The Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath) Is a Novel Copper-containing Three-subunit Enzyme: isolation and charactization

The particulate methane monooxygenase (pMMO) is known to be very difficult to study mainly due to its unusual activity instability in vitro. By cultivating Methylococcus capsulatus (Bath) under methane stress conditions and high copper levels in the growth medium, membranes highly enriched in the pMMO with exceptionally stable activity can be isolated from these cells. Purified and active pMMO can be subsequently obtained from these membrane preparations using protocols in which an excess of reductants and anaerobic conditions were maintained during membrane solubilization by dodecyl beta-D-maltoside and purification by chromatography. The pMMO was found to be the major constituent in these membranes, constituting 60-80% of total membrane proteins. The dominant species of the pMMO was found to consist of three subunits, alpha, beta, and gamma, with an apparent molecular mass of 45, 26, and 23 kDa, respectively. A second species of the pMMO, a proteolytically processed version of the enzyme, was found to be composed of three subunits, alpha', beta, and gamma, with an apparent molecular mass of 35, 26, and 23 kDa, respectively. The alpha and alpha' subunits from these two forms of the pMMO contain identical N-terminal sequences. The gamma subunit, however, exhibits variation in its N-terminal sequence. The pMMO is a copper-containing protein only and shows a requirement for Cu(I) ions. Approximately 12-15 Cu ions per 94-kDa monomeric unit were observed. The pMMO is sensitive to dioxygen tension. On the basis of dioxygen sensitivity, three kinetically distinct forms of the enzyme can be distinguished. A slow but air-stable form, which is converted into a "pulsed" state upon direct exposure to atmospheric oxygen pressure, is considered as type I pMMO. This form was the subject of our pMMO isolation effort. Other forms (types II and III) are deactivated to various extents upon exposure to atmospheric dioxygen pressure. Under inactivating conditions, these unstable forms release protons to the buffer (~10 H+/94-kDa monomeric unit) and eventually become completely inactive

Adaptive Disturbance Torque Estimation for Orbiting Spacecraft Using Recursive Least-Squares Methods

This paper develops a novel disturbance torque estimator for an orbiting spacecraft by using the adaptive least-squares parameter estimation technique. The disturbance estimation is first formulated as an adaptive least-squares minimization problem using a set of polynomial functions and then integrated with the feedback momentum estimator. The covariance update law with a variable forgetting factor is used, and it is shown that the convergent rate for estimation errors can be made at the same level as the forgetting factor. The proposed approach is particularly suited for orbiting small or microsatellite applications, where the momentum management capacity is often limited. The onboard estimated disturbance torque input can then be used as a part of control resource for spacecraft momentum management. The simulation results demonstrate the efficacy of the proposed concept

Adaptive Modal Identification for Flutter Suppression Control

In this paper, we will develop an adaptive modal identification method for identifying the frequencies and damping of a flutter mode based on model-reference adaptive control (MRAC) and least-squares methods. The least-squares parameter estimation will achieve parameter convergence in the presence of persistent excitation whereas the MRAC parameter estimation does not guarantee parameter convergence. Two adaptive flutter suppression control approaches are developed: one based on MRAC and the other based on the least-squares method. The MRAC flutter suppression control is designed as an integral part of the parameter estimation where the feedback signal is used to estimate the modal information. On the other hand, the separation principle of control and estimation is applied to the least-squares method. The least-squares modal identification is used to perform parameter estimation

Fuzzy Model-based Pitch Stabilization and Wing Vibration Suppression of Flexible Wing Aircraft.

This paper presents a fuzzy nonlinear controller to regulate the longitudinal dynamics of an aircraft and suppress the bending and torsional vibrations of its flexible wings. The fuzzy controller utilizes full-state feedback with input constraint. First, the Takagi-Sugeno fuzzy linear model is developed which approximates the coupled aeroelastic aircraft model. Then, based on the fuzzy linear model, a fuzzy controller is developed to utilize a full-state feedback and stabilize the system while it satisfies the control input constraint. Linear matrix inequality (LMI) techniques are employed to solve the fuzzy control problem. Finally, the performance of the proposed controller is demonstrated on the NASA Generic Transport Model (GTM)

Genetic mapping of QTL conditioning resistance to soybean cyst nematode in PI464925B

