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Background The role of the epithelial cell adhesion molecule (EpCAM) in cancer is still unclear. EpCAM cleavage through regulated intramembrane proteolysis results in fragments which interact with both oncogenic and tumor suppressive pathways. Additionally, the EpCAM molecule itself is used as a descriptive therapeutic target in urothelial cancer (UC), while data on its actual tumor specificity remain limited. Methods Samples from diagnostic formalin-fixed paraffin-embedded (FFPE) UC tissue and fresh-frozen UC cells were immunoblotted and used for qualitative characterization of five different EpCAM fragments. These expression patterns were quantified across a cohort of 76 samples with 52 UC and 24 normal urothelial samples. Cell viability effects of the extracellular EpEX fragment were assessed in the UC cell lines T24 and HT1376. Results The proteolytic EpCAM fragments could be identified in clinical FFPE tissue specimens too. Neither overall nor fragment-specific EpCAM expression showed relevant tumor specificity. EpEX and its deglycosylated variant showed an inverse relationship across healthy and tumor tissue with a decrease of deglycosylated EpEX in tumors. However, extracellular EpEX did not show a relevant effect in vitro. Conclusions EpCAM should not be regarded as tumor-specific in UC without patient-specific predictive testing. EpCAM fragment patterns indicate cancer-specific changes and could be involved in its complex tumor-biological role.

Objective Congenital defects of androgen synthesis or action in 46,XY individuals can result in impaired virilisation, despite the apparent testicular development. In a recent case, report of a young adult with complete androgen insensitivity syndrome (CAIS), tumourous gonadal tissue was shown to express HSD17B3 in Sertoli cells (SCs) and not in Leydig cells (LCs). This expression pattern differs from the typical adult human testis and resembles a foetal mouse testis, suggesting an underlying testicular development and function defect. Here, we investigate the effect of altered androgen signalling in gonads from five 46,XY individuals with defects in androgen synthesis or action. Methods Gonadal tissue sections from four patients with CAIS, one with CYP17A1 deficiency, and one control were immunostained for LC developmental and steroidogenic markers. The expression of some of these markers during development was investigated by reanalysing previously published single-cell RNA sequencing (scRNA-seq) data from normal human testicular tissues. Results All gonadal tissues from the patients show an exclusive expression of HSD17B3 in SCs and an expression of the foetal/immature LC marker DLK1 in a subset of LCs, suggesting an androgen-dependent differentiation defect of adult SCs and LCs. Furthermore, reanalysis of scRNA-seq data reveals an expression of HSD17B3 in foetal and neonatal SCs that is downregulated in adult SCs. Conclusions Androgen signalling may affect the differentiation of adults, but possibly not foetal SCs or LCs, and may induce a shift of testosterone production from the tubular compartment in the foetal phase to the interstitial compartment in the adult phase.

We present the case of a patient with granulomatous endocarditis of the mitral valve leading to severe valve stenosis caused by granulomatosis with polyangiitis. Endocarditis is a rare complication of granulomatosis with polyangiitis that may be misdiagnosed as infectious endocarditis or, as in our case, thrombotic lesions. (Level of Difficulty: Intermediate.), Graphical abstract

Zusammenfassung Hintergrund und Zielsetzung Die radikale Zystektomie (RCX) ist die kurative Standardbehandlung für muskelinvasive Harnblasenkarzinome. Aufgrund des operativen Ausmaßes und der damit verbundenen komplexen stationären Betreuung ist die RCX mit hoher Morbidität und Mortalität verbunden. Ziel war es, Daten aus dem klinischen Alltag zur gegenwärtigen Praxis des perioperativen Managements zu erfassen. Methoden Mitglieder der DGU und der German Society of Residents in Urology beantworteten anonym einen 24 Fragen umfassenden Online-Fragebogen. Gegenstand waren sowohl klinisch relevante Aspekte der gegenwärtigen EAU/DGU-Leitlinien als auch kontrovers diskutierte klinische Fragen. Ergebnisse Durch die Teilnehmer konnten mindestens 19 % aller infrage kommenden deutschen Zentren erfasst werden. Mehrheitlich wird das präoperative Staging mittels CT-Urografie und CT-Thorax durchgeführt. Die meistgenutzte Antibiotikaprophylaxe umfasst ein Drittgenerations-Cephalosporin plus Metronidazol (46 %), welche im Median für 5 Tage verabreicht werden. Harnleiterschienen (HS) werden im Mittel beim Ileumconduit am 11. und bei der Neoblase am 12. Tag nach RCX entfernt. Gemessen an Surrogatparametern wie oraler Darmvorbereitung, Beginn des Kostaufbaus und Einsatz einer Magensonde sind aktuelle Fast-Track-Konzepte noch nicht allgemein verbreitet ( Schlussfolgerungen Die meisten Aspekte des perioperativen Managements werden leitliniengerecht durchgeführt, so z. B. Staging-Untersuchungen, Einsatz von Antibiotika und Entfernung der HS. Andere klinische Fragestellungen wie Durchführung einer Bildgebung vor Entfernung der HS und Fast-Track-Konzepte werden unterschiedlich gehandhabt, obwohl zunehmende Evidenz in diesen Bereichen existiert. Leitlinienadhärenz war nicht mit Zentrumsgröße oder Anzahl der Eingriffe assoziiert.

