Novel educational training program for community pharmacists

There is little evidence that workshops alone have a lasting impact on the day-to-day practice of participants. The current paper examined a strategy to increase generalization and maintenance of skills in the natural environment using pseudo-patients and immediate performance feedback to reinforce skills acquisition. A random half of pharmacies (N=30) took part in workshop training aimed at optimizing consumers' use of nonprescription analgesic products. Pharmacies in the training group also received performance feedback on their adherence to the recommended protocol. Feedback occurred immediately after a pseudo-patient visit in which confederates posed as purchasers of analgesics, and combined positive and corrective elements. Trained pharmacists were significantly more accurate at identifying people who misused the medication (P<0.001). The trained pharmacists were more likely than controls to use open-ended questions (P<0.001), assess readiness to change problematic use (P <0.001), and to deliver a brief intervention that was tailored to the person's commitment to alter his/her usage (P <0.001). Participants responded to the feedback positively. Results were consistent with the hypothesis that when workshop is combined with on-site performance feedback, it enhances practitioners' adherence to protocols in the natural setting.

Comparison of methods for processing drinking water samples for the isolation of Myocbacterium avium and intracellulare

Several protocols for isolation of mycobacteria from water exist, but there is no established standard method. This study compared methods of processing potable water samples for the isolation of Mycobacterium avium and Mycobacterium intracellulare using spiked sterilized water and tap water decontaminated using 0.005% cetylpyridinium chloride (CPC). Samples were concentrated by centrifugation or filtration and inoculated onto Middlebrook 7H10 and 7H11 plates and Lowenstein-Jensen slants and into mycobacterial growth indicator tubes with or without polymyxin, azlocillin, nalidixic acid, trimethoprim, and amphotericin B. The solid media were incubated at 32°C, at 35°C, and at 35°C with CO2 and read weekly. The results suggest that filtration of water for the isolation of mycobacteria is a more sensitive method for concentration than centrifugation. The addition of sodium thiosulfate may not be necessary and may reduce the yield. Middlebrook M7H10 and 7H11 were equally sensitive culture media. CPC decontamination, while effective for reducing growth of contaminants, also significantly reduces mycobacterial numbers. There was no difference at 3 weeks between the different incubation temperatures.

The manipulation of apoptosis-related genes to generate resistance to Fusarium wilt and water stress in banana

