Identification, functional expression and kinetic analysis of two primary amine oxidases from Rhodococcus opacus

Two native copper-containing amine oxidases (EC 1.4.3.21) have been isolated from Rhodococcus opacus and reveal phenotypic plasticity and catalytic activity with respect to structurally diverse natural and synthetic amines. Altering the amine growth substrate has enabled tailored and targeted oxidase upreg-ulation, which with subsequent treatment by precipitation, ion exchange and gel filtration, achieved a 90–150 fold purification. MALDI-TOF mass spectrometric and genomic analysis has indicated multiple gene activation with complex biodegradation pathways and regulatory mechanisms. Additional post-purification characterisation has drawn on the use of carbonyl reagent and chelating agent inhibitors. Michaelis–Menten kinetics for common aliphatic and aromatic amine substrates and several structural analogues demonstrated a broad specificity and high affinity with Michaelis constants (K M) ranging from 0.1 to 0.9 mM for C 1 –C 5 aliphatic mono-amines and <0.2 mM for a range of aromatic amines. Potential exploitation of the enzymatic versatility of the two isolated oxidases in biosensing and bioprocessing is discussed.

Port-Hamiltonian control of fully actuated underwater vehicles

This chapter presents a novel control strategy for trajectory tracking of underwater marine vehicles that are designed using port-Hamiltonian theory. A model for neutrally buoyant underwater vehicles is formulated as a PHS, and then the tracking controller is designed for the horizontal plane-surge, sway and yaw. The control design is done by formulating the error dynamics as a set-point regulation port-Hamiltonian control problem. The control design is formulated in two steps. In the first step, a static-feedback tracking controller is designed, and the second step integral action is added. The global asymptotic stability of the closed loop system is proved and the performance of the controller is illustrated using a model of an open-frame offshore underwater vehicle.

Integrated approach for diagnostics and prognostics of HP LNG pump based on health state probability estimation

Effective machine fault prognostic technologies can lead to elimination of unscheduled downtime and increase machine useful life and consequently lead to reduction of maintenance costs as well as prevention of human casualties in real engineering asset management. This paper presents a technique for accurate assessment of the remnant life of machines based on health state probability estimation technique and historical failure knowledge embedded in the closed loop diagnostic and prognostic system. To estimate a discrete machine degradation state which can represent the complex nature of machine degradation effectively, the proposed prognostic model employed a classification algorithm which can use a number of damage sensitive features compared to conventional time series analysis techniques for accurate long-term prediction. To validate the feasibility of the proposed model, the five different level data of typical four faults from High Pressure Liquefied Natural Gas (HP-LNG) pumps were used for the comparison of intelligent diagnostic test using five different classification algorithms. In addition, two sets of impeller-rub data were analysed and employed to predict the remnant life of pump based on estimation of health state probability using the Support Vector Machine (SVM) classifier. The results obtained were very encouraging and showed that the proposed prognostics system has the potential to be used as an estimation tool for machine remnant life prediction in real life industrial applications.

Residential balconies with road traffic noise : a combined source image and radiosity model

Several significant studies have been made in recent decades toward understanding road traffic noise and its effects on residential balconies. These previous studies have used a variety of techniques such as theoretical models, scale models and measurements on real balconies. The studies have considered either road traffic noise levels within the balcony space or inside an adjacent habitable room or both. Previous theoretical models have used, for example, simplified specular reflection calculations, boundary element methods (BEM), adaptations of CoRTN or the use of Sabine Theory. This paper presents an alternative theoretical model to predict the effects of road traffic noise spatially within the balcony space. The model includes a specular reflection component by calculating up to 10 orders of source images. To account for diffusion effects, a two compartment radiosity component is utilised. The first radiosity compartment is the urban street, represented as a street with building facades on either side. The second radiosity compartment is the balcony space. The model is designed to calculate the predicted road traffic noise levels within the balcony space and is capable of establishing the effect of changing street and balcony geometries. Screening attenuation algorithms are included to determine the effects of solid balcony parapets and balcony ceiling shields.

