Investigating COMT Influence on the Proactive-Reactive Stress Coping Axis in Zebrafish

Individuals of the same species often display differences in correlated suites of behaviors which are made conspicuous when challenges – stressful, fear-inducing, etc. – are presented. In many species, a specific suite of behaviors (risk-aversion, aggression, exploration, learning, and memory) characterize an alternative set of stress coping styles (proactive and reactive). Such behaviors are regulated in the brain by specific neurotransmitters along with proteins that regulate them. One neurotransmitter regulator protein, catechol-O-methyltransferase (COMT) shows higher baseline whole-brain expression in proactive relative to reactive animals. However, it is not known whether its expression is a cause or a consequence of the stress coping style. In this ongoing study, I am testing the hypothesis that alteration of comta activity will lead to changes in stress coping behavior. Specifically, I attempted to knockout comtain zebrafish embryos using CRISPR/Cas9 with the intent of comparing stress-related behaviors and comtabrain-gene expression between mutant and control individuals in each stress coping style. I predict knocking out comtawill result in proactive zebrafish displaying similar stress-related behaviors as their reactive conspecifics and that reactive individuals will display elevated stress-related behaviors compared to controls. Across 155 breeding attempts, I found that approximately 23.75% (285/1200) of treatment embryos survived to 72 hours post-fertilization. High resolution melt analysis revealed 78.25% (18/23) to have variants in the target region, of which 11/18 (84.6%) showed successful mutations by Sanger sequencing. I intend to generate an F1 line in which to compare stress-related behaviors and comtabrain-gene expression by in situ hybridization

Towards Identifying and closing Gaps in Assurance of autonomous Road vehicleS - a collection of Technical Notes Part 1

This report provides an introduction and overview of the Technical Topic Notes (TTNs) produced in the Towards Identifying and closing Gaps in Assurance of autonomous Road vehicleS (Tigars) project. These notes aim to support the development and evaluation of autonomous vehicles. Part 1 addresses: Assurance-overview and issues, Resilience and Safety Requirements, Open Systems Perspective and Formal Verification and Static Analysis of ML Systems. Part 2: Simulation and Dynamic Testing, Defence in Depth and Diversity, Security-Informed Safety Analysis, Standards and Guidelines

Towards Identifying and closing Gaps in Assurance of autonomous Road vehicleS - a collection of Technical Notes Part 2

This report provides an introduction and overview of the Technical Topic Notes (TTNs) produced in the Towards Identifying and closing Gaps in Assurance of autonomous Road vehicleS (Tigars) project. These notes aim to support the development and evaluation of autonomous vehicles. Part 1 addresses: Assurance-overview and issues, Resilience and Safety Requirements, Open Systems Perspective and Formal Verification and Static Analysis of ML Systems. This report is Part 2 and discusses: Simulation and Dynamic Testing, Defence in Depth and Diversity, Security-Informed Safety Analysis, Standards and Guidelines

The Infocus Hard X-ray Telescope: Pixellated CZT Detector/Shield Performance and Flight Results

The CZT detector on the Infocus hard X-ray telescope is a pixellated solid-state device capable of imaging spectroscopy by measuring the position and energy of each incoming photon. The detector sits at the focal point of an 8m focal length multilayered grazing incidence X-ray mirror which has significant effective area between 20--40 keV. The detector has an energy resolution of 4.0keV at 32keV, and the Infocus telescope has an angular resolution of 2.2 arcminute and a field of view of about 10 arcminutes. Infocus flew on a balloon mission in July 2001 and observed Cygnus X-1. We present results from laboratory testing of the detector to measure the uniformity of response across the detector, to determine the spectral resolution, and to perform a simple noise decomposition. We also present a hard X-ray spectrum and image of Cygnus X-1, and measurements of the hard X-ray CZT background obtained with the SWIN detector on Infocus.Comment: To appear in the proceedings of the SPIE conference "Astronomical Telescopes and Instrumentation", #4851-116, Kona, Hawaii, Aug. 22-28, 2002. 12 pages, 9 figure

Novel cyclic di-GMP effectors of the YajQ protein family control bacterial virulence