Bananas are susceptible to a diverse range of biotic and abiotic stresses, many of which cause serious production constraints worldwide. One of the most destructive banana diseases is Fusarium wilt caused by the soil-borne fungus, Fusarium oxysporum f. sp. cubense (Foc). No effective control strategy currently exists for this disease which threatens global banana production. Although disease resistance exists in some wild bananas, attempts to introduce resistance into commercially acceptable bananas by conventional breeding have been hampered by low fertility, long generation times and association of poor agronomical traits with resistance genes. With the advent of reliable banana transformation protocols, molecular breeding is now regarded as a viable alternative strategy to generate disease-resistant banana plants. Recently, a novel strategy involving the expression of anti-apoptosis genes in plants was shown to result in resistance against several necrotrophic fungi. Further, the transgenic plants showed increased resistance to a range of abiotic stresses. In this thesis, the use of anti-apoptosis genes to generate transgenic banana plants with resistance to Fusarium wilt was investigated. Since water stress is an important abiotic constraint to banana production, the resistance of the transgenic plants to water stress was also examined. Embryogenic cell suspensions (ECS) of two commercially important banana cultivars, Grand Naine (GN) and Lady Finger (LF), were transformed using Agrobacterium with the anti-apoptosis genes, Bcl-xL, Bcl-xL G138A, Ced-9 and Bcl- 2 3’ UTR. An interesting, and potentially important, outcome was that the use of anti-apoptosis genes resulted in up to a 50-fold increase in Agrobacterium-mediated transformation efficiency of both LF and GN cells over vector controls. Regenerated plants were subjected to a complete molecular characterisation in order to detect the presence of the transgene (PCR), transcript (RT-PCR) and gene product (Western blot) and to determine the gene copy number (Southern blot). A total of 36 independently-transformed GN lines (8 x Bcl-xL, 5 x Bcl-xL G138A, 15 x Ced-9 and 8 x Bcl-2 3’ UTR) and 41 independently-transformed LF lines (8 x Bcl-xL, 7 x BclxL G138A, 13 x Ced-9 and 13 x Bcl-2 3’ UTR) were identified. The 41 transgenic LF lines were multiplied and clones from each line were acclimatised and grown under glasshouse conditions for 8 weeks to allow monitoring for phenotypic abnormalities. Plants derived from 3 x Bcl-xL, 2 x Ced-9 and 5 x Bcl-2 3’ UTR lines displayed a variety of aberrant phenotypes. However, all but one of these abnormalities were off-types commonly observed in tissue-cultured, non-transgenic banana plants and were therefore unlikely to be transgene-related. Prior to determining the resistance of the transgenic plants to Foc race 1, the apoptotic effects of the fungus on both wild-type and Bcl-2 3’ UTR-transgenic LF banana cells were investigated using rapid in vitro root assays. The results from these assays showed that apoptotic-like cell death was elicited in wild-type banana root cells as early as 6 hours post-exposure to fungal spores. In contrast, these effects were attenuated in the root cells of Bcl-2 3’ UTR-transgenic lines that were exposed to fungal spores. Thirty eight of the 41 transgenic LF lines were subsequently assessed for resistance to Foc race 1 in small-plant glasshouse bioassays. To overcome inconsistencies in rating the internal (vascular discolouration) disease symptoms, a MatLab-based computer program was developed to accurately and reliably assess the level of vascular discolouration in banana corms. Of the transgenic LF banana lines challenged with Foc race 1, 2 x Bcl-xL, 3 x Ced-9, 2 x Bcl-2 3’ UTR and 1 x Bcl-xL G138A-transgenic line were found to show significantly less external and internal symptoms than wild-type LF banana plants used as susceptible controls at 12 weeks post-inoculation. Of these lines, Bcl-2 3’ UTR-transgenic line #6 appeared most resistant, displaying very mild symptoms similar to the wild-type Cavendish banana plants that were included as resistant controls. This line remained resistant for up to 23 weeks post-inoculation. Since anti-apoptosis genes have been shown to confer resistance to various abiotic stresses in other crops, the ability of these genes to confer resistance against water stress in banana was also investigated. Clonal plants derived from each of the 38 transgenic LF banana plants were subjected to water stress for a total of 32 days. Several different lines of transgenic plants transformed with either Bcl-xL, Bcl-xL G138A, Ced-9 or Bcl-2 3’ UTR showed a delay in visual water stress symptoms compared with the wild-type control plants. These plants all began producing new growth from the pseudostem following daily rewatering for one month. In an attempt to determine whether the protective effect of anti-apoptosis genes in transgenic banana plants was linked with reactive oxygen species (ROS)-associated programmed cell death (PCD), the effect of the chloroplast-targeting, ROS-inducing herbicide, Paraquat, on wild-type and transgenic LF was investigated. When leaf discs from wild-type LF banana plants were exposed to 10 ìM Paraquat, complete decolourisation occurred after 48 hours which was confirmed to be associated with cell death and ROS production by trypan blue and 3,3-diaminobenzidine (DAB) staining, respectively. When leaf discs from the transgenic lines were exposed to Paraquat, those derived from some lines showed a delay in decolourisation, suggesting only a weak protective effect from the transgenes. Finally, the protective effect of anti-apoptosis genes against juglone, a ROS-inducing phytotoxin produced by the causal agent of black Sigatoka, Mycosphaerella fijiensis, was investigated. When leaf discs from wild-type LF banana plants were exposed to 25 ppm juglone, complete decolourisation occurred after 48 hours which was again confirmed to be associated with cell death and ROS production by trypan blue and DAB staining, respectively. Further, TdT-mediated dUTP nick-end labelling (TUNEL) assays on these discs suggested that the cell death was apoptotic. When leaf discs from the transgenic lines were exposed to juglone, discs from some lines showed a clear delay in decolourisation, suggesting a protective effect. Whether these plants are resistant to black Sigatoka is unknown and will require future glasshouse and field trials. The work presented in this thesis provides the first report of the use of anti-apoptosis genes as a strategy to confer resistance to Fusarium wilt and water stress in a nongraminaceous monocot, banana. Such a strategy may be exploited to generate resistance to necrotrophic pathogens and abiotic stresses in other economically important crop plants.

