Urology

Urodynamic Effects of Volume-adjustable Balloons for Treatment of Postprostatectomy Urinary Incontinence (Article)

Utomo, E. — 2013-02-26T00:00:00Z

Objective: To evaluate the urodynamic changes in patients treated with Adjustable Continence Therapy for men (ProACT) for postprostatectomy incontinence and to explore the clinical and urodynamic preimplantation parameters as predictors of clinical outcome. Materials and Methods: Patients underwent urodynamic studies before and after ProACT implantation. ProACT was considered successful if patients used none or 1 dry precautionary pad and nonsuccessful if the patient reported ≥1 wet pad/d. The pre- and postimplantation assessments were retrospectively compared within and between the success and nonsuccess groups. Multivariate logistic regression analysis was performed to investigate the association between the preimplantation variables and the clinical outcomes of ProACT implantation. Results: A total of 49 patients were included, 37 with successful and 12 with nonsuccessful clinical outcome. Postimplantation urodynamic studies were performed a median of 9 months after ProACT implantation. In the successfully treated patients, maximum free flow rate, bladder contractility index, maximum of bladder contractility parameter W, and bladder voiding efficiency were significantly lower after implantation. The detrusor pressure at maximum flow rate, postvoid residual urine volume, and bladder outlet obstruction index were significantly higher. A longer duration of urinary incontinence, the use of >5 pads daily, and a smaller cystometric bladder capacity were all independently associated with nonsuccessful clinical outcome after ProACT implantation. Conclusion: ProACT implantation with successful clinical outcome resulted in greater urethral resistance during voiding and reduced bladder contraction strength. A longer duration of incontinence, the use of >5 pads daily, and a smaller cystometric bladder capacity were independent predictors of unsuccessful clinical outcomes, suggesting ProACT implantation should be considered sooner, rather than later, after conservative treatment of postprostatectomy incontinence has failed.

Selective detection of aggressive prostate cancer (Article)

Schröder, F.H. — 2011-07-19T00:00:00Z

Selectively identifying aggressive prostate cancer will reduce unnecessary testing and overdiagnosis. Multivariate models can be of assistance but require proper validation and users must be aware of the setting from which the models were derived. See also pages 000-000.

Do androgens control the uptake of 18F-FDG, 11C-choline and 11C-acetate in human prostate cancer cell lines? (Article)

Emonds, K.M. — 2011-07-06T00:00:00Z

Purpose: The aim of this study was to evaluate the impact of androgen ablation therapy in different prostate cancer (PCa) cell lines-reflecting different stages of the disease-on18F-fluorodeoxyglucose (FDG),11C-choline and11C-acetate uptake. Methods: Uptake experiments were performed in androgen-sensitive (LNCaP, PC346C) and independent cell lines (22Rv1, PC346DCC, PC-3) as well as in a benign prostatic hyperplasia (BPH-1) cell line. Tracer uptake was assessed under androgen ablation. Results of the cancer cell lines were normalized to those of BPH-1. To evaluate the effect of androgen on the uptake of18F-FDG,11C-choline and11C-acetate in PCa cell lines, 10-8M R1881, 10-10M R1881, the combination of 10-10M R1881 plus 10-6M Casodex or 10-6M Casodex alone were added in parallel cell cultures 1 day before uptake experiments. Uptake in androgen-supplemented cell cultures was compared to the uptake under androgen deprivation. Uptake was corrected for cell number using protein content. Results: Compared to BPH-1, a higher18F-FDG uptake was observed only in PC346C cells, whereas a higher11C-choline and markedly increased11C-acetate uptake was seen in all cancer cell lines. Androgens significantly modulated the uptake of18F-FDG in LNCaP, PC346C and 22Rv1 cells, and of11C-choline in the PC346C and 22Rv1 cell line. No androgenic effect on11C-choline and18F-FDG uptake was observed in PC-3 and PC346DCC cells.11C-Acetate uptake was independent of androgen status in all PCa cell lines studied. Conclusion:18F-FDG uptake in PCa cell lines showed the highest variability and strongest androgen effect, suggesting its poor potential for metabolic imaging of advanced PCa. In contrast to18F-FDG and11C-choline,11C-acetate uptake was unaffected by androgens and thus11C-acetate seems best for monitoring PCa progression.

