The Cellular Environment Stabilizes Adenine Riboswitch RNA Structure

There are large differences between the intracellular environment and the conditions widely used to study RNA structure and function in vitro. To assess the effects of the crowded cellular environment on RNA, we examined the structure and ligand-binding function of the adenine riboswitch aptamer domain in healthy, growing Escherichia coli cells at single-nucleotide resolution on the minute timescale using SHAPE. The ligand-bound aptamer structure is essentially the same in cells and in buffer at 1 mM Mg2+, the approximate Mg2+ concentration we measured in cells. In contrast, the in-cell conformation of the ligand-free aptamer is much more similar to the fully folded ligand-bound state. Even adding high Mg2+ concentrations to the buffer used for in vitro analyses did not yield the conformation observed for the free aptamer in cells. The cellular environment thus stabilizes the aptamer significantly more than does Mg2+ alone. Our results show that the intracellular environment has a large effect on RNA structure that ultimately favors highly organized conformations

Environmental quality objective of principles and standards for planning, The

August 1975.Submitted to the Water Resources Planning Fellowship Steering Committee, Colorado State University, in fulfillment of requirements for NR 795 Special Study in Planning.Includes bibliographical references (pages 62-63)

Proliferated LEO Architecture Enabling Beyond Line of Sight Fires (pLEO BLOS Fires)

NPS NRP Project PosterChief of Naval Operations, ADM Gilday, and Commandant of the Marine Corps, Gen Berger, directed modernization efforts across the Naval Services. The concept of Naval Operational Architecture (NOA) enables the development of additional beyond-line-of-sight (BLOS) targeting and fires delivered from widely distributed points within and outside a near-peer's weapons engagement zones (WEZ). Research into proliferated low-earth orbiting communications satellites (pLEO) reveal an intersection with the Joint All Domain Command and Control (JADC2) priority and the Navy and Marine components leveraging efforts such as Task Force Overmatch (DEVSECOPS). There is an immediate need to develop requirements and analyze alternate architectures for delivering BLOS precision fires in future contested environments in the context of Great Power Competition. Faculty experience with space mission architecture design will be leveraged to advise students in achieving the following objectives: 1) Research and develop operational, functional objective and threshold requirements for a proliferated LEO systems within the JADC2 framework to enable BLOS precision fires. 2) Analyze alternative architectures for the requirements which enable BLOS precision fires in a contested environment. A phased approach will be used to meet the research objectives. First, a capabilities or system requirements-like document will be generated to include validation criteria and in-phase lessons learned. Second, standard aerospace modeling/simulation techniques will be used to define the mission and architecture requirements and analyze the alternatives for architectures enabling BLOS precision fires through a contested environment to include in-phase lessons learned. Third, individual student thesis research will occur to completion/graduation of the students and any final closing actions through the remainder of the project period.Marine Corps Capabilities Development Directorate (CDD), DC CD&IHQMC Combat Development and Integration (CD&I)This research is supported by funding from the Naval Postgraduate School, Naval Research Program (PE 0605853N/2098). https://nps.edu/nrpChief of Naval Operations (CNO)Approved for public release. Distribution is unlimited.

Proliferated LEO Architecture Enabling Beyond Line of Sight Fires (pLEO BLOS Fires)

NPS NRP Executive SummaryChief of Naval Operations, ADM Gilday, and Commandant of the Marine Corps, Gen Berger, directed modernization efforts across the Naval Services. The concept of Naval Operational Architecture (NOA) enables the development of additional beyond-line-of-sight (BLOS) targeting and fires delivered from widely distributed points within and outside a near-peer's weapons engagement zones (WEZ). Research into proliferated low-earth orbiting communications satellites (pLEO) reveal an intersection with the Joint All Domain Command and Control (JADC2) priority and the Navy and Marine components leveraging efforts such as Task Force Overmatch (DEVSECOPS). There is an immediate need to develop requirements and analyze alternate architectures for delivering BLOS precision fires in future contested environments in the context of Great Power Competition. Faculty experience with space mission architecture design will be leveraged to advise students in achieving the following objectives: 1) Research and develop operational, functional objective and threshold requirements for a proliferated LEO systems within the JADC2 framework to enable BLOS precision fires. 2) Analyze alternative architectures for the requirements which enable BLOS precision fires in a contested environment. A phased approach will be used to meet the research objectives. First, a capabilities or system requirements-like document will be generated to include validation criteria and in-phase lessons learned. Second, standard aerospace modeling/simulation techniques will be used to define the mission and architecture requirements and analyze the alternatives for architectures enabling BLOS precision fires through a contested environment to include in-phase lessons learned. Third, individual student thesis research will occur to completion/graduation of the students and any final closing actions through the remainder of the project period.Marine Corps Capabilities Development Directorate (CDD), DC CD&IHQMC Combat Development and Integration (CD&I)This research is supported by funding from the Naval Postgraduate School, Naval Research Program (PE 0605853N/2098). https://nps.edu/nrpChief of Naval Operations (CNO)Approved for public release. Distribution is unlimited.

