Will Oslo be the Next Ottawa? The Cluster-munitions Debate

More than a decade has passed since the monumental Ottawa Mine Ban Convention was opened for signature in December 1997. Now, with the adoption of the text of the Convention on Cluster Munitions in May 2008, the global community is closer than ever to an international agreement prohibiting the use of cluster munitions. A review of the key issues underpinning the debate on cluster munitions follows

DRONES FOR IMPROVING TRAFFIC SAFETY IN RITI COMMUNITIES IN WASHINGTON STATE

Transportation and traffic safety is a primary concern in Rural, Isolated, Tribal, or Indigenous (RITI) communities in Washington (WA) State. Parallel to this, while emerging technologies (e.g., connected/autonomous vehicles, drones) have been developed and tested in addressing traffic safety issues, they are often not widely shared in RITI communities for various reasons. Compared with other technological advances, drone technologies have been rapidly improved and can be flexibly applied to multiple fields, including engineering, agriculture and disaster managements. The goal of this study is to explore and synthesize the opportunities, challenges and scenarios that drone technologies can assist to resolve traffic safety related issues and concerns in RITI communities. Through the outreach activities with the outer Pacific Coast in WA state, it is found that the principal concern within these communities are disaster management and mitigation since they are facing the threat of coastal erosion, earthquake and tsunami. Thus, the emergency management and hazard mitigation becomes the major way to further explore drone applications in the selected communities. To achieve this, we reviewed the current state of the drone technologies, conducted surveys from National Guard and coastal communities in WA, including City of Westport, South Beach Region, Grays Harbor County, Shoalwater Bay Tribe, and Quinault Indian Nation, to better understand their current needs, challenges and issues. Ultimately, recommendations of drone applications under specific scenarios are provided based upon the integration of drone technologies with community safety needs.TABLE OF CONTENTS Disclaimer....................................................................................................................................................... i Technical Report Documentation Page ........................................................................................................ ii SI* (Modern Metric) Conversion Factors ..................................................................................................... iii List of Figures ................................................................................................................................................ v List of Tables ................................................................................................................................................ vi Executive Summary ....................................................................................................................................... 1 CHAPTER 1. Introduction .......................................................................................................................... 2 1.1. General Background ...................................................................................................................... 2 1.2. Problem Statement ....................................................................................................................... 3 CHAPTER 2. Literature review of drone technology and Application ....................................................... 5 2.1. State of the Art Drone Technologies............................................................................................. 5 2.2. Practice of drones in Washington State ........................................................................................ 6 CHAPTER 3. Outreach activities ................................................................................................................ 7 3.1. Community Engagement for Coastal Resilience Studio ................................................................ 8 3.1.1. Westport/South Beach Workshop – 11/2018 (27) ............................................................. 10 3.2. WA National Guard Workshop – 6/2019 .................................................................................... 11 CHAPTER 4. Online survey ...................................................................................................................... 14 CHAPTER 5. results and recommendationS ............................................................................................ 15 5.1. Outreach Activity Findings .......................................................................................................... 15 5.1.1. Transportation and Traffic Safety in Westport ................................................................... 15 5.1.2. Learning from WA National Guard Workshop .................................................................... 17 5.2. Online Survey Results.................................................................................................................. 18 5.3. Recommendation of drone applications .................................................................................... 18 CHAPTER 6. conclusion and future study ............................................................................................... 24 Reference .................................................................................................................................................... 25 Appendix A: UAV (Drones) technologies ...................................................................................................... 1 Appendix B: Workshop report for Westport Coastal Resilience................................................................... 1 Appendix C: Online Survey ............................................................................................................................ 1 Appendix D: Executive Summary of Westport Costal Resilience Report ...................................................... 1 Appendix E: Transportation Chapter of Westport Costal Resilience Report ................................................

Photoaffinity labeling with cholesterol analogues precisely maps a cholesterol-binding site in voltage-dependent anion channel-1

Voltage-dependent anion channel-1 (VDAC1) is a highly regulated β-barrel membrane protein that mediates transport of ions and metabolites between the mitochondria and cytosol of the cell. VDAC1 co-purifies with cholesterol and is functionally regulated by cholesterol, among other endogenous lipids. Molecular modeling studies based on NMR observations have suggested five cholesterol-binding sites in VDAC1, but direct experimental evidence for these sites is lacking. Here, to determine the sites of cholesterol binding, we photolabeled purified mouse VDAC1 (mVDAC1) with photoactivatable cholesterol analogues and analyzed the photolabeled sites with both top-down mass spectrometry (MS), and bottom-up MS paired with a clickable, stable isotope-labeled tag, FLI-tag. Using cholesterol analogues with a diazirine in either the 7 position of the steroid ring (LKM38) or the aliphatic tail (KK174), we mapped a binding pocket in mVDAC1 localized to Thr83 and Glu73, respectively. When Glu73 was mutated to a glutamine, KK174 no longer photolabeled this residue, but instead labeled the nearby Tyr62 within this same binding pocket. The combination of analytical strategies employed in this work permits detailed molecular mapping of a cholesterol-binding site in a protein, including an orientation of the sterol within the site. Our work raises the interesting possibility that cholesterol-mediated regulation of VDAC1 may be facilitated through a specific binding site at the functionally important Glu73 residue

Conformational transitions of the sodium-dependent sugar transporter, vSGLT.