Conversion of neutral C2COC2 to C4CO. potential interstellar molecules

Both [C4CO]−· and [C2COC2]−· are formed in the ion source of a VG ZAB 2HF mass spectrometer by the respective processes HO− + Me3Si–CC–CC–CO–CMe3 → [C4CO]−· + Me3SiOH + Me3C·, and Me3Si–CC–CO–CC–SiMe3 + SF6 + e → [C2COC2]−· + 2Me3SiF + SF4. The second synthetic pathway involves a double desilylation reaction similar to that first reported by Squires. The two radical anion isomers produce different and characteristic charge reversal spectra upon collisional activation. In contrast, following collision induced charge stripping, both radical anions produce neutral C4CO as evidenced by the identical neutralisation reionisation (−NR+) spectra. The exclusive rearrangement of C213COC2 to C413CO indicates that 12C–O bond formation is not involved in the reaction. Ab initio calculations (at the RCCSD(T)/aug-cc-pVDZ//B3LYP/6-31G∗ level of theory) have been used to investigate the reaction coordinates on the potential surfaces for both singlet and triplet rearrangements of neutral C2COC2. Singlet C2COC2 is less stable than singlet C4CO by 78.8 kcal mol−1 and requires only 8.5 kcal mol−1 of additional energy to effect conversion to C4CO by a rearrangement sequence involving three C–C ring opening/cyclisation steps.

Total energy shaping of a class of underactuated Port-Hamiltonian Systems using a new set of closed-loop potential shape variables

This paper proposes a method for designing set-point regulation controllers for a class of underactuated mechanical systems in Port-Hamiltonian System (PHS) form. A new set of potential shape variables in closed loop is proposed, which can replace the set of open loop shape variables-the configuration variables that appear in the kinetic energy. With this choice, the closed-loop potential energy contains free functions of the new variables. By expressing the regulation objective in terms of these new potential shape variables, the desired equilibrium can be assigned and there is freedom to reshape the potential energy to achieve performance whilst maintaining the PHS form in closed loop. This complements contemporary results in the literature, which preserve the open-loop shape variables. As a case study, we consider a robotic manipulator mounted on a flexible base and compensate for the motion of the base while positioning the end effector with respect to the ground reference. We compare the proposed control strategy with special cases that correspond to other energy shaping strategies previously proposed in the literature.

A framework for testing robust autonomy of UAS during design and certification

As the number of Uninhabited Airborne Systems (UAS) proliferates in civil applications, industry is increasingly putting pressure on regulation authorities to provide a path for certification and allow UAS integration into regulated airspace. The success of this integration depends on developments in improved UAS reliability and safety, regulations for certification, and technologies for operational performance and safety assessment. This paper focusses on the last topic and describes a framework for quantifying robust autonomy of UAS, which quantifies the system's ability to either continue operating in the presence of faults or safely shut down. Two figures of merit are used to evaluate vehicle performance relative to mission requirements and the consequences of autonomous decision making in motion control and guidance systems. These figures of merit are interpreted within a probabilistic framework, which extends previous work in the literature. The valuation of the figures of merit can be done using stochastic simulation scenarios during both vehicle development and certification stages with different degrees of integration of hardware-in-the-loop simulation technology. The objective of the proposed framework is to aid in decision making about the suitability of a vehicle with respect to safety and reliability relative to mission requirements.

An assessment of the geographical scale of recurrent gene flow in wild populations of two species of Mekong River carps (Henicorhynchus spp.)

The Mekong is the most productive river fishery in the world, and such as, the Mekong River Basin (MRB) is very important to very large human populations across the region as a source of revenue (through fishing and marketing of aquatic resources products) and as the major source for local animal protein. Threats to biodiversity in the MRB, either to the fishery sector itself or to other sectors are a major concern, even though currently, fisheries across this region are still very productive. If not managed properly however, fish population declines will cause significant economic impact and affect livelihoods of local people and will have a major impact on food security and nutrition. Biodiversity declines will undoubtedly affect food security, income and socio-economic status of people in the MRB that depend on aquatic resources. This is an indicator of unsustainable development and hence should be avoided. Genetic diversity (biodiversity) that can be measured using techniques based on DNA markers; refers to variation within and among populations within the same species or reproductive units. In a population, new genetic variation is generated by sexual recombination contributed by individuals with mutations in genes and chromosomes. Over time, populations of a species that are not reproducing together will diverge as differential impacts of selection and genetic drift change their genetic attributes. For mud carp (Henicorhynchus spp.), understanding the status of breeding units in the MRB will be important for their long term persistence, sustainability and for implementing effective management strategies. Earlier analysis of stock structure in two economically important mud carp species (Henicorhynchus siamensis and H. lobatus) in the MRB completed with mtDNA markers identified a number of populations of both species where gene flow had apparently been interrupted or reduced but applying these data directly to management unit identification is potentially compromised because information was only available about female dispersal patterns. The current study aimed to address this problem and to fully assess the extent of current gene flow (nDNA) and reproductive exchange among selected wild populations of two species of carp (Henicorhynchus spp.) of high economic importance in the MRB using combined mtDNA and nDNA markers. In combination, the data can be used to define effective management units for each species. In general, nDNA diversity for H. lobatus (with average allelic richness (A) 7.56 and average heterozygosity (Ho) 0.61) was very similar to that identified for H. siamensis (A = 6.81 and Ho = 0.75). Both mud carp species show significant but low FST estimates among populations as a result of lower genetic diversity among sampled populations compared with genetic diversity within populations that may potentially mask any 'real' population structure. Overall, population genetic structure patterns from mtDNA and nDNA in both Henicorhynchus species were largely congruent. Different population structures however, were identified for the two Henicorhynchus species across the same geographical area. Apparent co-similarity in morphology and co-distribution of these two relatively closely related species does not apparently imply parallel evolutionary histories. Differences in each species population structure likely reflect historical drainage rearrangement of the Mekong River. The data indicate that H. siamensis is likely to have occupied the Mekong system for much longer than has H. lobatus in the past. Two divergent stocks were identified for H. lobatus in the MRB below the Khone Falls while a single stock had been evident in the earlier mtDNA study. This suggests that the two Henicorhynchus species may possess different life history traits and that different patterns of gene flow has likely influenced modern genetic structure in these close congeners. In combination, results of the earlier mtDNA and the current study have implications for effective management of both Henicorhynchus species across the MRB. Currently, both species are essentially treated as a single management unit in this region. This strategy may be appropriate for H. lobatus as a single stock was evident in the main stream of the MRB, but may not be appropriate for H. siamensis as more than a single stock was identified across the same range for this species. Management strategies should consider this difference to conserve overall biodiversity (local discrete populations) and this will include maintaining natural habitat and migration pathways, provision of fish sanctuaries (refuges) and may also require close monitoring of any stock declines, a signal that may require effective recovery strategies.

