An improved microtiter plate assay to monitor the oxidative burst in monocot and dicot plant cell suspension cultures

BACKGROUND: A screening method for elicitor and priming agents does not only allow detecting new bioactive substances, it can also be used to understand structure–function relationships of known agents by testing different derivatives of them. This can not only provide new lead compounds for the development of novel, more environment-benign, bio-based agro-chemicals, it may eventually also lead to a better understanding of defense mechanisms in plants. Reactive oxygen species (ROS) are sensitive indicators of these mechanisms but current assay formats are not suitable for multiplex screening, in particularly not in the case of monocot systems. RESULTS: Here we describe continuous monitoring of ROS in 96-well microtiter plates using the chemiluminescent probe L012, a luminol derivative producing chemiluminescence when oxidised by ROS like hydrogen peroxide, superoxide, or hydroxyl radical that can thus be used as an indicator for these ROS. We were able to measure ROS in both monocot (Oryza sativa) and dicot (Medicago truncatula) cell suspension cultures and record dose dependencies for the carbohydrate elicitors and priming agents ulvan and chitosan at low substrate concentrations (0.3–2.5 µg/ml). The method was optimized in terms of cell density, L012 concentration, and pre-incubation time. In contrast to the single peak observed using a cuvette luminometer, the improved method revealed a double burst in both cell systems during the 90-min measuring period, probably due to the detection of multiple ROS rather than only H2O2. CONCLUSION: We provide a medium throughput screening method for monocot and dicot suspension-cultured cells that enables direct comparison of monocot and dicot plant systems regarding their reaction to different signaling molecules.<br

Magnetic hopfions in solids

Hopfions are an intriguing class of string-like solitons, named according to a classical topological concept classifying three-dimensional direction fields. The search of hopfions in real physical systems is going on for nearly half a century, starting with the seminal work of Faddeev. But so far realizations in solids are missing. Here, we present a theory that identifies magnetic materials featuring hopfions as stable states without the assistance of confinement or external fields. Our results are based on an advanced micromagnetic energy functional derived from a spin-lattice Hamiltonian. Hopfions appear as emergent particles of the classical Heisenberg model. Magnetic hopfions represent three-dimensional particle-like objects of nanometre-size dimensions opening the gate to a new generation of spintronic devices in the framework of a truly three-dimensional architecture. Our approach goes beyond the conventional phenomenological models. We derive material-realistic parameters that serve as concrete guidance in the search of magnetic hopfions bridging computational physics with materials science

Intermodulation electrostatic force microscopy for imaging surface photo-voltage

We demonstrate an alternative to Kelvin Probe Force Microscopy for imaging surface potential. The open-loop, single-pass technique applies a low-frequency AC voltage to the atomic force microscopy tip while driving the cantilever near its resonance frequency. Frequency mixing due to the nonlinear capacitance gives intermodulation products of the two drive frequencies near the cantilever resonance, where they are measured with high signal to noise ratio. Analysis of this intermodulation response allows for quantitative reconstruction of the contact potential difference. We derive the theory of the method, validate it with numerical simulation and a control experiment, and we demonstrate its utility for fast imaging of the surface photo-voltage on an organic photo-voltaic material.Comment: 4 pages, 3 figures, peer-reviewed, preprin

Abort Trigger False Positive and False Negative Analysis Methodology for Threshold-Based Abort Detection

This paper describes a quantitative methodology for bounding the false positive (FP) and false negative (FN) probabilities associated with a human-rated launch vehicle abort trigger (AT) that includes sensor data qualification (SDQ). In this context, an AT is a hardware and software mechanism designed to detect the existence of a specific abort condition. Also, SDQ is an algorithmic approach used to identify sensor data suspected of being corrupt so that suspect data does not adversely affect an AT's detection capability. The FP and FN methodologies presented here were developed to support estimation of the probabilities of loss of crew and loss of mission for the Space Launch System (SLS) which is being developed by the National Aeronautics and Space Administration (NASA). The paper provides a brief overview of system health management as being an extension of control theory; and describes how ATs and the calculation of FP and FN probabilities relate to this theory. The discussion leads to a detailed presentation of the FP and FN methodology and an example showing how the FP and FN calculations are performed. This detailed presentation includes a methodology for calculating the change in FP and FN probabilities that result from including SDQ in the AT architecture. To avoid proprietary and sensitive data issues, the example incorporates a mixture of open literature and fictitious reliability data. Results presented in the paper demonstrate the effectiveness of the approach in providing quantitative estimates that bound the probability of a FP or FN abort determination

