In-depth characterization of the fluorescent signal of HyPer, a probe for hydrogen peroxide, in bacteria exposed to external oxidative stress

Genetically encoded, fluorescent biosensors have been developed to probe the activities of various signaling molecules inside cells ranging from changes in intracellular ion concentrations to dynamics of lipid second messengers. HyPer is a member of this class of biosensors and is the first to dynamically respond to hydrogen peroxide (H[subscript 2]O[subscript 2]), a reactive oxygen species that functions as a signaling molecule. However, detailed characterization of HyPer's signal is not currently available within the context of bacteria exposed to external oxidative stress, which occurs in the immunological response of higher organisms against invasive pathogenic bacteria. Here, we performed this characterization, specifically in Escherichia coli exposed to external H[subscript 2]O[subscript 2]. We found that the temporal behavior of the signal does not correspond exactly to peroxide concentration in the system as a function of time and expression of the sensor decreases the peroxide scavenging activity of the cell. We also determined the effects of cell number, both before and after normalization of externally added H[subscript 2]O[subscript 2] to the number of cells. Finally, we report quantitative characteristics of HyPer's signal in this context, including the dynamic range of the signal, the signal-to-noise ratio, and the half saturation constant. These parameters show statistically meaningful differences in signal between two commonly used strains of E. coli, demonstrating how signal can vary with strain. Taken together, our results establish a systematic, quantitative framework for researchers seeking to better understand the role of H[subscript 2]O[subscript 2] in the immunological response against bacteria, and for understanding potential differences in the details of HyPer's quantitative performance across studies.Massachusetts Institute of Technology. James H. Ferry Fund for Innovation in Research EducationNational Science Foundation (U.S.) (NSF Graduate Research Fellowship)Massachusetts Institute of Technology (Joseph R. Mares endowed chair in chemical engineering)Burroughs Wellcome Fund (Career Award at the Scientific Interface

SAtlas: Spherical Versions of the Atlas Stellar Atmosphere Program

Context: The current stellar atmosphere programs still cannot match some fundamental observations of the brightest stars, and with new techniques, such as optical interferometry, providing new data for these stars, additional development of stellar atmosphere codes is required. Aims: To modify the open-source model atmosphere program Atlas to treat spherical geometry, creating a test-bed stellar atmosphere code for stars with extended atmospheres. Methods: The plane-parallel Atlas has been changed by introducing the necessary spherical modifications in the pressure structure, in the radiative transfer and in the temperature correction. Results: Several test models show that the spherical program matches the plane-parallel models in the high surface gravity regime, and matches spherical models computed by Phoenix and by MARCS in the low gravity case.Comment: 10 pages, 10 figures, Accepted for publication in A&

Monitoring the action of redox-directed cancer therapeutics using a human peroxiredoxin-2-based probe

Redox cancer therapeutics target the increased reliance on intracellular antioxidant systems and enhanced susceptibility to oxidant-induced stress of some cancer cells compared to normal cells. Many of these therapeutics are thought to perturb intracellular levels of the oxidant hydrogen peroxide (H[subscript 2]O[subscript 2]), a signaling molecule that modulates a number of different processes in human cells. However, fluorescent probes for this species remain limited in their ability to detect the small perturbations induced during successful treatments. We report a fluorescent sensor based upon human peroxiredoxin-2, which acts as the natural indicator of small H[subscript 2]O[subscript 2] fluctuations in human cells. The new probe reveals peroxide-induced oxidation in human cells below the detection limit of current probes, as well as peroxiredoxin-2 oxidation caused by two different redox cancer therapeutics in living cells. This capability will be useful in elucidating the mechanism of current redox-based therapeutics and in developing new ones.National Science Foundation (U.S.). Graduate Research Fellowship ProgramSamsung FellowshipMassachusetts Institute of Technology (Haas Family Fellowship in Chemical Engineering

Vibrational Enhancement of the Effective Donor - Acceptor Coupling

The paper deals with a simple three sites model for charge transfer phenomena in an one-dimensional donor (D) - bridge (B) - acceptor (A) system coupled with vibrational dynamics of the B site. It is found that in a certain range of parameters the vibrational coupling leads to an enhancement of the effective donor - acceptor electronic coupling as a result of the formation of the polaron on the B site. This enhancement of the charge transfer efficiency is maximum at the resonance, where the effective energy of the fluctuating B site coincides with the donor (acceptor) energy.Comment: 5 pages, 3 figure

Superconductivity coexisting with phase-separated static magnetic order in (Ba,K)Fe2_{2}As2_{2}, (Sr,Na)Fe2_{2}As2_{2} and CaFe2_{2}As2_{2}

The recent discovery and subsequent developments of FeAs-based superconductors have presented novel challenges and opportunities in the quest for superconducting mechanisms in correlated-electron systems. Central issues of ongoing studies include interplay between superconductivity and magnetism as well as the nature of the pairing symmetry reflected in the superconducting energy gap. In the cuprate and RE(O,F)FeAs (RE = rare earth) systems, the superconducting phase appears without being accompanied by static magnetic order, except for narrow phase-separated regions at the border of phase boundaries. By muon spin relaxation measurements on single crystal specimens, here we show that superconductivity in the AFe$_{2}$As$_{2}$ (A = Ca,Ba,Sr) systems, in both the cases of composition and pressure tunings, coexists with a strong static magnetic order in a partial volume fraction. The superfluid response from the remaining paramagnetic volume fraction of (Ba$_{0.5}$K$_{0.5}$)Fe$_{2}$As$_{2}$ exhibits a nearly linear variation in T at low temperatures, suggesting an anisotropic energy gap with line nodes and/or multi-gap effects.Comment: 14 pages 7 figures (4 for main text and 3 for on-line supplementary documents

