Gamma-tocotrienol as an effective agent in targeting prostate cancer stem cell-like population

Emerging evidence supports that prostate cancer originates from a rare sub-population of cells, namely prostate cancer stem cells (CSCs). Conventional therapies for prostate cancer are believed to mainly target the majority of differentiated tumor cells but spare CSCs, which may account for the subsequent disease relapse after treatment. Therefore, successful elimination of CSCs may be an effective strategy to achieve complete remission from this disease. Gamma-tocotrienols (-T3) is one of the vitamin-E constituents which have been shown to have anticancer effects against a wide-range of human cancers. Recently, we have reported that -T3 treatment not only inhibits prostate cancer cell invasion but also sensitizes the cells to docetaxel-induced apoptosis, suggesting that -T3 may be an effective therapeutic agent against advanced stage prostate cancer. Here, we demonstrate for the first time that -T3 can down-regulate the expression of prostate CSC markers (CD133/CD44) in androgen independent (AI) prostate cancer cell lines (PC-3 & DU145), as evident from western blotting analysis. Meanwhile, the spheroid formation ability of the prostate cancer cells was significantly hampered by -T3 treatment. In addition, pre-treatment of PC-3 cells with -T3 was found to suppress tumor initiation ability of the cells. More importantly, while CD133-enriched PC-3 cells were highly resistant to docetaxel treatment, these cells were as sensitive to -T3 treatment as the CD133-depleted population. Our data suggest that -T3 may be an effective agent in targeting prostate CSCs, which may account for its anticancer and chemosensitizing effects reported in previous studies.

Optimal track access rights allocation for agent negotiation in an open railway market

In open railway access markets, a train service provider (TSP) negotiates with an infrastructure provider (IP) for track access rights. This negotiation has been modeled by a multi-agent system (MAS) in which the IP and TSP are represented by separate software agents. One task of the IP agent is to generate feasible (and preferably optimal) track access rights, subject to the constraints submitted by the TSP agent. This paper formulates an IP-TSP transaction and proposes a branch-and-bound algorithm for the IP agent to identify the optimal track access rights. Empirical simulation results show that the model is able to emulate rational agent behaviors. The simulation results also show good consistency between timetables attained from the proposed methods and those derived by the scheduling principles adopted in practice.

Conflict resolution through negotiation in a railway open access market : a multi-agent system approach

Open access reforms to railway regulations allow multiple train operators to provide rail services on a common infrastructure. As railway operations are now independently managed by different stakeholders, conflicts in operations may arise, and there have been attempts to derive an effective access charge regime so that these conflicts may be resolved. One approach is by direct negotiation between the infrastructure manager and the train service providers. Despite the substantial literature on the topic, few consider the benefits of employing computer simulation as an evaluation tool for railway operational activities such as access pricing. This article proposes a multi-agent system (MAS) framework for the railway open market and demonstrates its feasibility by modelling the negotiation between an infrastructure provider and a train service operator. Empirical results show that the model is capable of resolving operational conflicts according to market demand.

Modelling issues on the railway resource management process using multi-agent system

