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Description

Nutrient availability and ratios can play an important role in shaping microbial communities of freshwater ecosystems. The Cuatro Ciénegas Basin (CCB) in Mexico is a desert oasis where, perhaps paradoxically, high microbial diversity coincides with extreme oligotrophy. To better understand the effects of nutrients on microbial communities in CCB, a

Nutrient availability and ratios can play an important role in shaping microbial communities of freshwater ecosystems. The Cuatro Ciénegas Basin (CCB) in Mexico is a desert oasis where, perhaps paradoxically, high microbial diversity coincides with extreme oligotrophy. To better understand the effects of nutrients on microbial communities in CCB, a mesocosm experiment was implemented in a stoichiometrically imbalanced pond, Lagunita, which has an average TN:TP ratio of 122 (atomic). The experiment had four treatments, each with five spatial replicates – unamended controls and three fertilization treatments with different nitrogen:phosphorus (N:P) regimes (P only, N:P = 16 and N:P = 75 by atoms). In the water column, quantitative PCR of the 16S rRNA gene indicated that P enrichment alone favored proliferation of bacterial taxa with high rRNA gene copy number, consistent with a previously hypothesized but untested connection between rRNA gene copy number and P requirement. Bacterial and microbial eukaryotic community structure was investigated by pyrosequencing of 16S and 18S rRNA genes from the planktonic and surficial sediment samples. Nutrient enrichment shifted the composition of the planktonic community in a treatment-specific manner and promoted the growth of previously rare bacterial taxa at the expense of the more abundant, potentially endemic, taxa. The eukaryotic community was highly enriched with phototrophic populations in the fertilized treatment. The sediment microbial community exhibited high beta diversity among replicates within treatments, which obscured any changes due to fertilization. Overall, these results showed that nutrient stoichiometry can be an important factor in shaping microbial community structure.

ContributorsLee, Zarraz (Author) / Poret-Peterson, Amisha (Author) / Siefert, Janet L. (Author) / Kaul, Drishti (Author) / Moustafa, Ahmed (Author) / Allen, Andrew E. (Author) / Dupont, Chris L. (Author) / Eguiarte, Luis E. (Author) / Souza, Valeria (Author) / Elser, James (Author) / College of Liberal Arts and Sciences (Contributor) / School of Life Sciences (Contributor) / School of Earth and Space Exploration (Contributor)
Created2017-05-30
Description

Cell-sediment separation methods can potentially enable determination of the elemental composition of microbial communities by removing the sediment elemental contribution from bulk samples. We demonstrate that a separation method can be applied to determine the composition of prokaryotic cells. The method uses chemical and physical means to extract cells from

Cell-sediment separation methods can potentially enable determination of the elemental composition of microbial communities by removing the sediment elemental contribution from bulk samples. We demonstrate that a separation method can be applied to determine the composition of prokaryotic cells. The method uses chemical and physical means to extract cells from benthic sediments and mats. Recovery yields were between 5% and 40%, as determined from cell counts. The method conserves cellular element contents to within 30% or better, as assessed by comparing C, N, P, Mg, Al, Ca, Ti, Mn, Fe, Ni, Cu, Zn, and Mo contents in Escherichia coli. Contamination by C, N, and P from chemicals used during the procedure was negligible. Na and K were not conserved, being likely exchanged through the cell membrane as cations during separation. V, Cr, and Co abundances could not be determined due to large (>100%) measurement uncertainties. We applied this method to measure elemental contents in extremophilic communities of Yellowstone National Park hot springs. The method was generally successful at separating cells from sediment, but does not discriminate between cells and detrital biological or noncellular material of similar density. This resulted in Al, Ti, Mn, and Fe contamination, which can be tracked using proxies such as metal:Al ratios. With these caveats, we present the first measurements, to our knowledge, of the elemental abundances of a chemosynthetic community. The communities have C:N ratios typical of aquatic microorganisms, are low in P, and their metal abundances vary between hot springs by orders of magnitude.