Several algorithms for the normalization of proteomic data are currently available, each based on a priori assumptions. Among these is the extent to which differential expression (DE) can be present in the dataset. This factor is usually unknown in explorative biomarker screens. Simultaneously, the increasing depth of proteomic analyses often requires the selection of subsets with a high probability of being DE to obtain meaningful results in downstream bioinformatical analyses. Based on the relationship of technical variation and (true) biological DE of an unknown share of proteins, we propose the "Normics" algorithm: Proteins are ranked based on their expression level-corrected variance and the mean correlation with all other proteins. The latter serves as a novel indicator of the non-DE likelihood of a protein in a given dataset. Subsequent normalization is based on a subset of non-DE proteins only. No a priori information such as batch, clinical, or replicate group is necessary. Simulation data demonstrated robust and superior performance across a wide range of stochastically chosen parameters. Five publicly available spike-in and biologically variant datasets were reliably and quantitively accurately normalized by Normics with improved performance compared to standard variance stabilization as well as median, quantile, and LOESS normalizations. In complex biological datasets Normics correctly determined proteins as being DE that had been cross-validated by an independent transcriptome analysis of the same samples. In both complex datasets Normics identified the most DE proteins. We demonstrate that combining variance analysis and data-inherent correlation structure to identify non-DE proteins improves data normalization. Standard normalization algorithms can be consolidated against high shares of (one-sided) biological regulation. The statistical power of downstream analyses can be increased by focusing on Normics-selected subsets of high DE likelihood.

ObjectivesFormalin-fixed paraffin-embedded (FFPE) tissue is the standard material for diagnostic pathology but poses relevant hurdles to accurate protein extraction due to cross-linking and chemical alterations. While numerous extraction protocols and chemicals have been described, systematic comparative analyses are limited. Various parameters were thus investigated in their qualitative and quantitative effects on protein extraction (PE) efficacy. Special emphasis was put on preservation of membrane proteins (MP) as key subgroup of functionally relevant proteins.MethodsUsing the example of urothelial carcinoma, FFPE tissue sections were subjected to various deparaffinization, protein extraction and antigen retrieval protocols and buffers as well as different extraction techniques. Performance was measured by protein concentration and western blot analysis of cellular compartment markers as well as liquid chromatography-coupled mass spectrometry (LC–MS).ResultsCommercially available extraction buffers showed reduced extraction of MPs and came at considerably increased costs. On-slide extraction did not improve PE whereas several other preanalytical steps could be simplified. Systematic variation of temperature and exposure duration demonstrated a quantitatively relevant corridor of optimal antigen retrieval.ConclusionsPreanalytical protein extraction can be optimized at various levels to improve unbiased protein extraction and to reduce time and costs.

