Yes / Platinum drugs are widely used for cancer treatment. Other precious metals are promising, but their clinical progress depends on achieving different mechanisms of action to overcome Pt-resistance. Here, we evaluate 13 organo-Os complexes: 16-electron sulfonyl-diamine catalysts [(η6-arene)Os(N,N′)], and 18-electron phenylazopyridine complexes [(η6-arene)Os(N,N’)Cl/I]+ (arene = p-cymene, biphenyl, or terphenyl). Their antiproliferative activity does not depend on p21 or p53 status, unlike cisplatin, and their selective potency toward cancer cells involves the generation of reactive oxygen species. Evidence of such a mechanism of action has been found both in vitro and in vivo. This work appears to provide the first study of osmium complexes in the zebrafish model, which has been shown to closely model toxicity in humans. A fluorescent osmium complex, derived from a lead compound, was employed to confirm internalization of the complex, visualize in vivo distribution, and confirm colocalization with reactive oxygen species generated in zebrafish. / Wellcome Trust (grant no. 107691/Z/15/Z), ERC (grant nos. 247450, 324594), Science City (AWM and ERDF), WCPRS and Bruker Daltonics (Studentship for JPCC), Mike and Enfys Bagguley, and EPSRC (Studentship for HEB, and grant no. EP/F034210/1).

The base-catalysed reaction of 4H-pyran-4-one with a range of nucleophiles, namely diethyl malonate, ethyl acetoacetate, nitromethane, acetylacetone and ethyl cyanoacetate, was developed as a reliable, high yielding method for the preparation of para-substituted phenols. The methodology was extended to include the use of the substituted pyranones, maltol, 2,6-dimethyl-4H-pyran-4-one and diethyl chelidonate. Reactions were studied using conventional heating methods and microwave irradiation. Microwave irradiation had definite beneficial effects, with improved yields, reduced reaction times and cleaner reaction profiles. The potential of this methodology was examined for the regioselective placement of ¹³C-atoms into benzene rings using ¹³C-labelled nucleophiles or ¹³C-labelled 4H-pyran-4-ones. [3,5-13C₂]4H-Pyran-4-one and [2,6-13C₂]4H-pyran-4-one were prepared from various ¹³C-labelled versions of triethyl orthoformate and acetone. This methodology was applied to the synthesis of [1,3,5-¹³C₃]gallic acid, via the base-catalysed reaction of [3,5-¹³C₂]4H-pyran-4-one with diethyl [2-¹³C]malonate, followed by subsequent transformations to yield [1,3,5-¹³C₃]gallic acid. The preparation of [2-¹³C]phloroglucinol was carried out via [2-¹³C]resorcinol, with regioselective placement of a single ¹³C-atom into the aromatic ring. This was accomplished from non-aromatic precursors, with the source of the ¹³C-atom being [¹³C]methyl iodide. The key step in this synthesis was the introduction of the third hydroxyl group, which was achieved using a modified iridium-catalysed C-H activation/borylation/oxidation procedure. The scope of an existing C-H activation/borylation reaction was modified and expanded to include a range of protected resorcinol derivatives. A catalyst system was developed which allowed high conversion to the intermediate arylboronic acids, followed by oxidation using aqueous Oxone® to yield the corresponding phenols. Finally, to demonstrate the potential of these new methods for application in the synthesis of isotopically labelled natural products and polyphenols, the syntheses of ¹³C-labelled anthocyanins were studied. A route was developed that could be applied to the synthesis of either cyanidin-3-glucoside or delphinidin-3-glucoside. Only the final coupling/cyclisation step to yield the desired anthocyanin targets remains to be carried out.

