Persister Cell Control Mechanisms in Uropathogenic Escherichia coli

Persister cells are a subpopulation of bacteria that demonstrate high tolerance to antibiotics, but revert to sensitivity after antibiotics are removed. The mechanism for induction of the persister cell state and antibiotic tolerance is not completely understood but likely occurs through the establishment of dormancy. Some of the suggested mechanisms for persister cell formation in Escherichia coli include: toxin­-antitoxin systems, starvation, gene regulation by (p)ppGpp, and stochastic formation. In our study we examine the mechanisms behind persistence in the E. coli strain CFT073, a uropathogenic isolate, which forms elevated levels of persisters compared to the laboratory strain MG1655. However, CFT073 lacks many of the type II toxin­-antitoxin pairs associated with modulating persister cell formation. In addition, global stress response is impaired in the CFT073 isolate used in these studies, since it contains a five base pair insertion in the rpoS gene, which encodes RpoS, the master regulator of the global stress response. This insertion results in the expression of truncated RpoS. We compared several CFT073 strains mutated at the rpoS locus for the ability to form persister cells, as well as a strain deleted for Lon, a protease important for modulating toxin­-antitoxin function. To identify additional regulators of persister cell formation in CFT073, we performed minitransposon mutagenesis to isolate mutant strains, and screened mutants for persister cell formation. As many pathogenic bacterial species, including CFT073, cause recalcitrant infections attributable to persister cell activity, these studies will identify additional mechanisms underlying the development of bacterial persistence in these organisms

Effects of the 8 psia / 32% O2 Atmosphere on the Human in the Spaceflight Environment

Extravehicular activity (EVA) is at the core of a manned space exploration program. There are elements of exploration that may be safely and effectively performed by robots, but there are critical elements of exploration that will require the trained, assertive, and reasoning mind of a human crewmember. To effectively use these skills, NASA needs a safe, effective, and efficient EVA component integrated into the human exploration program. The EVA preparation time should be minimized and the suit pressure should be low to accommodate EVA tasks without undue fatigue, physical discomfort, or suit-related trauma. Commissioned in 2005, the Exploration Atmospheres Working Group (EAWG) had the primary goal of recommending to NASA an internal environment that allowed efficient and repetitive EVAs for missions that were to be enabled by the former Constellation Program. At the conclusion of the EAWG meeting, the 8.0 psia and 32% oxygen (O2) environment were recommended for EVA intensive phases of missions. As a result of selecting this internal environment, NASA gains the capability for efficient EVA with low risk of decompression sickness (DCS), but not without incurring additional negative stimulus of hypobaric hypoxia to the already physiologically challenging spaceflight environment. This paper provides a literature review of the human health and performance risks associated with the 8 psia/32% O2 environment. Of most concern are the potential effects on the central nervous system including increased intracranial pressure, visual impairment, sensorimotor dysfunction, and oxidative damage. Other areas of focus include validation of the DCS mitigation strategy, incidence and treatment of acute mountain sickness (AMS), development of new exercise countermeasures protocols, effective food preparation at 8 psia, assurance of quality sleep, and prevention of suit-induced injury. As a first effort, the trade space originally considered in the EAWG was re-evaluated looking for ways to decrease the hypoxic dose by further enriching the O2% or increasing the pressure. After discussion with the NASA engineering and materials community, it was determined that the O2 could be enriched from 32% to 34% and the pressure increased from 8.0 to 8.2 psia without significant penalty. These two small changes increase alveolar O2 pressure by 11 mmHg, which is expected to significantly benefit crewmembers. The 8.2/34 environment (inspired O2 pressure = 128 mmHg) is also physiologically equivalent to the staged decompression atmosphere of 10.2 psia / 26.5% O2 (inspired O2 pressure = 127 mmHg) used on 34 different shuttle missions for approximately a week each flight. Once decided, the proposed internal environment, if different than current experience, should be evaluated through appropriately simulated research studies. In many cases, the human physiologic concerns can be investigated effectively through integrated multi-discipline ground-based studies. Although missions proposing to use an 8.2/34 environment are still years away, it is recommended that these studies begin early enough to ensure that the correct decisions pertaining to vehicle design, mission operational concepts, and human health countermeasures are appropriately informed

Radio Astronomy

Contains reports on three research projects.National Aeronautics and Space Administration (Grant NsG-419)National Aeronautics and Space Administration (Contract NSR-22-009-120)Lincoln Laboratory Purchase Order No. 74