Sodium-dependent transporters couple the flow of Na+ ions down their electrochemical potential gradient to the uphill transport of various ligands. Many of these transporters share a common core structure composed of a five-helix inverted repeat and deliver their cargo utilizing an alternating-access mechanism. A detailed characterization of inward-facing conformations of the Na+-dependent sugar transporter from Vibrio parahaemolyticus (vSGLT) has previously been reported, but structural details on additional conformations and on how Na+ and ligand influence the equilibrium between other states remains unknown. Here, double electron-electron resonance spectroscopy, structural modeling, and molecular dynamics are utilized to deduce ligand-dependent equilibria shifts of vSGLT in micelles. In the absence and presence of saturating amounts of Na+, vSGLT favors an inward-facing conformation. Upon binding both Na+ and sugar, the equilibrium shifts toward either an outward-facing or occluded conformation. While Na+ alone does not stabilize the outward-facing state, gating charge calculations together with a kinetic model of transport suggest that the resting negative membrane potential of the cell, absent in detergent-solubilized samples, may stabilize vSGLT in an outward-open conformation where it is poised for binding external sugars. In total, these findings provide insights into ligand-induced conformational selection and delineate the transport cycle of vSGLT

Investigation of Drone Applications to Improve Traffic Safety in RITI Communities

Transportation and traffic safety is a primary concern among the Rural, Isolated, Tribal, or Indigenous (RITI) communities in the U.S. Although emerging technologies (e.g., connected and autonomous vehicles, drones) have been developed and tested in addressing traffic safety issues, they are often not widely shared in RITI communities for various reasons. This research aims to explore, understand, and synthesize the opportunities and challenges of applying drone technologies to alleviate or resolve traffic safety and emergency related issues within RITI communities. The project team first sent out online surveys to communities on the outer Pacific coast of Washington State and selected the City of Westport as the study area based on the feedback. A pilot study using drones for mapping and sensing in Westport was then conducted, followed by two community meetings to explore potential drone applications. With the three outreach activities, it was found that the current need in the communities was education on drones, including training for remote pilot certification (drone license) and drone operations. Findings of this research will help guide the project team to set up specific drone-related programs in the Westport area in future research

Stochastic steps in secondary active sugar transport.

Secondary active transporters, such as those that adopt the leucine-transporter fold, are found in all domains of life, and they have the unique capability of harnessing the energy stored in ion gradients to accumulate small molecules essential for life as well as expel toxic and harmful compounds. How these proteins couple ion binding and transport to the concomitant flow of substrates is a fundamental structural and biophysical question that is beginning to be answered at the atomistic level with the advent of high-resolution structures of transporters in different structural states. Nonetheless, the dynamic character of the transporters, such as ion/substrate binding order and how binding triggers conformational change, is not revealed from static structures, yet it is critical to understanding their function. Here, we report a series of molecular simulations carried out on the sugar transporter vSGLT that lend insight into how substrate and ions are released from the inward-facing state of the transporter. Our simulations reveal that the order of release is stochastic. Functional experiments were designed to test this prediction on the human homolog, hSGLT1, and we also found that cytoplasmic release is not ordered, but we confirmed that substrate and ion binding from the extracellular space is ordered. Our findings unify conflicting published results concerning cytoplasmic release of ions and substrate and hint at the possibility that other transporters in the superfamily may lack coordination between ions and substrate in the inward-facing state

Protonation state of glutamate 73 regulates the formation of a specific dimeric association of mVDAC1.

The voltage-dependent anion channel (VDAC) is the most abundant protein in the outer mitochondrial membrane and constitutes the primary pathway for the exchange of ions and metabolites between the cytosol and the mitochondria. There is accumulating evidence supporting VDAC's role in mitochondrial metabolic regulation and apoptosis, where VDAC oligomerization has been implicated with these processes. Herein, we report a specific pH-dependent dimerization of murine VDAC1 (mVDAC1) identified by double electron-electron resonance and native mass spectrometry. Intermolecular distances on four singly spin-labeled mVDAC1 mutants were used to generate a model of the low-pH dimer, establishing the presence of residue E73 at the interface. This dimer arrangement is different from any oligomeric state previously described, and it forms as a steep function of pH with an apparent pKa of 7.4. Moreover, the monomer-dimer equilibrium affinity constant was determined using native MS, revealing a nearly eightfold enhancement in dimerization affinity at low pH. Mutation of E73 to either alanine or glutamine severely reduces oligomerization, demonstrating the role of protonated E73 in enhancing dimer formation. Based on these results, and the known importance of E73 in VDAC physiology, VDAC dimerization likely plays a significant role in mitochondrial metabolic regulation and apoptosis in response to cytosolic acidification during cellular stress

