Ground Testing of the EMCS Seed Cassette for Biocompatibility with the Cellular Slime Mold, Dictyostelium Discoideum

The European Modular Cultivation System, EMCS, was developed by ESA for plant experiments. To expand the use of flight verified hardware for various model organisms, we performed ground experiments to determine whether ARC EMCS Seed Cassettes could be adapted for use with cellular slime mold for future space flight experiments. Dictyostelium is a cellular slime mold that can exist both as a single-celled independent organism and as a part of a multicellular colony which functions as a unit (pseudoplasmodium). Under certain stress conditions, individual amoebae will aggregate to form multicellular structures. Developmental pathways are very similar to those found in Eukaryotic organisms, making this a uniquely interesting organism for use in genetic studies. Dictyostelium has been used as a genetic model organism for prior space flight experiments. Due to the formation of spores that are resistant to unfavorable conditions such as desiccation, Dictyostelium is also a good candidate for use in the EMCS Seed Cassettes. The growth substratum in the cassettes is a gridded polyether sulfone (PES) membrane. A blotter beneath the PES membranes contains dried growth medium. The goals of this study were to (1) verify that Dictyostelium are capable of normal growth and development on PES membranes, (2) develop a method for dehydration of Dictyostelium spores with successful recovery and development after rehydration, and (3) successful mock rehydration experiments in cassettes. Our results show normal developmental progression in two strains of Dictyostelium discoideum on PES membranes with a bacterial food source. We have successfully performed a mock rehydration of spores with developmental progression from aggregation to slug formation, and production of morphologically normal spores within 9 days of rehydration. Our results indicate that experiments on the ISS using the slime mold, Dictyostelium discoideum could potentially be performed in the flight verified hardware of the EMCS ARC Seed Cassettes

Utilizing ARC EMCS Seedling Cassettes as Highly Versatile Miniature Growth Chambers for Model Organism Experiments

The aim of our ground testing was to demonstrate the capability of safely putting specific model organisms into dehydrated stasis, and to later rehydrate and successfully grow them inside flight proven ARC EMCS seedling cassettes. The ARC EMCS seedling cassettes were originally developed to support seedling growth during space flight. The seeds are attached to a solid substrate, launched dry, and then rehydrated in a small volume of media on orbit to initiate the experiment. We hypothesized that the same seedling cassettes should be capable of acting as culture chambers for a wide range of organisms with minimal or no modification. The ability to safely preserve live organisms in a dehydrated state allows for on orbit experiments to be conducted at the best time for crew operations and more importantly provides a tightly controlled physiologically relevant growth experiment with specific environmental parameters. Thus, we performed a series of ground tests that involved growing the organisms, preparing them for dehydration on gridded Polyether Sulfone (PES) membranes, dry storage at ambient temperatures for varying periods of time, followed by rehydration. Inside the culture cassettes, the PES membranes were mounted above blotters containing dehydrated growth media. These were mounted on stainless steel bases and sealed with plastic covers that have permeable membrane covered ports for gas exchange. The results showed we were able to demonstrate acceptable normal growth of C.elegans (nematodes), E.coli (bacteria), S.cerevisiae (yeast), Polytrichum (moss) spores and protonemata, C.thalictroides (fern), D.discoideum (amoeba), and H.dujardini (tardigrades). All organisms showed acceptable growth and rehydration in both petri dishes and culture cassettes initially, and after various time lengths of dehydration. At the end of on orbit ISS European Modular Cultivation System experiments the cassettes could be frozen at ultra-low temperatures, refrigerated, or chemically preserved before being returned to Earth for analyses. Our results suggest that with protocol modifications and future verification testing we can utilize the versatile EMCS to conduct tightly controlled experiments inside our culture cassettes for a wide variety of organisms. These physiological experiments would be designed to answer questions at the molecular level about the specific stress responses of space flight

Structural basis of human kinesin-8 function and inhibition

Kinesin motors play diverse roles in mitosis and are targets for anti-mitotic drugs. The clinical significance of these motors emphasizes the importance of understanding the molecular basis of their function. Equally, investigations into the modes of inhibition of these motors provide crucial information about their molecular mechanisms. Kif18A regulates spindle microtubules through its dual functionality – microtubule-based stepping and regulation of microtubule dynamics. We investigated the mechanism of Kif18A and its inhibition by the small molecule BTB-1. The Kif18A motor domain drives ATP-dependent plus-end microtubule gliding, and undergoes conformational changes consistent with canonical mechanisms of plus-end directed motility. The Kif18A motor domain also depolymerises microtubule plus and minus ends. BTB-1 inhibits both microtubule-based Kif18A activities. A reconstruction of BTB-1-bound, microtubule-bound Kif18A, in combination with computational modelling, identified an allosteric BTB-1 binding site near loop5, where it blocks the ATP-dependent conformational changes we characterised. Strikingly, BTB-1 binding is close to that of well-characterised Kif11 inhibitors that block tight microtubule binding, whereas BTB-1 traps Kif18A on the microtubule. Our work highlights a general mechanism of kinesin inhibition in which small molecule binding near loop5 prevents a range of conformational changes, blocking motor function