Many infrastructure and necessity systems such as electricity and telecommunication in Europe and the Northern America were used to be operated as monopolies, if not state-owned. However, they have now been disintegrated into a group of smaller companies managed by different stakeholders. Railways are no exceptions. Since the early 1980s, there have been reforms in the shape of restructuring of the national railways in different parts of the world. Continuous refinements are still conducted to allow better utilisation of railway resources and quality of service. There has been a growing interest for the industry to understand the impacts of these reforms on the operation efficiency and constraints. A number of post-evaluations have been conducted by analysing the performance of the stakeholders on their profits (Crompton and Jupe 2003), quality of train service (Shaw 2001) and engineering operations (Watson 2001). Results from these studies are valuable for future improvement in the system, followed by a new cycle of post-evaluations. However, direct implementation of these changes is often costly and the consequences take a long period of time (e.g. years) to surface. With the advance of fast computing technologies, computer simulation is a cost-effective means to evaluate a hypothetical change in a system prior to actual implementation. For example, simulation suites have been developed to study a variety of traffic control strategies according to sophisticated models of train dynamics, traction and power systems (Goodman, Siu and Ho 1998, Ho and Yeung 2001). Unfortunately, under the restructured railway environment, it is by no means easy to model the complex behaviour of the stakeholders and the interactions between them. Multi-agent system (MAS) is a recently developed modelling technique which may be useful in assisting the railway industry to conduct simulations on the restructured railway system. In MAS, a real-world entity is modelled as a software agent that is autonomous, reactive to changes, able to initiate proactive actions and social communicative acts. It has been applied in the areas of supply-chain management processes (García-Flores, Wang and Goltz 2000, Jennings et al. 2000a, b) and e-commerce activities (Au, Ngai and Parameswaran 2003, Liu and You 2003), in which the objectives and behaviour of the buyers and sellers are captured by software agents. It is therefore beneficial to investigate the suitability or feasibility of applying agent modelling in railways and the extent to which it might help in developing better resource management strategies. This paper sets out to examine the benefits of using MAS to model the resource management process in railways. Section 2 first describes the business environment after the railway 2 Modelling issues on the railway resource management process using MAS reforms. Then the problems emerge from the restructuring process are identified in section 3. Section 4 describes the realisation of a MAS for railway resource management under the restructured scheme and the feasible studies expected from the model.

A prioritised fuzzy constraint satisfaction approach to model agent negotiation for railway scheduling

With the recent regulatory reforms in a number of countries, railways resources are no longer managed by a single party but are distributed among different stakeholders. To facilitate the operation of train services, a train service provider (SP) has to negotiate with the infrastructure provider (IP) for a train schedule and the associated track access charge. This paper models the SP and IP as software agents and the negotiation as a prioritized fuzzy constraint satisfaction (PFCS) problem. Computer simulations have been conducted to demonstrate the effects on the train schedule when the SP has different optimization criteria. The results show that by assigning different priorities on the fuzzy constraints, agents can represent SPs with different operational objectives.

The development of Lactococcus lactis as an antimicrobial agent

Non-pathogenic lactic acid bacteria are economically important Gram-positive bacteria used extensively in the food industry. Due to their “generally regarded as safe” status, certain species from the genera Lactobacillus and Lactococcus are also considered desirable as candidates for the production and secretion of recombinant proteins, particular those with therapeutic applications. The hypothesis examined by this thesis is that Lactococcus lactis can be modified to be an effective antimicrobial agent. Therefore, the aims of this thesis were to investigate the optimisation of the expression, secretion and/or activities of potential heterologous antimicrobial proteins by the model lactic acid bacterium, Lactococcus lactis subsp. cremoris MG1363. L. lactis strains were engineered to express and secrete the recombinant CyuC surface protein from Lactobacillus reuteri BR11, and a fusion protein consisting of CyuC and lysostaphin using the Sep promoter and secretion signal. CyuC has been characterised as a cystine-binding protein, but has also been demonstrated to have fibronectin binding activity. Lysostaphin is a bacteriolytic enzyme with specific activity against the Gram-positive pathogen, Staphylococcus aureus. These modified L. lactis strains were then investigated to see if they had the ability to inhibit the adhesion of S. aureus to host extracellular matrix (ECM) proteins. It was observed that the cell extracts of the L. lactis strain with the vector only (pGhost9:ISS1) was able to inhibit the adhesion of S. aureus to fibronectin, whilst the cell extracts of the L. lactis strain expressing lysostaphin was able to inhibit adhesion to keratin. Finally, this thesis has identified specific lactococcal genes that affect the secretion of lysostaphin through the use of random transposon mutagenesis. Ten mutants with higher lysostaphin activity contained insertions in four different genes encoding: (i) an uncharacterised putative transmembrane protein (llmg_0609), (ii) an enzyme catalysing the first step in peptidoglycan biosynthesis (murA2), (iii) a homolog of the oxidative defence regulator (trmA), and (iv) an uncharacterised putative enzyme involved in ubiquinone biosynthesis (llmg_2148). The higher lysostaphin activity observed in these mutants was found to be due to higher amounts of lysostaphin being secreted. The findings of this thesis contribute to the development of this organism as an antimicrobial agent and also to our understanding of L. lactis genetics.

