Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium

The recognition of the potential efficacy of plasmid DNA (pDNA) molecules as vectors in the treatment and prevention of emerging diseases has birthed the confidence to combat global pandemics. This is due to the close-to-zero safety concern associated with pDNA vectors compared to viral vectors in cell transfection and targeting. Considerable attention has been paid to the potential of pDNA vectors but comparatively less thought has been given to the practical challenges in producing large quantities to meet current rising demands. A pilot-scale fermentation scheme was developed by employing a stoichiometrically-designed growth medium whose exceptional plasmid yield performance was attested in a shake flask environment for pUC19 and pEGFP-N1 transformed into E. coliDH5α and E. coliJM109, respectively. Batch fermentation of E. coliDH5α-pUC19 employing the stoichiometric medium displayed a maximum plasmid volumetric and specific yield of 62.6 mg/L and 17.1 mg/g (mg plasmid/g dry cell weight), respectively. Fed-batch fermentation of E. coliDH5α-pUC19 on a glycerol substrate demonstrated one of the highest ever reported pilot-scale plasmid specific yield of 48.98 mg/g and a volumetric yield of 0.53 g/L. The attainment of high plasmid specific yields constitutes a decrease in plasmid manufacturing cost and enhances the effectiveness of downstream processes by reducing the proportion of intracellular impurities. The effect of step-rise temperature induction was also considered to maximize ColE1-origin plasmid replication.

Bio-hydrogen production from food waste through anaerobic fermentation

In order to protect our planet and ourselves from the adverse effects of excessive CO2 emissions and to prevent an imminent non-renewable fossil fuel shortage and energy crisis, there is a need to transform our current ‘fossil fuel dependent’ energy systems to new, clean, renewable energy sources. The world has recognized hydrogen as an energy carrier that complies with all the environmental quality and energy security, demands. This research aimed at producing hydrogen through anaerobic fermentation, using food waste as the substrate. Four food waste substrates were used: Rice, fish, vegetable and their mixture. Bio-hydrogen production was performed in lab scale reactors, using 250 mL serum bottles. The food waste was first mixed with the anaerobic sewage sludge and incubated at 37°C for 31 days (acclimatization). The anaerobic sewage sludge was then heat treated at 80°C for 15 min. The experiment was conducted at an initial pH of 5.5 and temperatures of 27, 35 and 55°C. The maximum cumulative hydrogen produced by rice, fish, vegetable and mixed food waste substrates were highest at 37°C (Rice =26.97±0.76 mL, fish = 89.70±1.25 mL, vegetable = 42.00±1.76 mL, mixed = 108.90±1.42 mL). A comparative study of acclimatized (the different food waste substrates were mixed with anaerobic sewage sludge and incubated at 37°C for 31days) and non-acclimatized food waste substrate (food waste that was not incubated with anaerobic sewage sludge) showed that acclimatized food waste substrate enhanced bio-hydrogen production by 90 - 100%.

Surface engineering of reduced graphene oxide for controllable ambipolar flash memories

Tunable charge-trapping behaviors including unipolar charge trapping of one type of charge carrier and ambipolar trapping of both electrons and holes in a complementary manner is highly desirable for low power consumption multibit flash memory design. Here, we adopt a strategy of tuning the Fermi level of reduced graphene oxide (rGO) through self-assembled monolayer (SAM) functionalization and form p-type and n-type doped rGO with a wide range of manipulation on work function. The functionalized rGO can act as charge-trapping layer in ambipolar flash memories, and a dramatic transition of charging behavior from unipolar trapping of electrons to ambipolar trapping and eventually to unipolar trapping of holes was achieved. Adjustable hole/electron injection barriers induce controllable Vth shift in the memory transistor after programming operation. Finally, we transfer the ambipolar memory on flexible substrates and study their charge-trapping properties at various bending cycles. The SAM-functionalized rGO can be a promising candidate for next-generation nonvolatile memories.

A framework for generating realistic traffic for distributed denial-of-service attacks and flash events

An intrinsic challenge associated with evaluating proposed techniques for detecting Distributed Denial-of-Service (DDoS) attacks and distinguishing them from Flash Events (FEs) is the extreme scarcity of publicly available real-word traffic traces. Those available are either heavily anonymised or too old to accurately reflect the current trends in DDoS attacks and FEs. This paper proposes a traffic generation and testbed framework for synthetically generating different types of realistic DDoS attacks, FEs and other benign traffic traces, and monitoring their effects on the target. Using only modest hardware resources, the proposed framework, consisting of a customised software traffic generator, ‘Botloader’, is capable of generating a configurable mix of two-way traffic, for emulating either large-scale DDoS attacks, FEs or benign traffic traces that are experimentally reproducible. Botloader uses IP-aliasing, a well-known technique available on most computing platforms, to create thousands of interactive UDP/TCP endpoints on a single computer, each bound to a unique IP-address, to emulate large numbers of simultaneous attackers or benign clients.

