The transformation of banana with potential virus resistance genes

One approach to reducing the yield losses caused by banana viral diseases is the use of genetic engineering and pathogen-derived resistance strategies to generate resistant cultivars. The development of transgenic virus resistance requires an efficient banana transformation method, particularly for commercially important 'Cavendish' type cultivars such as 'Grand Nain'. Prior to this study, only two examples of the stable transformation of banana had been reported, both of which demonstrated the principle of transformation but did not characterise transgenic plants in terms of the efficiency at which individual transgenic lines were generated, relative activities of promoters in stably transformed plants, and the stability of transgene expression. The aim of this study was to develop more efficient transformation methods for banana, assess the activity of some commonly used and also novel promoters in stably transformed plants, and transform banana with genes that could potentially confer resistance to banana bunchy top nanovirus (BBTV) and banana bract mosaic potyvirus (BBrMV). A regeneration system using immature male flowers as the explant was established. The frequency of somatic embryogenesis in male flower explants was influenced by the season in which the inflorescences were harvested. Further, the media requirements of various banana cultivars in respect to the 2,4-D concentration in the initiation media also differed. Following the optimisation of these and other parameters, embryogenic cell suspensions of several banana (Musa spp.) cultivars including 'Grand Nain' (AAA), 'Williams' (AAA), 'SH-3362' (AA), 'Goldfinger' (AAAB) and 'Bluggoe' (ABB) were successfully generated. Highly efficient transformation methods were developed for both 'Bluggoe' and 'Grand Nain'; this is the first report of microprojectile bombardment transformation of the commercially important 'Grand Nain' cultivar. Following bombardment of embryogenic suspension cells, regeneration was monitored from single transfom1ed cells to whole plants using a reporter gene encoding the green fluorescent protein (gfp). Selection with kanamycin enabled the regeneration of a greater number of plants than with geneticin, while still preventing the regeneration of non-transformed plants. Southern hybridisation confirmed the neomycin phosphotransferase gene (npt II) was stably integrated into the banana genome and that multiple transgenic lines were derived from single bombardments. The activity, stability and tissue specificity of the cauliflower mosaic virus 358 (CaMV 35S) and maize polyubiquitin-1 (Ubi-1) promoters were examined. In stably transformed banana, the Ubi-1 promoter provided approximately six-fold higher p-glucuronidase (GUS) activity than the CaMV 35S promoter, and both promoters remained active in glasshouse grown plants for the six months they were observed. The intergenic regions ofBBTV DNA-I to -6 were isolated and fused to either the uidA (GUS) or gfjJ reporter genes to assess their promoter activities. BBTV promoter activity was detected in banana embryogenic cells using the gfp reporter gene. Promoters derived from BBTV DNA-4 and -5 generated the highest levels of transient activity, which were greater than that generated by the maize Ubi-1 promoter. In transgenic banana plants, the activity of the BBTV DNA-6 promoter (BT6.1) was restricted to the phloem of leaves and roots, stomata and root meristems. The activity of the BT6.1 promoter was enhanced by the inclusion of intron-containing fragments derived from the maize Ubi-1, rice Act-1, and sugarcane rbcS 5' untranslated regions in GUS reporter gene constructs. In transient assays in banana, the rice Act-1 and maize Ubi-1 introns provided the most significant enhancement, increasing expression levels 300-fold and 100-fold, respectively. The sugarcane rbcS intron increased expression about 10-fold. In stably transformed banana plants, the maize Ubi-1 intron enhanced BT6.1 promoter activity to levels similar to that of the CaMV 35S promoter, but did not appear to alter the tissue specificity of the promoter. Both 'Grand Nain' and 'Bluggoe' were transformed with constructs that could potentially confer resistance to BBTV and BBrMV, including constructs containing BBTV DNA-1 major and internal genes, BBTV DNA-5 gene, and the BBrMV coat protein-coding region all under the control of the Ubi-1 promoter, while the BT6 promoter was used to drive the npt II selectable marker gene. At least 30 transgenic lines containing each construct were identified and replicates of each line are currently being generated by micropropagation in preparation for virus challenge.

