C5b-9 membrane attack complex formation and extracellular vesicle shedding in Barrett's esophagus and esophageal adenocarcinoma

The early complement components have emerged as mediators of pro-oncogenic inflammation, classically inferred to cause terminal complement activation, but there are limited data on the activity of terminal complement in cancer. We previously reported elevated serum and tissue C9, the terminal complement component, in esophageal adenocarcinoma (EAC) compared to the precursor condition Barrett’s Esophagus (BE) and healthy controls. Here, we investigate the level and cellular fates of the terminal complement complex C5b-9, also known as the membrane attack complex. Punctate C5b-9 staining and diffuse C9 staining was detected in BE and EAC by multiplex immunohistofluorescence without corresponding increase of C9 mRNA transcript. Increased C9 and C5b-9 staining were observed in the sequence normal squamous epithelium, BE, low- and high-grade dysplasia, EAC. C5b-9 positive esophageal cells were morphologically intact, indicative of sublytic or complement-evasion mechanisms. To investigate this at a cellular level, we exposed non-dysplastic BE (BAR-T and CP-A), high-grade dysplastic BE (CP-B and CP-D) and EAC (FLO-1 and OE-33) cell lines to the same sublytic dose of immunopurified human C9 (3 µg/ml) in the presence of C9-depleted human serum. Cellular C5b-9 was visualized by immunofluorescence confocal microscopy. Shed C5b-9 in the form of extracellular vesicles (EV) was measured in collected conditioned medium using recently described microfluidic immunoassay with capture by a mixture of three tetraspanin antibodies (CD9/CD63/CD81) and detection by surface-enhanced Raman scattering (SERS) after EV labelling with C5b-9 or C9 antibody conjugated SERS nanotags. Following C9 exposure, all examined cell lines formed C5b-9, internalized C5b-9, and shed C5b-9+ and C9+ EVs, albeit at varying levels despite receiving the same C9 dose. In conclusion, these results confirm increased esophageal C5b-9 formation during EAC development and demonstrate capability and heterogeneity in C5b-9 formation and shedding in BE and EAC cell lines following sublytic C9 exposure. Future work may explore the molecular mechanisms and pathogenic implications of the shed C5b-9+ EV

Modellierung von Helium-Atomstrahlen und ihr Einsatz zur Plasmadiagnostik der Tokamakrandschicht

Die vorliegende Arbeit befasst sich mit der Modellierung von Helium-Atomstrahlen zur Plasmadiagnostik. Hierzu wurde ein Computerprogramm entwickelt, das bei gegebenen Anfangsbedingungen die Besetzungsdichte in einem Atomstrahl berechnet. Durch Vergleich mit Messungen eines Atomstrahl-Diagnostikexperimentes mit Helium, das am Tokamak TEXTOR des Forschungszentrums Juelich installiert war, konnte das Modell ueberprueft werden. Die Arbeit gliedert sich in mehrere Abschnitte. Im ersten Teil wird auf die Grundlagen der Plasmadiagnostik mit Atomstrahlen und die Modellierung eingegangen. Der zweite Abschnitt beschreibt das thermische Atomstrahlexperiment in Juelich. Anschliessend werden Ergebnisse der Modellierung und Vergleiche mit TEXTOR-Messungen dargestellt. Den Abschluss der Arbeit bilden eine Zusammenfassung und Diskussion der erzielten Ergebnisse. (orig.)The dissertation is about modelling of helium atomic beams for plasma diagnostics. A computer code developed for the study computes for given initial conditions the population density in an atomic beam, thus modelling the conditions and interactions to be expected. The model has been verified by comparative analysis with the measurements of an atomic beam diagnostic experiment with helium performed at the TEXTOR tokamak of Juelich Research Center. The dissertation is divided into several chapters. The first discusses the fundamentals of plasma diagnostics using atomic beams, and subsequent modelling. The second explains the thermal atomic beam experiment at Juelich, with the following chapter presenting the comparative evaluation of the author's measurements and model with the results of the TEXTOR measurements. The final chapter summarizes and discusses the results obtained. (orig.)SIGLEAvailable from TIB Hannover: H94B1146 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekDEGerman

Complications and survival after hybrid and fully minimally invasive oesophagectomy

Background: Minimally invasive oesophagectomy (MIO) is reported to produce fewer respiratory complications than open oesophagectomy. This study assessed differences in postoperative complications between MIO and hybrid MIO (HMIO) employing thoracoscopy and laparotomy, along with the influence of co-morbidities on postoperative outcomes. Methods: Patients with oesophageal cancer undergoing three-stage MIO or three-stage HMIO between 1999 and 2018 were identified from a prospectively developed database, which included patient demographics, co-morbidities, preoperative therapies, and cancer stage. The primary outcome was postoperative complications in the two groups. Secondary outcomes included duration of operation, blood transfusion requirement, duration of hospital stay, and overall survival. Results: There were 828 patients, of whom 722 had HMIO and 106 MIO, without significant baseline differences. Median duration of operation was longer for MIO (325 versus 289 min; P < 0.001), but with less blood loss (median 250 versus 300 ml; P < 0.001) and a shorter hospital stay (median 12 versus 13 days; P = 0.006). Respiratory complications were not associated with operative approach (31.1 versus 35.2 per cent for MIO and HMIO respectively; P = 0.426). Anastomotic leak rates (10.4 versus 10.2 per cent) and 90-day mortality (1.0 versus 1.7 per cent) did not differ. Cardiac co-morbidity was associated with more medical and surgical complications. Overall survival was associated with AJCC stage and co-morbidities, but not operative approach. Conclusion: MIO had a small benefit in terms of blood loss and hospital stay, but not in operating time. Oncological outcomes were similar in the two groups. Postoperative complications were associated with pre-existing cardiorespiratory co-morbidities rather than operative approach