AITPM Email Newsletter Volume 0905, June 2009 : President's Message

President’s Message Hello fellow AITPM members, Due to three colliding forces of nature I find myself writing this month’s message from home – today, I am still getting over a persistent virus that seemed to set in just after returning from Singapore a couple of weeks ago, which my diabetes won’t let me get away with too easily (no Kermit the Frog swine flu jokes please). Combine this with a very wet day in Brisbane – in fact the wettest for 20 years (how can we complain, except for flash flooding?). And in Queensland today is a state school teachers’ strike, so one half of our brood is over watching TV. Family snapshots aside, the biggest news for our industry of late is the $8.5 billion announced in the Federal Budget for transport infrastructure projects; many “shovel ready”, but some – and fortunate for our profession – desktop ready. This newsletter provides nationwide coverage on the transport infrastructure aspects of the Federal Budget. We’ll need a bit more time to carefully look at the ensuing State Budgets’ announcements. Regarding the federal budget announcements, I am pleased to see serious attention being paid to upgrading the M1 system – I hope to see a motorway standard facility connecting Adelaide to at least Rockhampton in my lifetime. But some other important roads are of course missing out in this particular budget. Various levels of commitment are being made to urban passenger rail – some project significant while others planning significant. Enhancement of suburban rail is important across the capitals and Australia’s medium sized cities such as the Gold Coast and Newcastle. Not much on road safety initiatives jumped out at me, but I believe it is implied in the large road projects and in some of the detail elsewhere. I do believe it’s about time a ‘Vision Zero’ style policy is adopted at the National level, since any death is unacceptable on the road, just as it is in any other workplace. So, overall some good news on building transport infrastructure to keep the economy purring during this recession, and strongly supporting it during future boom times. The other edge to the sword, of course, is that we tax payers will be paying a considerable amount for borrowings for these projects over a long period of time. I close again in reminding everyone again that AITPM’s flagship event, the 2009 AITPM National Conference, Traffic Beyond Tomorrow, is being held in Adelaide from 5 to 7 August. www.aitpm.com has all of the details about how to register, sponsor a booth, session, etc. Best regards all, Jon Bunker

Improved image contrast of the bone-muscle interface with 3T MRI compared to 1.5T MRI [Abstract]

Virtual 3D models of long bones are increasingly being used for implant design and research applications. The current gold standard for the acquisition of such data is Computed Tomography (CT) scanning. Due to radiation exposure, CT is generally limited to the imaging of clinical cases and cadaver specimens. Magnetic Resonance Imaging (MRI) does not involve ionising radiation and therefore can be used to image selected healthy human volunteers for research purposes. The feasibility of MRI as alternative to CT for the acquisition of morphological bone data of the lower extremity has been demonstrated in recent studies [1, 2]. Some of the current limitations of MRI are long scanning times and difficulties with image segmentation in certain anatomical regions due to poor contrast between bone and surrounding muscle tissues. Higher field strength scanners promise to offer faster imaging times or better image quality. In this study image quality at 1.5T is quantitatively compared to images acquired at 3T. --------- The femora of five human volunteers were scanned using 1.5T and 3T MRI scanners from the same manufacturer (Siemens) with similar imaging protocols. A 3D flash sequence was used with TE = 4.66 ms, flip angle = 15° and voxel size = 0.5 × 0.5 × 1 mm. PA-Matrix and body matrix coils were used to cover the lower limb and pelvis respectively. Signal to noise ratio (SNR) [3] and contrast to noise ratio (CNR) [3] of the axial images from the proximal, shaft and distal regions were used to assess the quality of images from the 1.5T and 3T scanners. The SNR was calculated for the muscle and bone-marrow in the axial images. The CNR was calculated for the muscle to cortex and cortex to bone marrow interfaces, respectively. --------- Preliminary results (one volunteer) show that the SNR of muscle for the shaft and distal regions was higher in 3T images (11.65 and 17.60) than 1.5T images (8.12 and 8.11). For the proximal region the SNR of muscles was higher in 1.5T images (7.52) than 3T images (6.78). The SNR of bone marrow was slightly higher in 1.5T images for both proximal and shaft regions, while it was lower in the distal region compared to 3T images. The CNR between muscle and bone of all three regions was higher in 3T images (4.14, 6.55 and 12.99) than in 1.5T images (2.49, 3.25 and 9.89). The CNR between bone-marrow and bone was slightly higher in 1.5T images (4.87, 12.89 and 10.07) compared to 3T images (3.74, 10.83 and 10.15). These results show that the 3T images generated higher contrast between bone and the muscle tissue than the 1.5T images. It is expected that this improvement of image contrast will significantly reduce the time required for the mainly manual segmentation of the MR images. Future work will focus on optimizing the 3T imaging protocol for reducing chemical shift and susceptibility artifacts.