ContributorsNeveu, Marc (Author) / Poret-Peterson, Amisha (Author) / Lee, Zarraz (Author) / Anbar, Ariel (Author) / Elser, James (Author) / College of Liberal Arts and Sciences (Contributor) / School of Earth and Space Exploration (Contributor) / School of Life Sciences (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-07-01
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Description
Intense femtosecond x-ray pulses from free-electron laser sources allow the imaging of individual particles in a single shot. Early experiments at the Linac Coherent Light Source (LCLS) have led to rapid progress in the field and, so far, coherent diffractive images have been recorded from biological specimens, aerosols, and quantum

Intense femtosecond x-ray pulses from free-electron laser sources allow the imaging of individual particles in a single shot. Early experiments at the Linac Coherent Light Source (LCLS) have led to rapid progress in the field and, so far, coherent diffractive images have been recorded from biological specimens, aerosols, and quantum systems with a few-tens-of-nanometers resolution. In March 2014, LCLS held a workshop to discuss the scientific and technical challenges for reaching the ultimate goal of atomic resolution with single-shot coherent diffractive imaging. This paper summarizes the workshop findings and presents the roadmap toward reaching atomic resolution, 3D imaging at free-electron laser sources.
ContributorsAquila, A. (Author) / Barty, A. (Author) / Bostedt, C. (Author) / Boutet, S. (Author) / Carini, G. (Author) / dePonte, D. (Author) / Drell, P. (Author) / Doniach, S. (Author) / Downing, K. H. (Author) / Earnest, T. (Author) / Elmlund, H. (Author) / Elser, V. (Author) / Guhr, M. (Author) / Hajdu, J. (Author) / Hastings, J. (Author) / Hau-Riege, S. P. (Author) / Huang, Z. (Author) / Lattman, E. E. (Author) / Maia, F. R. N. C. (Author) / Marchesini, S. (Author) / Ourmazd, A. (Author) / Pellegrini, C. (Author) / Santra, R. (Author) / Schlichting, I. (Author) / Schroer, C. (Author) / Spence, John (Author) / Vartanyants, I. A. (Author) / Wakatsuki, S. (Author) / Weis, W. I. (Author) / Williams, G. J. (Author) / College of Liberal Arts and Sciences (Contributor) / Department of Physics (Contributor)
Created2015-04-21
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Description
The advent and application of the X-ray free-electron laser (XFEL) has uncovered the structures of proteins that could not previously be solved using traditional crystallography. While this new technology is powerful, optimization of the process is still needed to improve data quality and analysis efficiency. One area is sample heterogeneity,

The advent and application of the X-ray free-electron laser (XFEL) has uncovered the structures of proteins that could not previously be solved using traditional crystallography. While this new technology is powerful, optimization of the process is still needed to improve data quality and analysis efficiency. One area is sample heterogeneity, where variations in crystal size (among other factors) lead to the requirement of large data sets (and thus 10–100 mg of protein) for determining accurate structure factors. To decrease sample dispersity, we developed a high-throughput microfluidic sorter operating on the principle of dielectrophoresis, whereby polydisperse particles can be transported into various fluid streams for size fractionation. Using this microsorter, we isolated several milliliters of photosystem I nanocrystal fractions ranging from 200 to 600 nm in size as characterized by dynamic light scattering, nanoparticle tracking, and electron microscopy. Sorted nanocrystals were delivered in a liquid jet via the gas dynamic virtual nozzle into the path of the XFEL at the Linac Coherent Light Source. We obtained diffraction to ∼4 Å resolution, indicating that the small crystals were not damaged by the sorting process. We also observed the shape transforms of photosystem I nanocrystals, demonstrating that our device can optimize data collection for the shape transform-based phasing method. Using simulations, we show that narrow crystal size distributions can significantly improve merged data quality in serial crystallography. From this proof-of-concept work, we expect that the automated size-sorting of protein crystals will become an important step for sample production by reducing the amount of protein needed for a high quality final structure and the development of novel phasing methods that exploit inter-Bragg reflection intensities or use variations in beam intensity for radiation damage-induced phasing. This method will also permit an analysis of the dependence of crystal quality on crystal size.
ContributorsAbdallah, Bahige (Author) / Zatsepin, Nadia (Author) / Roy Chowdhury, Shatabdi (Author) / Coe, Jesse (Author) / Conrad, Chelsie (Author) / Dorner, Katerina (Author) / Sierra, Raymond G. (Author) / Stevenson, Hilary P. (Author) / Camacho Alanis, Fernanda (Author) / Grant, Thomas D. (Author) / Nelson, Garrett (Author) / James, Daniel (Author) / Calero, Guillermo (Author) / Wachter, Rebekka (Author) / Spence, John (Author) / Weierstall, Uwe (Author) / Fromme, Petra (Author) / Ros, Alexandra (Author) / Department of Chemistry and Biochemistry (Contributor) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2015-08-19
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Description
Open-ended evolution (OEE) is relevant to a variety of biological, artificial and technological systems, but has been challenging to reproduce in silico. Most theoretical efforts focus on key aspects of open-ended evolution as it appears in biology. We recast the problem as a more general one in dynamical systems theory,

