Tuberculosis (TB) is a deadly infectious lung disease caused by the pathogenic bacterium Mycobacterium tuberculosis (Mtb). The identification of macrophage signaling proteins exploited by Mtb during infection will enable the development of alternative host-directed therapies (HDT) for TB. HDT strategies will boost host immunity to restrict the intracellular replication of Mtb and therefore hold promise to overcome antimicrobial resistance, a growing crisis in TB therapy. Protein Kinase R (PKR) is a key host sensor that functions in the cellular antiviral response. However, its role in defense against intracellular bacterial pathogens is not clearly defined. Herein, we demonstrate that expression and activation of PKR is upregulated in macrophages infected with Mtb. Immunological profiling of human THP-1 macrophages that overexpress PKR (THP-PKR) showed increased production of IP-10 and reduced production of IL-6, two cytokines that are reported to activate and inhibit IFNy-dependent autophagy, respectively. Indeed, sustained expression and activation of PKR reduced the intracellular survival of Mtb, an effect that could be enhanced by IFNy treatment. We further demonstrate that the enhanced anti-mycobacterial activity of THP-PKR macrophages is mediated by a mechanism dependent on selective autophagy as indicated by increased levels of LC3-II that colocalize with intracellular Mtb. Consistent with this mechanism, inhibition of autophagolysosome maturation with bafilomycin A1 abrogated the ability of THP-PKR macrophages to limit replication of Mtb, whereas pharmacological activation of autophagy enhanced the anti-mycobacterial effect of PKR overexpression. As such, PKR represents a novel and attractive host target for development of HDT for TB, and our data suggest value in the design of more specific and potent activators of PKR.

京都大学 / 新制・課程博士 / 博士(情報学) / 甲第23315号 / 情博第751号 / 新制||情||128(附属図書館) / 京都大学大学院情報学研究科社会情報学専攻 / (主査)教授 緒方 広明, 教授 黒田 知宏, 教授 楠見 孝 / 学位規則第4条第1項該当 / Doctor of Informatics / Kyoto University / DFAM

Self-directed work groups are a growing phenomenon in the field of organisational psychology (Kauffeld, 2006; Neck & Manz, 1994). While much is known about what factors affect information sharing in these kinds of groups, little is known about why these factors have an impact and how they relate to each other. Through the concept of hidden profiles (tasks that by nature have shared and unshared information), this study explored the information sharing and group decision making processes and aimed to illuminate the group processes involved. The study employed a qualitative, ideographic approach where case studies were used. The sample consisted of twenty four undergraduate and postgraduate students studying at the University of the Witwatersrand divided into groups of four members each. A group task to rank the best candidate for a job was self-designed based on the theory and design utilized by Stasser and Titus (1985; 1987). This was first completed individually and then as a group – the group discussion was filmed and coded using a self-developed observation rubric. Participants also completed a self-developed post-task questionnaire regarding their perspectives of various aspects of the decision making process. The analysis was carried out using frequency counts and thematic content analysis. It was found that all the groups discussed more shared information and more unshared negative information was discussed than unshared positive information. Information sharing increased when there was debate about which pieces of information were relevant. In most cases, group members were motivated to share information because they wanted to have their opinion heard. Although the majority of the sample stated that they did not withhold any information, there was evidence of strategic information sharing. Group 5 made a decision that was closest to the ideal decision. Characteristics of this group that could have contributed to this included: long duration of discussion; high number of talking turns; respecting each other’s talking turns; moderate levels of disagreement; no obvious role of leader; moderately high levels of group familiarity; diversity in race not gender; similar educational backgrounds and a norm of critical evaluation. The study found that the interactions between factors that were perceived to affect the information sharing and decision-making (such as duration of discussion, number of talking turns, group familiarity, competitive aspects, group composition and group roles) were more interwoven than previously thought.