Serine Proteolytic Pathway Activation Reveals an Expanded Ensemble of Wound Response Genes in Drosophila

After injury to the animal epidermis, a variety of genes are transcriptionally activated in nearby cells to regenerate the missing cells and facilitate barrier repair. The range and types of diffusible wound signals that are produced by damaged epidermis and function to activate repair genes during epidermal regeneration remains a subject of very active study in many animals. In Drosophila embryos, we have discovered that serine protease function is locally activated around wound sites, and is also required for localized activation of epidermal repair genes. The serine protease trypsin is sufficient to induce a striking global epidermal wound response without inflicting cell death or compromising the integrity of the epithelial barrier. We developed a trypsin wounding treatment as an amplification tool to more fully understand the changes in the Drosophila transcriptome that occur after epidermal injury. By comparing our array results with similar results on mammalian skin wounding we can see which evolutionarily conserved pathways are activated after epidermal wounding in very diverse animals. Our innovative serine protease-mediated wounding protocol allowed us to identify 8 additional genes that are activated in epidermal cells in the immediate vicinity of puncture wounds, and the functions of many of these genes suggest novel genetic pathways that may control epidermal wound repair. Additionally, our data augments the evidence that clean puncture wounding can mount a powerful innate immune transcriptional response, with different innate immune genes being activated in an interesting variety of ways. These include puncture-induced activation only in epidermal cells in the immediate vicinity of wounds, or in all epidermal cells, or specifically in the fat body, or in multiple tissues

Nondestructive Evaluation of Standing Trees With a Stress Wave Method

The primary objective of this study was to investigate the usefulness of a stress wave technique for evaluating wood strength and stiffness of young-growth western hemlock and Sitka spruce in standing trees. A secondary objective was to determine if the effects of silvicultural practices on wood quality can be identified using this technique. Stress wave measurements were conducted on 168 young-growth western hemlock and Sitka spruce trees. After in situ measurements, a 0.61-m-long bole section in the test span was taken from 56 felled trees to obtain small, clear wood specimens. Stress wave and static bending tests were then performed on these specimens to determine strength and stiffness. Results of this study indicate that in situ stress wave measurements could provide relatively accurate and reliable information that would enable nondestructive evaluation of wood properties in standing trees. The mean values of stress wave speed and dynamic modulus of elasticity for trees agreed with those determined from small, clear wood specimens. Statistical regression analyses revealed good correlations between stress wave properties of trees and static bending properties of small, clear wood specimens obtained from the trees. Regression models showed statistical significance at the 0.01 confidence level. Results of this study also demonstrate that the effect of silvicultural practices on wood properties can be identified with the stress wave properties of trees. This indicates that this nondestructive stress wave technique can be used to track property changes in trees and help determine how forests could be managed to meet desired wood and fiber qualities

Progress achieved in restricting the marketing of high-fat, sugary and salty food and beverage products to children