Introduction: Guideline recommendations are meant to help minimize morbidity and to improve the care of nonmuscle invasive bladder cancer (NMIBC) patients but studies have suggested an underuse of guideline-recommended care. The aim of this study was to evaluate the level of adherence of German and Austrian urologists to German guideline recommendations. Methods: A survey of 27 items evaluating diagnostic and therapeutic recommendations (15 cases of strong consensus and 6 cases of consensus) for NMIBC was administered among 14 urologic training courses. Survey construction and realization followed the checklist for reporting results of internet e-surveys and was approved by an internal review board. Results: Between January 2018 and June 2019, a total of 307 urologists responded to the questionnaire, with a mean response rate of 71%. The data showed a weak role of urine cytology (54%) for initial diagnostics although it is strongly recommended by the guideline. The most frequently used supporting diagnostic tool during transurethral resection of the bladder was hexaminolevulinate (95%). Contrary to the guideline recommendation, 38% of the participants performed a second resection in the case of pTa low-grade NMIBC. Correct monitoring of Bacille Calmette-Guérin (BCG) response with cystoscopy and cytology was performed by only 34% of the urologists. Conclusions: We found a discrepancy between certain guideline recommendations and daily routine practice concerning the use of urine cytology for initial diagnostics, instillation therapy with a low monitoring rate of BCG response, and follow-up care with unnecessary second resection after pTa low-grade NMIBC in particular. Our survey showed a moderate overall adherence rate of 73%. These results demonstrate the need for sharpening awareness of German guideline recommendations by promoting more intense education of urologists to optimize NMIBC care thus decreasing morbidity and mortality rates.

Introduction For risk stratification of non-muscle invasive bladder cancer (NMIBC), the depth of stromal invasion can be further classified, where the lamina muscularis mucosae (MM) serves as a reference structure. While the overall identifiability of MM in standard transurethral specimens is low, en bloc resection may help in identification and overall orientation. The aims of this study were to report the detection rate of MM in en bloc resected bladder tumors (ERBT) and to provide real-world information on tissue stability and preservation of en bloc architecture during recovery and processing for histopathologic evaluation. Methods Thirty-four ERBT specimens were histologically re-evaluated with regard to MM detectability and structure as well as the presence of en bloc architecture and further histologic features. Associations with tumor size and energy source and within histologic parameters were assessed by standard Pearson's chi-squared analyses and Cramér’s V effect size testing (V). Results The first parameter assessed was MM detection rate. In 19 out of 34 samples (56%) MM was detectable: scattered in 9 cases (26%), interrupted in 8 cases (24%) and continuous in 2 cases (6%). The second parameter assessed was preservation of en bloc architecture. In 11 out of 34 samples (32%), en bloc architecture could not be confirmed, and these samples served as a reference group for the detection of MM. Preservation of en bloc architecture was associated with an increased MM detection rate (MM in en bloc preserved 16/23, 70% vs. non-preserved 3/11, 27%; p = 0.020; V = 0.398) and with tumor size (p = 0.005; V = 0.595). Medium-sized tumors (1.1–2 cm) were best preserved. The choice of energy source did not show relevant association with en bloc architecture (p = n.s.). Conclusions In line with recent publications, ERBT increases the MM detection rate considerably. However, a third of the ERBT specimens lost en bloc architecture during sample recovery and processing. Tumor size is a relevant factor, with optimal architecture preservation between 1 and 2 cm. Optimizing resection techniques, recovery, transport, and diagnostic processing of ERBT samples is warranted to verify the diagnostic value of MM-based substaging.