'It would be easier if she’d died’: young people with parents with dementia articulating inadmissible stories

In the U.K. context where the emphasis is (quite rightly) on living well with dementia, on positivity and enabling approaches, it can be difficult for researchers to investigate and report negative experiences. Failing to re-present perceptions and experiences as they are lived, however, does a serious disservice to the research endeavor and can prevent policy and service development and positive change. In this article, we present some stories told by participants in an Alzheimer’s Society (United Kingdom) Funded project uniquely investigating the perceptions and experiences of children and young people who have a parent with dementia. Sometimes the stories were not easy to hear, especially when they challenged dominant master narratives around dementia. We discuss our view that when the young people we spoke with told us how things were for them, we were ethically bound to respect and disseminate their accounts

“Prophetic, Dreaming on the Mounds of Heaven”: Schelling and Hölderlin and the Madness of Prophetic Time

Hölderlin, it must be said, was always ahead of his time! His was an experience of Heracleitean time, as the later hymns would conceive it, one in which the liveliness within poetry is transfigured into the ever-living fire of prophetic time and logos. What does it mean to speak of the prophetic? What does this strange word, wrenched from the comforts of institutional religion as usual—Nietzsche’s “monotonotheism”— give us to hear? In a time of diminishing returns for the traditional regimes of religion, how does the promise of the prophetic still speak to us

Actitudes y percepciones de los docentes universitarios en el proceso de adopción de una innovación. La iniciativa OpenCourseWare en la Universidad de Valencia (España)

El proyecto OpenCourseWare, repositorio de recursos educativos en abierto, se inició en la Universidad de Valencia en 2008 y en 2009, sólo incluía 10 asignaturas, ninguna de ellas del área de ciencias de la salud. El objetivo de este trabajo es investigar las actitudes de los docentes de esta área así como sus percepciones con relación a este repositorio para determinar cuáles son las estrategias necesarias para extender su uso. Se parte de la teoría de la difusión de las innovaciones de Rogers (2003) y de un enfoque cualitativo a través del muestreo teórico, entrevistas semiestructuradas y codificación abierta. Los resultados indican la existencia de dos grandes grupos de profesores. Los adoptantes tempranos con una predisposición alta a participar en la iniciativa y con una percepción positiva de sus atributos y los mainstream o corriente principal más reacios a participar y con una percepción negativa respecto a alguno de sus atributos. La identificación de las características de estos dos grandes grupos marca las líneas de acción y de investigación futura en el ámbito del marketing y difusión, formación, sensibilización y motivación para extender el uso del OCW. The OpenCourseWare project, an open learning resource repository, was set up at the University of Valencia between 2008 and 2009. It initially consisted of only 10 subjects, none of which was in the field of health sciences. The objective of this research is to investigate the attitudes and perceptions of teaching staff in the field of health sciences with regard to the project in order to identify strategies to increase its use and expand its coverage. The diffusion of innovations theory as propounded by Rogers is used, along with a qualitative approach using theoretical sampling, semi-structured interviews and open coding. The results indicate the existence of two categories of teaching staff. Firstly, those who adopt an innovation early, have a marked inclination to participate in the initiative and a positive perception of the characteristics of the innovation. Secondly, a larger category of those more reluctant to participate in it and with a negative perception of one or more of its characteristics. Identifying the nature of these two groups provides guidelines for future action and research in the marketing and distribution of such projects, and the training, awareness and motivation required in order to extend the use of OCW

A journey to client and therapist mutuality in person-centered psychotherapy: a case study

This aim of this case study was to build theory on the development of client–therapist mutuality in person-centered psychotherapy. A case study focusing on a 42-year-old female client who had presented for therapy following trauma within interpersonal relationships has been used. A reflective, theory-building, case study method was adopted that used data gathered from verbatim session notes and research interviews between the therapist (first author) and research supervisor (second author). Three primary therapeutic processes that contributed to the development of mutuality are discussed. First, the development of mutual empathy in the relationship; second, strategies for disconnection and staying out of relationship are identified. Third, client agency and mutuality is explored. In conclusion the study proposes that mutuality is a key construct within person-centered psychotherapy and develops as a natural consequence of the presence of Rogers’ therapeutic conditions

Abiotic conditions in cephalopod (Sepia officinalis) eggs: embryonic development at low pH and high pCO2

Low pO(2) values have been measured in the perivitelline fluids (PVF) of marine animal eggs on several occasions, especially towards the end of development, when embryonic oxygen consumption is at its peak and the egg case acts as a massive barrier to diffusion. Several authors have therefore suggested that oxygen availability is the key factor leading to hatching. However, there have been no measurements of PVF pCO(2) so far. This is surprising, as elevated pCO(2) could also constitute a major abiotic stressor for the developing embryo. As a first attempt to fill this gap in knowledge, we measured pO(2), pCO(2) and pH in the PVF of late cephalopod (Sepia officinalis) eggs. We found linear relationships between embryo wet mass and pO(2), pCO(2) and pH. pO(2) declined from > 12 kPa to less than 5 kPa, while pCO(2) increased from 0.13 to 0.41 kPa. In the absence of active accumulation of bicarbonate in the PVF, pH decreased from 7.7 to 7.2. Our study supports the idea that oxygen becomes limiting in cephalopod eggs towards the end of development; however, pCO(2) and pH shift to levels that have caused significant physiological disturbances in other marine ectothermic animals. Future research needs to address the physiological adaptations that enable the embryo to cope with the adverse abiotic conditions in their egg environment