Uterine and Tubal Lavage for Earlier Cancer Detection Using an Innovative Catheter: A Feasibility and Safety Study

Objectives Poor survival of high-grade serous pelvic cancer is caused by a lack of effective screening measures. The detection of exfoliated cells from high-grade serous pelvic cancer, or precursor lesions, is a promising concept for earlier diagnosis. However, collecting those cells in the most efficient way while fulfilling all requirements for a screening approach is a challenge. We introduce a new catheter for uterine and tubal lavage (UtL) and the clinical evaluation of its performance. Methods/Materials In study I, the clinical feasibility of the UtL using the new catheter was examined in 93 patients admitted for gynecologic surgery under general anesthesia. In study II, the safety of the UtL procedure was assessed. The pain during and after the UtL performed under local anesthesia was rated on a visual analog scale by 22 healthy women. Results In study I, the UtL was carried out successfully in 92 (98.9%) of 93 cases by 16 different gynecologists. It was rated as easy to perform in 84.8% of patients but as rather difficult in cancer patients (odds ratio, 5.559;95% confidence interval, 1.434-21.546;P = 0.007). For benign conditions, dilatation before UtL was associated with menopause status (odds ratio, 4.929;95% confidence interval, 1.439-16.884;P = 0.016). In study II, the pain during UtL was rated with a median visual analog scale score of 1.6. During a period of 4 weeks after UtL, none of the participants had to use medication or developed symptoms requiring medical attention. The UtL took 6.5 minutes on average. The amount of extracted DNA was above the lower limit for a sensitive, deep-sequencing mutation analysis in all cases. Conclusions Our studies demonstrate that the UtL, using the new catheter, is a safe, reliable, and well-tolerated procedure, which does not require elaborate training. Therefore, UtL fulfils all prerequisites to be used in a potential screening setting

Treatment Strategies and Outcome of the Exstrophy–Epispadias Complex in Germany: Data From the German CURE-Net

Introduction: To evaluate the impact of reconstructive strategies and post-operative management on short- and long-term surgical outcome and complications of classical bladder exstrophy (CBE) patients' comprehensive data of the multicenter German-wide Network for Congenital Uro-Rectal malformations (CURE-Net) were analyzed. Methods: Descriptive analyses were performed between 34 prospectively collected CBE patients born since 2009, median 3 months old [interquartile range (IQR), 2–4 months], and 113 cross-sectional patients, median 12 years old (IQR, 6–21 years). Results: The majority of included individuals were males (67%). Sixty-eight percent of the prospectively observed and 53% of the cross-sectional patients were reconstructed using a staged approach (p = 0.17). Although prospectively observed patients were operated on at a younger age, the post-operative management did not significantly change in the years before and after 2009. Solely, in prospectively observed patients, peridural catheters were used significantly more often (p = 0.017). Blood transfusions were significantly more frequent in males (p = 0.002). Only half of all CBE individuals underwent inguinal hernia repair. Cross-sectional patients after single-stage reconstructions showed more direct post-operative complications such as upper urinary tract dilatations (p = 0.0021) or urinary tract infections (p = 0.023), but not more frequent renal function impairment compared to patients after the staged approach (p = 0.42). Continence outcomes were not significantly different between the concepts (p = 0.51). Self-reported continence data showed that the majority of the included CBE patients was intermittent or continuous incontinent. Furthermore, subsequent consecutive augmentations and catheterizable stomata did not significantly differ between the two operative approaches. Urinary diversions were only reported after the staged concept. Conclusions: In this German multicenter study, a trend toward the staged concept was observed. While single-stage approaches tended to have initially more complications such as renal dilatation or urinary tract infections, additional surgery such as augmentations and stomata appeared to be similar after staged and single-stage reconstructions in the long term

Recent developments in cell-based ENS regeneration – a short review

Therapeutic options to treat neurogenic motility disorders of the gastrointestinal tract are usually limited to symptomatic treatment. The capacity of the enteric nervous system (ENS) to regenerate and the fact that progenitor cells of the enteric nervous system reside in the postnatal and adult gut led to the idea to develop cell-based strategies to treat ENS related disorders. This short review focuses on recent developments in cell-based ENS regeneration, discussing advantages and disadvantages of various cell sources, functional impact of transplanted cells and highlights the challenges of translation of small animal studies to human application