Demonstration of cellulosic ethanol production from sugarcane bagasse in Australia : the Mackay Renewable Biocommodities Pilot Plant

The ready availability of sugarcane bagasse at an existing industrial facility and the potential availability of extra fibre through trash collection make sugarcane fibre the best candidate for early stage commercialisation of cellulosic ethanol technologies. The commercialisation of cellulosic ethanol technologies in the sugar industry requires both development of novel technologies and the assessment of these technologies at a pre-commercial scale. In 2007, the Queensland University of Technology (QUT) received funding from the Australian and Queensland Governments to construct a pilot research and development facility for the production of bioethanol and other renewable biocommodities from biomass including sugarcane bagasse. This facility has been built on the site of the Racecourse Sugar Mill in Mackay, Queensland and is known as the Mackay Renewable Biocommodities Pilot Plant (MRBPP). This research facility is capable of processing cellulosic biomass by a variety of pretreatment technologies and includes equipment for enzymatic saccharification, fermentation and distillation to produce ethanol. Lignin and fermentation co-products can also be produced in the pilot facility.

Visceral hedonic rhetoric

Emotional responses can incite and entice consumers to select a particular product from a row of similar items and thus have a considerable impact on purchase decisions. Consequently, more and more companies are challenging designers to address the emotional impact of their work and to design for emotion and consumerproduct relationships. Furthermore, the creation of emotional attachment to one’s possessions is one way of approaching a sustainable consumer-product relationship. The aim of this research is to gain a deeper understanding of the instantaneous emotional attachment that consumers form with products and its subsequent implications for product development. The foci of the study are visceral design, consumer hedonics and product rhetoric. Studied in a conglomerate they become an area of new investigation: visceral hedonic rhetoric. In this context, the term “visceral hedonic rhetoric” is defined as the properties of a product that persuasively elicit the pursuit of pleasure at an instinctual level of cognition. This study explores visceral hedonic rhetoric evident in the design of interactive products and resides within the context of emotional design research. It employs an empirical approach to understand how consumers respond hedonically on a visceral level to rhetoric in products. Specifically, it examines visceral hedonic responses given by thirty participants to the stimuli of six mobile telephones, six Mp3 players and six USB memory flash drives. The study findings demonstrate a hierarchy of visceral hedonic rhetoric evident in interactive products. This hierarchy of visceral hedonic attributes include: colour, size, shape, intrigue, material, perceived usability, portability, perceived function, novelty, analogy, brand, quality, texture and gender. However, it is the interrelationships between these visceral hedonic attributes that are the most significant findings of this research. Certain associations were revealed between product attribute combinations and consumer perception. The most predominant of these were: gender bias associated with colour selection; the creation of intrigue through a vibrant attention-grabbing colour; perceived ease of use and function; product confidence as a result of brand familiarity and perceived usability; analogous association through familiarity with similar objects and shapes; and the association of longevity with quality, novelty or recent technology. A significant outcome of the research is the distillation of visceral hedonic rhetoric design principles, and a tool to assist designers in harnessing the full potential of visceral hedonic rhetoric. This study contributes to the identification of the emerging research field of visceral hedonic rhetoric. Application of this study’s findings has the potential to provide a hedonic consumer-product relationship that is more meaningful, less disposable and more sustainable. This theory of visceral hedonic rhetoric is not only a significant contribution to design knowledge but is also generally transferable to other research domains, as later suggested in future research avenues.

