Increasing the Analytical Accessibility of Multishell and Diffusion Spectrum Imaging Data Using Generalized Q-Sampling Conversion

Many diffusion MRI researchers, including the Human Connectome Project (HCP), acquire data using multishell (e.g., WU-Minn consortium) and diffusion spectrum imaging (DSI) schemes (e.g., USC-Harvard consortium). However, these data sets are not readily accessible to high angular resolution diffusion imaging (HARDI) analysis methods that are popular in connectomics analysis. Here we introduce a scheme conversion approach that transforms multishell and DSI data into their corresponding HARDI representations, thereby empowering HARDI-based analytical methods to make use of data acquired using non-HARDI approaches. This method was evaluated on both phantom and in-vivo human data sets by acquiring multishell, DSI, and HARDI data simultaneously, and comparing the converted HARDI, from non-HARDI methods, with the original HARDI data. Analysis on the phantom shows that the converted HARDI from DSI and multishell data strongly predicts the original HARDI (correlation coefficient > 0.9). Our in-vivo study shows that the converted HARDI can be reconstructed by constrained spherical deconvolution, and the fiber orientation distributions are consistent with those from the original HARDI. We further illustrate that our scheme conversion method can be applied to HCP data, and the converted HARDI do not appear to sacrifice angular resolution. Thus this novel approach can benefit all HARDI-based analysis approaches, allowing greater analytical accessibility to non-HARDI data, including data from the HCP

The Roles of Membrane Rafts in CD32A-Mediated Phagocytosis

Membrane rafts are highly dynamic heterogeneous sterol- and sphingolipid-rich micro-domains on cell surfaces. They are generally believed to provide residency for cell surface molecules (e.g., adhesion and signaling molecules) and scaffolding to facilitate the functions of these molecules such as membrane trafficking, receptor transport, cell signaling, and endocytosis.
The governing, or overall hypothesis, for this project is that membrane rafts provide residency for Fc[gamma]RIIA (CD32A) on K562 cells, and that by doing so they provide a platform from which Fc[gamma]RIIA initiate or carry out their functions, which include migration, signaling, phagocytic synapse formation, and internalization of IgG opsonized targets.
Using immuno-fluorescent laser scanning confocal microscopy and reflection interference microscopy (RIM), we studied the spatial and temporal distributions of membrane rafts and surface receptors, signaling molecules, and cell organelles during the formation of phagocytic contact areas. K562 cells, which naturally express CD32A, a cell surface receptor for the Fc portion of Immuno-globulin G(IgG), was chosen as a model for neutrophils. An opsonized target was modeled using a glass supported lipid bilayer reconstituted with IgG. CD32A was found to cluster and co-localize with membrane rafts. Placing the K562 cells on the lipid bilayer triggered a process of contact area formation that includes binding between receptors and ligands, their recruitment to the contact area, a concurrent membrane raft movement to and concentration in the contact area, and transport of CD32A, IgG, and membrane rafts to the Golgi complex. Characterization of these processes was performed using agents known to disrupt detergent resistant membranes (DRMs), dissolve actin microfilaments, and inhibit myosin motor activity, which abolished the CD32A clusters and prevented the contact area formation. 
The relevance to phagocytosis of contact area formation between K562 cells and lipid bilayers was demonstrated using micro-beads coated with a lipid bilayer reconstituted with IgG as the opsonized target instead of the glass supported planar lipid bilayer. Disruption of membrane rafts, salvation of the actin cytoskeleton, and inhibition of myosin II activity were found to inhibit phagocytosis.
These data suggest membrane rafts play several important roles in CD32A mediated phagocytosis including pre-clustering CD32A, transport of CD32A to the phagocytic cup, and transport of the opsonized target towards the Golgi complex. Here we have provided evidence that membrane rafts serve as platforms which are used to cluster CD32A and transport CD32A along the actin cytoskeleton to the site of phagocytic synapse formation thus allowing for the quick assembly of a phagocytic synapse.
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Public Relations and Promotion in Film: How It’s Done and Why It’s Important

Films have been huge revenue drivers in the entertainment industry. However, not all films succeed with high profit margins. Success can be contributed to aspects such as the writing, the production quality, or the acting within a film. These aspects contribute to how audiences, critics and media receive a movie. Unfortunately, these aspects typically overshadow one of the most important functions contributing to a film’s success: promotion and public relations. For my capstone, I researched how films are promoted, branded, and distributed through public relations functions. Research largely relied on secondary research, with primary research added for deeper concepts. After basic knowledge was attained, the theories and concepts were applied in an additional creative component included for the capstone. This component attempted to create promotional materials in line with industry requirements, giving readers and the writer a tangible example to see film communications theory