In May 2010, 192 Member States endorsed Resolution WHA63.14 to restrict the marketing of food and non-alcoholic beverage products high in saturated fats, trans fatty acids, free sugars and/or salt to children and adolescents globally. We examined the actions taken between 2010 and early 2016 – by civil society groups, the World Health Organization (WHO) and its regional offices, other United Nations (UN) organizations, philanthropic institutions and transnational industries – to help decrease the prevalence of obesity and diet-related noncommunicable diseases among young people. By providing relevant technical and policy guidance and tools to Member States, WHO and other UN organizations have helped protect young people from the marketing of branded food and beverage products that are high in fat, sugar and/or salt. The progress achieved by the other actors we investigated appears variable and generally less robust. We suggest that the progress being made towards the full implementation of Resolution WHA63.14 would be accelerated by further restrictions on the marketing of unhealthy food and beverage products and by investing in the promotion of nutrient-dense products. This should help young people meet government-recommended dietary targets. Any effective strategies and actions should align with the goal of WHO to reduce premature mortality from noncommunicable diseases by 25% by 2025 and the aim of the UN to ensure healthy lives for all by 2030

Mass fractionation of noble gases in synthetic methane hydrate : implications for naturally occurring gas hydrate dissociation

This paper is not subject to U.S. copyright. The definitive version was published in Chemical Geology 339 (2013): 242-250, doi:10.1016/j.chemgeo.2012.09.033.As a consequence of contemporary or longer term (since 15 ka) climate warming, gas hydrates in some settings may presently be dissociating and releasing methane and other gases to the ocean–atmosphere system. A key challenge in assessing the impact of dissociating gas hydrates on global atmospheric methane is the lack of a technique able to distinguish between methane recently released from gas hydrates and methane emitted from leaky thermogenic reservoirs, shallow sediments (some newly thawed), coal beds, and other sources. Carbon and deuterium stable isotopic fractionation during methane formation provides a first-order constraint on the processes (microbial or thermogenic) of methane generation. However, because gas hydrate formation and dissociation do not cause significant isotopic fractionation, a stable isotope-based hydrate-source determination is not possible. Here, we investigate patterns of mass-dependent noble gas fractionation within the gas hydrate lattice to fingerprint methane released from gas hydrates. Starting with synthetic gas hydrate formed under laboratory conditions, we document complex noble gas fractionation patterns in the gases liberated during dissociation and explore the effects of aging and storage (e.g., in liquid nitrogen), as well as sampling and preservation procedures. The laboratory results confirm a unique noble gas fractionation pattern for gas hydrates, one that shows promise in evaluating modern natural gas seeps for a signature associated with gas hydrate dissociation.Partial support for this research was provided by Interagency Agreements DE-FE0002911 and DE-NT0006147 between the U.S. Geological Survey Gas Hydrates Project and the U.S. Department of Energy's Methane Hydrates Research and Development Program

The causative–instrumental syncretism

Causative and applicative morphemes have been central in work on the morphosyntax of argument structure. However, several genetically unrelated languages use a single, syncretic form for both functions, which complicates the traditional view that a causative adds a new subject and an applicative adds a new object. In this paper, I propose an analysis of a morphological syncretism found in the Bantu language Kinyarwanda where the morphological causative and instrumental applicative are both realized by the morpheme –ish. I argue for Kinyarwanda that both causation and the introduction of an instrument are analyzable as two outgrowths of the same semantic notion of introducing a new link into the causal chain described by the verb. The different causative and instrumental readings derive from underspecification of the position of the new link in the causal chain, although its placement is restricted via general constraints on possible event types as well as constraints on verb meaning and argument realization. This analysis provides an explanation for the presence of the causative–instrumental syncretism as well as provides insight into the interface between verb meaning and valency-changing morphology

Design and Construction Solutions in the Accurate Realization of NCSX Magnetic Fields

The National Compact Stellarator Experiment, NCSX, is being constructed at the Princeton Plasma Physics Laboratory (PPPL) in partnership with the Oak Ridge national Laboratory. The goal of NCSX is to provide the understanding necessary to develop an attractive, disruption free, steady state compact stellaratorbased reactor design. This paper describes the recently revised designs of the critical interfaces between the modular coils, the construction solutions developed to meet assembly tolerances, and the recently revised trim coil system that provides the required compensation to correct for the “as built” conditions and to allow flexibility in the disposition of as-built conditions. In May, 2008, the sponsor decided to terminate the NCSX project due to growth in the project’s cost and schedule estimates. However significant technical challenges in design and construction were overcome, greatly reducing the risk in the remaining work to complete the project