Abstract only availableSoybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is estimated to cause the greatest yield losses to soybean [Glycine max (L.) Merr.] of any pest worldwide. It has been determined that host plant resistance is the most cost-effective and environmentally conscious method of controlling SCN. Phenotypic resistance appears to be quantitative and few cultivars exhibit resistance to one or more races of SCN. Identification of genetically resistant lines will be needed to compensate for various environmental SCN populations. Plant introductions (PIs) from the USDA Soybean Germplasm Collection have been screened for resistance to SCN and relatively few sources have been identified as new sources of SCN resistance. A wild soybean PI464925B (Glycine soja Siebold & Zucc.) is a soybean plant introduction from China shown to have resistance to SCN race 3. In this study, PI464925B was crossed with the SCN susceptible cultivar 'Hutcheson' to generate F1 hybrids. One hundred twenty-two F2 derived F3 progenies were evaluated for reaction to SCN race 3 in a thermo-regulated waterbath (27±1 ºC) in the greenhouse at the University of Missouri for reaction to SCN race 3. DNA from leaf tissue of the parents and progeny was extracted and one hundred seventeen of the progenies were used for construction of linkage maps and location of the QTL(quantitative trait loci) by using SSR(simple sequence repeats) markers. Multiplex PCR was performed using fluorescent labeled primers with subsequent analysis on an ABI 3100 DNA sequencer to increase high-throughput of genetic mapping. Genemapper (v3.5) was used for automatic allele sizing and genotyping. Parental testing showed 201 polymorphic SSR markers (56%), providing an average genomic coverage of 12 cM between two markers. Among them, genotypic data from 113 labeled SSR markers on the F3 progeny were collected to analyze association with the SCN response. QTL locations and genetic contribution of the favored alleles will be discussed.Plant Genomics Internship @ M

Time to Use Dendrohydrological Data in Water Resources Management?

In this editorial, we argue that the water resources management community should move away from relying only on short instrumental record, and instead incorporate long term tree-ring data into informing water management

Population pharmacokinetics of meropenem administered as a prolonged infusion in children with cystic fibrosis

OBJECTIVES: Meropenem is frequently used to treat pulmonary exacerbations in children with cystic fibrosis (CF) in the USA. Prolonged-infusion meropenem improves the time that free drug concentrations remain above the MIC (fT> MIC) in adults, but data in CF children are sparse. We describe the population pharmacokinetics, tolerability and treatment burden of prolonged-infusion meropenem in CF children. METHODS: Thirty children aged 6-17 years with a pulmonary exacerbation received 40 mg/kg meropenem every 8 h; each dose was administered as a 3 h infusion. Pharmacokinetics were determined using population methods in Pmetrics. Monte Carlo simulation was employed to compare 0.5 with 3 h infusions to estimate the probability of pharmacodynamic target attainment (PTA) at 40% fT> MIC. NCT#01429259. RESULTS: A two-compartment model fitted the data best with clearance and volume predicted by body weight. Clearance and volume of the central compartment were 0.41 ± 0.23 L/h/kg and 0.30 ± 0.17 L/kg, respectively. Half-life was 1.11 ± 0.38 h. At MICs of 1, 2 and 4 mg/L, PTAs for the 0.5 h infusion were 87.6%, 70.1% and 35.4%, respectively. The prolonged infusion increased PTAs to >99% for these MICs and achieved 82.8% at 8 mg/L. Of the 30 children, 18 (60%) completed treatment with prolonged infusion; 5 did so at home without any reported burden. Nine patients were changed to a 0.5 h infusion when discharged home. CONCLUSIONS: In these CF children, meropenem clearance was greater compared with published values from non-CF children. Prolonged infusion provided an exposure benefit against pathogens with MICs ≥1 mg/L, was well tolerated and was feasible to administer in the hospital and home settings, the latter depending on perception and family schedule

Control-focused, nonlinear and time-varying modelling of dielectric elastomer actuators with frequency response analysis

Current models of dielectric elastomer actuators (DEAs) are mostly constrained to first principal descriptions that are not well suited to the application of control design due to their computational complexity. In this work we describe an integrated framework for the identification of control focused, data driven and time-varying DEA models that allow advanced analysis of nonlinear system dynamics in the frequency-domain. Experimentally generated input–output data (voltage-displacement) was used to identify control-focused, nonlinear and time-varying dynamic models of a set of film-type DEAs. The model description used was the nonlinear autoregressive with exogenous input structure. Frequency response analysis of the DEA dynamics was performed using generalized frequency response functions, providing insight and a comparison into the time-varying dynamics across a set of DEA actuators. The results demonstrated that models identified within the presented framework provide a compact and accurate description of the system dynamics. The frequency response analysis revealed variation in the time-varying dynamic behaviour of DEAs fabricated to the same specifications. These results suggest that the modelling and analysis framework presented here is a potentially useful tool for future work in guiding DEA actuator design and fabrication for application domains such as soft robotics