Mapping Topographic Structure in White Matter Pathways with Level Set Trees

Fiber tractography on diffusion imaging data offers rich potential for describing white matter pathways in the human brain, but characterizing the spatial organization in these large and complex data sets remains a challenge. We show that level set trees---which provide a concise representation of the hierarchical mode structure of probability density functions---offer a statistically-principled framework for visualizing and analyzing topography in fiber streamlines. Using diffusion spectrum imaging data collected on neurologically healthy controls (N=30), we mapped white matter pathways from the cortex into the striatum using a deterministic tractography algorithm that estimates fiber bundles as dimensionless streamlines. Level set trees were used for interactive exploration of patterns in the endpoint distributions of the mapped fiber tracks and an efficient segmentation of the tracks that has empirical accuracy comparable to standard nonparametric clustering methods. We show that level set trees can also be generalized to model pseudo-density functions in order to analyze a broader array of data types, including entire fiber streamlines. Finally, resampling methods show the reliability of the level set tree as a descriptive measure of topographic structure, illustrating its potential as a statistical descriptor in brain imaging analysis. These results highlight the broad applicability of level set trees for visualizing and analyzing high-dimensional data like fiber tractography output

Third-codon transversion rate-based _Nymphaea_ basal angiosperm phylogeny -- concordance with developmental evidence

Flowering plants (angiosperms) appeared on Earth rather suddenly approximately 130 million years ago and underwent a massive expansion in the subsequent 10-12 million years. Current molecular phylogenies have predominantly identified _Amborella_, followed by _Nymphaea_ (water lilies) or _Amborella_ plus _Nymphaea_, in the ANITA clade (_Amborella_, Nymphaeales, Illiciaceae, Trimeniaceae and Austrobaileyaceae) as the earliest angiosperm. However, developmental studies suggest that the earliest angiosperm had a 4-cell/4-nucleus female gametophyte and a diploid endosperm represented by _Nymphaea_, suggesting that _Amborella_, having an 8-cell/9-nucleus female gametophyte and a triploid endosperm, cannot be representative of the basal angiosperm. This evolution-development discordance is possibly caused by erroneous inference based on phylogenetic signals with low neutrality and/or high saturation. Here we show that the 3rd codon transversion (P3Tv), with high neutrality and low saturation, is a robust high-resolution phylogenetic signal for such divergences and that the P3Tv-based land plant phylogeny cautiously identifies _Nymphaea_, followed by _Amborella_, as the most basal among the angiosperm species examined in this study. This P3Tv-based phylogeny contributes insights to the origin of angiosperms with concordance to fossil and stomata development evidence

A Sustainability Evaluation and Dynamic Modeling Tool for Landscape and Urban Planning Policy Scenarios

Incorporating sustainability principles into urban development is often complex involving strong interaction between ecosystem components and development goals. As identified in the Brundtland Commissions report (UN, 1987), sustainability has gained much attention in planning aimed at balancing current needs without depleting resources and ecological services available for future generations. While the decision-making process is embedded in a social framework, political sustainability depends on collective decisions and citizens’ preferences related to public policies (Munda, 2006; Webster, 1998). In recent decade, the sustainability concept has been adopted in landscape and urban planning. Specific approaches include assessing abiotic, biotic, and cultural (ABC) resources in the landscapes for goals setting, defining and resolving spatial conflicts, developing and evaluating alternative scenarios, selecting a landscape plan, employing adaptive management, and closing the planning process loop by continuous interdisciplinary and public involvement (Ahern, 1999). There is a need for a simple and effective tool to model interaction among landscape components, to facilitate the decision-making process in the planning framework, and to evaluate alternative scenarios for sustainability. Urban policies are often path-dependent with past decisions having consequences that constrain allocation of resources in later times. In addition, the policies are selfreinforcing (Woodlief, 1998) and interacting with ecosystem services of ABC resources over time. For example, when cities implemented urban renewal policy in the 1940s, hundreds of low-income neighborhood blocks were cleared and thousands of acres of wetlands were filled for building housing and highway systems. The consequences of past decisions as observed today include inequitable distribution affecting low income and minority communities and extensive degradation of the environment. The varying impacts of a policy decision are not only dynamic over time but also involving interplay between the landscape and society. To develop and assess landscape and urban plans with sustainability criteria, there is a critical need for policy evaluation under alternative planning scenarios. Assessment of the state of resources over time can inform planners on shifts in ecosystem conditions in landscapes under a particular planning scenario. This will also enable planners to anticipate changes in the ecosystem health and mitigate negative impacts on resource allocation. Balancing multiple goals, incorporating constraints facing communities, and including public participation are essential for developing effective sustainable plans. A dynamic modelling and participatory approach can inform the public on landscape interactions, the nature of trade-offs between scenarios, and long-term trends in sustainability criteria. For example, modeling could reveal that sustainability may be decreasing over time as one resource is rapidly depleted under a planning scenario and negatively impact on other resources. In order to assess and incorporate trade-off relationships into the planning process with continuous public participation, we propose a dynamic ecosystem and policy evaluation framework for landscape and urban planning