Swept frequency biompedance analysis for the determination of body water compartments

Bioelectrical impedance analysis, (BIA), is a method of body composition analysis first investigated in 1962 which has recently received much attention by a number of research groups. The reasons for this recent interest are its advantages, (viz: inexpensive, non-invasive and portable) and also the increasing interest in the diagnostic value of body composition analysis. The concept utilised by BIA to predict body water volumes is the proportional relationship for a simple cylindrical conductor, (volume oc length2/resistance), which allows the volume to be predicted from the measured resistance and length. Most of the research to date has measured the body's resistance to the passage of a 50· kHz AC current to predict total body water, (TBW). Several research groups have investigated the application of AC currents at lower frequencies, (eg 5 kHz), to predict extracellular water, (ECW). However all research to date using BIA to predict body water volumes has used the impedance measured at a discrete frequency or frequencies. This thesis investigates the variation of impedance and phase of biological systems over a range of frequencies and describes the development of a swept frequency bioimpedance meter which measures impedance and phase at 496 frequencies ranging from 4 kHz to 1 MHz. The impedance of any biological system varies with the frequency of the applied current. The graph of reactance vs resistance yields a circular arc with the resistance decreasing with increasing frequency and reactance increasing from zero to a maximum then decreasing to zero. Computer programs were written to analyse the measured impedance spectrum and determine the impedance, Zc, at the characteristic frequency, (the frequency at which the reactance is a maximum). The fitted locus of the measured data was extrapolated to determine the resistance, Ro, at zero frequency; a value that cannot be measured directly using surface electrodes. The explanation of the theoretical basis for selecting these impedance values (Zc and Ro), to predict TBW and ECW is presented. Studies were conducted on a group of normal healthy animals, (n=42), in which TBW and ECW were determined by the gold standard of isotope dilution. The prediction quotients L2/Zc and L2/Ro, (L=length), yielded standard errors of 4.2% and 3.2% respectively, and were found to be significantly better than previously reported, empirically determined prediction quotients derived from measurements at a single frequency. The prediction equations established in this group of normal healthy animals were applied to a group of animals with abnormally low fluid levels, (n=20), and also to a group with an abnormal balance of extra-cellular to intracellular fluids, (n=20). In both cases the equations using L2/Zc and L2/Ro accurately and precisely predicted TBW and ECW. This demonstrated that the technique developed using multiple frequency bioelectrical impedance analysis, (MFBIA), can accurately predict both TBW and ECW in both normal and abnormal animals, (with standard errors of the estimate of 6% and 3% for TBW and ECW respectively). Isotope dilution techniques were used to determine TBW and ECW in a group of 60 healthy human subjects, (male. and female, aged between 18 and 45). Whole body impedance measurements were recorded on each subject using the MFBIA technique and the correlations between body water volumes, (TBW and ECW), and heighe/impedance, (for all measured frequencies), were compared. The prediction quotients H2/Zc and H2/Ro, (H=height), again yielded the highest correlation with TBW and ECW respectively with corresponding standard errors of 5.2% and 10%. The values of the correlation coefficients obtained in this study were very similar to those recently reported by others. It was also observed that in healthy human subjects the impedance measured at virtually any frequency yielded correlations not significantly different from those obtained from the MFBIA quotients. This phenomenon has been reported by other research groups and emphasises the need to validate the technique by investigating its application in one or more groups with abnormalities in fluid levels. The clinical application of MFBIA was trialled and its capability of detecting lymphoedema, (an excess of extracellular fluid), was investigated. The MFBIA technique was demonstrated to be significantly more sensitive, (P<.05), in detecting lymphoedema than the current technique of circumferential measurements. MFBIA was also shown to provide valuable information describing the changes in the quantity of muscle mass of the patient during the course of the treatment. The determination of body composition, (viz TBW and ECW), by MFBIA has been shown to be a significant improvement on previous bioelectrical impedance techniques. The merit of the MFBIA technique is evidenced in its accurate, precise and valid application in animal groups with a wide variation in body fluid volumes and balances. The multiple frequency bioelectrical impedance analysis technique developed in this study provides accurate and precise estimates of body composition, (viz TBW and ECW), regardless of the individual's state of health.

