The effect of the histological properties of tumors on transfection efficiency of electrically assisted gene delivery to solid tumors in mice

Uniform DNA distribution in tumors is a prerequisite step for high transfection efficiency in solid tumors. To improve the transfection efficiency of electrically assisted gene delivery to solid tumors in vivo, we explored how tumor histological properties affected transfection efficiency. In four different tumor types (B16F1, EAT, SA-1 and LPB), proteoglycan and collagen content was morphometrically analyzed, and cell size and cell density were determined in paraffin-embedded tumor sections under a transmission microscope. To demonstrate the influence of the histological properties of solid tumors on electrically assisted gene delivery, the correlation between histological properties and transfection efficiency with regard to the time interval between DNA injection and electroporation was determined. Our data demonstrate that soft tumors with larger spherical cells, low proteoglycan and collagen content, and low cell density are more effectively transfected (B16F1 and EAT) than rigid tumors with high proteoglycan and collagen content, small spindle-shaped cells and high cell density (LPB and SA-1). Furthermore, an optimal time interval for increased transfection exists only in soft tumors, this being in the range of 5-15 min. Therefore, knowledge about the histology of tumors is important in planning electrogene therapy with respect to the time interval between DNA injection and electroporation.

Gene electrotransfer into murine skeletal muscle: A systematic analysis of parameters for long-term gene expression

Skeletal muscle is an attractive target tissue for delivery of therapeutic genes, since it is well vascularized, easily accessible, and has a high capacity for protein synthesis. For efficient transfection in skeletal muscle, several protocols have been described, including delivery of low voltage electric pulses and a combination of high and low voltage electric pulses. The aim of this study was to determine the influence of different parameters of electrotransfection on short-term and long-term transfection efficiency in murine skeletal muscle, and to evaluate histological changes in the treated tissue. Different parameters of electric pulses, different time lags between plasmid DNA injection and application of electric pulses, and different doses of plasmid DNA were tested for electrotransfection of tibialis cranialis muscle of C57BI/6 mice using DNA plasmid encoding green fluorescent protein (GFP). Transfection efficiency was assessed on frozen tissue sections one week after electrotransfection using a fluorescence microscope and also noninvasively, followed by an in vivo imaging system using a fluorescence stereo microscope over a period of several months. Histological changes in muscle were evaluated immediately or several months after electrotransfection by determining infiltration of inflammatory mononuclear cells and presence of necrotic muscle fibers. The most efficient electrotransfection into skeletal muscle of C57BI/6 mice in our experiments was achieved when one high voltage (HV) and four low voltage (LV) electric pulses were applied 5 seconds after the injection of 30 μg of plasmid DNA. This protocol resulted in the highest short-term as well as long-term transfection. The fluorescence intensity of the transfected area declined after 2-3 weeks, but GFP fluorescence was still detectable 18 months after electrotransfection. Extensive inflammatory mononuclear cell infiltration was observed immediately after the electrotransfection procedure using the described parameters, but no necrosis or late tissue damage was observed. This study showed that electric pulse parameters, time lag between the injection of DNA and application of electric pulses, and dose of plasmid DNA affected the duration of transgene expression in murine skeletal muscle. Therefore, transgene expression in muscle can be controlled by appropriate selection of electrotransfection protocol.

Muscle gene electrotransfer is increased by the antioxidant tempol in mice

Electropermeabilization (EP) is an effective method of gene transfer into different tissues. During EP, reactive oxygen species (ROS) are formed, which could affect transfection efficiency. The role of generated ROS and the role of antioxidants in electrotransfer in myoblasts in vitro and in Musculus tibialis cranialis in mice were, therefore, investigated. We demonstrate in the study that during EP of C2C12 myoblasts, ROS are generated on the surface of the cells, which do not induce long-term genomic DNA damage. Plasmid DNA for transfection (pEGFP-N1), which is present outside the cells during EP, neutralizes the generated ROS. The ROS generation is proportional to the amplitude of the electric pulses and can be scavenged by antioxidants, such as vitamin C or tempol. When antioxidants were used during gene electrotransfer, the transfection efficiency of C2C12 myoblasts was statistically significantly increased 1.6-fold with tempol. Also in vivo, the transfection efficiency of M. tibialis cranialis in mice was statistically significantly increased 1.4-fold by tempol. The study indicates that ROS are generated on cells during EP and can be scavenged by antioxidants. Specifically, tempol can be used to improve gene electrotransfer into the muscle and possibly also to other tissues.

