Isoellipticines: novel compounds for leukaemia treatment

Ellipticine, an anticancer agent, has had limited clinical success due to low solubility and toxic side effects. To overcome these limitations, a panel of novel ellipticine isomers were designed and synthesised with the aim of evaluating their anti-cancer effects on selected cancer cell lines. A preliminary NCI 60-cell screen demonstrated that these isoellipticines displayed promising anti-tumour activity across a number of different cell types, particularly leukaemia cell lines. We consequently examined the effect of these derivatives in detail on the Acute Myeloid Leukaemia (AML) cell line, MV4-11. Cell cycle analyses revealed that the compounds had a range of distinctive cell cycle effects on MV4-11 cells. 7-Hydroxyisoellipticine showed the most promise with respect to cytostatic activity. We demonstrated that this compound inhibited proliferation of leukaemia cells by preventing cells from progressing from G2 phase. Our research suggests that this is mediated by an induction of reactive oxygen species (ROS), which in turn activates the DNA damage response pathway. More extensive research on the source of ROS generated by the most potent derivative, 7-formyl-10-methylisoellipticine showed that this compounds cytotoxicity is partially mediated by an induction of mitochondrial derived reactive oxygen species (ROS). We showed that 7-formyl-10-methylisoellipticine has synergistic effects when used in combination with the clinically used AML drug, daunorubicin, as well as DPI, a Nox inhibitor. Additionally, combination experiments with other drugs served to give us a deeper insight into 7- formyl-10-methylisoellipticine mechanism of action. 7-Formyl-10-methylisoellipticine also displayed promising in vivo results. Treatment resulted in a lack of toxicity, as measured by body weight changes and liver enzyme analyses. Most importantly, 7-formyl-10-methylisoellipticine demonstrated potent anti-tumour activity in the in vivo xenograft mouse model, implying the potential of isoellipticines as novel chemotherapeutic agents in the treatment of leukaemia. In summary, this study provides for the first time detailed cellular information on the potential use of isoellipticines as chemotherapeutic agents. Our study documents for the first time, the therapeutic potential of an isoellipticine compound in a subcutaneous AML cell-derived xenograft (CDX) model. By probing the mechanism of action of this novel compound class we have uncovered a potential clinical application in the field of adjuvant therapy. We anticipate that the recent research on ellipticine derivatives, such as this study, will lead the development of an ellipticine analogue that may be employed clinically.

Synthesis and evaluation of novel ellipticines as potential anti-cancer agents

Ellipticine is a natural product which possesses multimodal anti-cancer activity. This thesis encompasses the synthesis and biological evaluation of novel ellipticine and isoellipticine derivatives as anti-cancer agents. Expanding on previous work within the group utilising vinylmagnesium bromide, derivatisation of the C5 position of ellipticine was accomplished by reaction of a key ketolactam intermediate with Grignard reagents. Corresponding attempts to introduce diverse substitution at the C11 position were unsuccessful, although one novel C11 derivative was produced using an alkyllithium reagent. A panel of novel ellipticinium salts encompassing a range of substitutions at the N2, C9 and N6 positions were prepared. Extensive derivatisation of the N10 position of isoellipticine was undertaken for the first time. Novel substitution in the form of acid and methyl ester functionalities were introduced at the C7 position of isoellipticine while novel C7 aldehyde and alcohol derivatives were synthesised. A large panel of isoellipticinium salts were prepared with conditions adjusted for the reactivity of the alkyl halide. Novel coupling reactions to increase the yield of isoellipticine were attempted but proved unsuccessful. A panel of 54 novel derivatives was prepared and a multimodal analysis of their anti-cancer activity was conducted. The NCI 60-human tumour cell lines screen was a primary source of information on the in vitro activity of compounds with derivatives found to exert potent anticancer effects, with mean GI50 values as low as 1.01 μM across the full range of cancer types and as low as 16 nM in individual cell lines. A second in vitro screen in collaboration with researchers in the University of Nantes identified derivatives which could potently inhibit growth in a p53 mutant NSCLC cell line. The cell cycle effects of a selected panel of isoellipticines were studied in leukaemia cell lines by researchers in the Department of Biochemistry and Cell Biology, UCC. Emerging from this, the therapeutic potential of one of the derivatives in AML was then assessed in vivo in an AML xenograft mouse model, with tumour weight reduced by a factor of 7 in treated mice relative to control.

