Comparative analysis of Bricker versus Wallace ureteroenteric anastomosis and identification of predictors for postoperative ureteroenteric stricture.

PURPOSE: Ureteroenteric anastomosis after cystectomy is usually performed using the Bricker or Wallace technique. Deterioration of renal function is the most common long-term complication of urinary diversion (UD). To improve surgical care and optimize long-term renal function, we compared the Bricker and Wallace anastomotic techniques and identified risk factors for ureteroenteric strictures (UES) in patients after cystectomy. MATERIAL AND METHODS: Retrospective, monocentric analysis of 135 patients who underwent cystectomy with urinary diversion at the University Hospital Essen between January 2015 and June 2019. Pre- and postoperative renal function, relevant comorbidities, prior chemo- or radiotherapy, pathological findings, urinary diversion, postoperative complications, and ureteroenteric strictures (UES) were analyzed. RESULTS: Of all 135 patients, 69 (51.1%) underwent Bricker anastomosis and 66 (48.9%) Wallace anastomosis. Bricker and Wallace groups included 134 and 132 renal units, respectively. At a median follow-up of 14 (6-58) months, 21 (15.5%) patients and 30 (11.27%) renal units developed UES. We observed 22 (16.6%) affected renal units in Wallace versus 8 (5.9%) in Bricker group (p < 0.001). A bilateral stricture was most common in Wallace group (69.2%) (p < 0.001). Previous chemotherapy and 90-day Clavien-Dindo grade ≥ III complications were independently associated with stricture formation, respectively (OR 9.74, 95% CI 2-46.2, p = 0.004; OR 4.01, 95% CI 1.36-11.82, p = 0.013). CONCLUSION: The results of this study show no significant difference in ureteroenteric anastomotic techniques with respect to UES development regarding individual patients but suggest a higher risk of bilateral UES formation in patients undergoing Wallace anastomosis. This is reflected in the increased UES rate under consideration of the individual renal units.

[Predictive biomarkers in bladder cancer]. / Therapieprädiktive Biomarker des Harnblasenkarzinoms.

BACKGROUND: In the last few years, significant progress has been achieved in the therapeutic options for advanced urothelial bladder cancer. OBJECTIVES: The aim of this work was to give an overview of the status and future perspective of the therapeutic options in this setting. Its focus is on the discussion of tissue-based therapy-predictive markers, which are evaluated through (molecular) pathology and thereby strengthening the role of pathology itself. MATERIALS AND METHODS: Current (clinical study) data, the literature, and our own expertise were considered and summarized in the areas of therapy prediction of platinum-based chemotherapy, immunotherapy, and other therapeutic approaches. RESULTS AND CONCLUSIONS: Molecular subtypes exhibit a predictive value both in platinum-based chemotherapy as well as in immunotherapy. However, further work is required to elucidate the predictive role of molecular subtypes in both settings. Changes in the DNA damage repair enzyme (DDR) genes, ERCC2, and ERBB2 as well as differences in the expression of EMMPRIN, survivin, and HMGA2 show promising results as further markers of chemotherapy efficacy. In the prediction of immunotherapy success, this mainly relates to the evaluation of the tumor mutation burden (TMB), tumor neoantigen burden (TNB), APOBEC signatures (MSig1; 3A/3B), and CD8-positive T­effector cell signature. When using the fibroblast growth factor receptor (FGFR) inhibitor erdafitinib, which has not yet been approved in Germany, the evaluation of specific FGFR mutations and/or gene fusions by a companion diagnostic test is mandatory in the USA.

Urachal cancer-current concepts of a rare cancer.

