Characterization of the Extracellular Proteome of a Natural Microbial Community with an Integrated Mass Spectrometric / Bioinformatic Approach

Proteomics comprises the identification and characterization of the complete suite of expressed proteins in a given cell, organism or community. The coupling of high performance liquid chromatography (LC) with high throughput mass spectrometry (MS) has provided the foundation for current proteomic progression. The transition from proteomic analysis of a single cultivated microbe to that of natural microbial assemblages has required significant advancement in technology and has provided greater biological understanding of microbial community diversity and function. To enhance the capabilities of a mass spectrometric based proteomic analysis, an integrated approach combining bioinformatics with analytical preparations and experimental data collection was developed and applied. This has resulted in a deep characterization of the extracellular fraction of a community of microbes thriving in an acid mine drainage system. Among the notable features of this relatively low complexity community, they exist in a solution that is highly acidic (pH \u3c 1) and hot (temperature \u3e 40°C), with molar concentrations of metals. The extracellular fraction is of particular interest due to the potential to identify and characterize novel proteins that are critical for survival and interactions with the harsh environment. The following analyses have resulted in the specific identification and characterization of novel extracellular proteins. In order to more accurately identify which proteins are present in the extracellular space, a combined computational prediction and experimental identification of the extracellular fraction was performed. Among the hundreds of proteins identified, a highly abundant novel cytochrome was targeted and ultimately characterized through high performance MS. In order to achieve deep proteomic coverage of the extracellular fraction, a metal affinity based protein enrichment utilizing seven different metals was developed and employed resulting in novel protein identifications. A combined top down and bottom up analysis resulted in the characterization of the intact molecular forms of extracellular proteins, including the identification of post-translational modifications. Finally, in order to determine the effectiveness of current MS methodologies, a software package was designed to characterize the \u3e 100,000 mass spectra collected during an MS experiment, revealing that specific optimizations in the LC, MS and protein sequence database have a significant impact on proteomic depth

Recent Changes in Farm Structure: A Canada-U.S. Comparison

Following a series of bilateral and multilateral agreements, the past two decades have seen increased trade and investment liberalization between Canada and the United States in the agri-food sector. Changes in trade policy are one of several paths by which farm structure can change. This increased liberalization, together with the largest drop in Canadian farm numbers recorded by the Census of Agriculture in thirty years, has provided the impetus to review some aspects of farm structure. In particular, this article presents the latest Canadian and U.S. data on the number of farms by sales class, the concentration of sales and other production-related variables, and the distribution of income and receipts. We explore whether significant changes in the latter two elements of farm structure have occurred during this period of trade and investment liberalization.Crop Production/Industries, Farm Management, International Relations/Trade,

New perspectives in ecosystem services science as instruments to understand environmental securities

As societal demand for food, water and other life-sustaining resources grows, the science of ecosystem services (ES) is seen as a promising tool to improve our understanding, and ultimately the management, of increasingly uncertain supplies of critical goods provided or supported by natural ecosystems. This promise, however, is tempered by a relatively primitive understanding of the complex systems supporting ES, which as a result are often quantified as static resources rather than as the dynamic expression of human-natural systems. This article attempts to pinpoint the minimum level of detail that ES science needs to achieve in order to usefully inform the debate on environmental securities, and discusses both the state of the art and recent methodological developments in ES in this light. We briefly review the field of ES accounting methods and list some desiderata that we deem necessary, reachable and relevant to address environmental securities through an improved science of ES. We then discuss a methodological innovation that, while only addressing these needs partially, can improve our understanding of ES dynamics in data-scarce situations. The methodology is illustrated and discussed through an application related to water security in the semi-arid landscape of the Great Ruaha river of Tanzania. © 2014 The Author(s) Published by the Royal Society. All rights reserved

A High Temperature Calcium Vapor Cell for Spectroscopy on the 4s^2 1S0 to 4s4p 3P1 Intercombination Line

We have demonstrated a high temperature vapor cell for absorption spectroscopy on the Ca intercombination line. The cell uses a dual chamber design to achieve the high temperatures necessary for an optically dense vapor while avoiding the necessity of high temperature vacuum valves and glass-to-metal seals. We have observed over 50 percent absorption in a single pass through the cell. Although pressure broadening in the cell prevented us from performing saturated-absorption spectroscopy, the broadening resulted in higher signal-to-noise ratios by allowing us to probe the atoms with intensities much greater than the 0.2 uW/cm^2 saturation intensity of the unbroadened transition.Comment: 5 pages, 2 figures, submitted to Rev. Sci. Instru

The Status of Women In Missouri: A Comprehensive Report of Leading Indicators and Findings

