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ZUSAMMENFASSUNGDie molekularen Prinzipien bei der Transduktion von Schwerkraftänderungen in zelluläre Antwort- und Anpassungsprozesse sind bisher weitgehend unbekannt. Wir konnten in humanen Jurkat-T-Zellen zeigen, dass Gene bei veränderter Schwerkraft in Clusterstrukturen („gravity-responsive chromosomal regions“, GRCRs) differenziell reguliert werden. Durch Kombination mit Hochdurchsatz-Chromatin-Konformationsanalysen (Hi-C) konnte eine hochsignifikante Assoziation von GRCRs mit strukturellen 3D-Chromatinveränderungen identifiziert werden, die vor allem auf den kleinen Chromosomen (chr16–chr22) kolokalisieren. Wir fanden weiterhin Hinweise auf einen mechanistischen Zusammenhang zwischen Spleißprozessen und differenzieller Genexpression bei veränderter Schwerkraft. Somit haben wir erste Belege dafür gefunden, dass Änderungen der Schwerkraft in den Zellkern übertragen werden und dort 3D-Chromosomen-Konformationsänderungen hervorrufen, die mit einer schnellen Transkriptionsantwort verbunden sind. Wir vermuten, dass die schnelle genomische Antwort auf veränderte Gravitationskräfte in der Organisation des Chromatins spezifisch codiert ist.

International audience; The Advanced GAmma Tracking Array (AGATA) has been installed at Laboratori Nazionali di Legnaro (LNL), Italy. In this installation, AGATA will consist, at the beginning, of 13 AGATA triple clusters (ATCs) with an angular coverage of 1π, and progressively the number of ATCs will increase up to a 2π angular coverage. This setup will exploit both stable and radioactive ion beams delivered by the Tandem–PIAVE-ALPI accelerator complex and the SPES facility. The new implementation of AGATA at LNL will be used in two different configurations, firstly one coupled to the PRISMA large-acceptance magnetic spectrometer and lately a second one at Zero Degrees, along the beam line. These two configurations will allow us to cover a broad physics program, using different reaction mechanisms, such as Coulomb excitation, fusion-evaporation, transfer and fission at energies close to the Coulomb barrier. These setups have been designed to be coupled with a large variety of complementary detectors such as charged particle detectors, neutron detectors, heavy-ion detectors, high-energy γ-ray arrays, cryogenic and gasjet targets and the plunger device for lifetime measurements. We present in this paper the conceptual design, characteristics and performance figures of this implementation of AGATA at LNL.

ZUSAMMENFASSUNGZelluläre Prozesse werden durch Änderungen der Gravitationskraft in vielfältiger Weise dauerhaft beeinflusst. Die Möglichkeit der kontinuierlichen Beobachtung dieser Abläufe ist von großer Relevanz. Die Etablierung und Anwendung von hochauflösender Fluoreszenzmikroskopie im Rahmen des FLUMIAS-Experiments erlaubte kontinuierliche Aufnahmen von lebenden Immunzellen an Bord der Forschungsraketenmission TEXUS-54. Gravitationsbedingte Kurzzeitänderungen der Zellmorphologie sowie des Zellskeletts und assoziierte schnelle zelluläre Anpassungsvorgänge konnten in primären menschlichen Makrophagen identifiziert werden. Die hier vorgestellte Möglichkeit der Echtzeitanalyse kann das Wissen über dynamische zelluläre Reaktionen und Adaptationsprozesse an die Weltraumbedingungen enorm erweitern.

The mechanisms underlying gravity perception in mammalian cells are unknown. We have recently discovered that the transcriptome of cells in the immune system, which is the most affected system during a spaceflight, responds rapidly and broadly to altered gravity. To pinpoint potential underlying mechanisms, we compared gene expression and three-dimensional (3D) chromosomal conformational changes in human Jurkat T cells during the short-term gravitational changes in parabolic flight and suborbital ballistic rocket flight experiments. We found that differential gene expression in gravity-responsive chromosomal regions, but not differentially regulated single genes, are highly conserved between different real altered gravity comparisons. These coupled gene expression effects in chromosomal regions could be explained by underlying chromatin structures. Based on a high-throughput chromatin conformation capture (Hi-C) analysis in altered gravity, we found that small chromosomes (chr16–22, with the exception of chr18) showed increased intra- and interchromosomal interactions in altered gravity, whereby large chromosomes showed decreased interactions. Finally, we detected a nonrandom overlap between Hi-C-identified chromosomal interacting regions and gravity-responsive chromosomal regions (GRCRs). We therefore demonstrate the first evidence that gravitational force-induced 3D chromosomal conformational changes are associated with rapid transcriptional response in human T cells. We propose a general model of cellular sensitivity to gravitational forces, where gravitational forces acting on the cellular membrane are rapidly and mechanically transduced through the cytoskeleton into the nucleus, moving chromosome territories to new conformation states and their genes into more expressive or repressive environments, finally resulting in region-specific differential gene expression.