Urachal cancer is a rare but aggressive disease. In addition to the non-glandular tumors, non-cystic urachal adenocarcinomas are nowadays distinguished from the primary cystic variant. (Immunohistochemical) markers are only of minor differential diagnostic value and, therefore, the diagnosis is primarily established in a multidisciplinary approach. The non-cystic variant accounts for the majority of cases (83%), is more common in men (63%), shows a median age at diagnosis of 51 years and has a 5-year survival rate of about 50%. In organ-confined disease, usually a partial cystectomy of the tumor in the bladder dome, including the median umbilical ligament and umbilicus, is performed. In advanced stages, systemic therapy is needed while 5­fuorouracil (5-FU) containing regimes have been shown to be more effective. Due to the rarity of the tumor, targeted therapy approaches based on a biological rationale are becoming increasingly relevant. As molecular data are still sparse, we compiled and analyzed the largest urachal cancer cohort to date. In 31% of the cases, MAPK-/PI3K signaling pathway alterations were detected (especially in K-/NRAS) with implications for anti-EGFR therapy approaches. Further potentially therapeutic alterations were detected in FGFR1, MET, PDGFRA, and erbB2/HER2. Additionally, PD-L1 tumor cell expression (clone: 22C3) was demonstrated in 16% of cases, therefore making anti-PD-1/PD-L1 immuno-oncological approaches worth considering despite the absence of mismatch repair deficiency (MMR-d) and/or high microsatellite instability (MSI-h). Finally, urachal adenocarcinomas seem to be a distinct entity on the molecular level with closer resemblance to colorectal adenocarcinomas than to urothelial carcinomas.

[Predictive biomarkers in oncologic uropathology]. / Prädiktive Biomarker in der onkologischen Uropathologie.

BACKGROUND: Tumors of the genitourinary system are common. In recent years, our understanding of their molecular background and therefore the number of potential predictive biomarkers has massively increased. OBJECTIVES: The aim of the current work is to give an overview of recent (molecular) developments and predictive biomarkers in urologic oncology and to give a perspective of what might become relevant in the future of the field. MATERIAL AND METHODS: We considered the recent literature and study data and combined it with our own expertise in tumors of the urinary system, kidneys, and prostate. RESULTS AND CONCLUSIONS: The molecular subtypes of muscle-invasive urothelial bladder cancer (MIBC) hold a predictive and prognostic significance and correlate with clinicopathological features. Immune therapy with checkpoint inhibitors (CPI) has a major role in urothelial carcinoma (UC), but also in renal cell carcinoma and a subgroup of prostate cancers. The first-line use in UC is restricted to PD-L1-"positive" cases (≥IC2/3, CPS ≥ 10). Further predictive markers are currently under evaluation, while the predictive significance of tumor mutational burden (TMB) is under debate. In addition to a subgroup of renal cell carcinomas, a subgroup of prostate carcinomas with alterations in the DNA repair system might benefit from a customized therapy approach (PARP inhibitors, platin-containing chemotherapy). The multitude of potentially therapy-relevant molecular alterations and related predictive biomarkers calls for the implementation of sophisticated molecular analyses in daily routine. This will lead to an even more rapid dynamic in the field of genitourinary pathology.

[Urachal Cancer: an update of current molecular findings]. / Das Urachuskarzinom: Ein Update aktueller molekularer Ergebnisse.

BACKGROUND: Urachal cancer is a rare type of cancer, often following a clinically aggressive course. Due to its rarity, knowledge about its molecular background is still limited. In addition, no sufficiently reliable diagnostic markers are available. OBJECTIVES: The aim of the present study is to give an overview of our recent molecular projects on urachal cancer and to connect it with current literature in the field. MATERIALS AND METHODS: Three projects are introduced. The first project identified and validated diagnostic biomarkers in urachal adenocarcinomas compared to colorectal adenocarcinomas and primary adenocarcinomas of the bladder using various proteomic methods. In the second project, the most relevant differential diagnostic markers between urachal adenocarcinomas and colorectal adenocarcinomas compared to normal tissue (urachal remnants) were determined by analyzing a miRNA panel. Sequence analyses were performed in the third project. The focus was on molecular differences to colorectal adenocarcinomas and urothelial carcinomas. RESULTS AND CONCLUSIONS: We detected potential biomarker candidates for the immunohistochemical differential-diagnosis and generated a miRNA-based diagnostic scoring system with a potentially high differential-diagnostic significance. The sequence analyses data confirm the molecular autonomy of the urachal adenocarcinomas compared to other entities.

[Urachal cancer - current concepts of a rare cancer (German version)]. / Das Urachuskarzinom ­ aktuelle Konzepte einer seltenen Tumorerkrankung.