Open-ended evolution (OEE) is relevant to a variety of biological, artificial and technological systems, but has been challenging to reproduce in silico. Most theoretical efforts focus on key aspects of open-ended evolution as it appears in biology. We recast the problem as a more general one in dynamical systems theory, providing simple criteria for open-ended evolution based on two hallmark features: unbounded evolution and innovation. We define unbounded evolution as patterns that are non-repeating within the expected Poincare recurrence time of an isolated system, and innovation as trajectories not observed in isolated systems. As a case study, we implement novel variants of cellular automata (CA) where the update rules are allowed to vary with time in three alternative ways. Each is capable of generating conditions for open-ended evolution, but vary in their ability to do so. We find that state-dependent dynamics, regarded as a hallmark of life, statistically out-performs other candidate mechanisms, and is the only mechanism to produce open-ended evolution in a scalable manner, essential to the notion of ongoing evolution. This analysis suggests a new framework for unifying mechanisms for generating OEE with features distinctive to life and its artifacts, with broad applicability to biological and artificial systems.
Created2017-04-20
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Description
Toward the end of his career, Zewail developed strong interest in fast electron spectroscopy and imaging, a field to which he made important contributions toward his aim of making molecular movies free of radiation damage. We therefore compare here the atomistic mechanisms leading to destruction of protein samples in diffract-and-destroy

Toward the end of his career, Zewail developed strong interest in fast electron spectroscopy and imaging, a field to which he made important contributions toward his aim of making molecular movies free of radiation damage. We therefore compare here the atomistic mechanisms leading to destruction of protein samples in diffract-and-destroy experiments for the cases of high-energy electron beam irradiation and X-ray laser pulses. The damage processes and their time-scales are compared and relevant elastic, inelastic, and photoelectron cross sections are given. Inelastic mean-free paths for ejected electrons at very low energies in insulators are compared with the bioparticle size. The dose rate and structural damage rate for electrons are found to be much lower, allowing longer pulses, reduced beam current, and Coulomb interactions for the formation of smaller probes. High-angle electron scattering from the nucleus, which has no parallel in the X-ray case, tracks the slowly moving nuclei during the explosion, just as the gain of the XFEL (X-ray free-electron laser) has no parallel in the electron case. Despite reduced damage and much larger elastic scattering cross sections in the electron case, leading to not dissimilar elastic scattering rates (when account is taken of the greatly increased incident XFEL fluence), progress for single-particle electron diffraction is seen to depend on the effort to reduce emittance growth due to Coulomb interactions, and so allow formation of intense sub-micron beams no larger than a virus.
ContributorsSpence, John (Author) / College of Liberal Arts and Sciences (Contributor) / Department of Physics (Contributor)
Created2017-06-01
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Description

Cataclysmic Variables (CVs) are close binary star systems with one component a white dwarf (WD) and the other a larger cooler star that fills its Roche Lobe. The cooler star is losing mass through the inner Lagrangian point of the binary and some unknown fraction of this material is accreted

Cataclysmic Variables (CVs) are close binary star systems with one component a white dwarf (WD) and the other a larger cooler star that fills its Roche Lobe. The cooler star is losing mass through the inner Lagrangian point of the binary and some unknown fraction of this material is accreted by the WD. One consequence of the WDs accreting material, is the possibility that they are growing in mass and will eventually reach the Chandrasekhar Limit. This evolution could result in a Supernova Ia (SN Ia) explosion and is designated the Single Degenerate Progenitor (SD) scenario. This paper is concerned with the SD scenario for SN Ia progenitors. One problem with the single degenerate scenario is that it is generally assumed that the accreting material mixes with WD core material at some time during the accretion phase of evolution and, since the typical WD has a carbon-oxygen CO core, the mixing results in large amounts of carbon and oxygen being brought up into the accreted layers. The presence of enriched carbon causes enhanced nuclear fusion and a Classical Nova explosion.

Both observations and theoretical studies of these explosions imply that more mass is ejected than is accreted. Thus, the WD in a Classical Nova system is losing mass and cannot be a SN Ia progenitor. However, the composition in the nuclear burning region is important and, in new calculations reported here, the consequences to the WD of no mixing of accreted material with core material have been investigated so that the material involved in the explosion has only a Solar composition. WDs with a large range in initial masses and mass accretion rates have been evolved. I find that once sufficient material has been accreted, nuclear burning occurs in all evolutionary sequences and continues until a thermonuclear runaway (TNR) occurs and the WD either ejects a small amount of material or its radius grows to about 10[superscript 12] cm and the evolution is ended. In all cases where mass ejection occurs, the mass of the ejecta is far less than the mass of the accreted material. Therefore, all the WDs are growing in mass. It is also found that the accretion time to explosion can be sufficiently short for a 1.0M[subscript ⊙] WD that recurrent novae can occur on a low mass WD. This mass is lower than typically assumed for the WDs in recurrent nova systems. Finally, the predicted surface temperatures when the WD is near the peak of the explosion imply that only the most massive WDs will be significant X-ray emitters at this time.