Prostate cancer screening has no effect on prostate cancer specific mortality over 20 years of follow-up of Swedish men (Article)

Leeuwen, P.J. van — 2011-07-06T00:00:00Z

Gastrin-releasing peptide receptor-based targeting using bombesin analogues is superior to metabolism-based targeting using choline for in vivo imaging of human prostate cancer xenografts (Article)

Schroeder, R.P.J. — 2011-07-01T00:00:00Z

Purpose: Prostate cancer (PC) is a major health problem. Overexpression of the gastrin-releasing peptide receptor (GRPR) in PC, but not in the hyperplastic prostate, provides a promising target for staging and monitoring of PC. Based on the assumption that cancer cells have increased metabolic activity, metabolism-based tracers are also being used for PC imaging. We compared GRPR-based targeting using the68Ga-labelled bombesin analogue AMBA with metabolism-based targeting using18F-methylcholine (18F-FCH) in nude mice bearing human prostate VCaP xenografts. Methods: PET and biodistribution studies were performed with both68Ga-AMBA and18F-FCH in all VCaP tumour-bearing mice, with PC-3 tumour-bearing mice as reference. Scanning started immediately after injection. Dynamic PET scans were reconstructed and analysed quantitatively. Biodistribution of tracers and tissue uptake was expressed as percent of injected dose per gram tissue (%ID/g). Results: All tumours were clearly visualized using68Ga-AMBA.18F-FCH showed significantly less contrast due to poor tumour-to-background ratios. Quantitative PET analyses showed fast tumour uptake and high retention for both tracers. VCaP tumour uptake values determined from PET at steady-state were 6.7±1.4%ID/g (20-30 min after injection, N=8) for68Ga-AMBA and 1.6±0.5%ID/g (10-20 min after injection, N=8) for18F-FCH, which were significantly different (p<0.001). The results in PC-3 tumour-bearing mice were comparable. Biodistribution data were in accordance with the PET results showing VCaP tumour uptake values of 9.5±4.8%ID/g (N=8) for68Ga-AMBA and 2.1±0.4%ID/g (N=8) for18F-FCH. Apart from the GRPR-expressing organs, uptake in all organs was lower for68Ga-AMBA than for18F-FCH. Conclusion: Tumour uptake of68Ga-AMBA was higher while overall background activity was lower than observed for18F-FCH in the same PC-bearing mice. These results suggest that peptide receptor-based targeting using the bombesin analogue AMBA is superior to metabolism-based targeting using choline for scintigraphy of PC.

Prospective evaluation of operating characteristics of prostate cancer detection biomarkers (Article)

Roobol, M.J. — 2011-07-01T00:00:00Z

Screening for prostate cancer-The controversy continues, but can it be resolved? (Article)

Bul, M. — 2011-06-01T00:00:00Z

Background. In 2009, the European Randomized Study of Screening for Prostate Cancer (ERSPC) was one of two studies to report interim data on the effect of screening for prostate cancer (PC) on the disease specific mortality. Contradictory results caused considerable discussion and misunderstanding in secondary literature. Methods. This document is based on a non systematic review of recent evidence for and against screening for PC, specifically considering three recently published randomized screening trials [1â€"3]. Results. The ERSPC data are based on a core age group of 162 387 men, aged 55â€"69 years, who were identified through population registries in seven European countries. Men were randomized between a screening group that received screening at an average of once every four years and a control group. After a median follow-up of nine years, a reduction in the rate of death from PC by 20% was shown which increased to 31% after adjusting for non-compliance and contamination. Overdetection and subsequent overtreatment (with a number needed to treat (NNT) of 48) are considered to be the major down sides of screening. The recently published 14-year results have shown that these down sides strongly depend on the duration of follow-up. In response to the outcomes of the ERSPC, several points of discussion have been brought up by various authors concerning the usefulness of screening considering benefits, harms and costs, the methodology of the ERSPC and the interpretation of its outcomes. Important issues to address regarding PC screening are addressed. Conclusions. This paper sheds a light on the controversial points of the ERSPC as well as on the priority issues of PC screening. On July 2, 2010 the Swedish section of ERSPC (Göteborg screening trial) published their results with a median follow-up of 14 years. With longer follow-up the data confirm the trend seen in improvement of PC mortality and suggest much more favorable future outcomes also with respect to the NNT to prevent one PC death.