Efficient one-round key exchange in the standard model

We consider one-round key exchange protocols secure in the standard model. The security analysis uses the powerful security model of Canetti and Krawczyk and a natural extension of it to the ID-based setting. It is shown how KEMs can be used in a generic way to obtain two different protocol designs with progressively stronger security guarantees. A detailed analysis of the performance of the protocols is included; surprisingly, when instantiated with specific KEM constructions, the resulting protocols are competitive with the best previous schemes that have proofs only in the random oracle model.

Efficient one-round key exchange in the efficient one-round key exchange in the standard model

We consider one-round key exchange protocols secure in the standard model. The security analysis uses the powerful security model of Canetti and Krawczyk and a natural extension of it to the ID-based setting. It is shown how KEMs can be used in a generic way to obtain two different protocol designs with progressively stronger security guarantees. A detailed analysis of the performance of the protocols is included; surprisingly, when instantiated with specific KEM constructions, the resulting protocols are competitive with the best previous schemes that have proofs only in the random oracle model.

Efficient one-round key exchange in the standard model

We consider one-round key exchange protocols secure in the standard model. The security analysis uses the powerful security model of Canetti and Krawczyk and a natural extension of it to the ID-based setting. It is shown how KEMs can be used in a generic way to obtain two different protocol designs with progressively stronger security guarantees. A detailed analysis of the performance of the protocols is included; surprisingly, when instantiated with specific KEM constructions, the resulting protocols are competitive with the best previous schemes that have proofs only in the random oracle model.

The role of intermediaries in the implementation and development of asynchronous rural access

In Orissa state, India, the DakNet system supports asynchronous Internet communication between an urban hub and rural nodes. DakNet is noteworthy in many respects, not least in how the system leverages existing transport infrastructure. Wi-Fi transceivers mounted on local buses send and receive user data from roadside kiosks, for later transfer to/from the Internet via wireless protocols. This store-and-forward system allows DakNet to offer asynchronous communication capacity to rural users at low cost. The original ambition of the DakNet system was to provide email and SMS facilities to rural communities. Our 2008 study of the communicative ecology surrounding the DakNet system revealed that this ambition has now evolved – in response to market demand – to the extent that e-shopping (rather than email) has become the primary driver behind the DakNet offer.

Scheduling trains as a blocking parallel-machine job shop scheduling problem

In this paper, the train scheduling problem is modelled as a blocking parallel-machine job shop scheduling (BPMJSS) problem. In the model, trains, single-track sections and multiple-track sections, respectively, are synonymous with jobs, single machines and parallel machines, and an operation is regarded as the movement/traversal of a train across a section. Due to the lack of buffer space, the real-life case should consider blocking or hold-while-wait constraints, which means that a track section cannot release and must hold the train until next section on the routing becomes available. Based on literature review and our analysis, it is very hard to find a feasible complete schedule directly for BPMJSS problems. Firstly, a parallel-machine job-shop-scheduling (PMJSS) problem is solved by an improved shifting bottleneck procedure (SBP) algorithm without considering blocking conditions. Inspired by the proposed SBP algorithm, feasibility satisfaction procedure (FSP) algorithm is developed to solve and analyse the BPMJSS problem, by an alternative graph model that is an extension of the classical disjunctive graph models. The proposed algorithms have been implemented and validated using real-world data from Queensland Rail. Sensitivity analysis has been applied by considering train length, upgrading track sections, increasing train speed and changing bottleneck sections. The outcomes show that the proposed methodology would be a very useful tool for the real-life train scheduling problems

Information interventions for orienting patients and their carers to cancer care facilities (Protocol)

To assess the effects of information interventions which orient patients and their carers/family to a cancer care facility and the services available in the facility.