The Kyoto School and Confucianism: a Confucian reading of the philosophy of history and political thought of Masaaki Kosaka

This dissertation examines the philosophy of Masaaki Kōsaka (1900-1969) from the East Asian perspective of Confucianism, which I believe is the most appropriate moral paradigm for comprehending his political speculations. Although largely neglected in post-war scholarship, Kōsaka was a prominent member of the Kyoto School during the 1930s and 40s. This was a group of Japanese thinkers strongly associated with the philosophies of Kitarō Nishida and Hajime Tanabe. Kōsaka is now best known for his participation in the three Chūō Kōron symposia held in 1941 and 1942. These meetings have been routinely denounced by liberal historians due to the participants’ support for the Pacific War and the Co-Prosperity Sphere. However, many of these liberal portrayals have failed to take into account the full extent of the group’s resistance to the military junta of Hideki Tōjō. Adopting the methods and techniques of the empirical disciplines of academic history and Orientalism, I develop an interpretative framework that is more receptive to the political values that mattered to Kōsaka as a Confucian inspired intellectual. This has necessitated the rejection of moral history, which typically prioritises modern liberal values brought a priori to the historical record of wartime Japan, as well as recognition of the different ontological foundations that inform the unique political theories of the East Asian intellectual tradition. Reinforced by the prior research of Michel Dalissier and Graham Parkes, as well as my own reading of the Confucian canon, I adopt David Williams’s thesis of ‘Confucian Revolution’ as my principle schema of interpretation. This, I believe, is better able to reconcile Kōsaka’s support for the war with his strong condemnation of the imperialist practices of the Japanese military. Moreover, acknowledging the importance of Confucianism allows us to fully appreciate Kōsaka’s strong affinity for Kant’s practical metaphysics, Hegel’s political philosophy and Ranke’s historiography.

La déconstruction de l'onto-théologie par Jacques Derrida : détour littéraire et mise en relief de la matrice langagière comme "différance".

Le mouvement derridien de la différance marque la rupture avec l'affirmation de la métaphysique de la présence, avec l'autorité du signifié transcendantal. Dans cet univers mouvant de signifiants qui se renvoient perpétuellement les uns aux autres, la logique d'univocité se disloque. La "présence" n'est que fantomatique, s'esquissant au sein d'une chaîne ininterrompue de signifiants et se laissant toujours creuser par la marque d'un irréductible manque. Face au logocentrisme, corollaire de l'affirmation de la présence, l'écriture se veut siège et articulation de la trace, d'une origine qui ne peut être que raturée, véhicule d'une irrémédiable fêlure. La volet littéraire de la déconstruction a pour but de mettre en évidence le fonctionnement de l'"indécidabilité" du discours, soit une certaine ambivalence dans la signification qui caractérise tout texte. L'objectif principal de la présente recherche est de fournir une compréhension plus approfondie de la déconstruction en insistant sur l'ancrage langagier de tout texte. Le discours philosophique n'échappe ainsi pas au mécanisme différentiel du langage et de la dérive métaphorique. La parenté entre la perspective déconstructiviste derridienne et la conception mallarméenne du langage poétique semble frappante. La mise en oeuvre, par Mallarmé, d'une dislocation de l'espace textuel, son minutieux "creusement" du vers après renoncement à toute quête d'"Idéal", la mise en relief du leurre de l'appropriation langagière, voilà qui trouve un écho particulier dans les thèses derridiennes. La "mimésis" platonicienne se voit au travers du prisme de la "mimique" mallarméenne. La déconstruction poursuit son travail de "luxation" de l'oreille philosophique, insérant les philosophèmes dans la matrice langagière, les livrant ainsi au hasard du cheminement textuel et les confrontant à l'aporie. La philosophie n'a alors d'autre choix que d'abandonner ses prétentions transcendantales. La marche de la "différance" instaure une inexorable distance qui prive le sujet de tout rapport direct avec une origine assurée et lui ôte toute possibilité de maîtrise sur le monde. Au travers de la langue, se profile la question de l'altérité, de la relation dissymétrique qui nous lie à cet "autre", ce "tout-autre" qui nous fonde et nous constitue. L'accueil inconditionnel de cette altérité nous mènera à l'étude de la "religion", la déconstruction se tournant vers le "religieux" tout en effectuant un "retournement" habile de tout credo essentialiste.