This thesis examines how the ‘terrorist’ label affects those that are labelled by this designation, particularly with reference on a subsequent choice to use violence in the context of an ethno-nationalist conflict. Drawing on the PKK as a case study, the study asks: what effect did the labelling of the PKK as a ‘terrorist organisation’ by the Turkish government have on the use of violence by Kurds in the Turkish-Kurdish ethno-nationalist conflict? The invocation of the label terrorist in any conflict often means both the labeller and the labelled are predisposed to use violence. This study argues that this process of labelling leads the labeller and the labelled to frame one another as an existential threat. To date, the effects of using the label ‘terrorist’ in an ethno-nationalist conflict context remain relatively understudied in both social and political sciences. The period under analysis extends from 1992 to 2015, corresponding to the period during which the Turkish government continuously designated the PKK as ‘terrorist’. In conflict discourse, belligerents use demeaning labels against each other to gather support, legitimacy or simply to increase combatants’ morale. The study argues that the label terrorist is a constituent element of the conflict. The Turkish government uses the label terrorist as a tool to securitise the Kurdish-Turkish ethno-nationalist conflict. The Turkish government’s labelling of the PKK as ‘terrorist’ places the Kurdish issue in the broader framework of securitisation, a theory in International Relations. While securitising the Kurdish issue has bestowed more powers to the Turkish government to combat violence described as ‘terrorist’, the resolution of the ethno-nationalist conflict became increasingly more complex leading to protracted waves of violence. Analysing data collected through semi-structured qualitative interviews with Kurds from Turkey, the study reveals that the impact of the label terrorist is far more complex than previously assumed in the existing academic literature. The specific effects of the label terrorist on any given conflict, however, are the subject of an empirical question to be settled through rigorous research. Drawing on the Labelling Theory of Deviance fathered by Howard S. Becker and complemented by discourse analysis, this study finds that the application of the label terrorist against the PKK increases the perception of victimization among its wider Kurdish community. Secondly, the research demonstrates that the invocation of the label terrorist against the PKK places the group’s actors and sympathizers in a situation that makes it harder for them to engage in peaceful means of resolving the conflict. The interplay between these two consequential effects of victimisation and political exclusion leads to the conclusion that there is an indirect relationship between designating an ethno-nationalist armed group ‘terrorist’ and the choice to use violence.

Natural products as sources of novel therapeutic agents experienced a steady increase from around the turn of the twentieth century until it peaked in the 1970s and 1980s. However since this time pharmaceutical research in natural products has experienced a decline. Despite this trend the natural products industry now seems to be experiencing a revival of sorts. This thesis represents a continuation of the work on the isolation and structure elucidation of potential drug leads from terrestrial fungal sources that the natural products group at the University of Canterbury is engaged in. The known compound, pseurotin A (2.7) and two novel diastereomers, pseurotin A2 (2.8) and pseurotin A3 (2.9) were isolated from the extract of a Penicillium sp. of fungus collected from the foreshore of a beach in Vancouver, Canada. The absolute stereochemistry of pseurotin A2 and proposed absolute stereochemistry for A3 were elucidated using a combination of X-ray crystallography (A2 only), circular dichrosim, oxidative cleavage reactions, and J2-resoved 2D NMR experiments. The extract of an as yet unidentified endophytic fungus has yielded eight novel compounds related to the spirobisnaphthalene class of compounds. These eight compounds fall into to distinct groupings. The spiro-mamakones, distinguished by a structurally unprecedented oxygenated spiro-nonene skeleton, comprise five compounds, spiro-mamakones A-E (3.11, 3.15-3.18). In addition to these naturally occurring compounds, the semi-synthetic compounds, 4-oxo-spiro-mamakone A (3.12) and O-acetyl-spiro-mamakone A (3.21), were also synthesised. spiro-Mamakone A was found to be racemic, while X-ray crystallography and optical rotation revealed spiro-mamakone C (3.15) to be present as an enantiomeric mixture (4S*, 5S*, 9R*). Unfortunately the enantiomeric excess was unable to be elucidated. NOE experiments revealed spiro-mamakone B (3.16) to have the relative stereochemistry 4S*, 5S*, 9S*. The relative stereochemistry of spiro-mamakones D (3.17) (4S*, 5S*, 8S*, 9S*) and E (3.18) (4S*, 5S*, 8S*, 9R*) was proposed from comparison of coupling constant calculations from energy-minimised models with those of the experimentally determined values. The second group, comprising three novel compounds named the mamakunoic acids, mamakunoic acid A-C (3.8, 3.7, 3.10), are characterised by their acid substituted dihydro benzofuran system. The low yield obtained of these compounds, unfortunately prevented their stereochemical elucidation. In addition to structure elucidation, biosynthetic studies on spiro-mamakone A and mamakunoic acid B were also carried out. Analysis of the NMR spectra derived from spiro-mamakone A, labelled with isotopic acetate, revealed a situation complicated by the presence of isotopomers and racemisation, resulting in NMR spectra that were somewhat anomalous in appearance. These irregularities however, were resolved leading to the proposal that spiro-mamakone A was derived from a dihydroxynaphthalene (DHN) intermediate, which proceeds through to spiro-mamakone via an epoxide intermediate. Despite problems with purity and low yields of isotopically labelled mamakunoic acid B, it was proposed that like spiro-mamakone A, it proceeded via a DHN intermediate. The extract derived from a Malaysian Scleroderma sp. was found to contain a new dichlorinated pulvinic acid derivative, methyl-3',5'-dichloro-4,4'-di-O-methylatromentate (4.14), the structure of which was confirmed by X-ray crystallography. In addition three previously reported compounds, 4,4'-dimethoxyvulpinic acid (4.11), methyl-3'-chloro-4,4'-di-O-methylatromentate (4.12) and methyl-4,4'-dimethoxyvulpinate (4.13), were also isolated. The extract of another, as yet unidentified endophytic fungus was found to contain the new acetogenin, 1,5-dihydroxy-6-(2-hydroxyethyl)-3-methoxyacetophenone (5.7), differing from the known compound, 2,4-dihydroxy-6-(2-hydroxyethyl)-3-methoxyacetophenone (5.8) only by virtue of the substitution pattern. The structure of 5.7 was confirmed by X-ray crystallography. The implementation of efficient dereplication procedures is paramount for those working in the field of natural products. The recent advances that have been made in the dereplication process in the natural products group at the University of Canterbury are given using examples from this research and where necessary from other group members.