Multi-factor password-authenticated key exchange

We consider a new form of authenticated key exchange which we call multi-factor password-authenticated key exchange, where session establishment depends on successful authentication of multiple short secrets that are complementary in nature, such as a long-term password and a one-time response, allowing the client and server to be mutually assured of each other's identity without directly disclosing private information to the other party. Multi-factor authentication can provide an enhanced level of assurance in higher-security scenarios such as online banking, virtual private network access, and physical access because a multi-factor protocol is designed to remain secure even if all but one of the factors has been compromised. We introduce a security model for multi-factor password-authenticated key exchange protocols, propose an efficient and secure protocol called MFPAK, and provide a security argument to show that our protocol is secure in this model. Our security model is an extension of the Bellare-Pointcheval-Rogaway security model for password-authenticated key exchange and accommodates an arbitrary number of symmetric and asymmetric authentication factors.

Protein complementation assay as a display system for screening protein libraries in the intracellular environment

A wide range of screening strategies have been employed to isolate antibodies and other proteins with specific attributes, including binding affinity, specificity, stability and improved expression. However, there remains no high-throughput system to screen for target-binding proteins in a mammalian, intracellular environment. Such a system would allow binding reagents to be isolated against intracellular clinical targets such as cell signalling proteins associated with tumour formation (p53, ras, cyclin E), proteins associated with neurodegenerative disorders (huntingtin, betaamyloid precursor protein), and various proteins crucial to viral replication (e.g. HIV-1 proteins such as Tat, Rev and Vif-1), which are difficult to screen by phage, ribosome or cell-surface display. This study used the β-lactamase protein complementation assay (PCA) as the display and selection component of a system for screening a protein library in the cytoplasm of HEK 293T cells. The colicin E7 (ColE7) and Immunity protein 7 (Imm7) *Escherichia coli* proteins were used as model interaction partners for developing the system. These proteins drove effective β-lactamase complementation, resulting in a signal-to-noise ratio (9:1 – 13:1) comparable to that of other β-lactamase PCAs described in the literature. The model Imm7-ColE7 interaction was then used to validate protocols for library screening. Single positive cells that harboured the Imm7 and ColE7 binding partners were identified and isolated using flow cytometric cell sorting in combination with the fluorescent β-lactamase substrate, CCF2/AM. A single-cell PCR was then used to amplify the Imm7 coding sequence directly from each sorted cell. With the screening system validated, it was then used to screen a protein library based the Imm7 scaffold against a proof-of-principle target. The wild-type Imm7 sequence, as well as mutants with wild-type residues in the ColE7- binding loop were enriched from the library after a single round of selection, which is consistent with other eukaryotic screening systems such as yeast and mammalian cell-surface display. In summary, this thesis describes a new technology for screening protein libraries in a mammalian, intracellular environment. This system has the potential to complement existing screening technologies by allowing access to intracellular proteins and expanding the range of targets available to the pharmaceutical industry.

An ecological assessment of secondary poisoning risk in the Australian sugarcane industry

