Nucleosomes indicate the in vitro radiosensitivity of irradiated bronchoepithelial and lung cancer cells

Nucleosomes, which are typical cell death products, are elevated in the serum of cancer patients and are known to rapidly increase during radiotherapy. As both normal and malignant cells are damaged by irradiation, we investigated to which extent both cell types contribute to the release of nucleosomes. We cultured monolayers of normal bronchoepithelial lung cells (BEAS-2B, n = 18) and epithelial lung cancer cells (EPLC, n = 18), exposed them to various radiation doses (0, 10 and 30 Gy) and observed them for 5 days. Culture medium was changed every 24 h. Subsequently, nucleosomes were determined in the supernatant by the Cell Death Detection-ELISA(plus) ( Roche Diagnostics). Additionally, the cell number was estimated after harvesting the cells in a second preparation. After 5 days, the cell number of BEAS-2B cultures in the irradiated groups (10 Gy: median 0.03 x 10(6) cells/culture, range 0.02-0.08 x 10(6) cells/culture; 30 Gy: median 0.08 x 10(6) cells/culture, range 0.02-0.14 x 10(6) cells/culture) decreased significantly (10 Gy: p = 0.005; 30 Gy p = 0.005; Wilcoxon test) compared to the non-irradiated control group (median 4.81 x 10(6) cells/culture, range 1.50-9.54 x 10(6) cells/culture). Consistently, nucleosomes remained low in the supernatant of nonirradiated BEAS-2B. However, at 10 Gy, BEAS-2B showed a considerably increasing release of nucleosomes, with a maximum at 72 h ( before irradiation: 0.24 x 10(3) arbitrary units, AU, range 0.13-4.09 x 10(3) AU, and after 72 h: 1.94 x 10(3) AU, range 0.11-5.70 x 10(3) AU). At 30 Gy, the release was even stronger, reaching the maximum earlier (at 48 h, 11.09 x 10(3) AU, range 6.89-18.28 x 10(3) AU). In non-irradiated EPLC, nucleosomes constantly increased slightly. At 10 Gy, we observed a considerably higher release of nucleosomes in EPLC, with a maximum at 72 h (before irradiation: 2.79 x 10(3) AU, range 2.42-3.80 x 10(3) AU, and after 72 h: 7.16 x 10(3) AU, range 4.30-16.20 x 10(3) AU), which was more than 3.5 times higher than in BEAS-2B. At 30 Gy, the maximum (6.22 x 10(3) AU, range 5.13-9.71 x 10(3) AU) was observed already after 24 h. These results indicate that normal bronchoepithelial and malignant lung cancer cells contribute to the release of nucleosomes during irradiation in a dose-and time-dependent manner with cancer cells having a stronger impact at low doses. Copyright (C) 2004 S. Karger AG, Basel

EmoCycling – Analysen von Radwegen mittels Humansensorik und Wearable Computing

Radfahren erfreut sich einer zunehmenden Wertschätzung. Einerseits als neuer Lifestyle, andererseits als wichtiges Thema der städtischen Mobilitätsplanung: Bike-Sharing-Angebote, Radwegekonzepte und Förderung eines umweltfreundlichen Mobilitätsmix sind hierbei wichtige Stichworte. Daher fördern zunehmend mehr Städte den Ausbau der Radwege-Infrastruktur, um das Radfahren attraktiver zu gestalten. Wie stark Radfahren aber tatsächlich angenommen und praktiziert wird, hängt von ganz verschiedenen Faktoren ab: Verkehrslage, Quantität und Qualität der Infrastruktur, Topografie sowie das subjektive Sicherheitsempfinden z.B. an unübersichtlichen Kreuzungen beeinflussen die Verkehrsmittelwahl. Insbesondere die Erfassung und Analyse des subjektiven Sicherheitsempfindens stellt hierbei eine große Herausforderung dar – wird aber durch neue Methoden der Humansensorik (Exner et al. 2012) möglich. Entwicklungen in den Bereichen des Wearable Computing sowie der Geoinformatik ermöglichen es, das subjektive Sicherheitsempfinden während der Fahrt genauer zu analysieren. Anknüpfend an Projekte zur emotionalen Stadtkartierung (Höffken et al. 2008, Zeile et al. 2010) erfolgt ein Live-Monitoring der Probanden während der Fahrt. Mittels eines Sensorarmbands (Smartband) zur Erfassung psychophysiologischer Reaktionen des Körpers in Kombination mit Video-Kamera-Daten und GPS-Koordinaten wird der emotionale Zustand der Probanden sekundengenau gemessen. Dadurch lassen sich Emotionen, insbesondere Stress, interpretieren und auf einer Karte verorten sowie die Auslöser (Trigger) identifizieren. Zudem kann auf diese Weise der Verkehr kontinuierlich erfasst und in die Analyse mit aufgenommen werden, um Gefahrenstellen zu lokalisieren. Nach einer Einführung in das Thema Radfahren in der Untersuchungsgemeinde Kaiserslautern, gibt das Paper einen Überblick über den aktuellen Stand der Methodik, die Konzeptionierung der Teststrecken sowie die Methodik im konkreten Projekt EmoCycling. Darauf basierend werden die Ergebnisse des Projektes vorgestellt und daraus resultierende weiterführende Fragenstellungen aufgezeigt

