MALDI MS profiling of post-DRE urine samples highlights the potential of ß-microseminoprotein as a marker for prostatic diseases.

BACKGROUND: To use spectra acquired by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) from pre- and post-digital rectal examination (DRE) urine samples to search for discriminating peaks that can adequately distinguish between benign and malignant prostate conditions, and identify the peaks' underlying biomolecules. METHODS: Twenty-five participants with prostate cancer (PCa) and 27 participants with a variety of benign prostatic conditions as confirmed by a 10-core tissue biopsy were included. Pre- and post-DRE urine samples were prepared for MALDI MS profiling using an automated clean-up procedure. Following mass spectra collection and processing, peak mass and intensity were extracted and subjected to statistical analysis to identify peaks capable of distinguishing between benign and cancer. Logistic regression was used to combine markers to create a sensitive and specific test. RESULTS: A peak at m/z 10,760 was identified as ß-microseminoprotein (ß-MSMB) and found to be statistically lower in urine from PCa participants using the peak's average areas. By combining serum prostate-specific antigen (PSA) levels with MALDI MS-measured ß-MSMB levels, optimum threshold values obtained from Receiver Operator characteristics curves gave an increased sensitivity of 96% at a specificity of 26%. CONCLUSIONS: These results demonstrate that with a simple sample clean-up followed by MALDI MS profiling, significant differences of MSMB abundance were found in post-DRE urine samples. In combination with PSA serum levels, obtained from a classic clinical assay led to high classification accuracy for PCa in the studied sample set. Our results need to be validated in a larger multicenter prospective randomized clinical trial.

Identification of prostate-enriched proteins by in-depth proteomic analyses of expressed prostatic secretions in urine.

Urinary expressed prostatic secretion or "EPS-urine" is proximal tissue fluid that is collected after a digital rectal exam (DRE). EPS-urine is a rich source of prostate-derived proteins that can be used for biomarker discovery for prostate cancer (PCa) and other prostatic diseases. We previously conducted a comprehensive proteome analysis of direct expressed prostatic secretions (EPS). In the current study, we defined the proteome of EPS-urine employing Multidimensional Protein Identification Technology (MudPIT) and providing a comprehensive catalogue of this body fluid for future biomarker studies. We identified 1022 unique proteins in a heterogeneous cohort of 11 EPS-urines derived from biopsy negative noncancer diagnoses with some benign prostatic diseases (BPH) and low-grade PCa, representative of secreted prostate and immune system-derived proteins in a urine background. We further applied MudPIT-based proteomics to generate and compare the differential proteome from a subset of pooled urines (pre-DRE) and EPS-urines (post-DRE) from noncancer and PCa patients. The direct proteomic comparison of these highly controlled patient sample pools enabled us to define a list of prostate-enriched proteins detectable in EPS-urine and distinguishable from a complex urine protein background. A combinatorial analysis of both proteomics data sets and systematic integration with publicly available proteomics data of related body fluids, human tissue transcriptomic data, and immunohistochemistry images from the Human Protein Atlas database allowed us to demarcate a robust panel of 49 prostate-derived proteins in EPS-urine. Finally, we validated the expression of seven of these proteins using Western blotting, supporting the likelihood that they originate from the prostate. The definition of these prostatic proteins in EPS-urine samples provides a reference for future investigations for prostatic-disease biomarker studies.

Clinical collection and protein properties of expressed prostatic secretions as a source for biomarkers of prostatic disease.