Ethylenedione : An intrinsically short-lived molecule

Ethylenedione C2O2 is one of the elusive small molecules which have remained undetected even after numerous attempts with different experimental techniques, This is surprising, since theoretical studies predicted the triplet state of C2O2 to be stable towards spin-allowed dissociation and hence long-lived. Here we report a comprehensive study of charged and neutral ethylenedione by means of charge reversal and neutralization -reionization mass spectrometry. These experimental results, in conjunction with theoretical calculations, suggest that neutral ethylenedione is intrinsically short-lived rather than being elusive, Both the singlet and triplet states of C2O2 are predicted to dissociate rapidly into two ground-state CO molecules, and for the triplet species, this dissociation involves facile curve-crossing to the singlet surface within a few nanoseconds.

The importance of accurate beam data measurement for SRT/SRS treatment planning

Introduction This study investigated the sensitivity of calculated stereotactic radiotherapy and radiosurgery doses to the accuracy of the beam data used by the treatment planning system. Methods Two sets of field output factors were acquired using fields smaller than approximately 1 cm2, for inclusion in beam data used by the iPlan treatment planning system (Brainlab, Feldkirchen, Germany). One set of output factors were measured using an Exradin A16 ion chamber (Standard Imaging, Middleton, USA). Although this chamber has a relatively small collecting volume (0.007 cm3), measurements made in small fields using this chamber are subject to the effects of volume averaging, electronic disequilibrium and chamber perturbations. The second, more accurate, set of measurements were obtained by applying perturbation correction factors, calculated using Monte Carlo simulations according to a method recommended by Cranmer-Sargison et al. [1] to measurements made using a 60017 unshielded electron diode (PTW, Freiburg, Germany). A series of 12 sample patient treatments were used to investigate the effects of beam data accuracy on resulting planned dose. These treatments, which involved 135 fields, were planned for delivery via static conformal arcs and 3DCRT techniques, to targets ranging from prostates (up to 8 cm across) to meningiomas (usually more than 2 cm across) to arterioveinous malformations, acoustic neuromas and brain metastases (often less than 2 cm across). Isocentre doses were calculated for all of these fields using iPlan, and the results of using the two different sets of beam data were evaluated. Results While the isocentre doses for many fields are identical (difference = 0.0 %), there is a general trend for the doses calculated using the data obtained from corrected diode measurements to exceed the doses calculated using the less-accurate Exradin ion chamber measurements (difference\0.0 %). There are several alarming outliers (circled in the Fig. 1) where doses differ by more than 3 %, in beams from sample treatments planned for volumes up to 2 cm across. Discussion and conclusions These results demonstrate that treatment planning dose calculations for SRT/SRS treatments can be substantially affected when beam data for fields smaller than approximately 1 cm2 are measured inaccurately, even when treatment volumes are up to 2 cm across.