Cellular responses to micro- and hypergravity are rapid and complex and appear within the first few seconds of exposure. Transcriptomic analyses are a valuable tool to analyze these genome-wide cellular alterations. For a better understanding of the cellular dynamics upon altered gravity exposure, it is important to compare different time points. However, since most of the experiments are designed as endpoint measurements, the combination of cross-experiment meta-studies is inevitable. Microarray and RNA-Seq analyses are two of the main methods to study transcriptomics. In the field of altered gravity research, both methods are frequently used. However, the generation of these data sets is difficult and time-consuming and therefore the number of available data sets in this research field is limited. In this study, we investigated the comparability of microarray and RNA-Seq data and applied the results to a comparison of the transcriptomics dynamics between the hypergravity conditions during two real flight platforms and a centrifuge experiment to identify temporal adaptation processes. We performed a comparative study on an Affymetrix HTA2.0 microarray and a paired-end RNA-Seq data set originating from the same Jurkat T cell RNA samples from a short-term hypergravity experiment. The overall agreeability was high, with better sensitivity of the RNA-Seq analysis. The microarray data set showed weaknesses on the level of single upregulated genes, likely due to its normalization approach. On an aggregated level of biotypes, chromosomal distribution, and gene sets, both technologies performed equally well. The microarray showed better performance on the detection of altered gravity-related splicing events. We found that all initially altered transcripts fully adapted after 15 min to hypergravity and concluded that the altered gene expression response to hypergravity is transient and fully reversible. Based on the combined multiple-platform meta-analysis, we could demonstrate rapid transcriptional adaptation to hypergravity, the differential expression of the ATPase subunits ATP6V1A and ATP6V1D, and the cluster of differentiation (CD) molecules CD1E, CD2AP, CD46, CD47, CD53, CD69, CD96, CD164, and CD226 in hypergravity. We could experimentally demonstrate that it is possible to develop methodological evidence for the meta-analysis of individual data.

ZUSAMMENFASSUNGZelluläre Prozesse werden durch Änderungen der Gravitationskraft in vielfältiger Weise beeinflusst. In unseren Studien konnten wir in verschiedenen zellulären Systemen und in verschiedenen Forschungsplattformen (2D Klinostat, Parabelflüge, suborbitale Forschungsraketen, Internationale Raumstation) nachweisen, dass Reaktionen und Anpassungsprozesse bereits innerhalb von Sekunden bis Minuten nach Beginn der veränderten Schwerkraft auftreten. Diese schnellen Prozesse fanden sich in der Genexpression, der Zellzykluskontrolle, der Signaltransduktion, der Zytoskelettorganisation und des oxidativen Burst. Es stellt sich die Frage nach der Ursache der Transduktion einer unspezifischen Kraft in eine hochspezifische zelluläre Antwort, die in der Spezifität der zellulären Eigenschaften begründet liegen kann. Es ist denkbar, dass die Gravitationskraft der Erde die Chromatinarchitektur und deren Zugänglichkeit stabilisiert und eine homöostatische Bedingung für die Genexpression darstellt. Die Untersuchung mechanobiologischer Mechanismen der Genexpression in Schwerelosigkeit kann die Aufdeckung fundamentaler Prinzipien ermöglichen, wie mechanische Kräfte die Zellfunktion regulieren.