On the efficiency of impingers with fritted nozzle tip for collection of ultrafine particles

The aim of this study was to determine the collection efficiency of ultrafine particles into an impinger fitted with a fritted nozzle tip as a means to increase contact surface area between the aerosol and the liquid. The influence of liquid sampling volume, frit porosity and the nature of the sampling liquid was explored and it was shown that all impact on the collection efficiency of particles smaller than 220 nm. Obtained values for overall collection efficiency were substantially higher (~30–95%) than have been previously reported, mainly due to the high deposition of particles in the fritted nozzle tip, especially in case of finer porosity frits and smaller particles. Values for the capture efficiency of the solvent alone ranged from 20 to 45%, depending on the type and the volume of solvent. Additionally, our results show that airstream dispersion into bubbles improves particle trapping by the liquid and that there is a difference in collection efficiencies based on the nature and volume of the solvent used.

Final report : tasks 1 and 4: eco-efficiency assessment of building refurbishment

The main objective was to compare the environmental impacts of a building undergoing refurbishment both before and after the refurbishment and to assist in the design of the refurbishment with what is learned.

Re-life of buildings - decision support tools for maximising project efficiency

This paper describes the process adopted in developing an integrated decision support framework for planning of office building refurbishment projects, with specific emphasize on optimising rentable floor space, structural strengthening, residual life and sustainability. Expert opinion on the issues to be considered in a tool is being captured through the DELPHI process, which is currently ongoing. The methodology for development of the integrated tool will be validated through decisions taken during a case study project: refurbishment of CH1 building of Melbourne City Council, which will be followed through to completion by the research team. Current status of the CH1 planning will be presented in the context of the research project.

Sustainable construction for the future - the role of government in energy efficiency and sustainability in buildings

This paper will summarise the findings from a study that explored the link between dwelling design, or type, and energy efficiencies in sub-tropical climates. An increasing number of government and private sector development companies are initiating projects that aim to deliver enhanced environmental outcomes at both sub-divisional and dwelling levels. The study used AccuRate, a new thermal modelling tool developed by CSIRO that responds to the need to improve ventilation modelling. The study found that dwellings developed in conjunction with the Departments of Housing and Public Works have set the benchmark. It provides a snapshot of the energy efficiency of a range of dwelling types found in recent subdivisions. However, the trend toward increasing urban densities may reduce the likelihood that cooling breezes will be available to cool dwellings. The findings are relevant to regulators, designers and industry in all states interested in reducing the energy used to cool dwellings in summer.

Integrating eco-efficiency assessment of commercial buildings into the design process : LCADesign

The ability to assess a commercial building for its impact on the environment at the earliest stage of design is a goal which is achievable by integrating several approaches into a single procedure directly from the 3D CAD representation. Such an approach enables building design professionals to make informed decisions on the environmental impact of building and its alternatives during the design development stage instead of at the post-design stage where options become limited. The indicators of interest are those which relate to consumption of resources and energy, contributions to pollution of air, water and soil, and impacts on the health and wellbeing of people in the built environment as a result of constructing and operating buildings. 3D object-oriented CAD files contain a wealth of building information which can be interrogated for details required for analysis of the performance of a design. The quantities of all components in the building can be automatically obtained from the 3D CAD objects and their constituent materials identified to calculate a complete list of the amounts of all building products such as concrete, steel, timber, plastic etc. When this information is combined with a life cycle inventory database, key internationally recognised environmental indicators can be estimated. Such a fully integrated tool known as LCADesign has been created for automated ecoefficiency assessment of commercial buildings direct from 3D CAD. This paper outlines the key features of LCADesign and its application to environmental assessment of commercial buildings.

