DNA DAMAGE RESPONSE OF EX-VIVO PORCINE EYE LENSES IN ORGAN-CULTURE AND IN-VITRO CULTURED LENS EPITHELIAL CELLS TO IONIZING RADIATION.

Astronauts on space missions, especially on long-term missions to Moon or Mars have a higher risk for the expression of radiation late effects such as cancer or sub-capsular cortical eye lens opacities. This is due to higher dose and different patterns of cellular energy deposition from high-linear-energy-transfer (LET) components of galactic cosmic radiation in space than that of terrestrial low-LET radiation on Earth. The eye lens is considered to be a radiation sensitive organ with radiation induced cataract to occur with a threshold absorbed dose of 0.5 Gy of sparsely ionizing radiation. For terrestrial occupational radiation lens exposure limit is set to yearly 20 mSv by the International Commission on Radiological Protection (1). Doses perceived by astronauts are much higher: in average 150 mSv per year on the International Space Station (ISS) and 1.2 to 1.4 mSv per day on Apollo and Skylab missions (2)

Kabinenluftqualität mit Schwerpunkt auf Geruchsereignissen – Eine Literaturstudie

Einleitung: Während eines Fume and Smell Events (FSE) können Dämpfe oder Gerüche ins Cockpit oder in die Flugzeugkabine gelangen. Insbesondere Kontaminationen der Zapfluft mit Triebwerks- oder Hydrauliköl werden mit vom fliegenden Personal berichteten vor allem neurologischen und/oder respiratorischen Symptomen während und / oder nach einem FSE in Verbindung gebracht. Zudem besteht Besorgnis über eine mögliche chronische Exposition mit toxischen Substanzen in der Kabinenluft. Fragestellungen: In dieser Studie wurde wissenschaftliche Literatur zu Ursachen von Gerüchen, Dampf- und Rauchentwicklung in Flugzeugen, zu chemischen Substanzen, die bei FSE in die Kabine gelangen könnten, zu einer möglichen Exposition von fliegendem Personal bei FSE, und zu akuten und chronischen Symptomen, die nach FSE-Exposition von fliegendem Personal im Kontext mit dem „aerotoxischen Syndrom“ beschrieben wurden und möglichen Ursachen, recherchiert und ausgewertet. Methodik: Durch eine orientierende Literaturrecherche wurde Suchstrings definiert und für eine systematische Recherche in den Literaturdatenbanken Medline, Web of Science und Scopus verwendet. Nach Import in eine Literaturdatenbank folgte ein Abstract- und Volltextscreening, eine Zuordnung zu den Fragestellungen und eine Bewertung der Evidenzklasse der Publikationen, die in Evidenztabellen mit einer zusammenfassenden Bewertung der Evidenzlage aufgelistet wurden. Ergebnisse: Die Häufigkeit von Kabinenluftereignissen wurde auf 1 Ereignis pro 2000-15.000 Flüge geschätzt. FSE können sich in allen Flugphasen ereignen, mit einer Häufung während Steig- und Reiseflug. Als Ursachen wurden u.a. Probleme mit der „Auxiliary Power Unit“ (APU), den Triebwerken, der Klimaversorgung/Environmental Control System (ECS), elektrischen Systemen und Küchengeräten, Kaffeemaschinen und Öfen identifiziert. Messflüge ergaben, CO, Ozon, volatile organic compounds (VOCs) und semi-volatile organic compounds (SVOCs) und Organophosphate im Normalbetrieb und bei vereinzelten Geruchsereignissen aktuelle Richt- oder Grenzwerte nicht überschreiten. Erhöhte CO₂-Konzentrationen und Partikelzählraten traten in der Taxiphase auf. Während des Reiseflugs sind die Konzentrationen von Luftkontaminanten aufgrund der hohen Luftwechselrate in der Flugzeugkabine meist gering und steigen gelegentlich durch Service und Passagieraktivitäten an. Die Beschreibung des durch FSE-ausgelösten Symptomkomplexes beruht im Wesentlichen auf Fall- und Fragebogen-Studien. Die in Expositionsstudien und nach versehentlicher Exposition beschriebenen Symptome zeigen eine gute Übereinstimmung mit einem Teil der durch Fragebogenstudien ermittelten akuten Symptome. Biomarker-Studien ergaben keinen Hinweis auf eine Organophosphatvergiftung. Genetische Variationen bei der Entgiftung wurden in einer Risikoabschätzung mit einem Faktor von 4000 als theoretischer maximaler Unterschied zwischen empfindlichen und unempfindlichen Individuen berücksichtigt. Die berichteten chronischen Symptome sind unspezifisch und divers und es bleibt unklar, ob die Symptome beim fliegenden Personal häufiger vorkommen als in der Allgemeinbevölkerung oder in anderen Berufsgruppen. Wenige Studien wiesen eine Kontrollgruppe auf und zeigten, dass Müdigkeit, Schlafstörungen und Depressionen von Flugbegleiterinnen häufiger berichtet wurden als von der Kontrollgruppe. Vereinzelte Studien mit neurologischer Untersuchung und psychometrischen Tests zeigten eine geringfügige Verschlechterung weniger Parameter. Des Weiteren wurden in kleinen Studien bei Betroffenen Veränderungen des Gehirns mit bildgebenden Verfahren gefunden, oder eine Erhöhung Nervensystem-spezifischer Auto-Antikörper im Serum. Die Bedeutung dieser morphologischen Veränderungen und der Auto-Antikörper im Krankheitsprozess und für die Diagnostik ist noch unklar. Schlussfolgerungen: Es lässt sich vermuten, dass in seltenen Fällen einer starken Kontamination der Zapfluft mit (pyrolysierten) Ölen CO, CO₂, Aldehyde und Feinstaub und ggf. weitere Substanzen ansteigen und akute Symptome wie Augen-, Nasen- und Rachenreizung, Schmerzen / Engegefühl in der Brust, Schwindel, Übelkeit, Erbrechen und Kopfschmerzen direkt oder indirekt verursachen können. Die Datenlage lässt eine Kausalitätsbewertung für mögliche chronische Symptome nicht zu. Dafür wäre u.a. ein Expositionsmonitoring des fliegenden Personals nach Identifikation geeigneter Markersubstanzen im Vergleich zu einer Kontrollgruppe ohne Exposition gegenüber Kabinenluftkontaminanten erforderlich. Danksagung: Die Literaturstudie erfolgte mit freundlicher Unterstützung der BG Verkehr