Formalin fixation affects equilibrium partitioning of an ionic contrast agent-microcomputed tomography (EPIC-[mu]CT) imaging of osteochondral samples

Equilibrium Partitioning of an Ionic Contrast agent with microcomputed tomography (EPIC-[mu]CT) is a non-invasive technique to quantify and visualize the three-dimensional distribution of glycosaminoglycans (GAGs) in fresh cartilage tissue. However, it is unclear whether this technique is applicable to already fixed tissues. Therefore, this study aimed at investigating whether formalin fixation of bovine cartilage affects X-ray attenuation, and thus the interpretation of EPIC-[mu]CT data.Design Osteochondral samples (n = 24) were incubated with ioxaglate, an ionic contrast agent, for 22 h prior to [mu]CT scanning. The samples were scanned in both formalin-fixed and fresh conditions. GAG content was measured using a biochemical assay and normalized to wet weight, dry weight, and water content to determine potential reasons for differences in X-ray attenuation.Results The expected zonal distribution of contrast agent/GAGs was observed for both fixed and fresh cartilage specimens. However, despite no significant differences in GAG concentrations or physical properties between fixed and fresh samples, the average attenuation levels of formalin-fixed cartilage were 14.3% lower than in fresh samples.Conclusions EPIC-[mu]CT is useful for three-dimensional visualization of GAGs in formalin-fixed cartilage. However, a significant reduction in X-ray attenuation for fixed (compared to fresh) cartilage must be taken into account and adjusted for accordingly when quantifying GAG concentrations using EPIC-[mu]CT.

Train schedule coordination at an interchange station through Agent negotiation

In open railway markets, coordinating train schedules at an interchange station requires negotiation between two independent train operating companies to resolve their operational conflicts. This paper models the stakeholders as software agents and proposes an agent negotiation model to study their interaction. Three negotiation strategies have been devised to represent the possible objectives of the stakeholders, and they determine the behavior in proposing offers to the proponent. Empirical simulation results confirm that the use of the proposed negotiation strategies lead to outcomes that are consistent with the objectives of the stakeholders.

Queensland requirements (old and new) for the appointment of a real estate agent

In Moneywood Pty Ltd v Salamon Nominees Pty Ltd 1 the High Court of Australia considered an appeal from the Queensland Court of Appeal in relation to the correct interpretation of s76 (1)(c) Auctioneers and Agents Act 1971 (Qld). In paraphrase, s76(1)(c) provides that a real estate agent shall not be entitled to sue for or recover any commission unless “the engagement or appointment to act as …..real estate agent ….. in respect of such transaction is in writing signed by the person to be charged with such…..commission…..or the person’s agent or representative” (“the statutory requirement”).

The spermostatic and microbicidal actions of quinones and malemides : towards a dual-purpose contraceptive agent

There is an urgent need to develop safe, effective, dual-purpose contraceptive agents that combine the prevention of pregnancy with protection against sexually transmitted diseases. Here we report the identification of a group of compounds that on contact with human spermatozoa induce a state of “spermostasis,” characterized by the extremely rapid inhibition of sperm movement without compromising cell viability. These spermostatic agents were more active and significantly less toxic than the reagent in current clinical use, nonoxynol 9, giving therapeutic indices (ratio of spermostatic to cytotoxic activity) that were orders of magnitude greater than this traditional spermicide. Although certain compounds could trigger reactive oxygen species generation by spermatozoa, this activity was not correlated with spermostasis. Rather, the latter was associated with alkylation of two major sperm tail proteins that were identified as A Kinase-Anchoring Proteins (AKAP3 and AKAP4) by mass spectrometry. As a consequence of disrupted AKAP function, the abilities of cAMP to drive protein kinase A-dependent activities in the sperm tail, such as the activation of SRC and the consequent stimulation of tyrosine phosphorylation, were suppressed. Furthermore, analysis of microbicidal activity using Chlamydia muridarum revealed powerful inhibitory effects at the same low micromolar doses that suppressed sperm movement. In this case, the microbicidal action was associated with alkylation of Major Outer Membrane Protein (MOMP), a major chlamydial membrane protein. Taken together, these results have identified for the first time a novel set of cellular targets and chemical principles capable of providing simultaneous defense against both fertility and the spread of sexually transmitted disease.