The prostate gland secretes many proteins in a prostatic fluid that combines with seminal vesicle derived fluids to promote sperm activation and function. Proximal fluids of the prostate that can be collected clinically are seminal plasma and expressed-prostatic secretion (EPS) fluids. EPS represents the fluid being secreted by the prostate following a digital rectal prostate massage, which in turn can be collected in voided urine post-exam. This collection is not disruptive to a standard urological exam, and it can be repeatedly collected from men across all prostatic disease states. A direct EPS fluid can also be collected under anesthesia prior to prostatectomy. While multiple genetic assays for prostate cancer detection are being developed for the shed epithelial cell fraction of EPS urines, the remaining fluid that contains many prostate-derived proteins has been minimally characterized. Approaches to optimization and standardization of EPS collection consistent with current urological exam and surgical practices are described, and initial proteomic and glycomic evaluations of the of EPS fluid are summarized for prostate specific antigen and prostatic acid phosphatase. Continued characterization of the prostate specific protein components of EPS urine combined with optimization of clinical collection procedures should facilitate discovery of new biomarkers for prostate cancer.

Benefits of the iQ200 automated urine microscopy analyser in routine urinalysis.

BACKGROUND: Urine microscopic analysis is hampered by its lack in standardisation and semi-quantitative reports, resulting in limited reliability. Automation of urinalysis could overcome these problems. METHODS: We compared the performance of the iQ200 with traditional microscopy and strip analysis in routine urinalysis. A total of 1482 routine samples, positive in dipstick testing, were evaluated for erythrocytes, leukocytes, casts, dysmorphic erythrocytes and bacteria using the iQ200 and traditional microscopy. The results of 320 of these samples were linked to underlying urological pathology as well as results from bacterial culturing. RESULTS: Analytically, the iQ200 surpasses traditional microscopy. The identification of casts and dysmorphic erythrocytes in routine samples improves when using the iQ200, although the sub-classification of casts required well-trained technicians. The auto-classification of particles was least reliable for yeast and bacterial cocci. The quantitative reports, and therefore the use of precise cut-off points allowed earlier and improved detection of urinary tract pathology. CONCLUSIONS: The performance of the iQ200 is equal to traditional microscopy, but it strongly improves the reliability of urinalysis by standardisation, quantitative reports and improved workflow. From a clinical point of view, renewed attention and improvement of routine urinalysis aids in the efficient detection of renal and urinary tract pathology.

Bladder tumor antigen stat test in non-urothelial malignant urologic conditions.

BACKGROUND: The bladder tumor antigen stat is a simple and fast one-step immunochromatographic assay for the detection of bladder tumor-associated antigen in urine. OBJECTIVES: To evaluate the BTA stat in non-bladder cancer patients in order to identify the categories contributing to its low specificity. METHODS: A single voided urine sample was collected from 45 patients treated in the urology clinic for conditions not related to bladder cancer. Each urine sample was examined by the BTA stat test and cytology. RESULTS: The overall specificity of the BTA stat test was 44%, which was significantly lower than that of urine cytology, 90%. The false positive rates for the BTA stat test varied among the different clinical categories, being highest in cases of urinary tract calculi (90%), and benign prostatic hypertrophy (73%). Exclusion of these categories from data analysis improved BTA stat specificity to 66%. CONCLUSIONS: Clinical categories contributing to low BTA stat specificity can be identified, and their exclusion improves the specificity of this test.

Case report: recurrent hematuria and hematospermia due to prostatic telangiectasia in classic von Willebrand's disease.

A previously healthy 32-year-old man presented with recurrent exercise induced painless gross hematuria and hematospermia. An extensive evaluation demonstrated classic von Willebrand's disease associated with vascular telangiectasia of the prostate gland as the locus of hemorrhage. The bleeding resolved spontaneously and without recurrence. The coexistence of von Willebrand's disease and vascular telangiectasia has been described previously, although it is a rare occurrence. However, a review of the English literature revealed no report of vascular telangiectasia involving the prostate gland, and therefore is the subject of this report. The prostate gland now should be added to the list of viscera associated with vascular telangiectasia and von Willebrand's disease.

[Significance of prostatic acid phosphatase, gamma-seminoprotein and prostatic specific antigen in the urine. First report: the measurement of PAP, gamma-Sm and PA in the urine of patients with prostatic diseases].