BackgroundIn systemic lupus erythematosus (SLE), the complement system (CS) is activated and thought to occur primarily through the classical pathway (CP) [1]. Traditionally, when diagnosing SLE or assessing disease activity, measurement of low C3 or C4 are used as proxies for complement activation [2]. However, measurement of C3 and C4 does not differentiate which complement pathway initiated the activation (i.e., the lectin pathway (LP), the CP, or the alternative pathway) [1, 3].C1-esterase inhibitor (C1inh) is one of the key regulators of the CS. C1inh is the exclusive inhibitor of the active CP enzymes C1r and C1s [4], and the major inhibitor of active LP enzymes MASP-1 and MASP-2 [5]. A possible way of assessing complement activation through a specific pathway, is by measuring activated enzymes complexed with C1inh in plasma, as these complexes only exist after complement enzyme activation.ObjectivesOur aim was to investigate and unravel LP and CP complement activation in SLE, by measuring the protein complexes MASP1/C1inh (LP specific activation) and C1r/C1inh (CP specific activation). Furthermore, we aimed to investigate whether there is an association between complement activity, disease activity (SLEDAI) and disease manifestations (lupus nephritis (LN)).MethodsA cross sectional cohort of 150 patients with SLE fulfilling the 1997 ACR classification criteria for SLE were included from the out-patient clinic at the department of Rheumatology, Aarhus University Hospital (AUH), Denmark. Disease manifestations and disease activity using SLEDAI score was assessed at inclusion. Fifty healthy individuals included at the Blood Bank, AUH, were used as controls. Both C1s/C1inh and MASP1/C1inh complexes were measured in all samples using two newly developed sandwich ELISAs (C1s/C1inh: cat# HK399; MASP1/C1inh: Cat#3001, Hycult Biotech, Uden, The Netherlands). EDTA-samples from both SLE patients and controls were measured in duplicates.ResultsWhen comparing SLE patients to controls, we observed a difference in complement activation through the LP, where a lower mean MASP1/C1inh plasma concentration was observed (pC1s/C1inh concentrations were significantly increased in active SLE patients (SLEDAI >6) when compared to SLE patients with low disease activity (SLEDAI No differences in MASP1/C1inh plasma concentrations were observed between active SLE patients and patients with low disease activity (p=0.11), nor did we observe a significant correlation with disease activity (r=0.12, p=0.13). In active LN, plasma concentrations of MASP1/C1inh were significantly elevated compared to non-active LN (p=0.02).ConclusionOur data suggest that the CP and the LP is activated in SLE. CS is generally activated in active SLE disease, whereas activation of the LP might be more specific to particular disease manifestations like LN.Our findings warrant further research into activation of the specific complement pathways in relation to specific disease manifestations in SLE.References[1]Troldborg, A., et al., J Rheumatol, 2018.[2]Weinstein, A., et al., Curr Rheumatol Rep, 2021.[3]Atkinson, J.P., Clin Exp Rheumatol, 1989.[4]Sim, R.B., et al., FEBS Lett, 1979.[5]Paréj, K., et al., Mol Immunol, 2013.Figure 1.Measurements of plasma concentrations of C1r in complex with C1-inhibitor (C1r/C1inh) (a1-a4) and MASP1 in complex with C1-inhibitor (MASP1/C1inh) (b1-b4) in Systemic Lupus Erythematosus (SLE) patients and controls. A1 and b1 show measurements of SLE patients versus controls. A2 and b2 show measurements in SLE patients with active disease (SLEDAI >6) versus low disease activity (SLEDAIDisclosure of InterestsMads Christian Lamm Larsen: None declared, Anne Troldborg: None declared, Erik Toonen Employee of: ET is an employee of Hycult Biotech, Lisa Hurler: None declared, Zoltán Prohászka: None declared, László Cervenak: None declared, Annette G Hansen: None declared, Steffen Thiel: None declared

Objective To systematically review and meta-analyze randomized controlled trials (RCTs) comparing postoperative bladder retrofilling to passive filling after outpatient gynecologic surgery to evaluate effects on postoperative outcomes. Data sources We searched MEDLINE, PubMed, Cochrane Central Register of Controlled Trials (CENTRAL), EMBASE, and ClinicalTrials.gov from 1947 to August 2020. Methods of study selection Two reviewers screened 1,465 articles. We included RCTs that compared postoperative bladder retrofilling to passive filling in patients who underwent outpatient gynecologic surgery by any approach. The primary outcome was the time to first void. Secondary outcomes included time to discharge, postoperative urinary retention, urinary tract infection, and patient satisfaction. Mean differences and relative risks (RRs) were calculated for the meta-analysis. Risk of bias was assessed using the Cochrane Risk of Bias Tool. Tabulation, integration, and results We included eight studies with 1,173 patients. Bladder retrofilling in the operating room resulted in a significant decrease in the time to first void (mean difference -33.5 minutes; 95% CI -49.1 to -17.9, 4 studies, 403 patients) and time to discharge (mean difference -32.0 minutes; 95% CI -51.5 to -12.6, eight studies, 1,164 patients). Bladder retrofilling did not shorten time to discharge when performed in the postanesthetic care unit (mean difference -14.8 min; 95% CI -62.6 to 32.9, three studies, 258 patients) or after laparoscopic hysterectomy (mean difference -26.0 min; 95% CI -56.5 to 4.5, five studies, 657 patients). There were no differences in postoperative urinary retention (RR 0.77; 95% CI 0.45-1.30, five studies, 910 patients) or risk of urinary tract infection between the retrofill and passive fill groups (RR 0.50; 95% CI 0.14-1.77, four studies, 387 patients). Patient satisfaction was comparable between groups. Conclusion Retrofilling the bladder in the operating room after outpatient gynecologic surgery modestly reduces the time to first void and discharge with no increase in adverse events. Systematic review registration PROSPERO, CRD42020203692.

Physical review / C 102(5), 054304 (2020). doi:10.1103/PhysRevC.102.054304, Published by Inst., Woodbury, NY

A projectile Coulomb-excitation experiment was performed at the radioactive ion beam facility HIE-ISOLDE at CERN. The radioactive 140Nd and 142Sm ions were post accelerated to the energy of 4.62 MeV/A and impinged on a 1.45 mg/cm2-thin 208Pb target. The γ rays depopulating the Coulomb-excited states were recorded by the HPGe-array MINIBALL. The scattered charged particles were detected by a double-sided silicon strip detector in forward direction. Experimental γ-ray intensities were used for the determination of electromagnetic transition matrix elements. Preliminary results for the reduced transition strength of the B ( M 1 ; 2 3 + → 2 1 + ) = 0.35 ( 19 ) μ N 2 of 140Nd and a first estimation for 142Sm have been deduced using the Coulomb-excitation calculation software GOSIA. The 2 3 + states of 140Nd and 142Sm show indications of being the main fragment of the proton-neutron mixed-symmetry 2 1 , ms + state.

