Copper as a target for prostate cancer therapeutics: copper-ionophore pharmacology and altering systemic copper distribution

Copper-ionophores that elevate intracellular bioavailable copper display significant therapeutic utility against prostate cancer cells in vitro and in TRAMP (Transgenic Adenocarcinoma of Mouse Prostate) mice. However, the pharmacological basis for their anticancer activity remains unclear, despite impending clinical trails. Herein we show that intracellular copper levels in prostate cancer, evaluated in vitro and across disease progression in TRAMP mice, were not correlative with copper-ionophore activity and mirrored the normal levels observed in patient prostatectomy tissues (Gleason Score 7 & 9). TRAMP adenocarcinoma cells harbored markedly elevated oxidative stress and diminished glutathione (GSH)-mediated antioxidant capacity, which together conferred selective sensitivity to prooxidant ionophoric copper. Copper-ionophore treatments [CuII(gtsm), disulfiram & clioquinol] generated toxic levels of reactive oxygen species (ROS) in TRAMP adenocarcinoma cells, but not in normal mouse prostate epithelial cells (PrECs). Our results provide a basis for the pharmacological activity of copper-ionophores and suggest they are amendable for treatment of patients with prostate cancer. Additionally, recent in vitro and mouse xenograft studies have suggested an increased copper requirement by prostate cancer cells. We demonstrated that prostate adenocarcinoma development in TRAMP mice requires a functional supply of copper and is significantly impeded by altered systemic copper distribution. The presence of a mutant copper-transporting Atp7b protein (tx mutation: A4066G/Met1356Val) in TRAMP mice changed copper-integration into serum and caused a remarkable reduction in prostate cancer burden (64% reduction) and disease severity (grade), abrogating adenocarcinoma development. Implications for current clinical trials are discussed.

Benzodiazepine and pharmaceutical opioid misuse and their relationship to crime : an examination of illicit prescription drug markets in Melbourne, Hobart and Darwin : Victorian report

This report presents the findings of the major research project 'benzodiazepine and pharmaceutical opioid misuse and their relationship to crime', and is an examination of illicit prescription drug markets in Melbourne.

Gene expressed in hypothalamus of obese animals and pharmaceutical use

Describes a method that relates generally to a nucleic acid molecule which encodes a protein associated with the modulation of obesity, diabetes and metabolic energy levels. More particularly, the present method is directed to a nucleic acid molecule and a recombinant and purified naturally occurring protein encoded thereby and their use in therapeutic and diagnostic protocols for conditions such as obesity, diabetes and energy imbalance. The subject nucleic acid molecule and protein and their derivatives, homologues, analogues and mimetics are proposed as therapeutic and diagnostic agents for obesity, diabetes and energy imbalance.

Evolutionary explanations in the social and behavioral sciences : introduction and overview

Despite a growing acceptance of the value of evolutionary approaches to understanding the natural world there has been relatively little attention paid to evolutionary ideas in sociology, socio-cultural anthropology, and — of particular relevance for this special issue — criminology and forensic/correctional psychology. The aim of this paper is to provide an introductory overview of evolutionary approaches to human behavior with a focus on illuminating the role they can play in enriching our understanding of criminal and antisocial behavior. We begin with an overview of the main approaches to applying evolutionary theory to human behavior and we suggest that a pluralistic perspective is most likely to advance conceptual and empirical work in the field. We then turn to a brief discussion of some common, but misguided criticisms of this approach. Some of the more substantive conceptual and methodological issues that evolutionary approaches need to address are then explored. Finally, we engage with the broader issues that relate to the role of evolutionary explanations in the social and behavioral sciences.