Urachal cancer is a rare but aggressive disease. In addition to the non-glandular tumors, non-cystic urachal adenocarcinomas are nowadays distinguished from the primary cystic variant. (Immunohistochemical) markers are only of minor differential diagnostic value and, therefore, the diagnosis is primarily established in a multidisciplinary approach. The non-cystic variant accounts for the majority of cases (83%), is more common in men (63%), shows a median age at diagnosis of 51 years and has a 5-year survival rate of about 50%. In organ-confined disease, usually a partial cystectomy of the tumor in the bladder dome, including the median umbilical ligament and umbilicus, is performed. In advanced stages, systemic therapy is needed while 5­fuorouracil (5-FU) containing regimes have been shown to be more effective. Due to the rarity of the tumor, targeted therapy approaches based on a biological rationale are becoming increasingly relevant. As molecular data are still sparse, we compiled and analyzed the largest urachal cancer cohort to date. In 31% of the cases, MAPK-/PI3K signaling pathway alterations were detected (especially in K-/NRAS) with implications for anti-EGFR therapy approaches. Further potentially therapeutic alterations were detected in FGFR1, MET, PDGFRA, and erbB2/HER2. Additionally, PD-L1 tumor cell expression (clone: 22C3) was demonstrated in 16% of cases, therefore making anti-PD-1/PD-L1 immuno-oncological approaches worth considering despite the absence of mismatch repair deficiency (MMR-d) and/or high microsatellite instability (MSI-h). Finally, urachal adenocarcinomas seem to be a distinct entity on the molecular level with closer resemblance to colorectal adenocarcinomas than to urothelial carcinomas.

Application of light scattering tomography for Si(111) samples.

The detection of bulk micro-defects in Czochralski-grown silicon (Si) 〈100〉 wafers has significant importance in wafer quality control. Light Scattering Tomography (LST) is an industry standard technique for this purpose. This optical non-contact metrology requires destructive sample preparation: Samples have to be cleaved into half. One particular feature of the method is a dark field detection arrangement, which is achieved by separating the light detection part (microscope unit) from the illumination. Illumination is applied to the front surface of the sample, and the light scattered off of the defects is collected via the cleaved surface. The technique requires the perpendicularity of the cleaved surface to the front surface, which is fulfilled for Si(100) wafers. However, the nominally cleaved surface for Si(111) wafers is not perpendicular to the front surface but has an angle of 70.5°. This significant difference in cleavage results in the fact that Si(111) wafers cannot be measured by standard LST systems. Fortunately, the standard LST system can be modified by tilting the detection part under a proper angle allowing the measurements of Si(111) samples. In this article, we present this new technique in detail, showing the design and measurement capability of the new system. The measurement results are validated by a direct comparison to standard LST measurements on the same samples after proper sample preparation.

Bulk micro-defect detection with low-angle illumination.

The detection of oxygen precipitates, voids, and other defects is critical for semiconductor wafer makers. One of the industry standard techniques for detecting these Bulk Micro-Defects (BMDs) is Semilab's Light Scattering Tomograph (LST) system. In this measurement, unpatterned wafers are nominally cleaved in half. Illumination is applied to the front surface of the sample, and the light scattered off of the defects is collected via the cleaved surface. This technique had been limited to the measurement of unpatterned wafers, but device makers show significant interest in measuring BMD distributions on patterned wafers using scattering-based techniques. A pattern on the surface of the wafer can cause significant scattering, making the standard LST technique unsuitable for this task. We present a solution for patterned wafer BMD measurements by an addition of a low-angle illumination unit to the standard LST system. This new illumination unit focuses the light into the bulk of the wafer via the cleaved surface, which enables measurement on patterned samples. The new system is called "light scattering tomograph enhanced by low-angle illumination." Excellent correlation was found between the detected defect densities obtained by the low-angle and the standard LST illumination mode.

The prognostic value of serum MMP-7 levels in prostate cancer patients who received docetaxel, abiraterone, or enzalutamide therapy.