Data analysis of the Coulomb excitation experiment of the exotic 206Hg nucleus, recently performed at CERN’s HIE-ISOLDE facility, needs to account for the contribution to target excitation due to the strongly-present beam contaminant 130Xe. In this paper, the contamination subtraction procedure is presented.

8 pags., 9 figs., We measured absolute cross sections for neutron transfer channels populated in the Rb-94 + Pb-208 binary reaction. Cross sections have been extracted identifying directly the lead isotopes with the high efficiency MINIBALL gamma-ray array coupled to a particle detector combined with a radioactive Rb-94 beam delivered at E-lab = 6.2 MeV/nucleon by the HIE-ISOLDE facility. We observed sizable cross sections in the neutron-rich mass region, where the heavy partner acquires neutrons. A fair agreement between the measured cross sections with those from GRAZING calculations gives confidence in the cross-section predictions of more neutron-rich nuclei produced via a larger number of transferred nucleons., This work was partly supported by the HORIZON 2020 ENSAR2 Grant Agreement no. 654002, and Marie Skłodowska-Curie COFUND (EU-CERN) grant agreement no. 665779, and by STFC (U.K.). This work has been supported in part by the Croatian Science Foundation under Project no. 7194 and in part under Project no. IP-2018-01- 1257, and the German BMBF under contract 05P18PKCIA + Verbundprojekt 05P2018

The gravitational force has been constant throughout Earth’s evolutionary history. Since the cell nucleus is subjected to permanent forces induced by Earth’s gravity, we addressed the question, if gene expression homeostasis is constantly shaped by the gravitational force on Earth. We therefore investigated the transcriptome in force-free conditions of microgravity, determined the time frame of initial gravitational force-transduction to the transcriptome and assessed the role of cation channels. We combined a parabolic flight experiment campaign with a suborbital ballistic rocket experiment employing the human myelomonocytic cell line U937 and analyzed the whole gene transcription by microarray, using rigorous controls for exclusion of effects not related to gravitational force and cross-validation through two fully independent research campaigns. Experiments with the wide range ion channel inhibitor SKF-96365 in combination with whole transcriptome analysis were conducted to study the functional role of ion channels in the transduction of gravitational forces at an integrative level. We detected profound alterations in the transcriptome already after 20 s of microgravity or hypergravity. In microgravity, 99.43% of all initially altered transcripts adapted after 5 min. In hypergravity, 98.93% of all initially altered transcripts adapted after 75 s. Only 2.4% of all microgravity-regulated transcripts were sensitive to the cation channel inhibitor SKF-96365. Inter-platform comparison of differentially regulated transcripts revealed 57 annotated gravity-sensitive transcripts. We assume that gravitational forces are rapidly and constantly transduced into the nucleus as omnipresent condition for nuclear and chromatin structure as well as homeostasis of gene expression.

ZusammenfassungEchtzeitmessungen auf der Internationalen Raumstation (ISS) konnten belegen, dass Zellen prinzipiell in der Lage sind, sich ultraschnell an veränderte Schwerkraftverhältnisse zu adaptieren. Im Experiment TRIPLE LUX A wurde der Einfluss von Gravitationskräften zwischen 1 und 0 g auf die Oxidative-Burst-Reaktion von NR8383-Alveolarmakrophagen untersucht. Erstmals konnte durch Echtzeit- und On-orbit-Messungen auf der ISS eine schwerkraftabhängige Reaktion in Säugerzellen direkt bewiesen und verfolgt werden. Durch den Messaufbau gelang ebenso der erstmalige Nachweis einer ultraschnellen und nach 42 Sekunden erfolgten vollständigen Anpassung einer zellulären Reaktion an die Schwerelosigkeit.

ZusammenfassungAm Ende sah es wie Routine aus: Am 14. April 2015 um 20:10:41 UTC startete vom Cape Canaveral Launch Complex SLC-40 eine Space X Falcon 9 Rakete mit dem Dragon-Raumschiff (Abb. 1) zur sechsten „Cargo Resupply Mission“ (Space X CRS-6) zur Internationalen Raumstation ISS. Mit an Bord: Die komplette Experimenthardware, tiefgefrorene Zellen und Reagenzien des DLR-Experiments TRIPLE LUX A. Im Experiment sollte der Einfluss von Gravitationskräften zwischen 1 und 0 g auf die Oxidative-Burst-Reaktion von Makrophagenzellen untersucht werden, erstmals durch Echtzeit- und On-orbit-Messungen auf der ISS im BIOLAB des europäischen COLUMBUS-Moduls 1.

Sensitivity of the human immune system to microgravity has been supposed since the first Apollo missions and was demonstrated during several space missions in the past. In vitro experiments demonstrated that cells of the immune system are exceptionally sensitive to microgravity. Therefore, serious concerns arose whether spaceflight-associated immune system weakening ultimately precludes the expansion of human presence beyond Earth’s orbit. In human cells, gravitational forces may be sensed by an individual cell in the context of altered extracellular matrix mechanics, cell shape, cytoskeletal organization, or internal prestress in the cell–tissue matrix. The development of cellular mechanosensitivity and signal transduction was probably an evolutionary requirement to enable our cells to sense their individual microenvironment. Therefore it is possible that the same mechanisms, which enable human cells to sense and to cope with mechanical stress, are potentially dangerous in microgravity. This chapter reviews the most recent developments in investigation to elucidate the influence of microgravity on immune cell signaling and functions and hereby bridges the phenotypic changes to transcriptome and epigenetic regulators.