Étude de la survie cellulaire lors du processus de myélopoïèse induit par le facteur de transcription PU.1 et la cytokine GM-CSF

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

The user's view level of the GOQL graphical query language

The paper addresses issues related to the design of a graphical query mechanism that can act as an interface to any object-oriented database system (OODBS), in general, and the object model of ODMG 2.0, in particular. In the paper a brief literature survey of related work is given, and an analysis methodology that allows the evaluation of such languages is proposed. Moreover, the user's view level of a new graphical query language, namely GOQL (Graphical Object Query Language), for ODMG 2.0 is presented. The user's view level provides a graphical schema that does not contain any of the perplexing details of an object-oriented database schema, and it also provides a foundation for a graphical interface that can support ad-hoc queries for object-oriented database applications. We illustrate, using an example, the user's view level of GOQL

The Visual Representation of a Systematic Metaphor in a UK Media Text

Much has been written on the organizational power of metaphor in discourse, eg on metaphor ‘chains’ and ‘clusters’ of linguistic metaphor in discourse (Koller 2003, Cameron & Stelma 2004, Semino 2008) and the role of extended and systematic metaphor in organizing long stretches of language, even whole texts (Cameron et al 2009, Cameron & Maslen 2010, Deignan et al 2013, Semino et al 2013). However, at times, this work belies the intricacies of how a single metaphoric idea can impact on a text. The focus of this paper is a UK media article derived from a HM Treasury press release on alleviating poverty. The language of the article draws heavily on orientational (spatial) metaphors, particularly metaphors of movement around GOOD IS UP. Although GOOD IS UP can be considered a single metaphoric idea, the picture the reader builds up as they move line by line through this text is complex and multifaceted. I take the idea of “building up a picture” literally in order to investigate the schema of motion relating to GOOD IS UP. To do this, fifteen informants (Masters students at a London university), tutored in Cognitive Metaphor Theory, were asked to read the article and underline words and expressions they felt related to GOOD IS UP. The text was then read back to the informant with emphasis given to the words they had underlined, while they drew a pictorial representation of the article based on the meanings of these words, integrating their drawings into a single picture as they went along. I present examples of the drawings the informants produced. I propose that using Metaphor-led Discourse Analysis to produce visual material in this way offers useful insights into how metaphor contributes to meaning making at text level. It shows how a metaphoric idea, such as GOOD IS UP, provides the text producer with a rich and versatile meaning-making resource for constructing text; and gives a ‘mind-map’ of how certain aspects of a media text are decoded by the text receiver. It also offers a partial representation of the elusive, intermediate ‘deverbalized’ stage of translation (Lederer 1987), where the sense of the source text is held in the mind before it is transferred to the target language. References Cameron, L., R. Maslen, Z. Todd, J. Maule, P. Stratton & N. Stanley. 2009. ‘The discourse dynamic approach to metaphor and metaphor-led analysis’. Metaphor and Symbol, 24(2), 63-89. Cameron, L. & R. Maslen (eds). 2010. Metaphor Analysis: Research Practice in Applied Linguistics, Social Sciences and Humanities. London: Equinox. Cameron, L. & J. Stelma. 2004. ‘Metaphor Clusters in Discourse’. Journal of Applied Linguistics, 1(2), 107-136. Deignan, A., J. Littlemore & E. Semino. 2013. Figurative Language, Genre and Register. Cambridge: Cambridge University Press. Koller, V. 2003. ‘Metaphor Clusters, Metaphor Chains: Analyzing the Multifunctionality of Metaphor in Text’. metaphorik.de, 5, 115-134. Lederer, M. 1987. ‘La théorie interprétative de la traduction’ in Retour à La Traduction. Le Francais dans Le Monde. Semino, E. 2008. Metaphor in Discourse. Cambridge: Cambridge University Press. Semino, E., A. Deignan & J. Littlemore. 2013. ‘Metaphor, Genre, and Recontextualization’. Metaphor and Symbol. 28(1), 41-59.