Frequency Domain Characterization Of Sliding Mode Control Of An Inverter Used In DSTATCOM Application

In this paper, the commonly used switching schemes for sliding mode control of power converters is analyzed and designed in the frequency domain. Particular application of a distribution static compensator (DSTATCOM) in voltage control mode is investigated in a power distribution system. Tsypkin's method and describing function is used to obtain the switching conditions for the two-level and three-level voltage source inverters. Magnitude conditions of carrier signals are developed for robust switching of the inverter under carrier-based modulation scheme of sliding mode control. The existence of border collision bifurcation is identified to avoid the complex switching states of the inverter. The load bus voltage of an unbalanced three-phase nonstiff radial distribution system is controlled using the proposed carrier-based design. The results are validated using PSCAD/EMTDC simulation studies and through a scaled laboratory model of DSTATCOM that is developed for experimental verification

A ‘cost-per-wear’ model based on contact lens replacement frequency

Purpose: The aim was to construct and advise on the use of a cost-per-wear model based on contact lens replacement frequency, to form an equitable basis for cost comparison. ---------- Methods: The annual cost of professional fees, contact lenses and solutions when wearing daily, two-weekly and monthly replacement contact lenses is determined in the context of the Australian market for spherical, toric and multifocal prescription types. This annual cost is divided by the number of times lenses are worn per year, resulting in a ‘cost-per-wear’. The model is presented graphically as the cost-per-wear versus the number of times lenses are worn each week for daily replacement and reusable (two-weekly and monthly replacement) lenses.---------- Results: The cost-per-wear for two-weekly and monthly replacement spherical lenses is almost identical but decreases with increasing frequency of wear. The cost-per-wear of daily replacement spherical lenses is lower than for reusable spherical lenses, when worn from one to four days per week but higher when worn six or seven days per week. The point at which the cost-per-wear is virtually the same for all three spherical lens replacement frequencies (approximately AUD$3.00) is five days of lens wear per week. A similar but upwardly displaced (higher cost) pattern is observed for toric lenses, with the cross-over point occurring between three and four days of wear per week (AUD$4.80). Multifocal lenses have the highest price, with cross-over points for daily versus two-weekly replacement lenses at between four and five days of wear per week (AUD$5.00) and for daily versus monthly replacement lenses at three days per week (AUD$5.50).---------- Conclusions: This cost-per-wear model can be used to assist practitioners and patients in making an informed decision in relation to the cost of contact lens wear as one of many considerations that must be taken into account when deciding on the most suitable lens replacement modality.

The potential to mitigate global warming with no-tillage management is only realized when practised in the long term

No-tillage (NT) management has been promoted as a practice capable of offsetting greenhouse gas (GHG) emissions because of its ability to sequester carbon in soils. However, true mitigation is only possible if the overall impact of NT adoption reduces the net global warming potential (GWP) determined by fluxes of the three major biogenic GHGs (i.e. CO2, N2O, and CH4). We compiled all available data of soil-derived GHG emission comparisons between conventional tilled (CT) and NT systems for humid and dry temperate climates. Newly converted NT systems increase GWP relative to CT practices, in both humid and dry climate regimes, and longer-term adoption (>10 years) only significantly reduces GWP in humid climates. Mean cumulative GWP over a 20-year period is also reduced under continuous NT in dry areas, but with a high degree of uncertainty. Emissions of N2O drive much of the trend in net GWP, suggesting improved nitrogen management is essential to realize the full benefit from carbon storage in the soil for purposes of global warming mitigation. Our results indicate a strong time dependency in the GHG mitigation potential of NT agriculture, demonstrating that GHG mitigation by adoption of NT is much more variable and complex than previously considered, and policy plans to reduce global warming through this land management practice need further scrutiny to ensure success.

Vector control of a high power induction machine

The design and implementation of a high-power (2 MW peak) vector control drive is described. The inverter switching frequency is low, resulting in high-harmonic-content current waveforms. A block diagram of the physical system is given, and each component is described in some detail. The problem of commanded slip noise sensitivity, inherent in high-power vector control drives, is discussed, and a solution is proposed. Results are given which demonstrate the successful functioning of the system

Statistical modeling to determine resonant frequency information present in combustion chamber pressure signals

A statistical modeling method to accurately determine combustion chamber resonance is proposed and demonstrated. This method utilises Markov-chain Monte Carlo (MCMC) through the use of the Metropolis-Hastings (MH) algorithm to yield a probability density function for the combustion chamber frequency and find the best estimate of the resonant frequency, along with uncertainty. The accurate determination of combustion chamber resonance is then used to investigate various engine phenomena, with appropriate uncertainty, for a range of engine cycles. It is shown that, when operating on various ethanol/diesel fuel combinations, a 20% substitution yields the least amount of inter-cycle variability, in relation to combustion chamber resonance.