Gravitational stress in general and microgravity (µg) in particular are regarded as major stress factors responsible for immune system dysfunction in space. To assess the effects of alternating µg and hypergravity (hyper-g) on immune cells, the attachment of peripheral blood mononuclear cells (PBMCs) to adhesion molecules under flow conditions and the antigen-induced immune activation in whole blood were investigated in parabolic flight (PF). In contrast to hyper-g (1.8 g) and control conditions (1 g), flow and rolling speed of PBMCs were moderately accelerated during µg-periods which were accompanied by a clear reduction in rolling rate. Whole blood analyses revealed a “primed” state of monocytes after PF with potentiated antigen-induced pro-inflammatory cytokine responses. At the same time, concentrations of anti-inflammatory cytokines were increased and monocytes displayed a surface molecule pattern that indicated immunosuppression. The results suggest an immunologic counterbalance to avoid disproportionate immune responses. Understanding the interrelation of immune system impairing and enhancing effects under different gravitational conditions may support the design of countermeasures to mitigate immune deficiencies in space.

The FLUMIAS (Fluorescence-Microscopic Analyses System for Life-Cell-Imaging in Space) confocal laser spinning disk fluorescence microscope represents a new imaging capability for live cell imaging experiments on suborbital ballistic rocket missions. During the second pioneer mission of this microscope system on the TEXUS-54 suborbital rocket flight, we developed and performed a live imaging experiment with primary human macrophages. We simultaneously imaged four different cellular structures (nucleus, cytoplasm, lysosomes, actin cytoskeleton) by using four different live cell dyes (Nuclear Violet, Calcein, LysoBrite, SiR-actin) and laser wavelengths (405, 488, 561, and 642 nm), and investigated the cellular morphology in microgravity (10&minus, 4 to 10&minus, 5 g) over a period of about six minutes compared to 1 g controls. For live imaging of the cytoskeleton during spaceflight, we combined confocal laser microscopy with the SiR-actin probe, a fluorogenic silicon-rhodamine (SiR) conjugated jasplakinolide probe that binds to F-actin and displays minimal toxicity. We determined changes in 3D cell volume and surface, nuclear volume and in the actin cytoskeleton, which responded rapidly to the microgravity environment with a significant reduction of SiR-actin fluorescence after 4&ndash, 19 s microgravity, and adapted subsequently until 126&ndash, 151 s microgravity. We conclude that microgravity induces geometric cellular changes and rapid response and adaptation of the potential gravity-transducing cytoskeleton in primary human macrophages.

Here we report the successful first operation of FLUMIAS-DEA, a miniaturized high-resolution 3D fluorescence microscope on the International Space Station (ISS) by imaging two scientific samples in a temperature-constant system, one sample with fixed cells and one sample with living human cells. The FLUMIAS-DEA microscope combines features of a high-resolution 3D fluorescence microscope based on structured illumination microscope (SIM) technology with hardware designs to meet the requirements of a space instrument. We successfully demonstrated that the FLUMIAS technology was able to acquire, transmit, and store high-resolution 3D fluorescence images from fixed and living cells, allowing quantitative and dynamic analysis of subcellular structures, e.g., the cytoskeleton. The capability of real-time analysis methods on ISS will dramatically extend our knowledge about the dynamics of cellular reactions and adaptations to the space environment, which is not only an option, but a requirement of evidence-based medical risk assessment, monitoring and countermeasure development for exploration class missions.

A new device, the GALILEO plunger, has been developed for lifetime measurement studies using the GALILEO γ -ray detector array at the Laboratori Nazionali di Legnaro (LNL) in Legnaro, Italy. This plunger device holds two stretched metal foils in the beam path for the use of the recoil distance Doppler-shift technique in order to measure lifetimes of nuclear excited states in the range of 1 ps to 1 ns. The compact design allows one to combine different auxiliary detectors with the plunger device inside the target chamber and takes care of high γ -ray transparency. The design and control of the device are presented in this paper, together with lifetimes of excited states in 180Pt measured within the first in-beam commissioning run.

The immune system belongs to the most affected systems during spaceflight, and sensitivity of cells of the human immune system to reduced gravity has been confirmed in numerous studies in real and simulated microgravity. Immune system dysfunction during spaceflight represents a substantial risk for exploration class mission knowledge about the clinical, cellular, and genetic basis of immune system response, and adaptation to altered gravity will provide key information for appropriate risk management, efficient monitoring, and countermeasures against existing limiting factors for human health and performance during manned exploration of the solar system. In spite of the immune system dysregulation, studies indicate an adaptation reaction of the immune system to the new microgravity environment, at least for the T-cell system, starting after 2 weeks and continuing until 6 months or longer, reflected by cytokine concentrations in blood plasma or in stimulation assays. At the cellular level, rapid adaptation responses could be detected as early as after seconds until minutes in T cells and macrophages. Therefore, adaptive responses of cells and the whole organism could be expected under microgravity and altered gravity in general. Preventive countermeasures should therefore consider support and stabilization of the endogenous adaptation programs. Potential countermeasures for risk mitigation are summarized in this chapter. We assume that the immune systems not only have a significant adaptation potential when challenged with low gravitational environments but also provide interesting preventive and therapeutic options for long-term space missions.

