\u3ci\u3ePseudomonas syringae\u3c/i\u3e Type III Secretion System Targeting Signals and Novel Effectors Studied with a Cya Translocation Reporter

Pseudomonas syringae pv. tomato strain DC3000 is a pathogen of tomato and Arabidopsis. The hrp-hrcencoded type III secretion system (TTSS), which injects bacterial effector proteins (primarily called Hop or Avr proteins) into plant cells, is required for pathogenicity. In addition to being regulated by the HrpL alternative sigma factor, most avr or hop genes encode proteins with N termini that have several characteristic features, including (i) a high percentage of Ser residues, (ii) an aliphatic amino acid (Ile, Leu, or Val) or Pro at the third or fourth position, and (iii) a lack of negatively charged amino acids within the first 12 residues. Here, the well-studied effector AvrPto was used to optimize a calmodulin-dependent adenylate cyclase (Cya) reporter system for Hrp-mediated translocation of P. syringae TTSS effectors into plant cells. This system includes a cloned P. syringae hrp gene cluster and the model plant Nicotiana benthamiana. Analyses of truncated AvrPto proteins fused to Cya revealed that the N-terminal 16 amino acids and/or codons of AvrPto are sufficient to direct weak translocation into plant cells and that longer N-terminal fragments direct progressively stronger translocation. AvrB, tested because it is poorly secreted in cultures by the P. syringae Hrp system, was translocated into plant cells as effectively as AvrPto. The translocation of several DC3000 candidate Hop proteins was also examined by using Cya as a reporter, which led to identification of three new intact Hop proteins, designated HopPtoQ, HopPtoT1, and HopPtoV, as well as two truncated Hop proteins encoded by the naturally disrupted genes hopPtoS4::tnpA and hopPtoAG::tnpA. We also confirmed that HopPtoK, HopPtoC, and AvrPphEPto are translocated into plant cells. These results increased the number of Hrp system-secreted proteins in DC3000 to 40. Although most of the newly identified Hop proteins possess N termini that have the same features as the N termini of previously described Hop proteins, HopPtoV has none of these characteristics. Our results indicate that Cya should be a useful reporter for exploring multiple aspects of the Hrp system in P. syringae

A large outbreak of Clostridium difficile‐associated disease with an unexpected proportion of deaths and colectomies at a teaching hospital following increased Fluoroquinolone use

BACKGROUND AND OBJECTIVE: Fluoroquinolones have not been frequently implicated as a cause of Clostridium difficile outbreaks. Nosocornial C. difficile infections increased from 2.7 to 6.8 cases per 1,000 discharges (P < .001). During the first 2 years of the outbreak, there were 253 nosocomial C difficile infections; of these, 26 resulted in colectomy and 18 resulted in death. We conducted an investigation of a large C. difficile outbreak in our hospital to identify risk factors and characterize the outbreak

Middle to Late Pleistocene stability of the central East Antarctic Ice Sheet at the head of Law Glacier

Past behavior of outlet glaciers draining the East Antarctic Ice Sheet (EAIS) remains unresolved prior to Marine Isotope Stage 2 (MIS2). Study of blue ice moraines provides a relatively untapped approach to understand former EAIS activity. We focus on a blue ice moraine near Mount Achernar in the central Transantarctic Mountains, at the edge of the polar plateau. The well-preserved moraine consists of quasi-continuous or hummocky sediment ridges that form on top of upward-flowing, sublimating ice along the margin of Law Glacier. 10Be, 26Al, and 3He cosmogenic nuclide ages on boulders from the ridges are coherent and in general are progressively older with distance from the relatively clean ice of the Law Glacier margin. Moraines closest to the Law Glacier margin postdate MIS2; farther away, they date to the last glacial cycle, and with more distance they are hundreds of thousands of years old. We conclude that cosmogenic dating of some blue ice moraines can provide age limits for changes at the heads of outlet glaciers that drain the central East Antarctic Ice Sheet, including prior to MIS2. Furthermore, the geomorphological, cosmogenic nuclide, and sedimentological evidence imply that the East Antarctic polar plateau adjacent to the central Transantarctic Mountains has been relatively stable for at least 200 k.y

ATA Practice Guidelines for Live, On-Demand Primary and Urgent Care

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/140286/1/tmj.2015.0008.pd

Microbial Monitoring of Common Opportunistic Pathogens by Comparing Multiple Real-Time PCR Platforms for Potential Space Applications