Athletes series

The researcher was invited to photograph athletes in the lead-up to the 2006 Commonwealth Games held in Melbourne. She photographed four indigenous athletes, to produce a series of four large-scale cotton rag prints, 1 meter x 1 meter, printed onto photorag paper from digital files. “My photographic practice can be described as both political and spiritual, in the sense that as an Aboriginal Indigenous artist I take stock of the rationalising effect of the technologies I use, and create work that evokes nature and spirit. My methods often involve re-photographing or digitally re-working landscape photographs and adding historical or cultural icons of significance. Working with Indigenous athletes has been an honour and a pleasure. I admire the athletes’ passion and dedication to their chosen sport, and above all their humility, which seems a trait somewhat in contrast to what it takes to attain the highest levels of achievement. Indigenous athletes are wonderful role models for all Australians, and in making creative work that places their luminary presence with the land, I am aligning sportspeople with a deep sense of nature and spirit.” – Leah King-Smith. These works were commissioned by the National Portrait Gallery for the exhibition FLASH: Australian Athletes in Focus. The exhibition was a significant element in Melbourne2006 Festival, the cultural festival of the Commonwealth Games. The exhibition was prominently reviewed in Portrait: Magazine of Australian and International Portraiture and was subsequently remounted at Old Parliament House, Canberra (15 July to 12 November, 2006). One image was used for the front cover of Art Monthly, (March 2006).

The undesirable acetylation of cellulose by the acetate ion of 1-ethyl-3-methylimidazolium acetate

The ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate ([C2mim]OAc) is considered to be an inert solvent of cellulose and lignocellulosic biomass. Acetylation (1.7 % mol, or DS 0.017) of cellulose after dissolution in [C2mim]OAc (150 °C for 20 min), is demonstrated by compositional analysis, FTIR analysis and 13C NMR spectroscopy (in [C2min]OAc with 13C enriched acetate). This acetylation, in the absence of added acylating agents, has not been reported before and may limit [C2mim]OAc utility in industrial scale biomass processing, even at this low extent. For example, cellulose acetylation may contribute to IL loss in processes where the IL is recovered and reused and inhibit enzyme saccharification of cellulose in lignocellulosic biofuel production processes based on saccharification and fermentation.

Why the long face?

In this video, a male voice recites a script comprised entirely of jokes. Words flash on screen in time with the spoken words. Sometimes the two sets of words match, and sometimes they differ. This work examines processes of signification. It emphasizes disruption and disconnection as fundamental and generative operations in making meaning. Extending on post-structural and deconstructionist ideas, this work questions the relationship between written and spoken words. By deliberately confusing the signifying structures of jokes and narratives, it questions the sites and mechanisms of comprehension, humour and signification.

Then I let you

In this video, a male voice recites a teenage love poem. Words flash on screen in time with the spoken words. Sometimes the two sets of words match, and sometimes they differ. This work examines processes of signification. It emphasizes disruption and disconnection as fundamental and generative operations in making meaning. Extending on post-structural and deconstructionist ideas, this work questions the relationship between written and spoken words. By actively disrupting the sincerity of a teenage love poem, it questions the sites and mechanisms of comprehension, poetry and signification.

Assessing of circuit breaker restrike risks using computer simulation and wavelet analysis