Property damage crash equivalency factors to solve crash frequency-severity dilemma: Case study on South Korean rural roads

Safety interventions (e.g., median barriers, photo enforcement) and road features (e.g., median type and width) can influence crash severity, crash frequency, or both. Both dimensions—crash frequency and crash severity—are needed to obtain a full accounting of road safety. Extensive literature and common sense both dictate that crashes are not created equal, with fatalities costing society more than 1,000 times the cost of property damage crashes on average. Despite this glaring disparity, the profession has not unanimously embraced or successfully defended a nonarbitrary severity weighting approach for analyzing safety data and conducting safety analyses. It is argued here that the two dimensions (frequency and severity) are made available by intelligently and reliably weighting crash frequencies and converting all crashes to property-damage-only crash equivalents (PDOEs) by using comprehensive societal unit crash costs. This approach is analogous to calculating axle load equivalents in the prediction of pavement damage: for instance, a 40,000-lb truck causes 4,025 times more stress than does a 4,000-lb car and so simply counting axles is not sufficient. Calculating PDOEs using unit crash costs is the most defensible and nonarbitrary weighting scheme, allows for the simple incorporation of severity and frequency, and leads to crash models that are sensitive to factors that affect crash severity. Moreover, using PDOEs diminishes the errors introduced by underreporting of less severe crashes—an added benefit of the PDOE analysis approach. The method is illustrated with rural road segment data from South Korea (which in practice would develop PDOEs with Korean crash cost data).

Incorporating weather into regionwide safety planning prediction models

Predicting safety on roadways is standard practice for road safety professionals and has a corresponding extensive literature. The majority of safety prediction models are estimated using roadway segment and intersection (microscale) data, while more recently efforts have been undertaken to predict safety at the planning level (macroscale). Safety prediction models typically include roadway, operations, and exposure variables—factors known to affect safety in fundamental ways. Environmental variables, in particular variables attempting to capture the effect of rain on road safety, are difficult to obtain and have rarely been considered. In the few cases weather variables have been included, historical averages rather than actual weather conditions during which crashes are observed have been used. Without the inclusion of weather related variables researchers have had difficulty explaining regional differences in the safety performance of various entities (e.g. intersections, road segments, highways, etc.) As part of the NCHRP 8-44 research effort, researchers developed PLANSAFE, or planning level safety prediction models. These models make use of socio-economic, demographic, and roadway variables for predicting planning level safety. Accounting for regional differences - similar to the experience for microscale safety models - has been problematic during the development of planning level safety prediction models. More specifically, without weather related variables there is an insufficient set of variables for explaining safety differences across regions and states. Furthermore, omitted variable bias resulting from excluding these important variables may adversely impact the coefficients of included variables, thus contributing to difficulty in model interpretation and accuracy. This paper summarizes the results of an effort to include weather related variables, particularly various measures of rainfall, into accident frequency prediction and the prediction of the frequency of fatal and/or injury degree of severity crash models. The purpose of the study was to determine whether these variables do in fact improve overall goodness of fit of the models, whether these variables may explain some or all of observed regional differences, and identifying the estimated effects of rainfall on safety. The models are based on Traffic Analysis Zone level datasets from Michigan, and Pima and Maricopa Counties in Arizona. Numerous rain-related variables were found to be statistically significant, selected rain related variables improved the overall goodness of fit, and inclusion of these variables reduced the portion of the model explained by the constant in the base models without weather variables. Rain tends to diminish safety, as expected, in fairly complex ways, depending on rain frequency and intensity.