Radical cystectomy (RCX) is the standard treatment for muscle-invasive and treatment-refractory non-invasive bladder cancer, but that is associated with high morbidity. We now survey current practice patterns on perioperative management among German urological departments of all sizes METHODS: Members of the German Association of Urology and the German Society of Residents in Urology (GeSRU) were contacted by email and asked to answer a 24-item online questionnaire covering clinically relevant aspects of current guidelines and controversies. Responses were obtained from at least 19 % of all German urological centers. About 60 % performed preoperative staging using CT urography and chest CT. The most common perioperative antibiotic prophylaxis was a third generation cephalosporin combined with metronidazole (46 %), administered for a median of 5 days. Stentograms for ileal conduit and neobladder are routinely performed in 38 % and 55 % of patients, respectively. Ureteral stents were usually removed 11 - 12 days after the procedure (ileal conduit and neobladder). Based on the surrogate parameters of preoperative bowel preparation, postoperative start of oral nutrition and use of nasogastric tube, fast-track concepts such as ERAS were not generally established ( 50 %). Robot-assisted cystectomy appears to be performed in 15 % of German urological centers and was associated with the number of performed cystectomies (p 0.001). Most aspects of perioperative management in cystectomy patients - staging diagnostics, use of antibiotics, stent removal - are performed in accordance with current guidelines. Other clinical questions such as stent imaging before removal and fast track concepts are handled heterogeneously. Guideline-adherence was not associated with hospital size or number of procedures performed. Die radikale Zystektomie (RCX) ist die kurative Standardbehandlung für muskelinvasive Harnblasenkarzinome. Aufgrund des operativen Ausmaßes und der damit verbundenen komplexen stationären Betreuung ist die RCX mit hoher Morbidität und Mortalität verbunden. Ziel war es, Daten aus dem klinischen Alltag zur gegenwärtigen Praxis des perioperativen Managements zu erfassen. Mitglieder der DGU und der German Society of Residents in Urology beantworteten anonym einen 24 Fragen umfassenden Online-Fragebogen. Gegenstand waren sowohl klinisch relevante Aspekte der gegenwärtigen EAU/DGU-Leitlinien als auch kontrovers diskutierte klinische Fragen. Durch die Teilnehmer konnten mindestens 19 % aller infrage kommenden deutschen Zentren erfasst werden. Mehrheitlich wird das präoperative Staging mittels CT-Urografie und CT-Thorax durchgeführt. Die meistgenutzte Antibiotikaprophylaxe umfasst ein Drittgenerations-Cephalosporin plus Metronidazol (46 %), welche im Median für 5 Tage verabreicht werden. Harnleiterschienen (HS) werden im Mittel beim Ileumconduit am 11. und bei der Neoblase am 12. Tag nach RCX entfernt. Gemessen an Surrogatparametern wie oraler Darmvorbereitung, Beginn des Kostaufbaus und Einsatz einer Magensonde sind aktuelle Fast-Track-Konzepte noch nicht allgemein verbreitet ( 50 %). Die meisten Aspekte des perioperativen Managements werden leitliniengerecht durchgeführt, so z. B. Staging-Untersuchungen, Einsatz von Antibiotika und Entfernung der HS. Andere klinische Fragestellungen wie Durchführung einer Bildgebung vor Entfernung der HS und Fast-Track-Konzepte werden unterschiedlich gehandhabt, obwohl zunehmende Evidenz in diesen Bereichen existiert. Leitlinienadhärenz war nicht mit Zentrumsgröße oder Anzahl der Eingriffe assoziiert.

The use of readily available mHealth apps on smartphones and tablets is increasing worldwide, including the field of urology. Access to knowledge, data collection and communication are thus simplified and accelerated. A wide range of health apps exists for the public, patients, health professionals and medical education. In particular, in the field of urology, several high quality mHealth contents have been created in recent years. Nevertheless, there is still need for improvement and development.The ubiquity of health apps might also bring particular risks. Healthcare professionals can use their private electronic devices when dealing with patients or patient data, and thus risk violations of patient and data protection rights.There are no universal quality standards or legal certification requirements for mHealth apps. Private (mostly commercial) app developers and providers possess quasi-proprietary rights concerning user data. Politics, scientific funding agencies and medical societies are therefore urgently needed to set standards and to promote and fund open-source key projects providing quality benchmarks within the field.Die Nutzung von Smartphones und Tablets und den darauf benutzten mHealth-Apps nimmt weltweit zu. Dies gilt ebenfalls für Gesundheits-Apps in der Urologie. Der Zugang zu Wissen, die Sammlung von Daten und die professionelle Kommunikation selbst können dadurch vereinfacht und beschleunigt werden.Das Spektrum an Gesundheits-Apps für Öffentlichkeit, Patienten, Gesundheitsberufe und dementsprechend auch Urologen ist groß. Insbesondere für Letztere entstanden in den vergangenen Jahren verschiedene, qualitativ hochwertige mHealth-Angebote. Dennoch besteht weiterhin großes Entwicklungs- und Verbesserungspotenzial und die Allgegenwärtigkeit von Gesundheits-Apps birgt auch Risiken. Beschäftigte in Gesundheitsberufen können zum Beispiel ihre privaten elektronischen Geräte in der Patientenversorgung nutzen und laufen so Gefahr, durch die Nutzung von Apps die Datenschutzrechte der Patienten zu verletzen.Es bestehen keine einheitlichen Qualitäts- oder Zertifizierungsstandards für mHealth-Apps. Die Datenhoheit liegt momentan bei den privaten, meist kommerziellen Anbietern der Apps. Politik sowie Fach- und Fördergesellschaften sind gefragt, hier Standards festzulegen und durch Finanzierung entsprechender Schlüsselprojekte Benchmarks bez. Inhalt und Qualität der Apps zu definieren.