Herbivore and Pathogen Damage on Grassland and Woodland Plants: A Test of The Herbivore Uncertainty Principle

Researchers can alter the behaviour and ecology of their study organisms by conducting such seemingly benign activities as non‐destructive measurements and observations. In plant communities, researcher visitation and measurement of plants may increase herbivore damage in some plant species while decreasing it in others. Simply measuring plants could change their competitive ability by altering the amount of herbivore damage that they suffer. Currently, however, there is only limited empirical evidence to support this `herbivore uncertainty principle\u27 (HUP). We tested the HUP by quantifying the amount of herbivore and pathogen damage in 13 plant species (\u3e 1400 individuals) at four different visitation intensities at Cedar Creek Natural History Area, Minnesota, USA. Altogether, we found very little evidence to support the HUP at any intensity of visitation. Researcher visitation did not alter overall plant herbivore damage or survival and we did not detect a significant visitation effect in any of the 13 species. Pathogen damage also did not significantly vary among visitation treatments, although there was some evidence that high visitation caused slightly higher pathogen damage. Based on our results, we question whether this phenomenon should be considered a `principle\u27 of plant ecology

Comparison of existing aneurysm models and their path forward

The two most important aneurysm types are cerebral aneurysms (CA) and abdominal aortic aneurysms (AAA), accounting together for over 80\% of all fatal aneurysm incidences. To minimise aneurysm related deaths, clinicians require various tools to accurately estimate its rupture risk. For both aneurysm types, the current state-of-the-art tools to evaluate rupture risk are identified and evaluated in terms of clinical applicability. We perform a comprehensive literature review, using the Web of Science database. Identified records (3127) are clustered by modelling approach and aneurysm location in a meta-analysis to quantify scientific relevance and to extract modelling patterns and further assessed according to PRISMA guidelines (179 full text screens). Beside general differences and similarities of CA and AAA, we identify and systematically evaluate four major modelling approaches on aneurysm rupture risk: finite element analysis and computational fluid dynamics as deterministic approaches and machine learning and assessment-tools and dimensionless parameters as stochastic approaches. The latter score highest in the evaluation for their potential as clinical applications for rupture prediction, due to readiness level and user friendliness. Deterministic approaches are less likely to be applied in a clinical environment because of their high model complexity. Because deterministic approaches consider underlying mechanism for aneurysm rupture, they have improved capability to account for unusual patient-specific characteristics, compared to stochastic approaches. We show that an increased interdisciplinary exchange between specialists can boost comprehension of this disease to design tools for a clinical environment. By combining deterministic and stochastic models, advantages of both approaches can improve accessibility for clinicians and prediction quality for rupture risk.Comment: 46 pages, 5 figure

Hip disability and osteoarthritis outcome score (HOOS) – validity and responsiveness in total hip replacement

BACKGROUND: The aim of the study was to evaluate if physical functions usually associated with a younger population were of importance for an older population, and to construct an outcome measure for hip osteoarthritis with improved responsiveness compared to the Western Ontario McMaster osteoarthritis score (WOMAC LK 3.0). METHODS: A 40 item questionnaire (hip disability and osteoarthritis outcome score, HOOS) was constructed to assess patient-relevant outcomes in five separate subscales (pain, symptoms, activity of daily living, sport and recreation function and hip related quality of life). The HOOS contains all WOMAC LK 3.0 questions in unchanged form. The HOOS was distributed to 90 patients with primary hip osteoarthritis (mean age 71.5, range 49–85, 41 females) assigned for total hip replacement for osteoarthritis preoperatively and at six months follow-up. RESULTS: The HOOS met set criteria of validity and responsiveness. It was more responsive than WOMAC regarding the subscales pain (SRM 2.11 vs. 1.83) and other symptoms (SRM 1.83 vs. 1.28). The responsiveness (SRM) for the two added subscales sport and recreation and quality of life were 1.29 and 1.65, respectively. Patients ≤ 66 years of age (range 49–66) reported higher responsiveness in all five subscales than patients >66 years of age (range 67–85) (Pain SRM 2.60 vs. 1.97, other symptoms SRM 3.0 vs. 1.60, activity of daily living SRM 2.51 vs. 1.52, sport and recreation function SRM 1.53 vs. 1.21 and hip related quality of life SRM 1.95 vs. 1.57). CONCLUSION: The HOOS 2.0 appears to be useful for the evaluation of patient-relevant outcome after THR and is more responsive than the WOMAC LK 3.0. The added subscales sport and recreation function and hip related quality of life were highly responsive for this group of patients, with the responsiveness being highest for those younger than 66