To study the significance of prostatic acid phosphatase (PAP), gamma-seminoprotein (gamma-Sm) and prostatic specific antigen (PA) in urine, we have determined the urinary levels of these proteins in women and infants, in patients without prostatic disease, in patients with benign prostatic hypertrophy, and in patients with prostatic adenocarcinoma. Women and infants were found to excrete little PAP (27.9 +/- 4.8 ng/mg) and undetectable levels of gamma-Sm except one case, and undetectable levels of PA in the urine. The excretion of PAP in patients with prostatic carcinoma who were either castrated, or treated with endocrine therapy was lower than the levels in women and infants, or the levels in patients without prostatic diseases, or the levels in patients with BPH. Urinary excretion levels of gamma-Sm and PA were undetectable in the patients with well-controlled prostatic carcinoma. The present study suggests that the determination of PAP, gamma-Sm and PA in the urine of patients with prostatic carcinoma may become a useful tool for monitoring of the primary locus of the carcinoma, but additional assays of urinary PAP, gamma-Sm and PA should be measured at regular intervals to be concluded.

DNA cytophotometry of voided urine sediment. Comparison with results of cytologic diagnosis and image analysis.

Measurements of nuclear DNA were performed in urothelial cells in 54 Feulgen-restained cytocentrifuge preparations of voided urine previously studied visually and with an image analysis system. The study included 30 patients with bladder tumors of various grades, 9 patients with prostatic disease and 15 control samples from normal donors. A number of additional control measurements were performed, including measurements in tissue samples of the 30 bladder tumors corresponding to the cytologic samples. It was documented that DNA can be measured in most urinary sediments. The diagnostic performance of the image analysis system reflected the DNA patterns in 47 of the 54 cases. In several instances, particularly in cases of prostatic disease, the image analysis system recognized abnormal DNA patterns in the absence of significant morphologic abnormalities in the urothelial cells. In seven cases, the image analysis findings failed to conform with the DNA patterns. The reasons for these surprising results are discussed, and future modifications of the image analysis system are proposed.

Disposable peak urinary flowmeter estimates lower urinary tract obstruction.

Peak urinary flow rate represents the highest flow rate achieved during a single urination and, as such, represents the patient's best effort at micturition. Peak flow rate, correlated with patient age and volume voided, effectively estimates lower urinary tract obstruction. The 63 normal and 368 abnormal male subjects urinated in privacy into a plastic sterile disposable device (the peakometer), which measured peak flow rate and volume voided. These data plus age, ultimate diagnosis and interval since last urination comprised our data base. Percentage distribution of diagnosis in this population was prostatic obstruction 47.3 per cent, stricture 19.3 per cent, normal 14.6 per cent, prostatitis 8.4 per cent, neurogenic bladder 2 per cent and miscellaneous 8.4 per cent. The average peak flow rate for normal male subjects reaches 27.6 ml. per second, which differs significantly from that for patients with prostatic obstruction--9.4 ml. per second, stricture--10.5 ml. per second, prostatitis--16.3 ml. per second and neurogenic bladder--13.9 ml. per second. The peak flow rate decreased progressively as the age of the subjects increased. We measured average decreases of 10 ml. per second peak flow for every 30 years after age 10. Peak flow rate increases as volume voided increases. Requirements of our measuring device combined with urodynamic responses caused us to select 100 ml. voided as the minimum acceptable volume. With volumes more than this any given individual may deviate plus or minus 10 per cent from the true mean peak flow depending upon volume voided. For practical purposes peak flow, age and volume must be considered to categorize voiding by peak flow rate. With these variables 2 graphs that compare peak flow, age and volume may be used to estimate voiding function for a given male patient. Comparison of peak flow rates, volume voided and voiding interval before and after surgical correction of obstruction documented significant increase in volume voided or in interval between voiding. Peak urinary flow rate measurement by this device predicted normality or abnormality with 90 to 95 per cent accuracy. Therefore, this represents a valid screening test but it does not in itself provide the diagnosis of abnormal urination.