Multiscale Toxicology- Building the Next Generation Tools for Toxicology

A Cooperative Research and Development Agreement (CRADA) was established between Battelle Memorial Institute (BMI), Pacific Northwest National Laboratory (PNNL), Oak Ridge National Laboratory (ORNL), Brookhaven National Laboratory (BNL), Lawrence Livermore National Laboratory (LLNL) with the goal of combining the analytical and synthetic strengths of the National Laboratories with BMI�s expertise in basic and translational medical research to develop a collaborative pipeline and suite of high throughput and imaging technologies that could be used to provide a more comprehensive understanding of material and drug toxicology in humans. The Multi-Scale Toxicity Initiative (MSTI), consisting of the team members above, was established to coordinate cellular scale, high-throughput in vitro testing, computational modeling and whole animal in vivo toxicology studies between MSTI team members. Development of a common, well-characterized set of materials for testing was identified as a crucial need for the initiative. Two research tracks were established by BMI during the course of the CRADA. The first research track focused on the development of tools and techniques for understanding the toxicity of nanomaterials, specifically inorganic nanoparticles (NPs). ORNL�s work focused primarily on the synthesis, functionalization and characterization of a common set of NPs for dissemination to the participating laboratories. These particles were synthesized to retain the same surface characteristics and size, but to allow visualization using the variety of imaging technologies present across the team. Characterization included the quantitative analysis of physical and chemical properties of the materials as well as the preliminary assessment of NP toxicity using commercially available toxicity screens and emerging optical imaging strategies. Additional efforts examined the development of high-throughput microfluidic and imaging assays for measuring NP uptake, localization, and toxicity in vitro. The second research track within the MSTI CRADA focused on the development of ex vivo animal models for examining druginduced cardiotoxicity. ORNL's role in the second track was limited initially, but was later expanded to include the development of microfluidic platforms that might facilitate the translation of Cardiac 'Microwire' technologies developed at the University of Toronto into a functional platform for drug screening and predictive assessment of cardiotoxicity via highthroughput measurements of contractility. This work was coordinated by BMI with the Centre for the Commercialization of Regenerative Medicine (CCRM) and the University of Toronto (U Toronto). This partnership was expanded and culminated in the submission of proposal to Work for Others (WFO) agencies to explore the development of a broader set of microphysiological systems, a so call human-on-a-chip, that could be used for toxicity screening and the evaluation of bio-threat countermeasures

Controlling antiferromagnetic domains in patterned La0.7Sr0.3FeO3 thin films

Transition metal oxide thin films and heterostructures are promising platforms to achieve full control of the antiferromagnetic (AFM) domain structure in patterned features as needed for AFM spintronic devices. In this work, soft x-ray photoemission electron microscopy was utilized to image AFM domains in micromagnets patterned into La0.7Sr0.3FeO3 (LSFO) thin films and La0.7Sr0.3MnO3 (LSMO)/LSFO superlattices. A delicate balance exists between magnetocrystalline anisotropy, shape anisotropy, and exchange interactions such that the AFM domain structure can be controlled using parameters such as LSFO and LSMO layer thickness, micromagnet shape, and temperature. In LSFO thin films, shape anisotropy gains importance only in micromagnets where at least one extended edge is aligned parallel to an AFM easy axis. In contrast, in the limit of ultrathin LSFO layers in the LSMO/LSFO superlattice, shape anisotropy effects dominate such that the AFM spin axes at micromagnet edges can be aligned along any in-plane crystallographic direction

A Trauma Patient Advocate Is a Valuable Addition to the Multidisciplinary Trauma Team: A Process Improvement Project

Any member of the MDTT could request a consultation and assistance from the TPA on an individual basis after identifying specific patient needs on rounds and meeting the criteria of one or more of the following: patients with complex care issues such as poly trauma, having multiple consultants or service lines involved in the care of the patient, difficult family situations such as dealing with estranged family, difficult family communication due to work and travel schedules, need for assistance in decision-making for discharge planning such as choosing a skilled nursing facility, or need for improved communication and coordination of medical care such as coordinating schedules for combined service line cases. [...]we determined that the addition of the TPA to our busy, urban Level I trauma service was well received by the MDTT, subjectively improved team communication and efficiency, and was perceived to increase patient and family engagement and satisfaction with the trauma experience