Mycobacterium genavense is regarded as the primary cause of mycobacteriosis in psittaciform and passeriform birds, which are commonly kept as pets. In humans, Mycobacterium genavense is especially pathogenic for young, old, pregnant and immunocompromised people (YOPIs). In birds, only few studies, mainly case reports, exist and there is still little e information about occurrence and relevance of this zoonotic pathogen. In this first pilot study concerning the prevalence of Mycobacterium genavense within flocks of naturally infected pet birds, real-time PCR examinations of 170 individual passeriform and psittaciform birds, including commonly kept budgerigars, lovebirds and zebra finches as well as gold finches and weaver finches, were conducted to determine the infection rate in six different aviaries. Antemortem examinations of faeces and cloacal swabs were compared with postmortem examinations of tissue samples to evaluate the reliability of antemortem diagnostics. Additional ophthalmologic examinations were performed to evaluate their diagnostic potential. Molecular examinations for viral co-infections, including circovirus, polyomavirus and adenovirus, were conducted to identify potential risk factors. PCR results revealed a detection prevalence of Mycobacterium genavense in the flocks varying from 3% to 91% based on postmortem testing, while antemortem diagnostics of faecal samples and swabs showed 64% discrepant (false negative) results. Ophthalmologic examinations were not useful in identifying infected birds within the flocks. Viral co-infections, especially with polyomavirus, were common. It has to be assumed that Mycobacterium genavense infections are widespread and underdiagnosed in companion birds. Viral infections might be an important risk factor. There is urgent need to improve antemortem diagnostics.

Projectile Coulomb excitation experiments were performed at HIE-ISOLDE at CERN with the radioactive ion beams of 140Nd and 142Sm. Ions with an energy of 4:62 MeV/A were impinging on a 1.45 mg/cm2 thick 208Pb target. The γ-rays depopulating the Coulomb-excited states were recorded by the HPGe-array MINIBALL and scattered particles were detected by a double-sided silicon strip detector. Experimental intensities were used for the determination of electromagnetic transition matrix elements. A preliminary result of the B(M1; 2+3 → 2+1) of 140Nd and an upper limit for the case of 142Sm are revealing the main fragments of the proton-neutron mixed-symmetry 2+1;ms states.

Background/Aims: Several limiting factors for human health and performance in microgravity have been clearly identified arising from the immune system, and substantial research activities are required in order to provide the basic information for appropriate integrated risk management. The gravity-sensitive nature of cells of the immune system renders them an ideal biological model in search for general gravity-sensitive mechanisms and to understand how the architecture and function of human cells is related to the gravitational force and therefore adapted to life on Earth. Methods: We investigated the influence of altered gravity in parabolic flight and 2D clinostat experiments on key proteins of activation and signaling in primary T lymphocytes. We quantified components of the signaling cascade 1.) in non-activated T lymphocytes to assess the “basal status” of the cascade and 2.) in the process of activation to assess the signal transduction. Results: We found a rapid decrease of CD3 and IL-2R surface expression and reduced p-LAT after 20 seconds of altered gravity in non-activated primary T lymphocytes during parabolic flight. Furthermore, we observed decreased CD3 surface expression, reduced ZAP-70 abundance and increased histone H3-acetylation in activated T lymphocytes after 5 minutes of clinorotation and a transient downregulation of CD3 and stable downregulation of IL-2R during 60 minutes of clinorotation. Conclusion: CD3 and IL-2R are downregulated in primary T lymphocytes in altered gravity. We assume that a gravity condition around 1g is required for the expression of key surface receptors and appropriate regulation of signal molecules in T lymphocytes.

ZusammenfassungDie Straußenhaltung in Deutschland verzeichnet weiterhin ein Wachstum. Strauße werden hierbei als landwirtschaftliche Nutztiere (Fleisch-, Leder-, Eierproduktion), als Schautiere, als Hobbytiere sowie in Tiergärten gehalten. In den letzten Jahren konnten hinsichtlich einer tierschutz- und artgerechten Haltung auf wissenschaftlicher Basis große Kenntniszugewinne erzielt werden, die auch für andere Straußenvögel wie Emus und Nandus von Relevanz sind. Beschrieben werden die zu berücksichtigenden rechtlichen Vorgaben sowie Empfehlungen zur praktischen Umsetzung.