Orion Entry Handling Qualities Assessments

The Orion Command Module (CM) is a capsule designed to bring crew back from the International Space Station (ISS), the moon and beyond. The atmospheric entry portion of the flight is deigned to be flown in autopilot mode for nominal situations. However, there exists the possibility for the crew to take over manual control in off-nominal situations. In these instances, the spacecraft must meet specific handling qualities criteria. To address these criteria two separate assessments of the Orion CM s entry Handling Qualities (HQ) were conducted at NASA s Johnson Space Center (JSC) using the Cooper-Harper scale (Cooper & Harper, 1969). These assessments were conducted in the summers of 2008 and 2010 using the Advanced NASA Technology Architecture for Exploration Studies (ANTARES) six degree of freedom, high fidelity Guidance, Navigation, and Control (GN&C) simulation. This paper will address the specifics of the handling qualities criteria, the vehicle configuration, the scenarios flown, the simulation background and setup, crew interfaces and displays, piloting techniques, ratings and crew comments, pre- and post-fight briefings, lessons learned and changes made to improve the overall system performance. The data collection tools, methods, data reduction and output reports will also be discussed. The objective of the 2008 entry HQ assessment was to evaluate the handling qualities of the CM during a lunar skip return. A lunar skip entry case was selected because it was considered the most demanding of all bank control scenarios. Even though skip entry is not planned to be flown manually, it was hypothesized that if a pilot could fly the harder skip entry case, then they could also fly a simpler loads managed or ballistic (constant bank rate command) entry scenario. In addition, with the evaluation set-up of multiple tasks within the entry case, handling qualities ratings collected in the evaluation could be used to assess other scenarios such as the constant bank angle maintenance case. The 2008 entry assessment was divided into two sections (see Figure 1). Entry I was the first, high speed portion of a lunar return and Entry II was the second, lower speed portion of a lunar return, which is similar (but not identical) to a typical ISS return

Theory of Circle Maps and the Problem of One-Dimensional Optical Resonator with a Periodically Moving Wall

We consider the electromagnetic field in a cavity with a periodically oscillating perfectly reflecting boundary and show that the mathematical theory of circle maps leads to several physical predictions. Notably, well-known results in the theory of circle maps (which we review briefly) imply that there are intervals of parameters where the waves in the cavity get concentrated in wave packets whose energy grows exponentially. Even if these intervals are dense for typical motions of the reflecting boundary, in the complement there is a positive measure set of parameters where the energy remains bounded.Comment: 34 pages LaTeX (revtex) with eps figures, PACS: 02.30.Jr, 42.15.-i, 42.60.Da, 42.65.Y

Synchrotron FTIR imaging of OH in quartz mylonites

Previous measurements of water in deformed quartzites using conventional Fourier transform infrared spectroscopy (FTIR) instruments have shown that water contents of larger grains vary from one grain to another. However, the non-equilibrium variations in water content between neighboring grains and within quartz grains cannot be interrogated further without greater measurement resolution, nor can water contents be measured in finely recrystallized grains without including absorption bands due to fluid inclusions, films, and secondary minerals at grain boundaries.Synchrotron infrared (IR) radiation coupled to a FTIR spectrometer has allowed us to distinguish and measure OH bands due to fluid inclusions, hydrogen point defects, and secondary hydrous mineral inclusions through an aperture of 10 µm for specimens > 40 µm thick. Doubly polished infrared (IR) plates can be prepared with thicknesses down to 4–8 µm, but measurement of small OH bands is currently limited by strong interference fringes for samples < 25 µm thick, precluding measurements of water within individual, finely recrystallized grains. By translating specimens under the 10 µm IR beam by steps of 10 to 50 µm, using a software-controlled x − y stage, spectra have been collected over specimen areas of nearly 4.5 mm2. This technique allowed us to separate and quantify broad OH bands due to fluid inclusions in quartz and OH bands due to micas and map their distributions in quartzites from the Moine Thrust (Scotland) and Main Central Thrust (Himalayas).Mylonitic quartzites deformed under greenschist facies conditions in the footwall to the Moine Thrust (MT) exhibit a large and variable 3400 cm−1 OH absorption band due to molecular water, and maps of water content corresponding to fluid inclusions show that inclusion densities correlate with deformation and recrystallization microstructures. Quartz grains of mylonitic orthogneisses and paragneisses deformed under amphibolite conditions in the hanging wall to the Main Central Thrust (MCT) exhibit smaller broad OH bands, and spectra are dominated by sharp bands at 3595 to 3379 cm−1 due to hydrogen point defects that appear to have uniform, equilibrium concentrations in the driest samples. The broad OH band at 3400 cm−1 in these rocks is much less common. The variable water concentrations of MT quartzites and lack of detectable water in highly sheared MCT mylonites challenge our understanding of quartz rheology. However, where water absorption bands can be detected and compared with deformation microstructures, OH concentration maps provide information on the histories of deformation and recovery, evidence for the introduction and loss of fluid inclusions, and water weakening processes

Radio Astronomy

Contains research objectives and reports on six research projects.National Aeronautics and Space Administration (Contract NSR-22-009-120)National Aeronautics and Space Administration (Grant NsG-419)Joint Services Electronics Programs (U. S. Army, U.S. Navy, and U.S. Air Force) under Contract DA 36-039-AMC-03200(E

Radio Astronomy

Contains reports on five research projects.National Aeronautics and Space Administration (Grant NsG-419)National Science Foundation (Grant GP-7046)National Aeronautics and Space Administration (Contract NSR-22-009-120)Joint Services Electronics Programs (U. S. Army, U. S. Navy, and U, S. Air Force, under Contract DA 28-043-AMC-02536(E)U. S. Navy (Office of Naval Research) under Contract N00014-67-A-0204-000

Report of Seminar on Estate Planning

Reports from the UK/CLE Seminar on Estate Planning held July 19-20, 1974

Second Annual Seminar on Estate Planning

Schedule and materials from the Second Annual Seminar on Estate Planning held by UK/CLE on July 18-19, 1975