Tumour markers in prostate cancer III: Biomarkers in urine (Article)

Roobol, M.J. — 2011-06-01T00:00:00Z

The serum PSA test still is the most important biomarker for the detection and follow-up of prostate cancer. PSA-based screening can reduce disease specific mortality but coinciding unnecessary testing and overdiagnosis warrant further research for more specific biomarkers. Numerous studies of both serum and urine-based prostate cancer biomarker candidates have been presented the last ten years. However, biomarkers for identifying the most aggressive subsets of this malignancy are still missing. Being non-invasive, urine-based tests might be suitable for both clinical and (mass) screening purposes, but also for prediction and to gain prognostic information. Protein-based, DNA-based and RNA-based urine biomarkers have been developed and tested. Protein markers in urine. Data on protein-based urine biomarkers (i.e. Annexin A3, matrix metalloproteinases and the urinary:serum PSA ratio) show up to now contradictory results and further studies are warranted to be able to assess their clinical value in which the cost aspect should not be overlooked. DNA markers in urine. Studies on DNA-based urine biomarkers focus on hypermethylation of gene panels with GSTP1 hypermethylation being the most promising individual marker. Larger prospective clinical studies of single markers and gene panels are however needed to validate their clinical utility. RNA markers in urine. RNA-based urine biomarkers are by far the most developed. The PCA3 test, the TMPRSS2â€"ERG fusion gene, transcript expression levels of GOLPH2, SPINK1 and their combination have been subject of many studies showing encouraging results. Conclusion. Up to now urine-based biomarkers represent a promising alternative or addition to serum-based biomarkers. Prospective studies in a multivariate setting, including larger sample sizes and avoiding attribution bias caused by preselection on the basis of serum PSA are however required.

The predictive value of immunohistochemical markers in untreated Wilms' tumour: are they useful? (Article)

Ghanem, M.A. — 2011-05-19T00:00:00Z

Purpose: This study reevaluates the potential role of different tumour markers as prognostic indicators in untreated nephroblastoma. Methods: Expression of a broad panel of tumour markers was investigated by means of immunohistochemical analysis in 43 WT patients. Patients were treated by radical nephrectomy and had a mean follow-up of 11.9 years. Results: Generally, all the tumour markers studied were expressed in normal kidney tissue and at variable levels in the three cell types of WT (blastema, epithelium and stroma). Immunoreactive blastemal (Bcl-X, Bcl-2 and CD44s) and epithelial (Bcl-X, Bcl-2 and MIB-1) cells were present in the majority of tumours. No correlation was found between their expression and pathological stages. Univariate analysis showed that blastemal WT-1, TGF-α, VEGF, MIB-1 and p27 Kip1 were indicative for clinical progression. In a multivariate analysis, WT-1 protein expression by blastemal cells was an independent prognostic marker for clinical progression. Conclusions: The blastemal WT-1, TGF-α, VEGF, MIB-1 and p27Kip1 expression correlate with clinical progression in untreated nephroblastoma. Therefore, their expression may be of value in identifying patients with a high propensity to develop distant metastases.

Lower sensitivity of screening mammography after previous benign breast surgery (Article)

Van Breest Smallenburg, V. — 2011-05-11T00:00:00Z

Few data are available on the effect of previous benign breast surgery on screening mammography accuracy. We determined whether sensitivity of screening mammography and tumor characteristics are different for women with and without previous benign breast surgery. We included a consecutive series of 317,398 screening mammograms of women screened between 1997 and 2008. During 2-year follow-up, clinical data, breast imaging, biopsy and surgery reports were collected from women with screen-detected or interval breast cancers. Screening sensitivity, tumor biology and tumor stages were compared between 168 women with breast cancer and prior ipsilateral benign breast surgery and 2,039 women with breast cancer but without previous ipsilateral, benign breast surgery. The sensitivity of screening mammography was significantly lower for women with prior surgery [64.3% (108/168) versus 73.4% (1,496/2,039), p = 0.01]. The concomitant increased interval cancer risk remained significant after logistic regression adjustment for age and breast density (OR = 1.5, 95% CI: 1.1-2.1). Comparing screen-detected cancers in women with and without prior breast surgery, no significant differences in estrogen receptor status (p = 0.56), mitotic activity (p = 0.17), proportions of large (T2+) tumors (p = 0.6) or lymph node positive tumors (p = 0.4) were found. Also for interval cancers, no differences were found in estrogen receptor status (p = 0.41), mitotic activity (p = 0.39), proportions of large tumors (p = 0.9) and lymph node positive tumors (p = 0.5) between women with and without prior breast surgery. We conclude that sensitivity of screening mammography is significantly lower in women with previous benign breast surgery than without, but tumor characteristics are comparable both for screen detected cancers and interval cancers. Copyright