Safety and clinical outcomes of rituximab therapy in patients with different autoimmune diseases: experience from a national registry (GRAID)

Introduction: Evidence from a number of open-label, uncontrolled studies has suggested that rituximab may benefit patients with autoimmune diseases who are refractory to standard-of-care. The objective of this study was to evaluate the safety and clinical outcomes of rituximab in several standard-of-care-refractory autoimmune diseases (within rheumatology, nephrology, dermatology and neurology) other than rheumatoid arthritis or non-Hodgkin's lymphoma in a real-life clinical setting. Methods: Patients who received rituximab having shown an inadequate response to standard-of-care had their safety and clinical outcomes data retrospectively analysed as part of the German Registry of Autoimmune Diseases. The main outcome measures were safety and clinical response, as judged at the discretion of the investigators. Results: A total of 370 patients (299 patient-years) with various autoimmune diseases (23.0% with systemic lupus erythematosus, 15.7% antineutrophil cytoplasmic antibody-associated granulomatous vasculitides, 15.1% multiple sclerosis and 10.0% pemphigus) from 42 centres received a mean dose of 2,440 mg of rituximab over a median (range) of 194 (180 to 1,407) days. The overall rate of serious infections was 5.3 per 100 patient-years during rituximab therapy. Opportunistic infections were infrequent across the whole study population, and mostly occurred in patients with systemic lupus erythematosus. There were 11 deaths (3.0% of patients) after rituximab treatment (mean 11.6 months after first infusion, range 0.8 to 31.3 months), with most of the deaths caused by infections. Overall (n = 293), 13.3% of patients showed no response, 45.1% showed a partial response and 41.6% showed a complete response. Responses were also reflected by reduced use of glucocorticoids and various immunosuppressives during rituximab therapy and follow-up compared with before rituximab. Rituximab generally had a positive effect on patient well-being (physician's visual analogue scale; mean improvement from baseline of 12.1 mm). Conclusions: Data from this registry indicate that rituximab is a commonly employed, well-tolerated therapy with potential beneficial effects in standard of care-refractory autoimmune diseases, and support the results from other open-label, uncontrolled studies

Genetic code expansion for multiprotein complex engineering

We present a baculovirus-based protein engineering method that enables site-specific introduction of unique functionalities in a eukaryotic protein complex recombinantly produced in insect cells. We demonstrate the versatility of this efficient and robust protein production platform, \u2018MultiBacTATAG\u2019, (i) for the fluorescent labeling of target proteins and biologics using click chemistries, (ii) for glycoengineering of antibodies, and (iii) for structure\u2013function studies of novel eukaryotic complexes using single-molecule F\uf6rster resonance energy transfer as well as site-specific crosslinking strategies

Untersuchungen von Zellmonoschichten mittels Elektrochemischer Rastermikroskopie

Die elektrochemische Rastermikroskopie (SECM) dient der Untersuchung der Topographie und lokalen Elektroaktivität mikrostrukturierter Proben. Seit ihrer Einführung in den 1980er Jahren hat die Technik enorme Akzeptanz im Bereich der Korrosionsforschung und Bioanalytik erfahren. In der SECM rastert eine Ultramikroelektrode (UME) die zu untersuchende Probe in geringem Abstand ab. Die aktiven Durchmesser der UME liegen dabei im Bereich weniger 100 nm. Die UMEs können dabei in Abhängig vom Aufbau der Elektroden amperometrisch, potentiometrisch oder impedimetrisch betrieben werden. Häufig werden in Natron-Kalk-Glas eingeschmolzene feine Platindrähte als UMEs verwendet. Die Einbettung jener UMEs kann durch eine nachgeschobene thermische Behandlung signifikant verbessert werden. Durch die Biokompatibilität der SECM und die intrinsisch-rasterförmige Bewegung der UME sind lebende Zellmonoschichten als Verbände von Zellen ideale Studienobjekte. Neben der hochauflösenden topographischen Abbildung von Zellmonoschichten kann die UME auch als Werkzeug zum gezielten Abtöten einzelner Zellen innerhalb eines Zellverbundes – ohne erkennbare Wirkung auf die Nachbarzellen – verwendet werden. Auch die in der Pharmakologie weit verbreiteten Transportstudien durch Zellmonoschichten liefern mittels SECM reproduzierbare und richtige Ergebnisse auf integraler Ebene. Zudem kann mittels SECM erstmalig die Diffusion von Modellsubstanzen auf zellulärer Ebene beobachtet und unterschieden werden