Cylindrical illumination with angular coupling for whole-prostate photoacoustic tomography

Current diagnosis of prostate cancer relies on histological analysis of tissue samples acquired by biopsy, which could benefit from real-time identification of suspicious lesions. Photoacoustic tomography has the potential to provide real-time targets for prostate biopsy guidance with chemical selectivity, but light delivered from the rectal cavity has been unable to penetrate to the anterior prostate. To overcome this barrier, a urethral device with cylindrical illumination is developed for whole-prostate imaging, and its performance as a function of angular light coupling is evaluated with a prostate-mimicking phantom

Enhanced killing of antibiotic-resistant bacteria enabled by massively parallel combinatorial genetics

New therapeutic strategies are needed to treat infections caused by drug-resistant bacteria, which constitute a major growing threat to human health. Here, we use a high-throughput technology to identify combinatorial genetic perturbations that can enhance the killing of drug-resistant bacteria with antibiotic treatment. This strategy, Combinatorial Genetics En Masse (CombiGEM), enables the rapid generation of high-order barcoded combinations of genetic elements for high-throughput multiplexed characterization based on next-generation sequencing. We created ~34,000 pairwise combinations of Escherichia coli transcription factor (TF) overexpression constructs. Using Illumina sequencing, we identified diverse perturbations in antibiotic-resistance phenotypes against carbapenem-resistant Enterobacteriaceae. Specifically, we found multiple TF combinations that potentiated antibiotic killing by up to 10[superscript 6]-fold and delivered these combinations via phagemids to increase the killing of highly drug-resistant E. coli harboring New Delhi metallo-beta-lactamase-1. Moreover, we constructed libraries of three-wise combinations of transcription factors with >4 million unique members and demonstrated that these could be tracked via next-generation sequencing. We envision that CombiGEM could be extended to other model organisms, disease models, and phenotypes, where it could accelerate massively parallel combinatorial genetics studies for a broad range of biomedical and biotechnology applications, including the treatment of antibiotic-resistant infections.National Institutes of Health (U.S.) (New Innovator Award DP2 OD008435)United States. Office of Naval ResearchEllison Medical Foundation (New Scholar in Aging Award)Henry L. and Grace Doherty Charitable Foundatio

Co-targeting HGF/cMET Signaling with MEK Inhibitors in Metastatic Uveal Melanoma.

Patients with metastatic uveal melanoma usually die within 1 year of diagnosis, emphasizing an urgent need to develop new treatment strategies. The liver is the most common site of metastasis. Mitogen-activated protein kinase kinase (MEK) inhibitors improve survival in V600 BRAF-mutated cutaneous melanoma patients but have limited efficacy in patients with uveal melanoma. Our previous work showed that hepatocyte growth factor (HGF) signaling elicits resistance to MEK inhibitors in metastatic uveal melanoma. In this study, we demonstrate that expression of two BH3-only family proteins, Bim-EL and Bmf, contributes to HGF-mediated resistance to MEK inhibitors. Targeting HGF/cMET signaling with LY2875358, a neutralizing and internalizing anti-cMET bivalent antibody, and LY2801653, a dual cMET/RON inhibitor, overcomes resistance to trametinib provided by exogenous HGF and by conditioned medium from primary hepatic stellate cells. We further determined that activation of PI3Kα/γ/δ isoforms mediates the resistance to MEK inhibitors by HGF. Combination of LY2801653 with trametinib decreases AKT phosphorylation and promotes proapoptotic PARP cleavage in metastatic uveal melanoma explants. Together, our data support the notion that selectively blocking cMET signaling or PI3K isoforms in metastatic uveal melanoma may break the intrinsic resistance to MEK inhibitors provided by factors from stromal cells in the liver