Created2014-02-25
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Description

The occurrence of nonliquid and liquid physical states of submicron atmospheric particulate matter (PM) downwind of an urban region in central Amazonia was investigated. Measurements were conducted during two intensive operating periods (IOP1 and IOP2) that took place during the wet and dry seasons of the GoAmazon2014/5 campaign. Air masses

The occurrence of nonliquid and liquid physical states of submicron atmospheric particulate matter (PM) downwind of an urban region in central Amazonia was investigated. Measurements were conducted during two intensive operating periods (IOP1 and IOP2) that took place during the wet and dry seasons of the GoAmazon2014/5 campaign. Air masses representing variable influences of background conditions, urban pollution, and regional- and continental-scale biomass burning passed over the research site. As the air masses varied, particle rebound fraction, an indicator of physical state, was measured in real time at ground level using an impactor apparatus. Micrographs collected by transmission electron microscopy confirmed that liquid particles adhered, while nonliquid particles rebounded. Relative humidity (RH) was scanned to collect rebound curves.

When the apparatus RH matched ambient RH, 95 % of the particles adhered as a campaign average. Secondary organic material, produced for the most part by the oxidation of volatile organic compounds emitted from the forest, produces liquid PM over this tropical forest. During periods of anthropogenic influence, by comparison, the rebound fraction dropped to as low as 60 % at 95 % RH. Analyses of the mass spectra of the atmospheric PM by positive-matrix factorization (PMF) and of concentrations of carbon monoxide, total particle number, and oxides of nitrogen were used to identify time periods affected by anthropogenic influences, including both urban pollution and biomass burning. The occurrence of nonliquid PM at high RH correlated with these indicators of anthropogenic influence. A linear model having as output the rebound fraction and as input the PMF factor loadings explained up to 70 % of the variance in the observed rebound fractions. Anthropogenic influences can contribute to the presence of nonliquid PM in the atmospheric particle population through the combined effects of molecular species that increase viscosity when internally mixed with background PM and increased concentrations of nonliquid anthropogenic particles in external mixtures of anthropogenic and biogenic PM.

ContributorsBateman, Adam P. (Author) / Gong, Zhaoheng (Author) / Harder, Tristan H. (Author) / de Sa, Suzane S. (Author) / Wang, Bingbing (Author) / Castillo, Paulo (Author) / China, Swarup (Author) / Liu, Yingjun (Author) / O'Brien, Rachel E. (Author) / Palm, Brett B. (Author) / Shiu, Hung-Wei (Author) / Cirino, Glauber G. (Author) / Thalman, Ryan (Author) / Adachi, Kouji (Author) / Alexander, M. Lizabeth (Author) / Artaxo, Paulo (Author) / Bertram, Allan K. (Author) / Buseck, Peter (Author) / Gilles, Mary K. (Author) / Jimenez, Jose L. (Author) / Laskin, Alexander (Author) / Manzi, Antonio O. (Author) / Sedlacek, Arthur (Author) / Souza, Rodrigo A. F. (Author) / Wang, Jian (Author) / Zaveri, Rahul (Author) / Martin, Scot T. (Author) / College of Liberal Arts and Sciences (Contributor) / School of Earth and Space Exploration (Contributor) / School of Molecular Sciences (Contributor)
Created2017-02-06
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Description
Current hard X-ray free-electron laser (XFEL) sources can deliver doses to biological macromolecules well exceeding 1 GGy, in timescales of a few tens of femtoseconds. During the pulse, photoionization can reach the point of saturation in which certain atomic species in the sample lose most of their electrons. This electronic