Patient and tumor characteristics of bilateral breast cancer at screening mammography in the Netherlands, a population-based study (Article)

Setz-Pels, W. — 2011-05-09T00:00:00Z

Few data are available on bilateral breast cancer in the screening population. The aim of this study was to determine patient and tumor characteristics of women with bilateral breast cancer at screening mammography. We included all 350,637 screening mammography examinations of women participating in a biennial screening program in a southern screening region of the Netherlands between May 1998 and January 2010. For referred women, all breast imaging reports, biopsy results, and surgery reports during one year after referral were collected. We compared patient and tumor characteristics of referred women with a diagnosis of bilateral breast cancer or unilateral breast cancer at workup. Bilateral or unilateral breast cancer had been diagnosed in respectively 40 (2.2%) and 1766 (97.8%) of 1806 referred women. Women with bilateral or unilateral breast cancer did not differ significantly in mean age, mammographic breast density, family history of breast cancer, or use of hormone replacement therapy. Compared with index cancers, contralateral cancers comprised significantly more lobular cancers (P = 0.02). Tumor size, mitotic activity, and estrogen receptor status were comparable for both groups, but contralateral cancers had a significantly lower risk of lymph node metastases (P = 0.03). Compared to unilateral breast cancer, contralateral malignancies in women with bilateral breast cancer comprised significantly more lobular cancers (P = 0.004) and lymph node negative cancers (P = 0.01). Contralateral breast cancers detected at screening comprise more lobular cancers and show less nodal involvement than index cancers or unilateral cancers. No differences are observed with respect to other patient and tumor characteristics.

Contemporary role of prostate cancer gene 3 in the management of prostate cancer (Article)

Roobol, M.J. — 2011-05-01T00:00:00Z

Purpose of Review: With increasing evidence that prostate-specific antigen (PSA)-based screening can reduce disease-specific mortality but coincides with unacceptable levels of unnecessary testing and the diagnosis of potentially nonlife-threatening disease, the need for new, more specific biomarkers is urgent. Within this context the role of the prostate cancer gene 3 (PCA3) test is evaluated. Recent Findings: Studies investigating the value of PCA3 as a diagnostic test virtually all show a beneficial effect as compared to PSA with respect to specificity. Beside the fact that most of these studies are subject to potential bias, the observed increased specificity was accompanied by relatively low sensitivities. Two studies, attempting to avoid selection bias as much as possible, show a marginal beneficial effect of the PCA3 test. Data on PCA3 as a staging tool for prostate cancer remain inconclusive. SUMMARY: The PCA3 test is not capable of replacing the PSA test in clinical practice and an appropriate cut-off level with acceptable performance characteristics is hard to define. Its value as a first-line diagnostic test is limited. The addition of PCA3 to risk assessment tools leads to an increase in predictive capability. Data relating to the accuracy of PCA3 on prostate cancer staging are contradictory and PCA3 as prognostic test should be subject of future studies.

Comparison of risk calculators from the Prostate Cancer Prevention Trial and the European Randomized Study of Screening for Prostate Cancer in a contemporary Canadian cohort (Article)

Trottier, G. — 2011-04-22T00:00:00Z

Patients And Methods: At Princess Margaret Hospital, 982 consecutive patients with PCPT-RC and ERSPC-RC covariables were prospectively catalogued before prostate biopsy for suspicion of prostate cancer (PCa). Receiver-operating characteristic (ROC) curves were generated for each calculator and prostate-specific antigen (PSA). Comparisons by area under the curve (AUC) and calibration plots were performed. Predictors of PCa were identified by univariable and multivariable logistic regression. Results: PCa was detected in 46% and high-grade (HG) PCa (Gleason ≥4) in 23% of subjects with a median PSA level of 6.02 ng/mL. Multivariable analysis identified transrectal ultrasonography nodule, prostate volume and PSA as the most important predictors of PCa and HG PCa. ROC curve analysis showed that the ERSPC-RC (AUC = 0.71) outperformed the PCPT-RC (AUC = 0.63) and PSA (AUC = 0.55), for PCa prediction, P < 0.001. The PCPT-RC was better calibrated in the higher prediction range (40-100%) than the ERSPC-RC, whereas the ERSPC-RC had better calibration and avoided more biopsies in the lower risk range (0-30%). Discrimination of the ERSPC-RC continued to be superior to the PCPT-RC when the cohort was stratified by different clinical variables. Conclusions: The ERSPC-RC had better discrimination for predicting PCa compared to the PCPT-RC in this Canadian cohort. Calibration would need to be improved to allow routine use of the ERSPC-RC in Canadian practice. What's known on the subject? and What does the study add? The European Randomized Study of Screening for Prostate Cancer risk calculator (ERSPC-RC) has been validated in a European population and shown to outperform the Prostate Cancer Prevention Trial risk calculator (PCPT-RC) for predicting prostate cancer. However, the ERSPC-RC has not been validated in North America where the PCPT-RC has been extensively validated. This study is the first to compare these calculators in non-European patient cohort showing better performance of the ERSPC-RC, but poor calibration. © 2011 THE AUTHORS. BJU INTERNATIONAL