Geometrical quadrupolar frustration in DyB4_4

Physical properties of DyB$_4$ have been studied by magnetization, specific heat, and ultrasonic measurements. The magnetic entropy change and the ultrasonic properties in the intermediate phase II indicate that the degeneracy of internal degrees of freedom is not fully lifted in spite of the formation of magnetic order. The ultrasonic attenuation and the huge softening of $C_{44}$ in phase II suggests existence of electric-quadrupolar (orbital) fluctuations of the 4$f$-electron. These unusual properties originate from the geometrical quadrupolar frustration.Comment: 4 pages, 4 figures, accepted for publication in Journal of the Physical Society of Japa

Evaluation of an Active Clearance Control System Concept

Reducing blade tip clearances through active tip clearance control in the high pressure turbine can lead to significant reductions in emissions and specific fuel consumption as well as dramatic improvements in operating efficiency and increased service life. Current engines employ scheduled cooling of the outer case flanges to reduce high pressure turbine tip clearances during cruise conditions. These systems have relatively slow response and do not use clearance measurement, thereby forcing cold build clearances to set the minimum clearances at extreme operating conditions (e.g., takeoff, reburst) and not allowing cruise clearances to be minimized due to the possibility of throttle transients (e.g., step change in altitude). In an effort to improve upon current thermal methods, a first generation mechanically-actuated active clearance control (ACC) system has been designed and fabricated. The system utilizes independent actuators, a segmented shroud structure, and clearance measurement feedback to provide fast and precise active clearance control throughout engine operation. Ambient temperature performance tests of this first generation ACC system assessed individual seal component leakage rates and both static and dynamic overall system leakage rates. The ability of the nine electric stepper motors to control the position of the seal carriers in both open- and closed-loop control modes for single and multiple cycles was investigated. The ability of the system to follow simulated engine clearance transients in closed-loop mode showed the system was able to track clearances to within a tight tolerance (0.001 in. error)

Application of Diagnostic Analysis Tools to the Ares I Thrust Vector Control System

The NASA Ares I Crew Launch Vehicle is being designed to support missions to the International Space Station (ISS), to the Moon, and beyond. The Ares I is undergoing design and development utilizing commercial-off-the-shelf tools and hardware when applicable, along with cutting edge launch technologies and state-of-the-art design and development. In support of the vehicle s design and development, the Ares Functional Fault Analysis group was tasked to develop an Ares Vehicle Diagnostic Model (AVDM) and to demonstrate the capability of that model to support failure-related analyses and design integration. One important component of the AVDM is the Upper Stage (US) Thrust Vector Control (TVC) diagnostic model-a representation of the failure space of the US TVC subsystem. This paper first presents an overview of the AVDM, its development approach, and the software used to implement the model and conduct diagnostic analysis. It then uses the US TVC diagnostic model to illustrate details of the development, implementation, analysis, and verification processes. Finally, the paper describes how the AVDM model can impact both design and ground operations, and how some of these impacts are being realized during discussions of US TVC diagnostic analyses with US TVC designers

Interaction imaging with amplitude-dependence force spectroscopy

Knowledge of surface forces is the key to understanding a large number of processes in fields ranging from physics to material science and biology. The most common method to study surfaces is dynamic atomic force microscopy (AFM). Dynamic AFM has been enormously successful in imaging surface topography, even to atomic resolution, but the force between the AFM tip and the surface remains unknown during imaging. Here, we present a new approach that combines high accuracy force measurements and high resolution scanning. The method, called amplitude-dependence force spectroscopy (ADFS) is based on the amplitude-dependence of the cantilever's response near resonance and allows for separate determination of both conservative and dissipative tip-surface interactions. We use ADFS to quantitatively study and map the nano-mechanical interaction between the AFM tip and heterogeneous polymer surfaces. ADFS is compatible with commercial atomic force microscopes and we anticipate its wide-spread use in taking AFM toward quantitative microscopy