Bis-(3 ',5 ') cyclic di-guanylate (cyclic di-GMP) is a key bacterial second messenger that is implicated in the regulation of many critical processes that include motility, biofilm formation and virulence. Cyclic di-GMP influences diverse functions through interaction with a range of effectors. Our knowledge of these effectors and their different regulatory actions is far from complete, however. Here we have used an affinity pull-down assay using cyclic di-GMP-coupled magnetic beads to identify cyclic di-GMP binding proteins in the plant pathogen Xanthomonas campestris pv. campestris (Xcc). This analysis identified XC_3703, a protein of the YajQ family, as a potential cyclic di-GMP receptor. Isothermal titration calorimetry showed that the purified XC_3703 protein bound cyclic di-GMP with a high affinity (K-d similar to 2 mu M). Mutation of XC_3703 led to reduced virulence of Xcc to plants and alteration in biofilm formation. Yeast two-hybrid and far-western analyses showed that XC_3703 was able to interact with XC_2801, a transcription factor of the LysR family. Mutation of XC_2801 and XC_3703 had partially overlapping effects on the transcriptome of Xcc, and both affected virulence. Electromobility shift assays showed that XC_3703 positively affected the binding of XC_2801 to the promoters of target virulence genes, an effect that was reversed by cyclic di-GMP. Genetic and functional analysis of YajQ family members from the human pathogens Pseudomonas aeruginosa and Stenotrophomonas maltophilia showed that they also specifically bound cyclic di-GMP and contributed to virulence in model systems. The findings thus identify a new class of cyclic di-GMP effector that regulates bacterial virulence

Approximating the inspiral of test bodies into Kerr black holes

We present a new approximate method for constructing gravitational radiation driven inspirals of test-bodies orbiting Kerr black holes. Such orbits can be fully described by a semi-latus rectum $p$, an eccentricity $e$, and an inclination angle $\iota$; or, by an energy $E$, an angular momentum component $L_z$, and a third constant $Q$. Our scheme uses expressions that are exact (within an adiabatic approximation) for the rates of change ($\dot{p}$, $\dot{e}$, $\dot{\iota}$) as linear combinations of the fluxes ($\dot{E}$, $\dot{L_z}$, $\dot{Q}$), but uses quadrupole-order formulae for these fluxes. This scheme thus encodes the exact orbital dynamics, augmenting it with approximate radiation reaction. Comparing inspiral trajectories, we find that this approximation agrees well with numerical results for the special cases of eccentric equatorial and circular inclined orbits, far more accurate than corresponding weak-field formulae for ($\dot{p}$, $\dot{e}$, $\dot{\iota}$). We use this technique to study the inspiral of a test-body in inclined, eccentric Kerr orbits. Our results should be useful tools for constructing approximate waveforms that can be used to study data analysis problems for the future LISA gravitational-wave observatory, in lieu of waveforms from more rigorous techniques that are currently under development.Comment: 15 pages, 5 figures, submitted to PR

Theory-assisted determination of nano-rippling and impurities in atomic resolution images of angle-mismatched bilayer graphene

Ripples and impurity atoms are universally present in 2D materials, limiting carrier mobility, creating pseudo–magnetic fields, or affecting the electronic and magnetic properties. Scanning transmission electron microscopy (STEM) generally provides picometer-level precision in the determination of the location of atoms or atomic 'columns' in the in-image plane (xy plane). However, precise atomic positions in the z-direction as well as the presence of certain impurities are difficult to detect. Furthermore, images containing moiré patterns such as those in angle-mismatched bilayer graphene compound the problem by limiting the determination of atomic positions in the xy plane. Here, we introduce a reconstructive approach for the analysis of STEM images of twisted bilayers that combines the accessible xy coordinates of atomic positions in a STEM image with density-functional-theory calculations. The approach allows us to determine all three coordinates of all atomic positions in the bilayer and establishes the presence and identity of impurities. The deduced strain-induced rippling in a twisted bilayer graphene sample is consistent with the continuum model of elasticity. We also find that the moiré pattern induces undulations in the z direction that are approximately an order of magnitude smaller than the strain-induced rippling. A single substitutional impurity, identified as nitrogen, is detected. The present reconstructive approach can, therefore, distinguish between moiré and strain-induced effects and allows for the full reconstruction of 3D positions and atomic identities

A luminous blue kilonova and an off-axis jet from a compact binary merger at z=0.1341

The recent discovery of a faint gamma-ray burst (GRB) coincident with the gravitational wave (GW) event GW 170817 revealed the existence of a population of low-luminosity short duration gamma-ray transients produced by neutron star mergers in the nearby Universe. These events could be routinely detected by existing gamma-ray monitors, yet previous observations failed to identify them without the aid of GW triggers. Here we show that GRB150101B was an analogue of GRB170817A located at a cosmological distance. GRB 150101B was a faint short duration GRB characterized by a bright optical counterpart and a long-lived X-ray afterglow. These properties are unusual for standard short GRBs and are instead consistent with an explosion viewed off-axis: the optical light is produced by a luminous kilonova component, while the observed X-rays trace the GRB afterglow viewed at an angle of ~13 degrees. Our findings suggest that these properties could be common among future electromagnetic counterparts of GW sources.Comment: 28 pages, 8 figures, 2 tables. Accepted for publicatio