Die Komplexität der Konstruktionen und Gebäudetechnik steigt seit Jahrzehnten stetig. Dies betrifft die Anforderungen an Standsicherheit, Wärme-, Feuchte-, Brand- und Schallschutz, Hygiene und Gesundheit wie auch den allgemeinen Nutzerkomfort. Das äußert sich in einer fast unüberblickbaren und weiter steigenden Zahl an Normen und Baugesetzen. Das damit anvisierte Ziel der Qualitätssicherung wird oft nicht erreicht: Die Folge der Komplexität ist eine hohe Fehlerquote in Planung und Ausführung sowie eine Überforderung von Bauherren und Nutzern. Hinsichtlich der Raumqualität ist der Standard in weiten Bereichen sogar gesunken: In der Abkehr von noch heute hochgeschätzten und flexibel nutzbaren Gebäuden der Gründerzeit entstanden seit der Nachkriegszeit auf den notwendigen Mindestmaßen basierende Gebäude. Die monofunktional angelegte Planung führte dazu, dass sich diese Typologien Veränderungen der Nutzung oder der Ansprüche nur schwierig anpassen konnten. Die Folge hiervon war und ist häufig der vorzeitige Umbau. Das Projekt soll vor diesem Hintergrund den Anfangspunkt zu einer neuen, gegenläufigen Entwicklung markieren und so einen wichtigen Impuls in der deutschen Bauwirtschaft setzen. Dem Vorhaben liegt die folgende Hypothese zugrunde: Wohngebäude mit hochwertiger und gleichzeitig suffizienter Architektur, robuster Baukonstruktion und reduzierter Gebäudetechnik sind - über einen Lebenszeitraum von 100 Jahren - bei besserer Aufenthaltsqualität sowohl üblichen Standardwohngebäuden als auch aktuellen Passivhäusern hinsichtlich Ökobilanz und Lebenszykluskosten überlegen. Eine neue Baukultur der Vereinfachung ist denkbar: Stringente und vielseitige Raumstrukturen, nutzungsflexible und hochwertig gestaltete Räume mit auskömmlicher Fläche (ca. 15-18 m²) und Höhe (ca. 2,60 m - 3,00 m), schichtenarme Bauteilaufbauten, zeitbeständige und haptisch ansprechende Oberflächen, einfache Anschlussdetails, Raumklimakonzepte mit reduzierter und anpassbarer Gebäudetechnik sind Bestandteile einer neuen Strategie. Diese zeichnet sich durch eine sehr ressourcenschonende Erstellung bei vertretbaren Kosten, eine dank hoher Qualität und Flexibilität lange Betriebs-phase bei geringem Instandhaltungsaufwand, moderaten Energieverbrauch für Heizung, Kühlung und Lüftung sowie hervorragende Recyclingfähigkeit aus. Ausgangspunkt der Untersuchungen sind die heute hochentwickelten Konstruktionsmaterialien Massivholz, Leichtbeton und hochwärmedämmendes Mauerwerk. Für diese Materialien wurden jeweils optimierte Konstruktionen, Raum- und Technikkonzepte, sowie Detaillösungen entwickelt und in einem Übersicht schaffenden Vergleich dargestellt.

The search for evidence of past or present life on Mars will require the detection of markers that indicate the presence of life. Because deoxyribonucleic acid (DNA) is found in all known living organisms, it is considered to be a ‘biosignature’ of life. The main function of DNA is the long-term storage of genetic information, which is passed on from generation to generation as hereditary material. The Polymerase Chain Reaction (PCR) is a revolutionary technique which allows a single fragment or a small number of fragments of a DNA molecule to be amplified millions of times, making it possible to detect minimal traces of DNA. The compactness of the contemporary PCR instruments makes routine sample analysis possible with a minimum amount of laboratory space. Furthermore the technique is effective, robust and straightforward. Our goal was to establish a routine for the detection of DNA from micro-organisms using the PCR technique during the EuroGeoMars simulation campaign. This took place at the Mars Society's Mars Desert Research Station (MDRS) in Utah in February 2009 (organized with the support of the International Lunar Exploration Working Group (ILEWG), NASA Ames and the European Space Research and Technology Centre (ESTEC)). During the MDRS simulation, we showed that it is possible to establish a minimal molecular biology lab in the habitat for the immediate on-site analysis of samples by PCR after sample collection. Soil and water samples were taken at different locations and soil depths. The sample analysis was started immediately after the crew returned to the habitat laboratory. DNA was isolated from micro-organisms and used as a template for PCR analysis of the highly conserved ribosomal DNA to identify representatives of the different groups of micro-organisms (bacteria, archaea and eukarya). The PCR products were visualized by agarose gel electrophoresis and documented by transillumination and digital imaging. The microbial diversity in the collected samples was analysed with respect to sampling depth and the presence or absence of vegetation. For the first time, we have demonstrated that it is possible to perform direct on-site DNA analysis by PCR at MDRS, a simulated planetary habitat in an extreme environment that serves as a model for preparation and optimization of techniques to be used for future Mars exploration.