A microfluidics and agent-based modeling framework for investigating spatial organization in bacterial colonies: The case of Pseudomonas Aeruginosa amd H1-type VI secretion interactions

The factors leading to changes in the organization of microbial assemblages at fine spatial scales are not well characterized or understood. However, they are expected to guide the succession of community development and function toward specific outcomes that could impact human health and the environment. In this study, we put forward a combined experimental and agent-based modeling framework and use it to interpret unique spatial organization patterns of H1-Type VI secretion system (T6SS) mutants of P. aeruginosa under spatial confinement. We find that key parameters, such as T6SS-mediated cell contact and lysis, spatial localization, relative species abundance, cell density and local concentrations of growth substrates and metabolites are influenced by spatial confinement. The model, written in the accessible programming language NetLogo, can be adapted to a variety of biological systems of interest and used to simulate experiments across a broad parameter space. It was implemented and run in a high-throughput mode by deploying it across multiple CPUs, with each simulation representing an individual well within a high-throughput microwell array experimental platform. The microfluidics and agent-based modeling framework we present in this paper provides an effective means by which to connect experimental studies in microbiology to model development. The work demonstrates progress in coupling experimental results to simulation while also highlighting potential sources of discrepancies between real-world experiments and idealized models

Microstencils to generate defined, multi-species patterns of bacteria

Citation: Timm, C. M., Hansen, R. R., Doktycz, M. J., Retterer, S. T., & Pelletier, D. A. (2015). Microstencils to generate defined, multi-species patterns of bacteria. Biomicrofluidics, 9(6). doi:10.1063/1.4935938Microbial communities are complex heterogeneous systems that are influenced by physical and chemical interactions with their environment, host, and community members. Techniques that facilitate the quantitative evaluation of how microscale organization influences the morphogenesis of multispecies communities could provide valuable insights into the dynamic behavior and organization of natural communities, the design of synthetic environments for multispecies culture, and the engineering of artificial consortia. In this work, we demonstrate a method for patterning microbes into simple arrangements that allow the quantitative measurement of growth dynamics as a function of their proximity to one another. The method combines parylene-based liftoff techniques with microfluidic delivery to simultaneously pattern multiple bacterial species with high viability using low-cost, customizable methods. Quantitative measurements of bacterial growth for two competing isolates demonstrate that spatial coordination can play a critical role in multispecies growth and structure. © 2015 AIP Publishing LLC

Modes of Multiple Star Formation

This paper argues that star forming environments should be classified into finer divisions than the traditional isolated and clustered modes. Using the observed set of galactic open clusters and theoretical considerations regarding cluster formation, we estimate the fraction of star formation that takes place within clusters. We find that less than 10% of the stellar population originates from star forming regions destined to become open clusters, confirming earlier estimates. The smallest clusters included in the observational surveys (having at least N=100 members) roughly coincide with the smallest stellar systems that are expected to evolve as clusters in a dynamical sense. We show that stellar systems with too few members N < N_\star have dynamical relaxation times that are shorter than their formation times (1-2 Myr), where the critical number of stars N_\star \approx 100. Our results suggest that star formation can be characterized by (at least) three principal modes: I. isolated singles and binaries, II. groups (N<N_\star), and III. clusters (N>N_\star). Many -- if not most -- stars form through the intermediate mode in stellar groups with 10<N<100. Such groups evolve and disperse much more rapidly than open clusters; groups also have a low probability of containing massive stars and are unaffected by supernovae and intense ultraviolet radiation fields. Because of their short lifetimes and small stellar membership, groups have relatively little effect on the star formation process (on average) compared to larger open clusters.Comment: accepted to The Astrophysical Journa

The formation of permanent soft binaries in dispersing clusters

Wide, fragile binary stellar systems are found in the galactic field, and have recently been noted in the outskirts of expanding star clusters in numerical simulations. Energetically soft, with semi-major axes exceeding the initial size of their birth cluster, it is puzzling how these binaries are created and preserved. We provide an interpretation of the formation of these binaries that explains the total number formed and their distribution of energies. A population of weakly bound binaries can always be found in the cluster, in accordance with statistical detailed balance, limited at the soft end only by the current size of the cluster and whatever observational criteria are imposed. At any given time, the observed soft binary distribution is predominantly a snapshot of a transient population. However, there is a constantly growing population of long-lived soft binaries that are removed from the detailed balance cycle due to the changing density and velocity dispersion of an expanding cluster. The total number of wide binaries that form, and their energy distribution, are insensitive to the cluster population; the number is approximately one per cluster. This suggests that a population composed of many dissolved small-N clusters will more efficiently populate the field with wide binaries than that composed of dissolved large-N clusters. Locally such binaries are present at approximately the 2% level; thus the production rate is consistent with the field being populated by clusters with a median of a few hundred stars rather than a few thousand.Comment: 10 pages, accepted to MNRA

