positive rate in the examination of urine cytology was 60% for initial tumors, 74% for recurrent tumors. The urine cytology was a significant method for the detection and monitoring of the patients with occupational urothelial tumors.(ABSTRACT TRUNCATED AT 250 WORDS)     

Positive urinary cytology after tumor resection: an indicator for concomitant carcinoma in situ

Concomitant urothelial atypia (grade II atypia or carcinoma in situ) is predictive of new tumor growth after transurethral tumor resection. Concomitant urothelial atypia can be demonstrated by pre-selected site mucosal biopsies. However, a number of patients have new tumors despite normal pre-selected site biopsies. To investigate whether urinary cytology is a better indicator for concomitant urothelial atypia than pre-selected site biopsies, we studied in bladder tumor patients the correlation between the findings of pre-selected site biopsies (8 per patient) at tumor resection and urinary cytology (2 per patient) after successful resection. Concomitant urothelial atypia was demonstrated by biopsies in 52 per cent of the patients, of whom 60 per cent had grade II atypia and 40 per cent had carcinoma in situ. All patients with concomitant carcinoma in situ in biopsies had positive cytology findings. Of the patients with concomitant grade II atypia in biopsies 15 per cent had negative cytology studies. In 48 per cent of the patients no urothelial atypia in pre-selected site biopsies was demonstrable. However, cytology was positive, that is neoplastic cells were present, in 64 per cent of these specimens (19 patients). Of the 19 patients 16 currently have had demonstrable urothelial atypia in pre-selected site mucosal biopsies at a later occasion. We conclude that urinary cytology seems to be a better indicator for the presence of concomitant urothelial atypia than pre-selected site mucosal biopsies and, therefore, it can be used as a screening procedure for patients without demonstrable concomitant carcinoma in situ at tumor resection.     

Methods and current significance of the evaluation of microscopic haematuria

Haematuria is the main symptom of malignant diseases of the urinary tract. Hence urine analysis for the detection of microscopic haematuria is an accepted diagnostic procedure in daily urologic practice. Until now there are neither international nor national agreements relating to the definition of microscopic haematuria, the choice of verification procedures and a diagnostic algorithm. As there are diverse reasons for microscopic haematuria the extent of continuative diagnostics should be adapted to the existence of risk factors for a clinically apparent disease. Low-risk patients with asymptomatic microscopic haematuria do not necessarily have to undergo primary cystoscopy if there are no pathological findings on urine cytology or ultrasound examination. Microhaematuria in high-risk patients should lead to a more intensive evaluation of the urinary tract, which should include cystoscopy and imaging of the upper urinary tract. In the diagnostics of microhaematuria you have to be aware of that intermittent bleeding is often characteristic of urothelial malignancies. Therefore, a single negative urine analysis should not lead to abandonment of further diagnostic procedures.     

Clinical evaluation of urinary NMP22 (nuclear matrix protein 22) bladder chek in the detection of patients with bladder cancer

The clinical usefulness of the nuclear matrix protein 22 (NMP22) Bladder Chek test as a novel urine marker in the detection of patients with bladder cancer was evaluated in comparison with the urinary NMP22 enzyme-linked immunosorbent assay (ELISA) and urinary cytology. A total of 40 patients with pathologically proven bladder cancer voided urine specimen before treatment. The urine samples were divided for NMP22 Bladder Chek test, NMP22 ELISA, and urinary cytology. In the 40 patients with bladder cancer, the overall positive rate was 62.5% for the NMP22 Bladder Chek test, 55% for the NMP22 ELISA test, and 27.5% for urine cytology. There was a significant difference between NMP22 Bladder Chek, NMP22 ELISA and cytology. The positive rate with the NMP22 Bladder Chek and NMP22 ELISA was higher in the patients with high grade and large-size (1 cm < or =) tumor. In 40 patients presenting with microhematuria without urothelial cancer, the false positive rate 12.5, 10, and 0% for NMP22 Bladder Chek, NMP22 ELISA, and urinary cytology. No significant difference was found with the test. In conclusion, the urine NMP22 Bladder Chek test provided a higher positive rate than the NMP22 ELISA test and urinary cytology. Therefore, the NMP22 Bladder Chek test may be clinically more useful as a tumor marker for the diagnosis of bladder cancer.     