Prostate Cancer Biomarkers to Improve Risk Stratification: Is Our Knowledge of Prostate Cancer Sufficient to Spare Prostate Biopsies Safely? (Article)

Roobol, M.J. — 2011-04-21T00:00:00Z

Identifying and characterizing "escapes"-men who develop metastases or die from prostate cancer despite screening (ERSPC, section Rotterdam) (Article)

Zhu, X.D. — 2011-04-06T00:00:00Z

We aim to identify and characterize "escapes," men who developed metastasis and/or died from prostate cancer (PCa) despite screening, in the framework of the novel international ESCAPE-project. With this knowledge, the ultimate goal is to improve screening strategy. In this article, we focus on the study cohort of the European Randomized Study of Screening for Prostate Cancer (ERSPC), section Rotterdam. In all, 21,210 men were randomized to the screening arm of whom 19,950 were actually screened. The screening interval was 4 years. Men with prostate-specific antigen ≥3.0 ng/ml were recommended to undergo lateralized sextant prostate biopsy. The follow-up was complete until January 1, 2009. Of 19,950 screened men, 2,317 were diagnosed with PCa. Of these cancers 1,946 were detected in a screening round and 371 during an interval. The median follow-up was 11.1 years for the whole cohort and 7.3 years for men diagnosed with PCa. In total, we identified 168 escapes among 2,317 cancers (7.3%) within our screening cohort of 19,950 men (0.8%). More than half of these escapes were found in the initial screening round (94 of 168). Possible mechanisms behind escaping are nonattending, inadequate screening tests, the relative long screening interval, the age cut-off at 75 years, and undertreatment. International cooperation is crucial to compare the escapes of our cohort with other study groups participating in the ESCAPE-project which have different, more aggressive screening strategies. Subsequently, we can achieve improvements of the current screening algorithm, which hopefully will further decrease PCa-specific mortality without increasing overdiagnosis and overtreatment. Copyright

Prostate Cancer Incidence and Disease-Specific Survival of Men with Initial Prostate-Specific Antigen Less Than 3.0 ng/ml Who Are Participating in ERSPC Rotterdam (Article)

Bul, M. — 2011-04-01T00:00:00Z

Background: The European Randomised Study of Screening for Prostate Cancer (ERSPC) applies a prostate-specific antigen (PSA) cut-off value ≥3.0 ng/ml as an indication for lateralised sextant biopsy. Objective: To analyse the incidence and disease-specific mortality for prostate cancer (PCa) in men with an initial PSA <3.0 ng/ml. Design, setting and participants: From November 1993 to December 1999, a total of 42 376 men identified from population registries in the Rotterdam region (55-74 yr of age) were randomised to an intervention or control arm. A total of 19 950 men were screened during the first screening round. Intervention: A PSA <3.0 ng/ml was below the biopsy threshold. PCa cases were identified at rescreens every 4 yr or as interval cancers. Measurements: Distribution of incidence, aggressiveness, and disease-specific mortality of PCa per PSA range was measured. Causes of death were evaluated by an independent committee, and follow-up was complete until 31 December 2008. Results and limitations: From 1993 to 2008, 915 PCa cases were diagnosed in 15 758 men (5.8%) with an initial PSA <3.0 ng/ml and a median age of 62.3 yr. Median overall follow-up was 11 yr. PCa incidence increased significantly with higher initial PSA levels. Aggressive PCa (clinical stage ≥T2c, Gleason score ≥8, PSA >20 ng/ml, positive lymph nodes, or metastases at diagnosis) was detected in 66 of 733 screen-detected PCa cases (9.0%) and 72 of 182 interval-detected PCa cases (39.6%). Twenty-three PCa deaths occurred in the total population (0.15%), with an increasing risk of PCa mortality in men with higher initial PSA values. Conclusions: The risk of PCa, aggressive PCa and PCa mortality in a screening population with initial PSA <3.0 ng/ml increases significantly with higher initial PSA levels. These results contribute to the risk stratification and individual management of men in PSA-based screening programmes.