OBJECTIVES: The rapidly changing treatment landscape in metastatic castration-resistant prostate cancer (mCRPC) calls for biomarkers to guide treatment decisions. We recently identified MMP-7 as a potential serum marker for the prediction of response and survival in mCRPC patients who received docetaxel (DOC) chemotherapy. Here, we aimed to test this finding in an independent patient cohort and in addition to explore the prognostic potential of serum MMP-7 in abiraterone (ABI) or enzalutamide (ENZA) treated patients. METHODS AND MATERIALS: MMP-7 levels were measured in 836 serum samples from 320 mCRPC patients collected before and during DOC (n = 95), ABI (n = 140), or ENZA (n = 85) treatment by using the ELISA method. Results were correlated with clinical and follow-up data. RESULTS: MMP-7 baseline levels were similar between the 3 treatment groups. In the ABI and ENZA cohorts, baseline MMP-7 levels were lower in patients with prior radical prostatectomy (P = 0.058 and P = 0.041, respectively). Baseline MMP-7 levels above the median were associated with shorter overall survival for the DOC (P = 0.001) and ENZA (P = 0.006) cohorts. Multivariable analyses in the DOC and ENZA cohorts revealed that high pretreatment MMP-7 level is an independent risk factor for patients' survival. In addition, in DOC-treated patients with high baseline MMP-7 level, marker decrease at the third DOC cycle was associated with improved survival. Patients with high baseline MMP-7 levels had better survival when treated with ABI compared to DOC or ENZA. CONCLUSIONS: We confirmed the prognostic value of pretreatment MMP-7 serum level and its changes as independent predictors of survival in DOC-treated mCRPC patients. In addition, high MMP-7 was a negative predictor in ENZA-treated but not in ABI-treated patients. These results warrant further research to confirm the predictive value of serum MMP-7 and to explore the potential mechanistic involvement of MMP-7 in DOC and ENZA resistance of mCRPC patients.

High-soluble CGA levels are associated with poor survival in bladder cancer.

Recently, a neuroendocrine-like molecular subtype has been discovered in muscle-invasive urothelial bladder cancer (BC). Chromogranin A (CGA) is a widely used tissue and serum marker in neuroendocrine tumors. Our aim was to evaluate serum CGA (sCGA) concentrations and their associations with clinical and follow-up data in BC and renal cell carcinoma (RCC). sCGA concentrations were analyzed in the following cohorts: (1) BC training set (n = 188), (2) BC validation set (n = 125), (3) RCC patients (n = 77), (4) healthy controls (n = 97). CGA immunohistochemistry and RT-qPCR analyses were performed in 20 selected FFPE and 29 frozen BC tissue samples. Acquired data were correlated with clinicopathological parameters including comorbidities with known effect on sCGA as well as with patients' follow-up data. sCGA levels were significantly higher in BC but not in RCC patients compared to healthy controls. High sCGA levels were independently associated with poor overall and disease-specific survival both in the BC training (P < 0.001, P = 0.002) and validation set (P = 0.009, P = 0.017). sCGA levels were inversely correlated with glomerulus filtrating rate (GFR) and linearly correlated with creatinine clearance and urea concentrations. These correlations were not related to the prognostic value of sCGA. Tissue CGA levels were low to absent independently of sCGA concentrations. Our results demonstrate elevated levels and an independent prognostic value for sCGA in BC but not in RCC. Despite the significant correlation between sCGA and GFR, the prognostic relevance of sCGA seems not related to impaired renal function or other comorbidities.

[The revival of "uroscopy": an easy way to evaluate the anastomotic region after radical retropubic prostatectomy]. / Die Wiedergeburt der "Harnschau" : Eine einfache Methode zur Beurteilung der Anastomose nach radikaler retropubischer Prostatektomie.