N-acetylcysteine for antioxidant therapy : pharmacology and clinical utility

Glutathione is an endogenous antioxidant and has a ubiquitous role in many of the body’s defences. Treatment with N -acetylcysteine (NAC) has been shown to increase levels of glutathione. NAC has been proposed as a treatment for several illnesses. Objectives : The efficacy and tolerability of NAC was examined across a range of conditions to evaluate the evidence supporting the use of NAC for each indication. Methods : A literature search was conducted using PubMed. Information was also collected from other online sources including the websites of the Therapeutic Goods Administration of Australia and the FDA. Results : Reports ranged from case studies to clinical trials. There is strong evidence to support the use of NAC for the treatment of paracetamol overdose and emerging evidence suggesting it has utility in psychiatric disorders, particularly schizophrenia and bipolar disorder. NAC is safe and well tolerated when administered orally but has documented risks with intravenous administration.

Irreversible and bivalent ligands for selected G-protein coupled receptors

Derivatives of pharmaceutical compounds that could potentially bind irreversibly to a hypertension controlling receptor were synthesised to provide tools for studying this receptor. Also compounds that simultaneously activate two cardiovascular receptors with opposing cellular mechanisms were synthesised. This resulted in a desired partial reduction in the activity of one receptor.

Phospholipase activity in human mesenteric ischaemia and infarction

Currently, diagnostic tests for mesenteric ischaemia and infarction are inadequate due to poor sensitivity and specificity. In addition, many potential markers appear too late to be clinically useful. At present, definitive diagnosis can only be made at the time of surgery, which is not ideal as surgery is often to be avoided in critically ill and elderly patients. A clinically useful, minimally invasive test is likely to decrease the currently very high mortality rate and allow monitoring of 'at risk' patients during their hospital stay. A two-dimensional electrophoresis based proteomic approach was undertaken to assess plasma protein differences between patients with surgically confirmed bowel infarction and control Intensive Care patients. The major protein differences were found to be members or variants of acute phase proteins. Serum amyloid A showed the largest difference between the two patient groups, and this protein was investigated in greater depth. An analysis was performed to compare the diagnostic ability of several commonly used indicators of critical illness and bowel infarction with serum amyloid A and phospholipase A2. Although none of the variables were ideal for clinical use, plasma phospholipase A2 activity showed the best discriminatory power, as determined by Receiver Operating Characteristic curves. From a review of the literature, phospholipase AI (PLA2) appeared to be increased in the bowel as a result of ischaemia and infarction. In one patient, matched tissues were obtained, and PLA2 activity was found to be significantly higher in infarcted bowel tissue compared to ischaemic bowel tissue. PLA2 activity was significantly greater in bowel lumen than tissue, suggesting that the protein was being released, and may enter the circulation. PLA2 activity was increased in the plasma of bowel infarction patients compared with control patients, though the difference was not significant. The phospholipase activity exhibited a number of similarities to typical phospholipase A2 proteins, but also showed a number of inconsistent characteristics. For this reason, we wished to identify the protein responsible for the increased phospholipase activity in infarcted human bowel. The PLA2 activity in human bowel could not be abolished by immunoprecipitation of the PLA2 isoforms IIA (well described in bowel) and V (a closely related isoform). To investigate these proteins, a native urea protein gel devised for snake venom phospholipase A2 was modified for use with mammalian phospholipase AI. The modified gel was used to show that the protein with phospholipase activity from infarcted gut was different from normal gut PLA2 and type IIA PLA2. A number of extensions were devised for these native gels and were found to be useful both in this investigation and for venom investigations. Protein purification was undertaken to identify the protein responsible for the increased phospholipase activity in infarcted bowel. Protein was purified from infarcted human bowel using a number of techniques that exploited unusual characteristics of the protein. The purification techniques each retained the native activity of the protein and the purification could therefore be monitored with a phospholipid hydrolysis assay at each stage. The protein identified by mass spectrometry was an excellent match for cyclophilin B, an inflammatory protein that had previously been identified in rat bowel at the mRNA level (Hasel et al, 1991, Kainer & Doris, 2000). As the purification progress had been monitored throughout with a phospholipid hydrolysis assay, cyclophilin B was an unexpected identification, as it is not known to have phospholipase activity. Cyclophilin B was removed from the highly purified samples via immunoprecipitation and this process abolished all phospholipase activity. The addition of cyclosporin A, (the pharmaceutical ligand of cyclophilin B), did not effect the phospholipase activity. Cyclophilin B protein was found in normal and infarcted human bowel using Western blotting. Cyclophilin B protein also appeared to be present in the bowel lumen and plasma of several patients with bowel infarction, but not in control patients. Immunohistochemistry confirmed the ubiquitous nature of cyclophilin B that had been reported by other groups. This project has investigated the use of two dimensional gel electrophoresis based proteomics to identify proteins present in the plasma of patients with confirmed bowel infarction and control intensive care patients. The major protein classes observed were members of the acute phase proteins, which highlights the need for pre-fractionation of plasma to identify lower abundance, disease associated proteins. A series of potential plasma markers were compared using Receiver Operating Characteristic Curves. Although no ideal marker was clear from this analysis, phospholipase activity appeared to warrant further investigation. Phospholipase activity was investigated in human infarcted bowel. Protein purification identified cyclophilin B as a bowel protein that showed unusual phospholipid hydrolysing activity. Cyclophilin B is a ubiquitous protein in intestinal cell types in both normal and infarcted tissue. There appears to be release of cyclophilin B into bowel lumen and plasma under conditions of mesenteric ischaemia and infarction.