Controlling condensation and frost growth with chemical micropatterns

Citation: Boreyko, J. B., Hansen, R. R., Murphy, K. R., Nath, S., Retterer, S. T., & Collier, C. P. (2016). Controlling condensation and frost growth with chemical micropatterns. Scientific Reports, 6, 15. doi:10.1038/srep19131In-plane frost growth on chilled hydrophobic surfaces is an inter-droplet phenomenon, where frozen droplets harvest water from neighboring supercooled liquid droplets to grow ice bridges that propagate across the surface in a chain reaction. To date, no surface has been able to passively prevent the in-plane growth of ice bridges across the population of supercooled condensate. Here, we demonstrate that when the separation between adjacent nucleation sites for supercooled condensate is properly controlled with chemical micropatterns prior to freezing, inter-droplet ice bridging can be slowed and even halted entirely. Since the edge-to-edge separation between adjacent supercooled droplets decreases with growth time, deliberately triggering an early freezing event to minimize the size of nascent condensation was also necessary. These findings reveal that inter-droplet frost growth can be passively suppressed by designing surfaces to spatially control nucleation sites and by temporally controlling the onset of freezing events

Development of transparent microwell arrays for optical monitoring and dissection of microbial communities

Citation: Halsted, M., Wilmoth, J. L., Briggs, P. A., Hansen, R. R., Briggs, D. P., Timm, A. C., & Retterer, S. T. (2016). Development of transparent microwell arrays for optical monitoring and dissection of microbial communities. Journal of Vacuum Science & Technology B, 34(6), 5. doi:10.1116/1.4962739Microbial communities are incredibly complex systems that dramatically and ubiquitously influence our lives. They help to shape our climate and environment, impact agriculture, drive business, and have a tremendous bearing on healthcare and physical security. Spatial confinement, as well as local variations in physical and chemical properties, affects development and interactions within microbial communities that occupy critical niches in the environment. Recent work has demonstrated the use of silicon based microwell arrays, combined with parylene lift-off techniques, to perform both deterministic and stochastic assembly of microbial communities en masse, enabling the high-throughput screening of microbial communities for their response to growth in confined environments under different conditions. The implementation of a transparent microwell array platform can expand and improve the imaging modalities that can be used to characterize these assembled communities. Here, the fabrication and characterization of a next generation transparent microwell array is described. The transparent arrays, comprised of SU-8 patterned on a glass coverslip, retain the ability to use parylene lift-off by integrating a low temperature atomic layer deposition of silicon dioxide into the fabrication process. This silicon dioxide layer prevents adhesion of the parylene material to the patterned SU-8, facilitating dry lift-off, and maintaining the ability to easily assemble microbial communities within the microwells. These transparent microwell arrays can screen numerous community compositions using continuous, high resolution, imaging. The utility of the design was successfully demonstrated through the stochastic seeding and imaging of green fluorescent protein expressing Escherichia coli using both fluorescence and brightfield microscopies. (C) 2016 American Vacuum Society

The Evolution of Wide Binary Stars

We study the orbital evolution of wide binary stars in the solar neighborhood due to gravitational perturbations from passing stars. We include the effects of the Galactic tidal field and continue to follow the stars after they become unbound. For a wide variety of initial semi-major axes and formation times, we find that the number density (stars per unit logarithmic interval in projected separation) exhibits a minimum at a few times the Jacobi radius r_J, which equals 1.7 pc for a binary of solar-mass stars. The density peak interior to this minimum arises from the primordial distribution of bound binaries, and the exterior density, which peaks at \sim 100--300 pc separation, arises from formerly bound binaries that are slowly drifting apart. The exterior peak gives rise to a significant long-range correlation in the positions and velocities of disk stars that should be detectable in large astrometric surveys such as GAIA that can measure accurate three-dimensional distances and velocities.Comment: 36 pages, 9 figures, accepted by MNRAS, typos correcte