A simultaneous equations model of crash frequency by collision type for rural intersections

Safety at roadway intersections is of significant interest to transportation professionals due to the large number of intersections in transportation networks, the complexity of traffic movements at these locations that leads to large numbers of conflicts, and the wide variety of geometric and operational features that define them. A variety of collision types including head-on, sideswipe, rear-end, and angle crashes occur at intersections. While intersection crash totals may not reveal a site deficiency, over exposure of a specific crash type may reveal otherwise undetected deficiencies. Thus, there is a need to be able to model the expected frequency of crashes by collision type at intersections to enable the detection of problems and the implementation of effective design strategies and countermeasures. Statistically, it is important to consider modeling collision type frequencies simultaneously to account for the possibility of common unobserved factors affecting crash frequencies across crash types. In this paper, a simultaneous equations model of crash frequencies by collision type is developed and presented using crash data for rural intersections in Georgia. The model estimation results support the notion of the presence of significant common unobserved factors across crash types, although the impact of these factors on parameter estimates is found to be rather modest.

Load frequency control for rural distributed generation

In rural low-voltage networks, distribution lines are usually highly resistive. When many distributed generators are connected to such lines, power sharing among them is difficult when using conventional droop control, as the real and reactive power have strong coupling with each other. A high droop gain can alleviate this problem but may lead the system to instability. To overcome4 this, two droop control methods are proposed for accurate load sharing with frequency droop controller. The first method considers no communication among the distributed generators and regulates the output voltage and frequency, ensuring acceptable load sharing. The droop equations are modified with a transformation matrix based on the line R/X ration for this purpose. The second proposed method, with minimal low bandwidth communication, modifies the reference frequency of the distributed generators based on the active and reactive power flow in the lines connected to the points of common coupling. The performance of these two proposed controllers is compared with that of a controller, which includes an expensive high bandwidth communication system through time-domain simulation of a test system. The magnitude of errors in power sharing between these three droop control schemes are evaluated and tabulated.

A practical implementation of a continuous isotropic spherical omnidirectional drive

This paper presents a continuous isotropic spherical omnidirectional drive mechanism that is efficient in its mechanical simplicity and use of volume. Spherical omnidirectional mechanisms allow isotropic motion, although many are limited from achieving true isotropic motion by practical mechanical design considerations. The mechanism presented in this paper uses a single motor to drive a point on the great circle of the sphere parallel to the ground plane, and does not require a gearbox. Three mechanisms located 120 degrees apart provide a stable drive platform for a mobile robot. Results show the omnidirectional ability of the robot and demonstrate the performance of the spherical mechanism compared to a popular commercial omnidirectional wheel over edges of varying heights and gaps of varying widths.

A two-Dimensional finite volume method for variable density flow and solute transport through saturated-unsaturated media

Extensive groundwater withdrawal has resulted in a severe seawater intrusion problem in the Gooburrum aquifers at Bundaberg, Queensland, Australia. Better management strategies can be implemented by understanding the seawater intrusion processes in those aquifers. To study the seawater intrusion process in the region, a two-dimensional density-dependent, saturated and unsaturated flow and transport computational model is used. The model consists of a coupled system of two non-linear partial differential equations. The first equation describes the flow of a variable-density fluid, and the second equation describes the transport of dissolved salt. A two-dimensional control volume finite element model is developed for simulating the seawater intrusion into the heterogeneous aquifer system at Gooburrum. The simulation results provide a realistic mechanism by which to study the convoluted transport phenomena evolving in this complex heterogeneous coastal aquifer.

Models of collective cell spreading with variable cell aspect ratio : a motivation for degenerate diffusion models

Continuum diffusion models are often used to represent the collective motion of cell populations. Most previous studies have simply used linear diffusion to represent collective cell spreading, while others found that degenerate nonlinear diffusion provides a better match to experimental cell density profiles. In the cell modeling literature there is no guidance available with regard to which approach is more appropriate for representing the spreading of cell populations. Furthermore, there is no knowledge of particular experimental measurements that can be made to distinguish between situations where these two models are appropriate. Here we provide a link between individual-based and continuum models using a multi-scale approach in which we analyze the collective motion of a population of interacting agents in a generalized lattice-based exclusion process. For round agents that occupy a single lattice site, we find that the relevant continuum description of the system is a linear diffusion equation, whereas for elongated rod-shaped agents that occupy L adjacent lattice sites we find that the relevant continuum description is connected to the porous media equation (pme). The exponent in the nonlinear diffusivity function is related to the aspect ratio of the agents. Our work provides a physical connection between modeling collective cell spreading and the use of either the linear diffusion equation or the pme to represent cell density profiles. Results suggest that when using continuum models to represent cell population spreading, we should take care to account for variations in the cell aspect ratio because different aspect ratios lead to different continuum models.