LCADesign : an integrated approach to automatic eco-efficiency assessment of commercial buildings

Buildings consume resources and energy, contribute to pollution of our air, water and soil, impact the health and well-being of populations and constitute an important part of the built environment in which we live. The ability to assess their design with a view to reducing that impact automatically from their 3D CAD representations enables building design professionals to make informed decisions on the environmental impact of building structures. Contemporary 3D object-oriented CAD files contain a wealth of building information. LCADesign has been designed as a fully integrated approach for automated eco-efficiency assessment of commercial buildings direct from 3D CAD. LCADesign accesses the 3D CAD detail through Industry Foundation Classes (IFCs) - the international standard file format for defining architectural and constructional CAD graphic data as 3D real-world objects - to permit construction professionals to interrogate these intelligent drawing objects for analysis of the performance of a design. The automated take-off provides quantities of all building components whose specific production processes, logistics and raw material inputs, where necessary, are identified to calculate a complete list of quantities for all products such as concrete, steel, timber, plastic etc and combines this information with the life cycle inventory database, to estimate key internationally recognised environmental indicators such as CML, EPS and Eco-indicator 99. This paper outlines the key modules of LCADesign and their role in delivering an automated eco-efficiency assessment for commercial buildings.

Assessing the energy efficiency of gear oils via the FZG test machine

Energy efficient lubricants are becoming increasingly popular. This is due to a global increase in environmental awareness combined with the potential of reducing operating costs. A new test method of evaluating the energy efficiency of gear oils has been described in this report. The method involves measuring the power required by an FZG test rig to run while using a particular test lubricant. For each oil that was being evaluated, the rig was run for 10 minutes at a load stage of 10. Six extreme pressure (EP) industrial gear oils of mineral base were tested. The difference in power requirements between the best and the worst performing oils was 2.77 and 3.24 kW, respectively. This equates to a 14.6% reduction in power, a significant amount if considered in relation to a high powered industrial machine. The oils of superior performance were noticed to run at reduced temperatures. They were also more expensive than the other products of lesser performance.

The innovation deficit in public services : the curious problem of too much efficiency and not enough waste and failure

It has long been recognised that government and public sector services suffer an innovation deficit compared to private or market-based services. This paper argues that this can be explained as an unintended consequence of the concerted public sector drive toward the elimination of waste through efficiency, accountability and transparency. Yet in an evolving economy this can be a false efficiency, as it also eliminates the 'good waste' that is a necessary cost of experimentation. This results in a systematic trade0off in the public sector between the static efficiency of minimizing the misuse of public resources and the dynamic efficiency of experimentation. this is inherently biased against risk and uncertainty and therein, explains why governments find service innovation so difficult. In the drive to eliminate static inefficiencies, many political systems have susequently overshot and stifled policy innovation. I propose the 'Red Queen' solution of adaptive economic policy.

Using a current source to improve efficiency of a plasma system

This paper presents the possibility of utilizing a current source topology instead of a voltage source as an efficient, flexible and reliable power supply for plasma applications. A buck-boost converter with a current controller has been used to transfer energy from an inductor to a plasma system. A control strategy has also been designed to satisfy all the desired purposes. The main concept behind this topology is to provide high dv/dt regardless of the switching speed of a power switch and to control the current level to properly transfer adequate energy to various plasma applications.

Impacts of vessel capacity reduction programs on the efficiency in fisheries. The case of Australia's multispecies in northern prawn fishery

Capacity reduction programs in the form of buybacks or decommissioning programs have had relatively widespread application in fisheries in the US, Europe and Australia. A common criticism of such programs is that they remove the least efficient vessels first, resulting in an increase in average efficiency of the remaining fleet. The effective fishing power of the fleet, therefore, does not decrease in proportion to the number of vessels removed. Further, reduced crowding may increase efficiency of the remaining vessels. In this paper, the effects of a buyback program on average technical efficiency in Australia’s Northern Prawn Fishery are examined using a multi-output distance function approach with an explicit inefficiency model. The results indicate that average efficiency of the remaining vessels was greater than that of the removed vessels, and that average efficiency of remaining vessels also increased as a result of reduced crowding.