La teoría de los diferenciales de Salomon Maimon, la pregunta quid juris y la posibilidad de la metafísica como ciencia

Este trabajo estudia la teoría de los diferenciales de Salomon Maimon como respuesta a la cuestión quid juris y a la vez como clave para la fundamentación de la posibilidad de la metafísica como ciencia. Se reconstruye primero la crítica de Maimon al tratamiento kantiano de la pregunta quid juris. Luego, se analiza la respuesta del propio Maimon a esa pregunta, para establecer finalmente cómo tal respuesta abre el camino para la explicación de la posibilidad de la metafísica como ciencia.

Training proposal in the degree of Master of Early Childhood Education for working the method of historical research in the classroom

Future teachers must be competent in creating educational settings, which provide tools to their students future they can develop a conscious mind, able to interpret their experiences, to make decisions and imagine innovative solutions to help you participate autonomously and responsible in society. This requires an educational system that allows them to integrate the subjective into a broader spatial and temporal context. La patrimonializatión of “Cultural artefacts” and oral history, the basis of which, are found in the active mind and links both the personal and the group experience, don’t only serve as a catalyst to achieving this goal, but rather, they facilitate the implementation of established practice in infant education. To gain this experience we offer the opportunity for students of their degree in Infant Education in the Public University of Navarre, training within the framework of social didactics, allowing students to learn about established practice from iconic, materials and oral sources in the Archive of Intangible Cultural Heritage of Navarra. The vidences points to their effectiveness and presented in a work in progress.

Mental logic and its difficulties with disjunction

The mental logic theory does not accept the disjunction introduction rule of standard propositional calculus as a natural schema of the human mind. In this way, the problem that I want to show in this paper is that, however, that theory does admit another much more complex schema in which the mentioned rule must be used as a previous step. So, I try to argue that this is a very important problem that the mental logic theory needs to solve, and claim that another rival theory, the mental models theory, does not have these difficulties.

Testing After Worked Example Study Does Not Enhance Delayed Problem-Solving Performance Compared to Restudy

Four experiments investigated whether the testing effect also applies to the acquisition of problem-solving skills from worked examples. Experiment 1 (n=120) showed no beneficial effects of testing consisting of isomorphic problem solving or example recall on final test performance, which consisted of isomorphic problem solving, compared to continued study of isomorphic examples. Experiment 2 (n=124) showed no beneficial effects of testing consisting of identical problem solving compared to restudying an identical example. Interestingly, participants who took both an immediate and a delayed final test outperformed those taking only a delayed test. This finding suggested that testing might become beneficial for retention but only after a certain level of schema acquisition has taken place through restudying several examples. However, experiment 2 had no control condition restudying examples instead of taking the immediate test. Experiment 3 (n=129) included such a restudy condition, and there was no evidence that testing after studying four examples was more effective for final delayed test performance than restudying, regardless of whether restudied/tested problems were isomorphic or identical. Experiment 4 (n=75) used a similar design as experiment 3 (i.e., testing/restudy after four examples), but with examples on a different topic and with a different participant population. Again, no evidence of a testing effect was found. Thus, across four experiments, with different types of initial tests, different problem-solving domains, and different participant populations, we found no evidence that testing enhanced delayed test performance compared to restudy. These findings suggest that the testing effect might not apply to acquiring problem-solving skills from worked examples

Inter-Laboratory Evaluation of a Next-Generation Sequencing Panel for Acute Myeloid Leukemia

INTRODUCTION: Acute myeloid leukemia (AML) is a heterogeneous clonal disorder often associated with dismal overall survival. The clinical diversity of AML is reflected in the range of recurrent somatic mutations in several genes, many of which have a prognostic and therapeutic value. Targeted next-generation sequencing (NGS) of these genes has the potential for translation into clinical practice. In order to assess this potential, an inter-laboratory evaluation of a commercially available AML gene panel across three diagnostic centres in the UK and Ireland was performed.

METHODS: DNA from six AML patient samples was distributed to each centre and processed using a standardised workflow, including a common sequencing platform, sequencing chips and bioinformatics pipeline. A duplicate sample in each centre was run to assess inter- and intra-laboratory performance.