Comparison of HLA-Class-I-antigenpresentation and geneexpression

In der vorliegenden Arbeit wurden aus zwei Nierenzellkarzinomen und zwei Kolonkarzinomen nach MHC-Klasse-I-Immunpräzipitation insgesamt 38 HLA-Klasse-I-gebundene Peptide massenspektrometrisch identifiziert. Zusätzlich wurde mit Hilfe von DNA-Chip-Technologie, quantitativer PCR und Laser Capture-Mikroskopie eine Expressionsanalyse eines Nierenzellkarzinoms durchgeführt und deren Ergebnisse mit den identifizierten HLA-Klasse-I-Liganden des selben Karzinoms vergleichen. So wurde die Expression im Tumorgewebe gegenüber dem Normalgewebe der gleichen Niere für ca. 7000 Gene überprüft. Verglichen mit den 22 gefundenen HLA-Klasse-I-Liganden des Nierenzellkarzinoms zeigte sich, daß sieben der den Peptiden zugehörigen Antigene im Tumor überexprimiert waren. Darunter waren u.a. die Nicotinamide-N-Methyltransferase (NNMT) (YYMIGEQKF) und das Protoonkogen c-Met (YVDPVITSI). Für sie konnte mittels quantitativer PCR von RNA verschiedener Körpergewebe nachgewiesen werden, daß ihre Expression - mit Ausnahme der Leber für NNMT - auf die Tumoren beschränkt ist bzw. sie dort stark überexprimiert sind. Somit wird angenommen, daß diese beiden Peptide mögliche Kandidaten für eine Tumorvakzinierung sind. Des weiteren wurde im untersuchten Nierenzellkarzinom ein Peptid eines vermeintlichen Frameshift-Proteins gefunden, für das die Nukleotidsequenz317-343 des DEAD/H box 3-Gens kodiert. Ob es sich dabei um ein tumorspezifisches Phänomen handelt, ist bis jetzt noch unklar. Für die restlichen Antigene der gefundenen Peptide konnte keine Überexpression in Tumoren nachgewiesen werden oder es war eine weite Verbreitung in normalen Körpergeweben offensichtlich oder unklar. Durch Einführung einer neuen MHC-Klasse-I-Immunpräzipitationsmethode mit Protein A-Sepharose wurde die Peptidausbeute bis auf das vierfache gesteigert. Es ist anzunehmen, daß der hier dargestellte Ansatz der Kombination von DNA-Expressionsanalyse und HLA-Klasse-I-Ligandenidentifikation und die daraus resultierende Möglichkeit des patientenspezifischen Vakzins die Tumorimmuntherapie entscheidend verbessern kann.In total 38 HLA-Class-I-bound peptides have been identified after MHC-Class-I-immunoprecepitation and mass-spectrometry from two renal cell carcinomas and two colon carcinomas. In addition gene-expression analysis of one renal cell carcinoma has been performed by DNA-microarray-technology, quantitative PCR and laser capture microscopy. The results of this expression analysis have been compared with identified MHC-Class-I-ligands of the same carcinoma. About 7000 Genes have been examined for expression in tumor-tissue and corresponding normal tissue of the same kidney. Compared with 22 HLA-Class-I-ligands identified from a renal cell carcinoma it could be shown that seven of the identified peptides have been overexpressed in tumor-tissue. For example peptides of Nicotine-N-Methyltransferase (NNMT) (YYMIGEQKF) and of the protooncogene c-Met (YVDPVITSI) have been identified. By quantitative PCR of different body-tissues we could show that expression of the corresponding genes was restricted to or overexpressed in tumor-tissue, exept NNMT, which was also expressed in normal liver-tissue. It has been concluded, that the two peptides could be potential candidates for tumor-vaccination. In the exmamined renal cell carcinoma a peptide of a potenial frameshift protein has been identified for which the DEAD/H box 3-Gene is coding. If this is a tumor-specific phenomenon is not clear. Other antigens, from which peptides have been identified did not show overexpression in tumor-tissue or were expressed in other normal tissue or their expression pattern has not yet examined. By a new MHC-Class-I-immunoprecipitation-technique a four time higher peptide-yield could be achived. In conclusion, the combination of DNA-expression analysis and HLA-Class-I-ligand-identification and the resulting possibility of tailoring patient specific vaccines could improve tumor-immunotherapy significantly