Prognostic Implications of Important Genetic Alterations in Prostate Cancer (Doctoral Thesis)

Boormans, J.L. — 2011-02-18T00:00:00Z

The prostate is a walnut-sized gland that is located caudally from the urinary bladder. It excretes fluid as a part of the semen of men. Prostatic neoplasia is common; in Western developed countries prostate cancer is the most common non-cutaneous malignancy in men and it is the second leading cause of all male cancer deaths. The detection of prostate cancer has markedly increased since the introduction of the serum prostatespecifi c antigen (PSA) in the late eighties. Although familial and hereditary prostate cancer occurs, sporadic cancers account for at least 85% of all prostate cancers. Age is the strongest risk factor for developing prostate cancer. From autopsy studies it is known that in the 6th decade already 55% of the male population has the disease, and more than 75% of men older than 85 years have cancer foci in their prostate. Prostate cancer is a very heterogeneous disease that can range from indolent, asymptomatic tumours in many patients to a rapidly fatal malignancy in some. At diagnosis, the majority of tumours are confined to the prostatic gland, named clinical stage cT1 and cT2 (see Table 1 for the Tumour, Node, Metastasis (TNM) classifi cation). Some tumours, however, already grow outside the prostatic capsule (cT3) or even invade the surrounding organs (cT4). Dissemination of the disease usually occurs to the regional pelvic lymph nodes and the axial skeleton. The latter will mainly cause pain, although neurological deficit due to compression of the myelum is also possible in severe cases. Diagnostic modalities to detect prostate cancer are digital rectal examination (DRE), measurement of the serum prostate-specific antigen (PSA), transrectal ultrasound of the prostate (TRUS) with subsequent ultrasound-guided prostate biopsies, and magnetic resonance imaging (MRI). Computerized tomography (CT) of the pelvis is reserved for detection of enlarged lymph nodes, whereas bone metastases can be detected by nuclear scintigraphy, CT, MRI or conventional X-ray of the skeleton. The serum PSA is a marker for prostate cancer. Although PSA lacks specificity, it is currently the most valid biomarker available, not only for diagnostic purposes but also for disease monitoring after treatment with curative intent. Since its introduction, PSA has been widely used as a screening tool to detect prostate cancer in asymptomatic men. Recent findings showed that PSA-based screening for prostate cancer resulted in a relative risk reduction of 20% to die from the disease. It must be noted, however, that at the time of the first statistical relevant difference between the intervention and control arm, it was shown that 48 patients had to be treated to prevent one death from prostate cancer. The findings by Schroder et al. show the difficulties that remain to correctly identify the indolent prostate tumours from the ones that need curative treatment. Therefore, prognostic markers that facilitate a risk stratification of prostate cancer patients and that are helpful for optimizing treatment decisions are still very much needed.

Overexpression of full-length ETV1 transcripts in clinical prostate cancer due to gene translocation (Article)

Gasi, D. — 2011-02-07T00:00:00Z

ETV1 is overexpressed in a subset of clinical prostate cancers as a fusion transcript with many different partners. However, ETV1 can also be overexpressed as a full-length transcript. Full-length ETV1 protein functions differently from truncated ETV1 produced by fusion genes. In this study we describe the genetic background of full-length ETV1 overexpression and the biological properties of different full-length ETV1 isoforms in prostate cancer. Break-apart FISH showed in five out of six patient samples with overexpression of full-length ETV1 a genomic rearrangement of the gene, indicating frequent translocation. We were able to study the rearrangements in more detail in two tumors. In the first tumor 59′-RACE on cDNA showed linkage of the complete ETV1 transcript to the first exon of a prostate-specific two exon ncRNA gene that maps on chromosome 14 (EST14). This resulted in the expression of both full-length ETV1 transcripts and EST14-ETV1 fusion transcripts. In chromosome spreads of a xenograft derived from the second prostate cancer we observed a complex ETV1 translocation involving a chromosome 7 fragment that harbors ETV1 and fragments of chromosomes 4 and 10. Further studies revealed the overexpression of several different full-length transcripts, giving rise to four protein isoforms with different N-terminal regions. Even the shortest isoform synthesized by full-length ETV1 stimulated in vitro anchorage-independent growth of PNT2C2 prostate cells. This contrasts the lack of activity of even shorter N-truncated ETV1 produced by fusion transcripts. Our findings that in clinical prostate cancer overexpression of full-length ETV1 is due to genomic rearrangements involving different chromosomes and the identification of a shortened biologically active ETV1 isoform are highly relevant for understanding the mechanism of ETV1 function in prostate cancer.