The status of women in Missouri reflects the status of women throughout the United States. Missouri women have the same opportunities, but also face similar challenges. The Institute of Public Policy, in concert with an academic advisory committee at the University of Missouri, has worked diligently to examine existing data, analyze actionable steps at the state level, and understand women's successes and challenges through a series of focus groups across the state. These focus groups gave the researchers the opportunity to hear from very engaged and diverse groups of women, and also a group of men

Trait-like Resting-State Brain Oscillations Predict Subsequent Problem-Solving Strategies

People use different processing strategies to solve problems. Previous research distinguishes between solving problems by analysis, that is, in a conscious, deliberate manner, versus by insight, in which the solution appears abruptly in awareness (the "Aha" phenomenon) after a period of unconscious processing. Prior work provides little evidence whether the tendency to solve problems using one or the other of these strategies constitutes a stable, trait-like cognitive style. We tested this hypothesis by assessing whether individuals evince a consistent preference for a particular solving strategy across days and types of problems and whether these cognitive styles have neural correlates. We recorded participants' resting-state electroencephalograms (EEGs) on 4 occasions, approximately once per week. At the end of the third and fourth sessions, participants attempted to solve a series of short verbal problems (compound remote associates during session 3 and anagrams during session 4). Based on participants' trial-by-trial reports of the manner in which they solved anagram problems, individuals were categorized as predominantly relying on an insight or an analytic solving strategy. The resting-state EEGs of these groups, recorded during previous sessions were compared. Participants in the analytic group showed greater EEG beta power over midline and right inferior-frontal regions compared to insightful participants; participants in the insightful group showed greater beta power over left superior parietal cortex compared to those in the analytic group. Group differences in solving strategy and resting-state EEGs assessed with anagram problems generalized to the compound remote associates problems. Overall, these results demonstrate the behavioral and neural consistency of these cognitive styles over both time and type of problem. The finding that insightful solvers exhibited a lower ratio of frontal to parietal neural activity supports the hypothesis that insightfulness results from chronic relative frontal hypoactivation and concomitant parietal disinhibition whereas analytic solvers exhibit chronic relative frontal hyperactivation and parietal inhibition.Ph.D., Psychology -- Drexel University, 201

Some parameters affecting respiratory control in Azotobacter vinelandii membranes

AbstractRCI :respiratory control index (state 3/state 4 rate of oxygen uptake)m-C1 CCP: carbonylcyanide m-chlorophenyl hydrazoneHQNO : 2-n-heptyl-4-hydroxyquinoline-N-oxidePIPES: piperazine-N,N′-bis 2-ethane sulphonic aci

Effects of Teaching Household Actions to Address Ocean Acidification on Student Knowledge and Attitudes

Ocean acidification (OA), the change in ocean chemistry due to increasing concentrations of anthropogenic carbon dioxide in the atmosphere, is an environmental problem that is an active area of scientific research yet remains largely outside of the public’s awareness. It is often assumed that if we raise OA awareness, then the public will support and take action to help mitigate the problem. This research project examines this assumption through the lens of educating high school students about OA. The research included three phases: (i) review of existing teaching resources on OA, (ii) development and refinement of a new OA curriculum based on strengths and gaps identified during the review process, and (iii) a longitudinal experiment testing the impacts to knowledge and attitudes of two approaches to teaching about OA. This study has implications for those engaging in OA outreach and education efforts specifically, and for environmental education campaigns in general. During this study, we found that at least 90 teaching resources focused on OA are already available. These resources provide teachers with multiple approaches to teaching about OA, yet do not adequately address the multiple impacts of OA nor teach students about ways to help address the problem. We developed our own curriculum that underwent four rounds of revisions before appearing in the form presented here. Our experiment found that our teaching intervention increased knowledge but that attitudinal changes, when present, did not persist over time. Despite this lack of attitude change, student attitudes were generally sufficient to support mitigation actions

In-situ synchrotron x-ray diffraction study of quartz deformation using the D-DIA apparatus

Deformation is a rock’s mechanical response to external parameters such as temperature and pressure. Knowledge of deformation is necessary to understand geodynamic processes. The behavior of rocks and minerals during deformation depends on the behavior of individual grains within those minerals as they interact. The mineral we are studying is quartz, which we chose since it is very well studied and is highly abundant. We use high-pressure high temperature experiments to simulate conditions of Earth’s interior and the D-DIA apparatus combined with synchrotron x-ray diffraction allows us to record data directly during the process of deformation. The data we collect is used to determine the macroscopic sample stress and tells us also how individual grains react to that stress depending on their orientation within the minera