In the last decades, a plethora of in vitro studies with living human cells contributed a vast amount of knowledge about cellular and molecular effects of microgravity. Previous studies focused mostly on the identification of gravity-responsive genes, whereas a multi-platform analysis at an integrative level, which specifically evaluates the extent and robustness of transcriptional response to an altered gravity environment was not performed so far. Therefore, we investigated the stability of gene expression response in non-activated human Jurkat T lymphocytic cells in different gravity environments through the combination of parabolic flights with a suborbital ballistic rocket and 2D clinostat and centrifuge experiments, using strict controls for excluding all possible other factors of influence. We revealed an overall high stability of gene expression in microgravity and identified olfactory gene expression in the chromosomal region 11p15.4 as particularly robust to altered gravity. We identified that classical reference genes ABCA5, GAPDH, HPRT1, PLA2G4A, and RPL13A were stably expressed in all tested gravity conditions and platforms, while ABCA5 and GAPDH were also known to be stably expressed in U937 cells in all gravity conditions. In summary, 10–20% of all transcripts remained totally unchanged in any gravitational environment tested (between 10−4 and 9 g), 20–40% remained unchanged in microgravity (between 10−4 and 10−2 g) and 97–99% were not significantly altered in microgravity if strict exclusion criteria were applied. Therefore, we suppose a high stability of gene expression in microgravity. Comparison with other stressors suggests that microgravity alters gene expression homeostasis not stronger than other environmental factors., Gene expression: potentially low-risk for long-term space missions Approximately 99% of genes in human lymphocytic cells have the same transcription activity on Earth as they do in microgravity environments. An international team led by Oliver Ullrich and Cora Thiel at the University of Zurich in Switzerland used a combination of parabolic aircraft and suborbital rocket flights and ground-based experiments to show how gene expression is stable through a range of gravity conditions. Screening of microarrays identified several reference genes that may serve as standardized controls for future immune cell trials. Their experiments also revealed that the chromosomal region associated with olfactory and taste receptor genes is particularly robust to altered gravity. Because altered transcripts were associated with fast cellular adaptation, the researchers predict that the risk of unusual gene behavior will be quite low during extended space deployments.

We investigated the dynamics of immediate and initial gene expression response to different gravitational environments in human Jurkat T lymphocytic cells and compared expression profiles to identify potential gravity-regulated genes and adaptation processes. We used the Affymetrix GeneChip® Human Transcriptome Array 2.0 containing 44,699 protein coding genes and 22,829 non-protein coding genes and performed the experiments during a parabolic flight and a suborbital ballistic rocket mission to cross-validate gravity-regulated gene expression through independent research platforms and different sets of control experiments to exclude other factors than alteration of gravity. We found that gene expression in human T cells rapidly responded to altered gravity in the time frame of 20 s and 5 min. The initial response to microgravity involved mostly regulatory RNAs. We identified three gravity-regulated genes which could be cross-validated in both completely independent experiment missions: ATP6V1A/D, a vacuolar H + -ATPase (V-ATPase) responsible for acidification during bone resorption, IGHD3-3/IGHD3-10, diversity genes of the immunoglobulin heavy-chain locus participating in V(D)J recombination, and LINC00837, a long intergenic non-protein coding RNA. Due to the extensive and rapid alteration of gene expression associated with regulatory RNAs, we conclude that human cells are equipped with a robust and efficient adaptation potential when challenged with altered gravitational environments.

The immune system is one of the most affected systems of the human body during space flight. The cells of the immune system are exceptionally sensitive to microgravity. Thus, serious concerns arise, whether space flight associated weakening of the immune system ultimately precludes the expansion of human presence beyond the Earth's orbit. For human space flight, it is an urgent need to understand the cellular and molecular mechanisms by which altered gravity influences and changes the functions of immune cells. The CELLBOX-PRIME (= CellBox-Primary Human Macrophages in Microgravity Environment) experiment investigated for the first time microgravity-associated long-term alterations in primary human macrophages, one of the most important effector cells of the immune system. The experiment was conducted in the U.S. National Laboratory on board of the International Space Station ISS using the NanoRacks laboratory and Biorack type I standard CELLBOX EUE type IV containers. Upload and download were performed with the SpaceX CRS-3 and the Dragon spaceship on April 18th, 2014 / May 18th, 2014. Surprisingly, primary human macrophages exhibited neither quantitative nor structural changes of the actin and vimentin cytoskeleton after 11 days in microgravity when compared to 1g controls. Neither CD18 or CD14 surface expression were altered in microgravity, however ICAM-1 expression was reduced. The analysis of 74 metabolites in the cell culture supernatant by GC-TOF-MS, revealed eight metabolites with significantly different quantities when compared to 1g controls. In particular, the significant increase of free fucose in the cell culture supernatant was associated with a significant decrease of cell surface-bound fucose. The reduced ICAM-1 expression and the loss of cell surface-bound fucose may contribute to functional impairments, e.g. the activation of T cells, migration and activation of the innate immune response. We assume that the surprisingly small and non-significant cytoskeletal alterations represent a stable "steady state" after adaptive processes are initiated in the new microgravity environment. Due to the utmost importance of the human macrophage system for the elimination of pathogens and the clearance of apoptotic cells, its apparent robustness to a low gravity environment is crucial for human health and performance during long-term space missions.