Microscopic Characterization of a Transposon-Induced Male-Sterile, Female-Sterile Mutant in Glycine max L.

Premise of research. A male-sterile, female-sterile mutant was discovered in a w4-m mutable line of Glycine max L. The mechanism of its sterility was not well understood. Therefore, different cytological and microscopic techniques were undertaken to better understand the process of mutant phenotype development. Molecular research indicated that mer3 was responsible for the sterility. Methodology. Macro images were collected of whole plants, flowers, anthers, pods, and ovules. Chromosome spreads from anthers at various meiotic stages were examined. Confocal scanning laser microscopy using optical sectioning was utilized on whole anthers and ovules at various developmental stages. Whole mature anthers and isolated pollen images were collected and studied with SEM. Pivotal results. In observations of the mutant, male cell development was found to begin normally and then digresses at metaphase I of meiosis, when abnormal segregation of chromosomes with reduced bivalent formation was observed. It was the abnormal formation of univalents and bivalents that led to male sterility. On the female side, the progression of development was arrested in the megagametophyte stage likely because of abnormal meiosis, leading to ovule abortion and female sterility. Conclusions. The G. max male-sterile, female-sterile mutant was shown to have the same phenotype of mer3 sterility already shown in Arabidopsis, rice, yeast, and some animal systems

Identification of iron(III) peroxo species in the active site of the superoxide reductase SOR from Desulfoarculus baarsii.

International audienceThe active site of superoxide reductase SOR consists of an Fe2+ center in an unusual [His4 Cys1] square-pyramidal geometry. It specifically reduces superoxide to produce H2O2. Here, we have reacted the SOR from Desulfoarculus baarsii directly with H2O2. We have found that its active site can transiently stabilize an Fe3+-peroxo species that we have spectroscopically characterized by resonance Raman. The mutation of the strictly conserved Glu47 into alanine results in a stabilization of this Fe3+-peroxo species, when compared to the wild-type form. These data support the hypothesis that the reaction of SOR proceeds through such a Fe3+-peroxo intermediate. This also suggests that Glu47 might serve to help H2O2 release during the reaction with superoxide

in silico trials verification validation and uncertainty quantification of predictive models used in the regulatory evaluation of biomedical products

Abstract Historically, the evidences of safety and efficacy that companies provide to regulatory agencies as support to the request for marketing authorization of a new medical product have been produced experimentally, either in vitro or in vivo. More recently, regulatory agencies started receiving and accepting evidences obtained in silico, i.e. through modelling and simulation. However, before any method (experimental or computational) can be acceptable for regulatory submission, the method itself must be considered "qualified" by the regulatory agency. This involves the assessment of the overall "credibility" that such a method has in providing specific evidence for a given regulatory procedure. In this paper, we describe a methodological framework for the credibility assessment of computational models built using mechanistic knowledge of physical and chemical phenomena, in addition to available biological and physiological knowledge; these are sometimes referred to as "biophysical" models. Using guiding examples, we explore the definition of the context of use, the risk analysis for the definition of the acceptability thresholds, and the various steps of a comprehensive verification, validation and uncertainty quantification process, to conclude with considerations on the credibility of a prediction for a specific context of use. While this paper does not provide a guideline for the formal qualification process, which only the regulatory agencies can provide, we expect it to help researchers to better appreciate the extent of scrutiny required, which should be considered early on in the development/use of any (new) in silico evidence

The Full-sky Astrometric Mapping Explorer -- Astrometry for the New Millennium

FAME is designed to perform an all-sky, astrometric survey with unprecedented accuracy. It will create a rigid astrometric catalog of 4x10^7 stars with 5 < m_V < 15. For bright stars, 5 < m_V < 9, FAME will determine positions and parallaxes accurate to < 50 microarcseconds, with proper motion errors < 50 microarcseconds/year. For fainter stars, 9 < m_V < 15, FAME will determine positions and parallaxes accurate to < 500 microarcseconds, with proper motion errors < 500 microarcseconds/year. It will also collect photometric data on these 4 x 10^7 stars in four Sloan DSS colors.Comment: 6 pages, 4 figures, to appear in "Working on the Fringe

On Mean Field Glassy Dynamics out of Equilibrium

We study the off equilibrium dynamics of a mean field disordered systems which can be interpreted both as a long range interaction spin glass and as a particle in a random potential. The statics of this problem is well known and exhibits a low temperature spin glass phase with continuous replica symmetry breaking. We study the equations of off equilibrium dynamics with analytical and numerical methods. In the spin glass phase, we find that the usual equilibrium dynamics (observed when the observation time is much smaller than the waiting time) coexists with an aging regime. In this aging regime, we propose a solution implying a hierarchy of crossovers between the observation time and the waiting time.Comment: LaTeX, LPTENS preprint 94/0