RESULTS: An average sample read depth of 2725X (range 629-5600) was achieved using six samples per chip, with some variability observed in the depth of coverage generated for individual samples and between centres. A total of 16 somatic mutations were detected in the six AML samples, with a mean of 2.7 mutations per sample (range 1-4) representing nine genes on the panel. 15/16 mutations were identified by all three centres. Allelic frequencies of the mutations ranged from 5.6 to 53.3 % (median 44.4 %), with a high level of concordance of these frequencies between centres, for mutations detected.

CONCLUSION: In this inter-laboratory comparison, a high concordance, reproducibility and robustness was demonstrated using a commercially available NGS AML gene panel and platform.

Selective FLT3 inhibition of FLT3-ITD+ acute myeloid leukaemia resulting in secondary D835Y mutation: a model for emerging clinical resistance patterns.

Acquired resistance to selective FLT3 inhibitors is an emerging clinical problem in the treatment of FLT3-ITD(+) acute myeloid leukaemia (AML). The paucity of valid pre-clinical models has restricted investigations to determine the mechanism of acquired therapeutic resistance, thereby limiting the development of effective treatments. We generated selective FLT3 inhibitor-resistant cells by treating the FLT3-ITD(+) human AML cell line MOLM-13 in vitro with the FLT3-selective inhibitor MLN518, and validated the resistant phenotype in vivo and in vitro. The resistant cells, MOLM-13-RES, harboured a new D835Y tyrosine kinase domain (TKD) mutation on the FLT3-ITD(+) allele. Acquired TKD mutations, including D835Y, have recently been identified in FLT3-ITD(+) patients relapsing after treatment with the novel FLT3 inhibitor, AC220. Consistent with this clinical pattern of resistance, MOLM-13-RES cells displayed high relative resistance to AC220 and Sorafenib. Furthermore, treatment of MOLM-13-RES cells with AC220 lead to loss of the FLT3 wild-type allele and the duplication of the FLT3-ITD-D835Y allele. Our FLT3-Aurora kinase inhibitor, CCT137690, successfully inhibited growth of FLT3-ITD-D835Y cells in vitro and in vivo, suggesting that dual FLT3-Aurora inhibition may overcome selective FLT3 inhibitor resistance, in part due to inhibition of Aurora kinase, and may benefit patients with FLT3-mutated AML.

Tyrosine kinase inhibitor insensitivity of non-cycling CD34+ human acute myeloid leukaemia cells with FMS-like tyrosine kinase 3 mutations.

The efficacy of tyrosine kinase (TK) inhibitors on non-cycling acute myeloid leukaemia (AML) cells, previously shown to have potent tumourigenic potential, is unknown. This pilot study describes the first attempt to characterize non-cycling cells from a small series of human FMS-like tyrosine kinase 3 (FLT3) mutation positive samples. CD34+ AML cells from patients with FLT3 mutation positive AML were cultured on murine stroma. In expansion cultures, non-cycling cells were found to retain CD34+ expression in contrast to dividing cells. Leukaemic gene rearrangements could be detected in non-cycling cells, indicating their leukaemic origin. Significantly, the FLT3-internal tandem duplication (ITD) mutation was found in the non-cycling fraction of four out of five cases. Exposure to the FLT3-directed inhibitor TKI258 clearly inhibited the growth of AML CD34+ cells in short-term cultures and colony-forming unit assays. Crucially, non-cycling cells were not eradicated, with the exception of one case, which exhibited exquisite sensitivity to the compound. Moreover, in longer-term cultures, TKI258-treated non-cycling cells showed no growth impairment compared to treatment-naive non-cycling cells. These findings suggest that non-cycling cells in AML may constitute a disease reservoir that is resistant to TK inhibition. Further studies with a larger sample size and other inhibitors are warranted.

Lo spettro di un anello e i suoi punti

L'elaborato tratta argomenti di geometria algebrica. Si analizzeranno nel dettaglio lo spettro di un anello commutativo con unità e la sua topologia e si vedrà come cambia la nozione di varietà algebrica con l'introduzione degli schemi. Verranno poi definiti i punti di uno schema (punti chiusi e punti generici) terminando la discussione con l'analisi delle varietà su un campo non algebricamente chiuso e del funtore dei punti associato ad uno schema.