No description available.

The chloride/proton antiporter ClC-5 is primarily expressed in the kidney where it aids in re-absorption of proteins from the glomerular filtrate. Functional disruption of ClC-5 causes Dent’s Disease – a renal condition characterized by proteinuria and kidney failure in a third of all cases. The majority of disease-causing mutations translate into premature truncations of the carboxy-terminal (Ct) region of ClC-5 and are predicted to disrupt the protein-protein interactions mediated by this domain. In this thesis, direct ATP binding to the two cystathionine β-synthase (CBS) domains of ClC-5 was demonstrated. ATP binding enhanced the global compactness of the ClC-5 Ct region likely through a clamping motion of the CBS domains around the nucleotide. Along with ATP, the sodium proton exchange regulatory factor 2 (NHERF2) also binds ClC-5; however, the molecular mechanism behind this interaction was unknown as ClC-5 lacked the PDZ binding motif traditionally localized at the Ct end of bait proteins. Here, we also identified a class I PDZ binding motif (657-660; TSII) within the internal sequence of ClC-5. Despite the buried position of this motif in the Ct peptide’s X-ray crystal structure (PDB: 2J9L), the high propensity of this region for dynamic flexibility prompted us to test whether it could mediate NHERF2 interactions. In support of this hypothesis, we demonstrated that the motif is transiently available to interact directly with NHERF2 in vivo and to enable an enhancement in receptor-mediated endocytosis in mammalian cells. Collectively, these results gave further evidence that the intracellular Ct region of ClC-5 serves as a hub to mediate interactions essential for its maturation, stability, and trafficking in renal epithelium, as well as providing further insights into the molecular basis of Dent’s Disease.

Les « Purple Acid Phosphatases » (PAPs) sont des phosphatases acides présentes dans le règne végétal et animal. Elles sont caractérisées à la fois par la présence de sept acides aminés conservés impliqués dans la structure d’un centre bimétallique et par leur couleur rose/violète lorsqu’elles se trouvent en solution. Chez les plantes, certaines PAPs présenteraient à la fois une activité phosphatase sur des métabolites (PEP, G6P…) et sur des phospho-peptides (glycosidases…), d’une part, et une activité peroxydase, de l’autre. Ces PAPs pourraient donc, non seulement être impliquées dans le métabolisme carboné, mais aussi dans la détoxication des espèces réactives de l’oxygène. On compte à ce jour 29 gènes codant pour des PAPs chez Arabidopsis thaliana , dont un certain nombre est induit par la carence en Pi et par le stress oxydatif ou la senescence. Parmi eux, le gène de l’AtPAP26 (At5g34850) présente de très fortes homologies de séquence avec des orthologues présents chez d’autres plantes. La protéine AtPAP26, qui présente in vitro une activité phosphatase sur le PEP et le G6P et une activité peroxydase, est sécrétée même si de nombreux indices laissent penser qu’elle pourrait également être localisée dans la vacuole.Notre objectif est d’étudier le rôle d’AtPAP26 dans le métabolisme et le stockage des sucres dans la vacuole. Notre hypothèse de départ est que la protéine AtPAP26 pourrait intervenir via son activité phosphatase dans un cycle de substrat vacuolaire impliquant le glucose-6-phosphate et le glucose. Nous avons montré que chez le mutant pap26, l’activité glucose-6-phosphatase, mesurée in vitro et in organello, chute de manière importante. Parallèlement, chez le mutant gpt2, le transport tonoplastique des hexoses phosphates diminue de façon significative. Des expériences de marquages isotopiques sur du tissu racinaire en culture ont montré qu’une part importante du glucose et du fructose ne provient pas de l’hydrolyse du saccharose et que AtPAP26 est en partie responsable de cette synthèse. Enfin, par une approche in-silico, nous avons modélisé le métabolisme des sucres dans un système compartimenté impliquant la vacuole et ses transporteurs. La confrontation du modèle avec deux jeux de données expérimentales (concentrations à l’état stationnaire et cinétiques de marquage) a permis de confirmer l’existence d’une activité hydrolytique sur les hexoses phosphates vacuolaires. / «Purple Acid Phosphatases» (PAPs) are acid phosphatases found both in animal and vegetal kingdoms. They are characterized by the presence of seven conserved amino acids involved in a di-metal center and by their pink/purple color in solution. In plants, some PAPs exhibit an acid phosphatase activity on various phospho-esters and on phospho-peptides. An alcalin peroxidase activity has been demonstrated in vitro. Therefore, PAPs could be implicated both in the carbon metabolism and in the scavenging of reactive oxygen species. To date, 29 different PAP-encoding genes were identified in the Arabidopsis thaliana genome. Some of them are induced by phosphate deprivation, oxidative burst or senescence. The gene encoding the protein AtPAP26 (At5g34850) exhibits a high sequence similarity with orthologous from other plants. The AtPAP26 protein, showing both a phosphatase activity on PEP, G6P and F6P, and a peroxidase activity, is one of the most abundant root-secreted acid phosphatases but also appears to be localized in the vacuole. Our aim is to study the implication of AtPAP26 in the metabolism and storage of sugars in the vacuole. Our first hypothesis is that AtPAP26, via its phosphatase activity, could be involved in a glucose / glucose-6-phosphate cycle localized in the vacuole. We showed that the glucose-6-phosphatase activity in the pap26 mutant is highly affected both in vitro and in organello compared to the wild type. At the same time, the hexose phosphate transport capacity of the vacuole is highly affected in the gpt2 mutant compared to the wild-type. Moreover, isotopic labeling experiments performed on cultured root tissues have shown that a significant part of the intracellular glucose and fructose pool does not originate from sucrose and that AtPAP26 is, at least in part, responsible for this synthesis. Finally, we build a kinetic model of sugar metabolism in a compartmented system which has been validated by using two independent experimental data sets (steady state concentrations and kinetics of labeling of soluble sugars). This in silico modeling approach confirmed the involvement of a hexose phosphatase activity localized in the vacuole.