A breaker restrike is an abnormal arcing phenomenon, leading to a possible breaker failure. Eventually, this failure leads to interruption of the transmission and distribution of the electricity supply system until the breaker is replaced. Before 2008, there was little evidence in the literature of monitoring techniques based on restrike measurement and interpretation produced during switching of capacitor banks and shunt reactor banks in power systems. In 2008 a non-intrusive radiometric restrike measurement method and a restrike hardware detection algorithm were developed by M.S. Ramli and B. Kasztenny. However, the limitations of the radiometric measurement method are a band limited frequency response as well as limitations in amplitude determination. Current restrike detection methods and algorithms require the use of wide bandwidth current transformers and high voltage dividers. A restrike switch model using Alternative Transient Program (ATP) and Wavelet Transforms which support diagnostics are proposed. Restrike phenomena become a new diagnostic process using measurements, ATP and Wavelet Transforms for online interrupter monitoring. This research project investigates the restrike switch model Parameter „A. dielectric voltage gradient related to a normal and slowed case of the contact opening velocity and the escalation voltages, which can be used as a diagnostic tool for a vacuum circuit-breaker (CB) at service voltages between 11 kV and 63 kV. During current interruption of an inductive load at current quenching or chopping, a transient voltage is developed across the contact gap. The dielectric strength of the gap should rise to a point to withstand this transient voltage. If it does not, the gap will flash over, resulting in a restrike. A straight line is fitted through the voltage points at flashover of the contact gap. This is the point at which the gap voltage has reached a value that exceeds the dielectric strength of the gap. This research shows that a change in opening contact velocity of the vacuum CB produces a corresponding change in the slope of the gap escalation voltage envelope. To investigate the diagnostic process, an ATP restrike switch model was modified with contact opening velocity computation for restrike waveform signature analyses along with experimental investigations. This also enhanced a mathematical CB model with the empirical dielectric model for SF6 (sulphur hexa-fluoride) CBs at service voltages above 63 kV and a generalised dielectric curve model for 12 kV CBs. A CB restrike can be predicted if there is a similar type of restrike waveform signatures for measured and simulated waveforms. The restrike switch model applications are used for: computer simulations as virtual experiments, including predicting breaker restrikes; estimating the interrupter remaining life of SF6 puffer CBs; checking system stresses; assessing point-on-wave (POW) operations; and for a restrike detection algorithm development using Wavelet Transforms. A simulated high frequency nozzle current magnitude was applied to an Equation (derived from the literature) which can calculate the life extension of the interrupter of a SF6 high voltage CB. The restrike waveform signatures for a medium and high voltage CB identify its possible failure mechanism such as delayed opening, degraded dielectric strength and improper contact travel. The simulated and measured restrike waveform signatures are analysed using Matlab software for automatic detection. Experimental investigation of a 12 kV vacuum CB diagnostic was carried out for the parameter determination and a passive antenna calibration was also successfully developed with applications for field implementation. The degradation features were also evaluated with a predictive interpretation technique from the experiments, and the subsequent simulation indicates that the drop in voltage related to the slow opening velocity mechanism measurement to give a degree of contact degradation. A predictive interpretation technique is a computer modeling for assessing switching device performance, which allows one to vary a single parameter at a time; this is often difficult to do experimentally because of the variable contact opening velocity. The significance of this thesis outcome is that it is a non-intrusive method developed using measurements, ATP and Wavelet Transforms to predict and interpret a breaker restrike risk. The measurements on high voltage circuit-breakers can identify degradation that can interrupt the distribution and transmission of an electricity supply system. It is hoped that the techniques for the monitoring of restrike phenomena developed by this research will form part of a diagnostic process that will be valuable for detecting breaker stresses relating to the interrupter lifetime. Suggestions for future research, including a field implementation proposal to validate the restrike switch model for ATP system studies and the hot dielectric strength curve model for SF6 CBs, are given in Appendix A.

The utilization of waste date seed as bio-oil and activated carbon by pyrolysis process

The renovation of biomass waste in the form of date seed waste into activated carbon and biofuel by fixed bed pyrolysis reactor has been focused in this study to obtain gaseous, liquid, and solid products. The date seed in particle form is pyrolysed in an externally heated fixed bed reactor with nitrogen as the carrier gas. The reactor is heated from 400◦C to 600◦C. A maximum liquid yield of 50wt.% and char of 30wt.% are obtained at a reactor bed temperature of 500◦C with a running time of 120 minutes. The oil is found to possess favorable flash point and reasonable density and viscosity. The higher calorific value is found to be 28.636 MJ/kg which is significantly higher than other biomass derived. Decolonization of 85–97% is recorded for the textile effluent and 75–90% for the tannery effluent, in all cases decreasing with temperature increase. Good adsorption capacity of the prepared activated carbon in case of diluted textile and tannery effluent was found.

Fixed bed pyrolysis of date seed waste for liquid oil production

The conversion of biomass waste in the form of date seed into pyrolysis oil by fixed bed pyrolysis reactor has been taken into consideration in this study. A fixed bed pyrolysis has been designed and fabricated for obtaining liquid fuel from these date seeds. The major component of the system are fixed bed pyrolysis reactor, liquid condenser and liquid collector. The date seed in particle form is pyrolysed in an externally heated 7.6 cm diameter and 46 cm high fixed bed reactor with nitrogen as the carrier gas. The reactor is heated by means of a biomass source cylindrical heater from 4000C to 6000C. The products are oil, char and gas. The reactor bed temperature, running time and feed particle size are considered as process parameters. The parameters are found to influence the product yield significantly. A maximum liquid yield of 50 wt.% is obtained at a reactor bed temperature of 5000 C for a feed size volume of 0.11- 0.20 cm3 with a running time of 120 minutes. The pyrolysis oil obtained at this optimum process conditions are analyzed for some fuel properties and compared with some other biomass derived pyrolysis oils and also with conventional fuels. The oil is found to possess favorable flash point and reasonable density and viscosity. The higher calorific value is found to be 28.636 MJ/kg which is significantly higher than other biomass derived pyrolysis oils.