Synthesis and determination of illicit drugs for improved forensic detection

This research describes the development of several innovative methodologies for the rapid and sensitive determination of analytes of forensic and social significance. The project incorporated separation technology and chemiluminescence (light from a chemical reaction) to determine the active compounds and excipients in illicit drugs and natural products including 'magic mushrooms'

Chiral and achiral precursers for extended and supramolecular compounds

Organometallic compounds are building blocks for materials with applications in catalysis, pharmaceutical production and molecular sensors. Research presented in this thesis focused on the design and synthesis of compounds with supramolecular architectures. Crystal engineering these new compounds provides the basis for the next generation of advanced materials required by industry.

Novel routes to new chiral rings and cages

The successful synthesis and characterisation of a series of new chiral cages was achieved through the use of a BINOL backbone linked to a variety of organotin carboxylates. These novel chiral cages may ultimately find use as catalysts in reaction syntheses for the pharmaceutical industry.

Recent insights into microbial catalases: isolation, production and purification

Catalase, an oxidoreductase enzyme, works as a detoxification system inside living cells against reactive oxygen species formed as a by-product of different metabolic reactions. The enzyme is found in a wide range of aerobic and anaerobic organisms. Catalase has also been employed in various analytical and diagnostic methods in the form of biosensors and biomarkers in addition to its other applications in textile, paper, food and pharmaceutical industries. New applications for catalases are constantly emerging thanks to their high turnover rate, distinct evolutionary origin, relatively simple and well-defined reaction mechanisms. The following review provides comprehensive information on isolation, production and purification of catalases with different techniques from various microbial sources along with their types, structure, mechanism of action and applications.

Preparation and characterization of chia seed protein isolate-chia seed gum complex coacervates

Chia seed protein isolate (CPI) and chia seed gum (CSG) were extracted and complex coacervation between these two was studied. The pH and the CPI-to-CSG ratio were optimized to obtain the highest yield of complex coacervates underpinned by zeta potential and turbidity values. CPI-CSG complex coacervates were found to form primarily due to electrostatic interaction and remained stable within a pH range of 2.1-2.9 at ambient temperature. The optimum pH and CPI-to-CSG ratio for complex coacervation was found to be 2.7 and 6:1, respectively. Spray dried complex coacervate particles possessed smoother surface morphology compared to the freeze dried ones. CPI-CSG complex coacervates demonstrated better thermal stability as compared to that of individual CPI and CSG. The crosslinking of these complex coacervates by transglutaminase further improved their thermal stability. Therefore, the crosslinked CPI-CSG complex coacervates will be able to better protect the oxygen and heat sensitive food and pharmaceutical ingredients.