The diploma thesis "The Implementation of rating of the audio-visual products by suitability for age groups in the Czech Republic" deals with issues of classification of audio- visual products in Czech medial space. At first, text analyses, what the audio-visual media product is, and what are the legal, psychological and economic arguments for its implementation. The history and current status of labelling in the Czech Republic is also described in this part. Included is also a description of the labelling status in other selected countries, namely United States of America and several countries of European Union. Next part of thesis deals with the Dutch labelling system Kijkwijzer, as it was recommended by experts as an example for future implementation of classification system in our country. It's labelling process and categories are described as well as reasons for why this system was chosen. Diploma thesis also describes the effort to implement this system in Czech Republic by the Audio-visual Department at the Ministry of Culture, and tries to find out why it hasn't been implemented yet. Last part is concerned with the representatives of Czech TV companies, mothers, a psychologist and member of Council for radio and television broadcasting and their opinions on existing classification of...

The diploma thesis "The Implementation of rating of the audio-visual products by suitability for age groups in the Czech Republic" deals with issues of classification of audio- visual products in Czech medial space. At first, text analyses, what the audio-visual media product is, and what are the legal, psychological and economic arguments for its implementation. The history and current status of labelling in the Czech Republic is also described in this part. Included is also a description of the labelling status in other selected countries, namely United States of America and several countries of European Union. Next part of thesis deals with the Dutch labelling system Kijkwijzer, as it was recommended by experts as an example for future implementation of classification system in our country. It's labelling process and categories are described as well as reasons for why this system was chosen. Diploma thesis also describes the effort to implement this system in Czech Republic by the Audio-visual Department at the Ministry of Culture, and tries to find out why it hasn't been implemented yet. Last part is concerned with the representatives of Czech TV companies, mothers, a psychologist and member of Council for radio and television broadcasting and their opinions on existing classification of...

The diploma thesis deals with regional products and their labelling schemes which have rarely been studied in the context of Czech geography. The thesis aims to identify the role of regional product labelling schemes in the process of institutionalizing regions (in the sense of Anssi Paasi) as well as to discover their contribution to the development of regions. The thesis is based on a theoretical discussion which includes the concept of region and regional identity formation, the issue of regional products as a part of the alternative food networks concept and the issue of symbolism and branding/labelling. The research consists of three parts. The first part provides a database of Czech labelling schemes and an assessment of regional product labelling in Czechia. In the second part, 22 labelling schemes united by the Association of Regional Brands are assessed from different perspectives (location, structural aspects etc.). The last part verifies and completes the findings using semi-structured interviews with key actors of the labelling schemes. The existence of the relation between labelling schemes and the institutionalization of regions was verified at each scale. Regional product labelling is particularly involved in the formation of the region's territorial shape and in the strengthening of...