Study on the diagnosis of urothelial cancer using multi-colour fluorescence in situ hybridization (FISH)--comparative analysis between FISH and cytology

To determine the diagnostic merit or demerit of genetic procedures using fluorescence in situ hybridization (FISH) for detecting both new and recurrent urothelial cancer, we analyzed the specimens from 81 out-patients with asymptomatic haematuria, aged over 40, in comparison with urine cytology. Of 10 with atypical cytology, 6 showed positive for FISH, and of these, 4 manifested urothelial cancer. FISH showed higher sensitivity in low/intermediate grade cases compared with cytology (FISH; 66.7% vs cytology; 11.1%). Of 15 primary bladder cancer, 4 showed recurrence, and all of these cases showed a positive FISH reaction, but only 1 in cytology. The sensitivity, specificity and accuracy of FISH tests were 81.2, 72.3 and 74.1%, respectively, and these of cytology were 37.5, 98.5 and 86.4% respectively. The FISH test was superior to cytology for sensitivity, but specificity and accuracy were inferior. The FISH tests could be a potent procedure for detecting urothelial cancer in cases of low/intermediate grade, atypical cytology and surveillance setting.     

尿纤维连接蛋白试纸诊断膀胱尿路上皮癌的价值

目的 研究尿纤维连接蛋白试纸（FN试纸）诊断膀胱尿路上皮癌的灵敏度和特异度,探讨其与膀胱尿路上皮癌各项临床指标之间的关系,为进一步无创性地诊断、随访膀胱尿路上皮癌提供依据.方法 运用自主研制的FN试纸测定膀胱尿路上皮癌患者、良性泌尿系疾病患者、正常体检人群尿液纤维连接蛋白.结果 FN试纸诊断膀胱尿路上皮癌的敏感度和特异度分别为72.94%和79.03%,其阳性预测值和阴性预测值分别为85.67%和68.06%;尿液细胞学诊断膀胱尿路上皮癌的特异度较高（100%）,但其敏感度仅为47.06%.结论 FN试纸检测对不同分化及浸润深度的膀胱尿路上皮癌有着较高的灵敏度与特异度,诊断价值高于尿液液基细胞学检查,但不能对膀胱肿瘤的分级以及病灶数量进行检测.     

Clinical study of urothelial tumors of the upper urinary tract. 1: Primary renal pelvic tumors

Forty primary renal pelvic tumors treated at our University between 1963 and 1981, were reviewed retrospectively. The conclusions of this study are as follows. Sex and age distribution of the patients were 30 males and 10 females (3: 1), and average age was 60.5 years old. The major symptoms were hematuria and flank pain; however, palpable mass was rare. The majority of patients were admitted to our clinic within 6 months from manifestation of symptoms. The major findings of IVP were non-functioning kidney and filling defect. The positive rate of urinary cytology was 46.7%. Total nephroureterectomy with bladder cuff was performed in 20 out of 32 cases. Histologically, 29 cases were transitional cell carcinoma and 4 cases were squamous cell carcinoma with renal calculi. Simultaneous urothelial tumors were seen in 10 cases, 3 in the ureter and 7 in the bladder. A subsequent ureteral tumor was found in one out of 12 cases in which ureters were resected incompletely, and 7 subsequent bladder tumors were found out of 32 cases receiving surgical treatment in the follow-up period. The 5-year survival rate by the actuarial method was 75.9%. Among several factors, grade and stage of the tumor were the most influencing factors for prognosis. An effective method of post-operative treatment could not be established.