Current hard X-ray free-electron laser (XFEL) sources can deliver doses to biological macromolecules well exceeding 1 GGy, in timescales of a few tens of femtoseconds. During the pulse, photoionization can reach the point of saturation in which certain atomic species in the sample lose most of their electrons. This electronic radiation damage causes the atomic scattering factors to change, affecting, in particular, the heavy atoms, due to their higher photoabsorption cross sections. Here, it is shown that experimental serial femtosecond crystallography data collected with an extremely bright XFEL source exhibit a reduction of the effective scattering power of the sulfur atoms in a native protein. Quantitative methods are developed to retrieve information on the effective ionization of the damaged atomic species from experimental data, and the implications of utilizing new phasing methods which can take advantage of this localized radiation damage are discussed.
ContributorsGalli, L. (Author) / Son, S.-K. (Author) / Klinge, M. (Author) / Bajt, S. (Author) / Barty, A. (Author) / Bean, R. (Author) / Betzel, C. (Author) / Beyerlein, K. R. (Author) / Caleman, C. (Author) / Doak, R. B. (Author) / Duszenko, M. (Author) / Fleckenstein, H. (Author) / Gati, C. (Author) / Hunt, B. (Author) / Kirian, R. A. (Author) / Liang, M. (Author) / Nanao, M. H. (Author) / Nass, K. (Author) / Oberthur, D. (Author) / Redecke, L. (Author) / Shoeman, R. (Author) / Stellato, F. (Author) / Yoon, C. H. (Author) / White, T. A. (Author) / Yefanov, O. (Author) / Spence, John (Author) / Chapman, H. N. (Author) / College of Liberal Arts and Sciences (Contributor) / Department of Physics (Contributor)
Created2015-04-29
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Description
CTB-MPR is a fusion protein between the B subunit of cholera toxin (CTB) and the membrane-proximal region of gp41 (MPR), the transmembrane envelope protein of Human immunodeficiency virus 1 (HIV-1), and has previously been shown to induce the production of anti-HIV-1 antibodies with antiviral functions. To further improve the design

CTB-MPR is a fusion protein between the B subunit of cholera toxin (CTB) and the membrane-proximal region of gp41 (MPR), the transmembrane envelope protein of Human immunodeficiency virus 1 (HIV-1), and has previously been shown to induce the production of anti-HIV-1 antibodies with antiviral functions. To further improve the design of this candidate vaccine, X-ray crystallography experiments were performed to obtain structural information about this fusion protein. Several variants of CTB-MPR were designed, constructed and recombinantly expressed in Escherichia coli. The first variant contained a flexible GPGP linker between CTB and MPR, and yielded crystals that diffracted to a resolution of 2.3 Å, but only the CTB region was detected in the electron-density map. A second variant, in which the CTB was directly attached to MPR, was shown to destabilize pentamer formation. A third construct containing a polyalanine linker between CTB and MPR proved to stabilize the pentameric form of the protein during purification. The purification procedure was shown to produce a homogeneously pure and monodisperse sample for crystallization. Initial crystallization experiments led to pseudo-crystals which were ordered in only two dimensions and were disordered in the third dimension. Nanocrystals obtained using the same precipitant showed promising X-ray diffraction to 5 Å resolution in femtosecond nanocrystallography experiments at the Linac Coherent Light Source at the SLAC National Accelerator Laboratory. The results demonstrate the utility of femtosecond X-ray crystallography to enable structural analysis based on nano/microcrystals of a protein for which no macroscopic crystals ordered in three dimensions have been observed before.
ContributorsLee, Ho-Hsien (Author) / Cherni, Irene (Author) / Yu, HongQi (Author) / Fromme, Raimund (Author) / Doran, Jeffrey (Author) / Grotjohann, Ingo (Author) / Mittman, Michele (Author) / Basu, Shibom (Author) / Deb, Arpan (Author) / Dorner, Katerina (Author) / Aquila, Andrew (Author) / Barty, Anton (Author) / Boutet, Sebastien (Author) / Chapman, Henry N. (Author) / Doak, R. Bruce (Author) / Hunter, Mark (Author) / James, Daniel (Author) / Kirian, Richard (Author) / Kupitz, Christopher (Author) / Lawrence, Robert (Author) / Liu, Haiguang (Author) / Nass, Karol (Author) / Schlichting, Ilme (Author) / Schmidt, Kevin (Author) / Seibert, M. Marvin (Author) / Shoeman, Robert L. (Author) / Spence, John (Author) / Stellato, Francesco (Author) / Weierstall, Uwe (Author) / Williams, Garth J. (Author) / Yoon, Chun Hong (Author) / Wang, Dingjie (Author) / Zatsepin, Nadia (Author) / Hogue, Brenda (Author) / Matoba, Nobuyuki (Author) / Fromme, Petra (Author) / Mor, Tsafrir (Author) / ASU Biodesign Center Immunotherapy, Vaccines and Virotherapy (Contributor) / Department of Chemistry and Biochemistry (Contributor) / College of Liberal Arts and Sciences (Contributor) / School of Life Sciences (Contributor) / Biodesign Institute (Contributor) / Infectious Diseases and Vaccinology (Contributor) / Department of Physics (Contributor)
Created2014-08-20