Congenital heart defects and biomarkers of methylation in children: A case-control study (Article)

Obermann-Borst, S.A. — 2011-02-01T00:00:00Z

Eur J Clin Invest 2011; 41 (2): 143-150Background Derangements in the maternal methylation pathway, expressed by global hypomethylation and hyperhomocysteinemia, are associated with the risk of having a child with a congenital heart defect (CHD). It is not known whether periconception exposure to these metabolic derangements contributes to chromosome segregation and metabolic programming of this pathway in the foetus.Design In a Dutch population-based case-control study of 143 children with CHD and 186 healthy children, we investigated S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), total homocysteine (tHcy), the vitamins folate and B12 and the functional single nucleotide polymorphisms in the folate gene MTHFR 677C>T and 1298A>C. Comparisons were made between cases and controls adjusting for age, medication, vitamin use and CHD family history.Results In the overall CHD group, the median concentrations of SAM (P = 0.011), folate in serum (P = 0.021) and RBC (P = 0.030) were significantly higher than in the controls. Subgroup analysis showed that this was mainly attributable to complex CHD with higher SAM (P < 0.001), SAH (P = 0.012) and serum folate (P = 0.010) independent of carriership of MTHFR polymorphisms. Highest concentrations of SAM, SAH and folate RBC were observed in complex syndromic CHD. The subgroup of children with Down syndrome, however, showed significantly higher SAH (P = 0.037) and significantly lower SAM:SAH ratio (P = 0.034) compared with other complex CHD, suggesting a state of global hypomethylation.Conclusion High concentrations of methylation biomarkers in very young children are associated with complex CHD. Down syndrome and CHD may be associated with a global hypomethylation status, which has to be confirmed in tissues and global DNA methylation in future studies.

Diagnosis of testicular carcinoma in situ'(intratubular and microinvasive)' seminoma and embryonal carcinoma using direct enzymatic alkaline phosphatase reactivity on frozen histological sections (Article)

Stoop, J.A. — 2011-02-01T00:00:00Z

Aims: Testis-sparing surgery might benefit quality of life, but can only be applied with histological examination for the presence of invasive germ cell tumour components, and the precursor carcinoma in situ (CIS). Currently, diagnosis is based on paraffin-embedded tissue, therefore a delay in further surgery is mainly unavoidable. The aim was to develop an intraoperative assessment technique using alkaline phosphatase as a marker. Methods and results: A total of 4093 snap-frozen samples and matched paraffin-embedded tissue of 1500 patients were included. Besides standard haematoxylin and eosin (H&E) staining, the direct enzymatic alkaline phosphatase reactivity (dAP) test (duration 15min) was applied on frozen sections, while H&E and immunohistochemistry for detection of OCT3/4, α-fetoprotein, human chorionic gonadotrophin (hCG) and cytokeratin was performed on the paraffin-embedded slides. Endothelial cells served as control for the dAP test. Positive staining was found in all CIS (n=965), seminoma (n=1035) and embryonal carcinoma (n=584), either intratubular, microinvasive or invasive. Differentiated non-seminomas (n=1238) showed variable staining. No staining was identified in spermatocytic seminomas (n=5), testicular lymphomas (n=42), testicular rhabdomyosarcomas (n=7), Leydig cell tumours (n=31), Sertoli-cell-only nodules (n=4), (epi) dermoid cyst (n=16), normal testicular parenchyma (n=116), testicular torsion (n=32) and inflammation of the epididymis (n=19). The dAP test results matched H&E-stained parallel sections, as well paraffin-embedded tissue analysis, including immunohistochemistry. Conclusions: The dAP test is an informative, reproducible and easy tool to diagnose CIS, (intratubular and microinvasive) seminoma and embryonal carcinoma on frozen tissue sections, being of great value in the context of sparing surgery.