Degradation of microbial fluorescence biosignatures by solar ultraviolet radiation on Mars

Recent and proposed robotic missions to Mars are equipped with implements to expose or excavate fresh material from beneath the immediate surface. Once brought into the open, any organic molecules or potential biosignatures of present or past life will be exposed to the unfiltered solar ultraviolet (UV) radiation and face photolytic degradation over short time courses. The key question, then, is what is the window of opportunity for detection of recently exposed samples during robotic operations? Detection of autofluorescence has been proposed as a simple method for surveying or triaging samples for organic molecules. Using a Mars simulation chamber we conduct UV exposures on thin frozen layers of two model microorganisms, the radiation-resistant polyextremophile Deinococcus radiodurans and the cyanobacterium Synechocystis sp. PCC 6803. Excitation–emission matrices (EEMs) are generated of the full fluorescence response to quantify the change in signal of different cellular fluorophores over Martian equivalent time. Fluorescence of Deinococcus cells, protected by a high concentration of carotenoid pigments, was found to be relatively stable over 32 h of Martian UV irradiation, with around 90% of the initial signal remaining. By comparison, fluorescence from protein-bound tryptophan in Synechocystis is much more sensitive to UV photodegradation, declining to 50% after 64 h exposure. The signal most readily degraded byUV irradiation is fluorescence of the photosynthetic pigments – diminished to only 35% after 64 h. This sensitivity may be expected as the biological function of chlorophyll and phycocyanin is to optimize the harvesting of light energy and so they are readily photobleached. A significant increase in a *450 nm emission feature is interpreted as accumulation of fluorescent cellular degradation products from photolysis. Accounting for diurnal variation in Martian sunlight, this study calculates that frozen cellular biosignatures would remain detectable by fluorescence for at least several sols; offering a sufficient window for robotic exploration operations

Magnetic shielding of exomoons beyond the circumplanetary habitable edge

ABSTRACT: With most planets and planetary candidates detected in the stellar habitable zone (HZ) being super-Earths and gas giants rather than Earth-like planets, we naturally wonder if their moons could be habitable. The first detection of such an exomoon has now become feasible, and due to observational biases it will be at least twice as massive as Mars. However, formation models predict that moons can hardly be as massive as Earth. Hence, a giant planet’s magnetosphere could be the only possibility for such a moon to be shielded from cosmic and stellar high-energy radiation. Yet, the planetary radiation belt could also have detrimental effects on exomoon habitability. Here we synthesize models for the evolution of the magnetic environment of giant planets with thresholds from the runaway greenhouse (RG) effect to assess the habitability of exomoons. For modest eccentricities, we find that satellites around Neptune-sized planets in the center of the HZ around K dwarf stars will either be in an RG state and not be habitable, or they will be in wide orbits where they will not be affected by the planetary magnetosphere. Saturn-like planets have stronger fields, and Jupiter-like planets could coat close-in habitable moons soon after formation. Moons at distances between about 5 and 20 planetary radii from a giant planet can be habitable from an illumination and tidal heating point of view, but still the planetary magnetosphere would critically influence their habitability