Levels of the enzyme α-methylacyl-CoA racemase (AMACR) are increased ca. 9-fold in prostate cancer cells. AMACR is a very promising novel drug target as reducing AMACR levels converts castrate-resistant prostate cancer cells to androgen-dependent cells which will respond to androgen-deprivation. Despite the importance of AMACR in prostate and other cancers, there are very limited numbers of AMACR inhibitors described to-date. This is mainly due to the absence of a high-throughput assay for the screening of inhibitors against AMACR. The active-site residues and catalytic mechanism of human AMACR are still unknown, which make the rational design of drugs targeting AMACR very difficult. A range of novel potential inhibitors were synthesised using a rational drug design approach to explore the structure-activity relationship (SAR) on the side-chains of AMACR inhibitors. Their potencies were assessed using the fluoride elimination assay based on 1H and 19F NMR. Potency, mode of binding and kinetic parameters of these inhibitors were assessed using the multi-well colorimetric assay, which is the first AMACR high-throughput continuous assay reported to-date. A site-directed mutagenesis study was carried out to identify the active-site residues and catalytic mechanism of human AMACR. His-122, Asp-152, Met-184 and Glu-237 were identified as potential active-site residues, so the cDNA was mutated and expressed. The activity of wild-type and mutant AMACR enzymes were assessed using the deuterium wash-in, fluoride elimination and multi-well colorimetric assays. Results from these assays showed that human AMACR does not operate using a ‘two-base’ mechanism. Instead, it operates using a ‘one-base’ mechanism, most likely via water molecules acting as intermediaries within the hydrogen-bondings network in the active site. The knowledge obtained from this research informs rational drug design for this castrate-resistant prostate cancer target.

Action control arises from the interaction of two anatomically distinct decision-making systems, namely goal-directed and habitual behaviour. Goal-directed behaviour is characterized by the consideration of future choices and respective outcomes whereas habitual responding is driven by stimulus-response associations. Response inhibition is essential for goal-directed behaviour and deficits are shown in impulsivity. We administered an acute clinically relevant dosage of the commonly used serotonin reuptake inhibitor escitalopram to sixty-six healthy volunteers in a double-blind, randomized, placebo-controlled design. We administered a large task battery in order to study the effect of escitalopram in several cognitive functions including response inhibition, learning and affective processing. We found dissociate effects on cognitive aspects possibly mediated by distinct cortico-striatal loops. Acute escitalopram administration had a beneficial effect on action cancellation, one aspect of inhibitory control, without any effect on action restraint or waiting impulsivity. The treatment resulted in impaired performance in a probabilistic reversal-learning task and increased sensitivity to misleading feedback thus leading to maladaptive performance. An extra-dimensional set shift impairment during an attention set shift task and a tendency towards impaired instrumental learning discrimination were also observed in the escitalopram group. Our results are discussed in the context of well-documented effects of the dopaminergic system and suggestions of opponent interaction of serotonin and dopamine.

A major change being advocated in education is that of making students more self-directing; that is, helping them become more responsible for their own learning. The focus of this investigation was on fifth grade students' use of self-directed learning strategies and self-directed perceptual skills. An experimental study was conducted using the nested design for analyzing data obtained from the Guglielmino Self-Directed Learning Readiness Scale, the Zimmerman and Martinez-Pons1 Self-Regulated Learning Interview Schedule, and the Bradley-Lane Self-Directing Perceptual Scale. One hundred fifty-two fifth graders were involved in the eight week study along with their six teachers. Both students and teachers were immersed in a module of training that included emphasis upon self-directing behaviors and learning strategies. Two striking findings emerged; namely, (a) in comparing the average number of learning strategies acquired after treatment, the experimental group (low, middle, and high IQ levels) scored 40%, 50% and 29% higher respectively, than did the control group; (b) in comparing the fifth grade students use of learning strategies it was found that most students nearly doubled the number of learning strategies they had previously acquired. Thus, it was concluded that children who do not have actual teaching of information or data concerning learning strategies will likely never acquire the same repertoire of skills that students acquire when exposed to this critical information in some specific, systematic fashion. A primary product developed for the purposes of this investigation was the Bradley-Lane Self-Directing Perceptual Scale—a 132-item Likert Scale designed to identify the self-perceptions of elementary and middle school students. After field-testing, a chi-square treatment was applied to each item of the Perceptual Scale resulting in a reliability of p<.01 for the majority (79%) of the test items, while an additional twelve items (9%) were found to be reliable at the .02 level of significance.