Rodenticide use in agriculture can lead to the secondary poisoning of avian predators. Currently the Australian sugarcane industry has two rodenticides, Racumin® and Rattoff®, available for in-crop use but, like many agricultural industries, it lacks an ecologically-based method of determining the potential secondary poisoning risk the use of these rodenticides poses to avian predators. The material presented in this thesis addresses this by: a. determining where predator/prey interactions take place in sugar producing districts; b. quantifying the amount of rodenticide available to avian predators and the probability of encounter; and c. developing a stochastic model that allows secondary poisoning risk under various rodenticide application scenarios to be investigated. Results demonstrate that predator/prey interactions are highly constrained by environmental structure. Rodents used crops that provided high levels of canopy cover and therefore predator protection and poorly utilised open canopy areas. In contrast, raptors over-utilised areas with low canopy cover and low rodent densities, but which provided high accessibility to prey. Given this pattern of habitat use, and that industry baiting protocols preclude rodenticide application in open canopy crops, these results indicate that secondary poisoning can only occur if poisoned rodents leave closed canopy crops and become available for predation in open canopy areas. Results further demonstrate that after in-crop rodenticide application, only a small proportion of rodents available in open areas are poisoned and that these rodents carry low levels of toxicant. Coupled with the low level of rodenticide use in the sugar industry, the high toxic threshold raptors have to these toxicants and the low probability of encountering poisoned rodents, results indicate that the risk of secondary poisoning events occurring is minimal. A stochastic model was developed to investigate the effect of manipulating factors that might influence secondary poisoning hazard in a sugarcane agro-ecosystem. These simulations further suggest that in all but extreme scenarios, the risk of secondary poisoning is also minimal. Collectively, these studies demonstrate that secondary poisoning of avian predators associated with the use of the currently available rodenticides in Australian sugar producing districts is minimal. Further, the ecologically-based method of assessing secondary poisoning risk developed in this thesis has broader applications in other agricultural systems where rodenticide use may pose risks to avian predators.

How can flexible learning centres use social networking to improve their ongoing educational assessments?

Most online assessment systems now incorporate social networking features, and recent developments in social media spaces include protocols that allow the synchronisation and aggregation of data across multiple user profiles. In light of these advances and the concomitant fear of data sharing in secondary school education this papers provides important research findings about generic features of online social networking, which educators can use to make sound and efficient assessments in collaboration with their students and colleagues. This paper reports on a design experiment in flexible educational settings that challenges the dichotomous legacy of success and failure evident in many assessment activities for at-risk youth. Combining social networking practices with the sociology of education the paper proposes that assessment activities are best understood as a negotiable field of exchange. In this design experiment students, peers and educators engage in explicit, "front-end" assessment (Wyatt-Smith, 2008) to translate digital artefacts into institutional, and potentiality economic capital without continually referring to paper based pre-set criteria. This approach invites students and educators to use social networking functions to assess “work in progress” and final submissions in collaboration, and in doing so assessors refine their evaluative expertise and negotiate the value of student’s work from which new criteria can emerge. The mobile advantages of web-based technologies aggregate, externalise and democratise this transparent assessment model for most, if not all, student work that can be digitally represented.

One-time-password-authenticated key exchange

To reduce the damage of phishing and spyware attacks, banks, governments, and other security-sensitive industries are deploying one-time password systems, where users have many passwords and use each password only once. If a single password is compromised, it can be only be used to impersonate the user once, limiting the damage caused. However, existing practical approaches to one-time passwords have been susceptible to sophisticated phishing attacks. ---------- We give a formal security treatment of this important practical problem. We consider the use of one-time passwords in the context of password-authenticated key exchange (PAKE), which allows for mutual authentication, session key agreement, and resistance to phishing attacks. We describe a security model for the use of one-time passwords, explicitly considering the compromise of past (and future) one-time passwords, and show a general technique for building a secure one-time-PAKE protocol from any secure PAKE protocol. Our techniques also allow for the secure use of pseudorandomly generated and time-dependent passwords.

Predicate-based key exchange

We provide the first description of and security model for authenticated key exchange protocols with predicate-based authentication. In addition to the standard goal of session key security, our security model also provides for credential privacy: a participating party learns nothing more about the other party's credentials than whether they satisfy the given predicate. Our model also encompasses attribute-based key exchange since it is a special case of predicate-based key exchange.---------- We demonstrate how to realize a secure predicate-based key exchange protocol by combining any secure predicate-based signature scheme with the basic Diffie-Hellman key exchange protocol, providing an efficient and simple solution.