DRONE TECHNOLOGY EDUCATION IN RURAL, ISOLATED, TRIBAL AND INDIGENOUS (RITI) COMMUNITIES

Transportation and traffic safety is a primary concern within Rural, Isolated, Tribal and Indigenous (RITI) communities in Washington State. Emerging technologies such as connected and autonomous vehicles, sensors and drones have been tested and developed to improve traffic safety, but these advances have largely been limited to urban areas. This project identified opportunities and challenges of adopting drone technologies in RITI communities, and explored context-sensitive applications to traffic safety and related goals. In three phases, the team conducted community workshops, online surveys and other outreach activities with state and county agencies responsible for emergency management and crisis response in coastal Tribal and non-tribal communities; a planning studio and Comprehensive Plan Update for the City of Westport and its surrounding South Beach community straddling two rural counties and including the Shoalwater Bay Indian Tribe; and a pilot educational program with the School District that serves it. To be effective in rural contexts, adoption of drone technology depends on a broadening of local skill development and needs to target diverse community goals. In short, it needs to be broadly embedded in the community. Taking this sociotechnical approach, we focused on long-term workforce development and designed and implemented an after-school program (October 2021 – June 2022) for Ocosta Junior High School students. The course taught students how to assemble and pilot drones and apply them to a variety of practical needs including public works inspection, search and rescue, and environmental monitoring of coastal flooding

Transfer of ion binding site from ether-à-go-go to Shaker: Mg2+ binds to resting state to modulate channel opening

In ether-à-go-go (eag) K+ channels, extracellular divalent cations bind to the resting voltage sensor and thereby slow activation. Two eag-specific acidic residues in S2 and S3b coordinate the bound ion. Residues located at analogous positions are ∼4 Å apart in the x-ray structure of a Kv1.2/Kv2.1 chimera crystallized in the absence of a membrane potential. It is unknown whether these residues remain in proximity in Kv1 channels at negative voltages when the voltage sensor domain is in its resting conformation. To address this issue, we mutated Shaker residues I287 and F324, which correspond to the binding site residues in eag, to aspartate and recorded ionic and gating currents in the presence and absence of extracellular Mg2+. In I287D+F324D, Mg2+ significantly increased the delay before ionic current activation and slowed channel opening with no readily detectable effect on closing. Because the delay before Shaker opening reflects the initial phase of voltage-dependent activation, the results indicate that Mg2+ binds to the voltage sensor in the resting conformation. Supporting this conclusion, Mg2+ shifted the voltage dependence and slowed the kinetics of gating charge movement. Both the I287D and F324D mutations were required to modulate channel function. In contrast, E283, a highly conserved residue in S2, was not required for Mg2+ binding. Ion binding affected activation by shielding the negatively charged side chains of I287D and F324D. These results show that the engineered divalent cation binding site in Shaker strongly resembles the naturally occurring site in eag. Our data provide a novel, short-range structural constraint for the resting conformation of the Shaker voltage sensor and are valuable for evaluating existing models for the resting state and voltage-dependent conformational changes that occur during activation. Comparing our data to the chimera x-ray structure, we conclude that residues in S2 and S3b remain in proximity throughout voltage-dependent activation

Statistics for Fission-Track Thermochronology

This chapter introduces statistical tools to extract geologically meaningful information from fission-track (FT) data using both the external detector and LA-ICP-MS methods. The spontaneous fission of 238U is a Poisson process resulting in large single-grain age uncertainties. To overcome this imprecision, it is nearly always necessary to analyse multiple grains per sample. The degree to which the analytical uncertainties can explain the observed scatter of the single-grain data can be visually assessed on a radial plot and objectively quantified by a chi-square test. For sufficiently low values of the chi-square statistic (or sufficiently high p values), the pooled age of all the grains gives a suitable description of the underlying ‘true’ age population. Samples may fail the chi-square test for several reasons. A first possibility is that the true age population does not consist of a single discrete age component, but is characterised by a continuous range of ages. In this case, a ‘random effects’ model can constrain the true age distribution using two parameters: the ‘central age’ and the ‘(over)dispersion’. A second reason why FT data sets might fail the chi-square test is if they are underlain by multimodal age distributions. Such distributions may consist of discrete age components, continuous age distributions, or a combination of the two. Formalised statistical tests such as chi-square can be useful in preventing overfitting of relatively small data sets. However, they should be used with caution when applied to large data sets (including length measurements) which generate sufficient statistical ‘power’ to reject any simple yet geologically plausible hypothesis