OBJECTIVE: The cystogram after retropubic radical prostatovesiculectomy (RRP) examines the condition of the anastomosis region. There is no clear temporal recommendation for the determination of urine extravasation in cases of early removal of the catheter. This study investigates the macroscopic and microscopic examination of the bladder catheter urine after RRP. PATIENTS AND METHODS: The catheter urine of 110 patients after RRP was macroscopically and microscopically examined and compared with the results of transrectal ultrasound and cystogram with regard to urine extravasation. RESULTS: The examination of cloudy or bloody catheter urine indicated an extravasation in 32 of 33 cases (positive predictive value 97%; sensitivity 74%), for clear urine, there was sufficient anastomosis in 66 of 77 cases without any extravasation (negative predictive value 86%; specificity 99%). By performing a microscopic urine analysis unremarkable anastomosis was predicted only in 23 of 67 cases, which leads to a specificity of 34%. This shows that microscopic urine sediment is an inappropriate instrument to evaluate the anastomotic region. CONCLUSION: In cases of macroscopic pathological urine findings, further diagnostics are not recommended; in 97% of the cases, extravasation is expected. The macroscopic urine examination sets a trend for other examinations like cystogram or dynamic transrectal ultrasound (dTRUS).

[Use of silicon chip technology to detect protein-based tumor markers in bladder cancer]. / Einsatz der Siliziumchiptechnologie zur Detektion von Tumormarkern auf Proteinbasis beim Harnblasenkarzinom.

The protein structure of human tumor tissue has a significant influence on the molecular attributes. It was demonstrated that the individual prognosis of tumor patients is among other things dependent on molecular tumor tissue characteristics.A promising marker is E-cadherin, an adhesion glycoprotein which plays a central role in the mediation of cell-cell contacts. Aberrant E-cadherin expressions were described in several tumors such as in bladder cancer. This was also found to be correlated with tumor invasion and survival. There are hardly any fast, quantitative and easily automated protein assays in everyday practice which can analyze several markers at the same time. With silicon chip technology we have a new detection and measurement method which makes it possible to give a quantitative analysis of numerous different proteins in tissue, urine, or serum in a few minutes.

HPLC and HPLC-MS analysis of urinary N(epsilon)-monomethyl-lysine.

Administration of (14)C-labelled L-deprenyl to rats results in the urinary elimination of a 14C-labelled compound. The 9-fluorenylmethoxycarbonyl chloride-reacted urine sample is fractionated by high-performance liquid chromatography (HPLC) on an octadecyl silica stationary phase. N(epsilon)-Monomethyl-lysine is identified in the fraction containing the majority of the radioactivity. Structural elucidation is carried out using HPLC-mass spectrometry in atmospheric pressure chemical ionization mode. Identification of the 14C-labelled fragment in Ne-monomethyl-lysine is an experimental proof that an N-methylated amino acid is generated by transmethylation from a well-known drug. This type of transmethylation may have basic importance in the positive side effects of certain drugs.

Presence and possible origin of epsilon(gamma-glutamyl)lysine isodipeptide in human plasma.

epsilon(gamma-glutamyl)lysine isodipeptide has been detected in normal human plasma by a sensitive HPLC technique in a concentration of 1.9-3.6 mumol/l. Incubation of in vitro clotted plasma at 37 degrees C for 12 h resulted in an increased amount of isodipeptide, and there was no further significant change when streptokinase was also present. Increased in vivo isodipeptide concentrations were also observed in hypercoagulable states and during fibrinolytic therapy.

HPLC analysis of metabolically produced formaldehyde.

Radiolabelled (-)-deprenyl is orally administered to rats, and urinary elimination of radiolabelled formaldehyde is detected. The separation is performed using high-performance liquid chromatography on octadecyl-silica stationary phase. Both the radioactivity and the UV absorbance of the dinitrophenylhydrazine formaldehyde peak are determined. Formaldehyde generation takes place by N-demethylation. Low levels of formaldehyde may have a beneficial role in counterbalancing the oxidative stress of the everyday person's life.

The hydrazine derivative aminoguanidine inhibits the reaction of tetrahydrofolic acid with hydroxymethylarginine biomolecule.