Because the International Space Station is a closed environment with rotations of astronauts and equipment that each introduce their own microbial flora, it is necessary to monitor the air, surfaces, and water for microbial contamination. Current microbial monitoring includes labor- and time-intensive methods to enumerate total bacterial and fungal cells, with limited characterization, during in-flight testing. Although this culture-based method is sufficient for monitoring the International Space Station, on future long-duration missions more detailed characterization will need to be performed during flight, as sample return and ground characterization may not be available. At a workshop held in 2011 at NASA's Johnson Space Center to discuss alternative methodologies and technologies suitable for microbial monitoring for these long-term exploration missions, molecular-based methodologies such as polymerase chain reaction (PCR) were recommended. In response, a multi-center (Marshall Space Flight Center, Johnson Space Center, Jet Propulsion Laboratory, and Kennedy Space Center) collaborative research effort was initiated to explore novel commercial-off-the-shelf hardware options for space flight environmental monitoring. The goal was to evaluate quantitative or semi-quantitative PCR approaches for low-cost in-flight rapid identification of microorganisms that could affect crew safety. The initial phase of this project identified commercially available platforms that could be minimally modified to perform nominally in microgravity. This phase was followed by proof-of-concept testing of the highest qualifying candidates with a universally available challenge organism, Salmonella enterica. The analysis identified two technologies that were able to perform sample-to-answer testing with initial cell sample concentrations between 50 and 400 cells. In addition, the commercial systems were evaluated for initial flight safety and readiness

A Space for Co-constructing Counter Stories Under Surveillance

Using our experiences as members of a participatory action research committee (from the City University of New York Graduate Center and the Bedford Hills Correctional Facility) documenting the impact of college in a maximum security prison, this essay illustrates the power of Participatory Action Research in the construction of counter stories. We raise for discussion a set of theoretical, methodological and ethical challenges that emerged from the co-production of counter stories under surveillance: the creation of a critical space for producing \u27counter knowledge\u27; the co-mingling of counter and dominant discourses, the negotiation of power over and within research in prison, and the opening of a dialogue between counter stories and public policy makers

Blueberry Progress Reports

The 1983 edition of the Blueberry Progress Reports was prepared for the Maine Blueberry Commission and the University of Maine Blueberry Advisory Committee by researchers with the Maine Agricultural Experiment Station and Maine Cooperative Extension Service at the University of Maine, Orono. Projects in this report include: 1. Introduction 2. Forest Tent Caterpillar in Blueberries 3. Control, Biology, and Ecology of Insects Affecting Lowbush Blueberries 4. Blueberry Diseases: Incidence and Control 5. Physiology and Culture of the Lowbush Blueberry 6. Weed Control in Lowbush Blueberry Fields 7. Product Development of Lowbush Blueberrie

The Safety Attitudes Questionnaire: psychometric properties, benchmarking data, and emerging research

BACKGROUND: There is widespread interest in measuring healthcare provider attitudes about issues relevant to patient safety (often called safety climate or safety culture). Here we report the psychometric properties, establish benchmarking data, and discuss emerging areas of research with the University of Texas Safety Attitudes Questionnaire. METHODS: Six cross-sectional surveys of health care providers (n = 10,843) in 203 clinical areas (including critical care units, operating rooms, inpatient settings, and ambulatory clinics) in three countries (USA, UK, New Zealand). Multilevel factor analyses yielded results at the clinical area level and the respondent nested within clinical area level. We report scale reliability, floor/ceiling effects, item factor loadings, inter-factor correlations, and percentage of respondents who agree with each item and scale. RESULTS: A six factor model of provider attitudes fit to the data at both the clinical area and respondent nested within clinical area levels. The factors were: Teamwork Climate, Safety Climate, Perceptions of Management, Job Satisfaction, Working Conditions, and Stress Recognition. Scale reliability was 0.9. Provider attitudes varied greatly both within and among organizations. Results are presented to allow benchmarking among organizations and emerging research is discussed. CONCLUSION: The Safety Attitudes Questionnaire demonstrated good psychometric properties. Healthcare organizations can use the survey to measure caregiver attitudes about six patient safety-related domains, to compare themselves with other organizations, to prompt interventions to improve safety attitudes and to measure the effectiveness of these interventions