Identification and characterisation of F3GT1 and F3GGT1, two glycosyltransferases responsible for anthocyanin biosynthesis in red-fleshed kiwifruit (Actinidia chinensis)

Much of the diversity of anthocyanins is due to the action of glycosyltransferases, which add sugar moieties to anthocyanidins. We identified two glycosyltransferases, F3GT1 and F3GGT1, from red-fleshed kiwifruit (Actinidia chinensis) that perform sequential glycosylation steps. Red-fleshed genotypes of kiwifruit accumulate anthocyanins mainly in the form of cyanidin 3-O-xylo-galactoside. Genes in the anthocyanin and flavonoid biosynthetic pathway were identified and shown to be expressed in fruit tissue. However, only the expression of the glycosyltransferase F3GT1 was correlated with anthocyanin accumulation in red tissues. Recombinant enzyme assays in vitro and in vivo RNA interference (RNAi) demonstrated the role of F3GT1 in the production of cyanidin 3-O-galactoside. F3GGT1 was shown to further glycosylate the sugar moiety of the anthocyanins. This second glycosylation can affect the solubility and stability of the pigments and modify their colour. We show that recombinant F3GGT1 can catalyse the addition of UDP-xylose to cyanidin 3-galactoside. While F3GGT1 is responsible for the end-product of the pathway, F3GT1 is likely to be the key enzyme regulating the accumulation of anthocyanin in red-fleshed kiwifruit varieties.

On evaluation of robust autonomy for uninhabited vehicles and systems

This paper discusses a method to quantify robust autonomy of Uninhabited Vehicles and Systems (UVS) in aerospace, marine, or land applications. Based on mission-vehicle specific performance criteria, we define an system utility function that can be evaluated using simulation scenarios for an envelope of environmental conditions. The results of these evaluations are used to compute a figure of merit or measure for operational efectiveness (MOE). The procedure is then augmented to consider faults and the performance of mechanisms to handle these faulty operational modes. This leads to a measure of robust autonomy (MRA). The objective of the proposed figures of merit is to assist in decision making about vehicle performance and reliability at both vehicle development stage (using simulation models) and at certification stage (using hardware-in-the-loop testing). Performance indices based on dynamic and geometric tasks associated with vehicle manoeuvring problems are proposed, and an example of a two- dimensional y scenario is provided to illustrate the use of the proposed figures of merit.

Hydrogen-bonded two- and three-dimensional polymeric structures in the ammonium salts of 3,5-dinitrobenzoic acid, 4-nitrobenzoic acid and 2,4-dichlorobenzoic acid

The structures of the ammonium salts of 3,5-dinitrobenzoic acid, NH4+ C7H3N2O6- (I), 4-nitrobenzoic acid, NH4+ C7H4N2O4- . 2H2O (II) and 2,4-dichlorobenzoic acid, NH4+ C7H3Cl2O2- . 0.5H2O (III), have been determined and their hydrogen-bonded structures are described. All salts form hydrogen-bonded polymeric structures, three-dimensional in (I) and two-dimensional in (II) and (III). With (I), a primary cation-anion cyclic association is formed [graph set R3/4(10)] through N-H...O hydrogen bonds, involving a carboxyl O,O' group on one side and a single carboxyl O-atom on the other. Structure extension involves both N-H...O hydrogen bonds to both carboxyl and nitro O-atom acceptors. With structure (II), the primary inter-species interactions and structure extension into layers lying parallel to (0 0 1) are through conjoined cyclic hydrogen-bonding motifs: R3/4(10) [one cation, a carboxyl (O,O') group and two water molecules] and centrosymmetric R2/4(8) [two cations and two water molecules]. The structure of (III) also has conjoined R3/4(10) and centrosymmetric R2/4(8) motifs in the layered structure but these differ in that he first involves one cation, a carboxyl (O,O') as well as a carboxyl (O) group and one water molecule, the second, two cations and two carboxyl O-groups. The layers lie parallel to (1 0 0). The structures of the salt hydrates (II) and (III) reported in this work, giving two-dimensional layered arrays through conjoined hydrogen-bonded nets provide further illustrations of a previously indicated trend among ammonium salts of carboxylic acids, but the anhydrous three-dimensional structure of (I) is inconsistent.

Drying kinetics, quality changes and shrinkage of Two grape varieties of Italy

Two varieties of grapes, white grape and red grape grown in the Campania region of Italy were selected for the study of drying characteristics, moisture diffusion, quality changes (colour) and shrinkage behaviour. Comparisons were made with treated and untreated grapes under constant drying condition of 50o C in a conventional drying system. This temperature was selected to represent farm drying conditions. Grapes were purchased from a local market from the same supplier to maintain the same size of grapes and same properties. An abrasive physical treatment was used as pretreatment. The drying curves were constructed and drying kinetics was calculated using several commonly available models. It was found that treated samples shows better drying characteristics than untreated samples. The objective of this study is to obtain drying kinetics which can be used to optimize the drying operations in grape drying.