Credible practice of modeling and simulation in healthcare: ten rules from a multidisciplinary perspective

The complexities of modern biomedicine are rapidly increasing. Thus, modeling and simulation have become increasingly important as a strategy to understand and predict the trajectory of pathophysiology, disease genesis, and disease spread in support of clinical and policy decisions. In such cases, inappropriate or ill-placed trust in the model and simulation outcomes may result in negative outcomes, and hence illustrate the need to formalize the execution and communication of modeling and simulation practices. Although verification and validation have been generally accepted as significant components of a model\u27s credibility, they cannot be assumed to equate to a holistic credible practice, which includes activities that can impact comprehension and in-depth examination inherent in the development and reuse of the models. For the past several years, the Committee on Credible Practice of Modeling and Simulation in Healthcare, an interdisciplinary group seeded from a U.S. interagency initiative, has worked to codify best practices. Here, we provide Ten Rules for credible practice of modeling and simulation in healthcare developed from a comparative analysis by the Committee\u27s multidisciplinary membership, followed by a large stakeholder community survey. These rules establish a unified conceptual framework for modeling and simulation design, implementation, evaluation, dissemination and usage across the modeling and simulation life-cycle. While biomedical science and clinical care domains have somewhat different requirements and expectations for credible practice, our study converged on rules that would be useful across a broad swath of model types. In brief, the rules are: (1) Define context clearly. (2) Use contextually appropriate data. (3) Evaluate within context. (4) List limitations explicitly. (5) Use version control. (6) Document appropriately. (7) Disseminate broadly. (8) Get independent reviews. (9) Test competing implementations. (10) Conform to standards. Although some of these are common sense guidelines, we have found that many are often missed or misconstrued, even by seasoned practitioners. Computational models are already widely used in basic science to generate new biomedical knowledge. As they penetrate clinical care and healthcare policy, contributing to personalized and precision medicine, clinical safety will require established guidelines for the credible practice of modeling and simulation in healthcare

Aging without disorder on long time scales

We study the Metropolis dynamics of a simple spin system without disorder, which exhibits glassy dynamics at low temperatures. We use an implementation of the algorithm of Bortz, Kalos and Lebowitz \cite{bortz}. This method turns out to be very efficient for the study of glassy systems, which get trapped in local minima on many different time scales. We find strong evidence of aging effects at low temperatures. We relate these effects to the distribution function of the trapping times of single configurations.Comment: 8 pages Revtex, 7 figures uuencoded (Revised version: the figures are now present

A tentative Replica Study of the Glass Transition

We propose a method to study quantitatively the glass transition in a system of interacting particles. In spite of the absence of any quenched disorder, we introduce a replicated version of the hypernetted chain equations. The solution of these equations, for hard or soft spheres, signals a transition to the glass phase. However the predicted value of the energy and specific heat in the glass phase are wrong, calling for an improvement of this method.Comment: 9 pages, four postcript figures attache

Large A-fiber activity is required for microglial proliferation and p38 MAPK activation in the spinal cord: different effects of resiniferatoxin and bupivacaine on spinal microglial changes after spared nerve injury

BACKGROUND: After peripheral nerve injury, spontaneous ectopic activity arising from the peripheral axons plays an important role in inducing central sensitization and neuropathic pain. Recent evidence indicates that activation of spinal cord microglia also contributes to the development of neuropathic pain. In particular, activation of p38 mitogen-activated protein kinase (MAPK) in spinal microglia is required for the development of mechanical allodynia. However, activity-dependent activation of microglia after nerve injury has not been fully addressed. To determine whether spontaneous activity from C- or A-fibers is required for microglial activation, we used resiniferatoxin (RTX) to block the conduction of transient receptor potential vanilloid subtype 1 (TRPV1) positive fibers (mostly C- and Adelta-fibers) and bupivacaine microspheres to block all fibers of the sciatic nerve in rats before spared nerve injury (SNI), and observed spinal microglial changes 2 days later. RESULTS: SNI induced robust mechanical allodynia and p38 activation in spinal microglia. SNI also induced marked cell proliferation in the spinal cord, and all the proliferating cells (BrdU+) were microglia (Iba1+). Bupivacaine induced a complete sensory and motor blockade and also significantly inhibited p38 activation and microglial proliferation in the spinal cord. In contrast, and although it produced an efficient nociceptive block, RTX failed to inhibit p38 activation and microglial proliferation in the spinal cord. CONCLUSION: (1) Blocking peripheral input in TRPV1-positive fibers (presumably C-fibers) is not enough to prevent nerve injury-induced spinal microglial activation. (2) Peripheral input from large myelinated fibers is important for microglial activation. (3) Microglial activation is associated with mechanical allodynia