UNLABELLED: Aminoguanidine (AG), a hydrazine derivative is known to inhibit the formation of Advanced Glycosylation Endproducts (AGE) and AG has been proposed as an agent in prophylaxis of diabetic complications. However, treatment with hydrazine produced liver and lung tumors by formation of N7- and O6-methylguanine in the DNA of rodents. The hydrazine derivative, isonicotinic acid hydrazide induced pulmonary tumors in mice. N(G)-hydroxymethyl-arginine (HMA) was synthesized by our research group and it showed anticancer effect against experimental tumors. HMA was found earlier in human blood and urine, and recently in many plants (in fruits and vegetables). We could demonstrate a reaction (pH = 7.5, 37 degrees C, 1h) between HMA and tetrahydrofolate (THF) producing N5,N10-methylene-tetrahydrofolate (CH2-THF), the coenzyme of thymidylate synthase (TS). In model experiments AG proved to react with formaldehyde (HCHO) and to eliminate the C1-fragment of HMA, but not that of CH2-THF. In the presence of AG burst chemiluminescence and a higher speed of the formylation and methylation reactions were found in the AG, HCHO, hydrogen peroxide (H2O2) and L-lysine system than without AG. CONCLUSIONS: HMA as a biomolecule is one of the compounds which are responsible for the endogenous HCHO level. The biochemical function of HMA may be the direct supply of C1-fragment for the folate cycle. AG can disturb the above function of HMA. The reaction between AG and HCHO seems to be dangerous for biological systems because of the possible presence of L-lysine and H2O2. The burst chemiluminescence indicates excited molecules with extreme high energy producing uncontrolled formylation and methylation reactions. Considering the results of the experiments with AG its use as a medicament seems to be questionable.

Determination of endogenous formaldehyde in plants (fruits) bound to L-arginine and its relation to the folate cycle, photosynthesis and apoptosis.

A very powerful nucleophilic reagent, hydralazine(1-hydrazino-phtalazine) proved to be suitable for determination of the endogenous formaldehyde level in biological samples. It was found that in different plants (vegetables, fruits, especially in red beet, cauliflower, kohlrabi, grapes) is a large amount of releasable endogenous formaldehyde (0.5-1.0 mM) bound to L-arginine mainly in the form of N(G)-trihydroxymethyl-L-arginine (TriHMA). N(G)-hydroxymethyl-L-arginines (HMA) were proved to transfer their hydroxymethyl groups to tetrahydrofolic acid producing N5,N10-methylene-tetrahydrofolate, the coenzyme of thymidylate synthase. HMA was found to inhibit the cell proliferation of HT-29 cell culture (human colon adenocarcinoma ATCC HT-B 38) causing apoptosis. Photosynthetic experiments produced confirmatory evidences that 14CH2O could be formed in photosynthesis already after 10 seconds of 14CO2 fixation in the seedlings of Zea mays L. (single cross) and the 14CH2O was immediately trapped by L-arginine mainly as TriHMA.

Analogies and differences in the excited reactions of formaldehyde and D-glucose.

The investigations proved that D-glucose (as reducing sugar) can easily be activated in a ternary system (L-lysine: D-glucose: H2O2) similarly to formaldehyde at 20 degrees C, in pH = 7.4 forming chemiluminescence (CL) and singlet oxygen. The kinetic investigation showed that: CL lasted many hours (permanent emission) and had no bell-shaped curve differently from other aldehydes e.g. formaldehyde. The reason of the effect is that D-glucose exists mainly in ring form in water solution (Haworth ring form) and the open form (the aldehyde group) is slowly liberated during the excited reaction. These excited reactions may be important in human organism, because D-glucose and lysyl residues of proteins occur permanently in human body and endogenous formaldehyde and H2O2 may be liberated there, too.

The clinical diagnostic significance of the investigation of receptor mediated pathway of LDL.

Some types of LDL-receptor investigations have been demonstrated which can also be carried out in routine clinical laboratory. Special attention has been layed on the testing of lymphocytes and monocytes which are available easier than tissue samples but, which are, however, according to our experiences appropriate for the LDL receptor analysis.

[Transitional cell carcinoma of the bladder: bladder-sparing therapy]. / Urothelkarzinom der Harnblase : Organerhaltende Therapiestrategien.

Transitional cell carcinoma of the bladder can - in the majority of cases - be safely treated by transurethral resection and bladder preservation. In case of more aggressive and genetically instable tumors, the effect of radical cystectomy depends on tumor volume. If complete resection of invasive tumors is also possible, the additional effect of radical cystectomy seems to be marginal. In patients with favorable tumor location and acceptable prostate parameters, prostate-sparing surgery seems to be oncologically safe with good quality of life.