Landsat-7 ETM+ Radiometric Calibration Status

Now in its 17th year of operation, the Enhanced Thematic Mapper + (ETM+), on board the Landsat-7 satellite, continues to systematically acquire imagery of the Earth to add to the 40+ year archive of Landsat data. Characterization of the ETM+ on-orbit radiometric performance has been on-going since its launch in 1999. The radiometric calibration of the reflective bands is still monitored using on-board calibration devices, though the Pseudo-Invariant Calibration Sites (PICS) method has proven to be an effect tool as well. The calibration gains were updated in April 2013 based primarily on PICS results, which corrected for a change of as much as -0.2%/year degradation in the worst case bands. A new comparison with the SADE database of PICS results indicates no additional degradation in the updated calibration. PICS data are still being tracked though the recent trends are not well understood. The thermal band calibration was updated last in October 2013 based on a continued calibration effort by NASA/Jet Propulsion Lab and Rochester Institute of Technology. The update accounted for a 0.31 W/sq m/ sr/micron bias error. The updated lifetime trend is now stable to within + 0.4K

Bacterial communities in penile skin, male urethra, and vaginas of heterosexual couples with and without bacterial vaginosis

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Microbiome 4 (2016): 16, doi:10.1186/s40168-016-0161-6.The epidemiology of bacterial vaginosis (BV) suggests it is sexually transmissible, yet no transmissible agent has been identified. It is probable that BV-associated bacterial communities are transferred from male to female partners during intercourse; however, the microbiota of sexual partners has not been well-studied. Pyrosequencing analysis of PCR-amplified 16S rDNA was used to examine BV-associated bacteria in monogamous couples with and without BV using vaginal, male urethral, and penile skin specimens. The penile skin and urethral microbiota of male partners of women with BV was significantly more similar to the vaginal microbiota of their female partner compared to the vaginal microbiota of non-partner women with BV. This was not the case for male partners of women with normal vaginal microbiota. Specific BV-associated species were concordant in women with BV and their male partners. In monogamous heterosexual couples in which the woman has BV, the significantly higher similarity between the vaginal microbiota and the penile skin and urethral microbiota of the male partner, supports the hypothesis that sexual exchange of BV-associated bacterial taxa is common.This work was supported by National Institute of Health Grant R01 AI079071-01A1

Good Laboratory Practices: Myers et al. Respond

Reproduced with permission from Environmental Health Perspectives. DOI:10.1289/ehp.0900884RMyers et al. respond to a letter written by Becker et al. regarding Myers' article "Why public health agencies cannot depend on Good Laboratory Practices as a criterion for selecting data: the case of bisphenol A.

A Note on Conserved Charges of Asymptotically Flat and Anti-de Sitter Spaces in Arbitrary Dimensions

The calculation of conserved charges of black holes is a rich problem, for which many methods are known. Until recently, there was some controversy on the proper definition of conserved charges in asymptotically anti-de Sitter (AdS) spaces in arbitrary dimensions. This paper provides a systematic and explicit Hamiltonian derivation of the energy and the angular momenta of both asymptotically flat and asymptotically AdS spacetimes in any dimension D bigger or equal to 4. This requires as a first step a precise determination of the asymptotic conditions of the metric and of its conjugate momentum. These conditions happen to be achieved in ellipsoidal coordinates adapted to the rotating solutions.The asymptotic symmetry algebra is found to be isomorphic either to the Poincare algebra or to the so(D-1, 2) algebra, as expected. In the asymptotically flat case, the boundary conditions involve a generalization of the parity conditions, introduced by Regge and Teitelboim, which are necessary to make the angular momenta finite. The charges are explicitly computed for Kerr and Kerr-AdS black holes for arbitrary D and they are shown to be in agreement with thermodynamical arguments.Comment: 27 pages; v2 : references added, minor corrections; v3 : replaced to match published version forthcoming in General Relativity and Gravitatio

Field Theoretical Quantum Effects on the Kerr Geometry

We study quantum aspects of the Einstein gravity with one time-like and one space-like Killing vector commuting with each other. The theory is formulated as a \coset nonlinear $\sigma$-model coupled to gravity. The quantum analysis of the nonlinear $\sigma$-model part, which includes all the dynamical degrees of freedom, can be carried out in a parallel way to ordinary nonlinear $\sigma$-models in spite of the existence of an unusual coupling. This means that we can investigate consistently the quantum properties of the Einstein gravity, though we are limited to the fluctuations depending only on two coordinates. We find the forms of the beta functions to all orders up to numerical coefficients. Finally we consider the quantum effects of the renormalization on the Kerr black hole as an example. It turns out that the asymptotically flat region remains intact and stable, while, in a certain approximation, it is shown that the inner geometry changes considerably however small the quantum effects may be.Comment: 16 pages, LaTeX. The hep-th number added on the cover, and minor typos correcte