Validation of biological recognition elements for signal transduction as first step in the development of whole cell biosensors

Choosing the proper combination of receptor element, cell type and measurable signal requires major consideration for developing cell-based biosensors. In order to use physiologically relevant cellular responses towards (geno)toxic conditions, information on the mechanism of action and of the expected outcome of exposure needs to be considered

Radiation Response of Murine Embryonic Stem Cells

To understand the mechanisms of disturbed differentiation and development by radiation, murine CGR8 embryonic stem cells (mESCs) were exposed to ionizing radiation and differentiated by forming embryoid bodies (EBs). The colony forming ability test was applied for survival and the MTT test for viability determination after X-irradiation. Cell cycle progression was determined by flow cytometry of propidium iodide-stained cells, and DNA double strand break (DSB) induction and repair by γH2AX immunofluorescence. The radiosensitivity of mESCs was slightly higher compared to the murine osteoblast cell line OCT-1. The viability 72 h after X-irradiation decreased dose-dependently and was higher in the presence of leukemia inhibitory factor (LIF). Cells exposed to 2 or 7 Gy underwent a transient G2 arrest. X-irradiation induced γH2AX foci and they disappeared within 72 h. After 72 h of X-ray exposure, RNA was isolated and analyzed using genome-wide microarrays. The gene expression analysis revealed amongst others a regulation of developmental genes (Ada, Baz1a, Calcoco2, Htra1, Nefh, S100a6 and Rassf6), downregulation of genes involved in glycolysis and pyruvate metabolism whereas upregulation of genes related to the p53 signaling pathway. X-irradiated mESCs formed EBs and differentiated toward cardiomyocytes but their beating frequencies were lower compared to EBs from unirradiated cells. These results suggest that X-irradiation of mESCs deregulate genes related to the developmental process. The most significant biological processes found to be altered by X-irradiation in mESCs were the development of cardiovascular, nervous, circulatory and renal system. These results may explain the X-irradiation induced-embryonic lethality and malformations observed in animal studies

Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies

The neuroblastoma cell line SH-SY5Y has been a well-established and very popular in vitro model in neuroscience for decades, especially focusing on neurodevelopmental disorders, such as Parkinson’s disease. The ability of this cell type to differentiate compared with other models in neurobiology makes it one of the few suitable models without having to rely on a primary culture of neuronal cells. Over the years, various, partly contradictory, methods of cultivation have been reported. This study is intended to provide a comprehensive guide to the in vitro cultivation of undifferentiated SH-SY5Y cells. For this purpose, the morphology of the cell line and the differentiation of the individual subtypes are described, and instructions for cell culture practice and long-term cryoconservation are provided. We describe the key growth characteristics of this cell line, including proliferation and confluency data, optimal initial seeding cell numbers, and a comparison of different culture media and cell viability during cultivation. Furthermore, applying an optimized protocol in a long-term cultivation over 60 days, we show that cumulative population doubling (CPD) is constant over time and does not decrease with incremental passage, enabling stable cultivation, for example, for recurrent differentiation to achieve the highest possible reproducibility in subsequent analyses. Therefore, we provide a solid guidance for future research that employs the neuroblastoma cell line SH-SY5

The Use of ProteoTuner Technology to Study Nuclear Factor κB Activation by Heavy Ions

Nuclear factor κB (NF-κB) activation might be central to heavy ion-induced detrimental processes such as cancer promotion and progression and sustained inflammatory responses. A sensitive detection system is crucial to better understand its involvement in these processes. Therefore, a DD-tdTomato fluorescent protein-based reporter system was previously constructed with human embryonic kidney (HEK) cells expressing DD-tdTomato as a reporter under the control of a promoter containing NF-κB binding sites (HEK-pNFκB-DD-tdTomato-C8). Using this reporter cell line, NF-κB activation after exposure to different energetic heavy ions (¹⁶O, 95 MeV/n, linear energy transfer—LET 51 keV/µm; ¹²C, 95 MeV/n, LET 73 keV/μm; ³⁶Ar, 95 MeV/n, LET 272 keV/µm) was quantified considering the dose and number of heavy ions hits per cell nucleus that double NF-κB-dependent DD-tdTomato expression. Approximately 44 hits of ¹⁶O ions and ≈45 hits of ¹²C ions per cell nucleus were required to double the NF-κB-dependent DD-tdTomato expression, whereas only ≈3 hits of ³⁶Ar ions were sufficient. In the presence of Shield-1, a synthetic molecule that stabilizes DD-tdTomato, even a single particle hit of ³⁶Ar ions doubled NF-κB-dependent DD-tdTomato expression. In conclusion, stabilization of the reporter protein can increase the sensitivity for NF-κB activation detection by a factor of three, allowing the detection of single particle hits’ effects