Lithium bis(amido)cuprates are an important class of bimetallic base, which can chemo- and regioselectively metalate aromatic compounds, via directed ortho cupration (DoCu). This thesis begins with an introduction to aspects of the chemistry of organolithium compounds, group 11 organometallic compounds and their lithium 'ate complexes. Examples of such synergic bases are presented and the introduction is concluded with a discussion of lithium bis(amido)cuprate bases, which along with their silver congeners, are the subject of this dissertation. In general, syntheses involve the addition of a lithium amide to a group 11 salt, resulting in the formation of a lithium bis(amido)cuprate or argentate. Structurally focussed work commences with the use of new amide ligands to develop heteroleptic bis(amido)cuprate systems. The reaction of mixtures of lithium amides with CuBr provides a series of novel Lipshutz-type and Gilman cuprates. Interesting structural features are uncovered, which are rationalised in terms of altered steric demands in the newly introduced amide ligands in these systems. CuSCN and CuOCN are investigated as inexpensive and safer alternatives to CuCN in cuprate formation. In the solid state, a series of Lipshutz-type cuprates (TMP)2Cu(SCN)Li2(L) (L = Et2O, THF, THP) are revealed, whose molecular conformations are infuenced by the identity of the Lewis base. However, in benzene solution, in situ conversion of Lipshutz-type to Gilman cuprate is found to occur. Moving to the synthetic setting, derivatisation of chloropyridines is attempted and gives functionalised halopyridines in 51-71 % yield. CuOCN is found to behave quite differently when reacted in the same way as CuSCN, whereby X-ray crystallography reveals structures in which Cu-Li substitution is apparent. The unique reactivity of CuOCN is interpreted with the aid of multinuclear NMR spectroscopy. A new route to Lipshutz-type cuprates is explored by the synthesis of (TMP)2Cu(OCN)Li2(THF) from Gilman cuprate and LiOCN. This avoids Cu-Li substitution. Meanwhile, reaction of lithium N,N-diisopropylamide with CuOCN also avoids metal disorder, to give a novel lithium cuprate-lithium amide adduct. Further advances in our understanding of group 11 'ate complexes are made by introducing silver as a spectroscopically active nucleus in the lithium argentates (TMP)2AgLi and (TMP)2Ag(CN)Li2(THF). In the solid state, these parallel the structures known for Gilman cuprate (TMP)2CuLi and Lipshutz cuprate (TMP)2Cu(CN)Li2(THF), respectively. In solution, NMR spectroscopy reveals features consistent with retention of these structures. Lastly, the formation of mixed Cu-Li aggregates from combining TMPLi and TMPCu in aromatic solvent are investigated. Surprising reactivity is uncovered, in which the aromatic solvent is metalated and incorporated into mixed-metal aggregates. This thesis concludes with a summary of the findings and suggestions for future work, including how the findings presented herein may be transformed into practical improvements to cuprate systems. In particular, the possibility that Gilman cuprate may be activated towards the metalation of aromatic substrates by the addition of sub-stoichiometric or catalytic amounts of a lithium salt additive is explored.

Esta tese objetiva caracterizar o escoamento provocado pela incidência de um jato vertical em um fundo plano, descrevendo assim as características médias e flutuantes das pressões a partir da investigação experimental. O modelo utilizado é composto por um jato retangular posicionado, perpendicularmente, a uma distância de 0,92 m do fundo plano de um canal. Utilizando transdutores de pressão instalados na posição de incidência do jato, adquiriu-se os valores de pressão “instantânea” a uma frequência de 128 Hz durante 10 minutos. Foram ensaiadas cinco (5) vazões: 25 l/s (Fr1 = 7,12), 30 l/s (Fr1 = 8,51), 36,6 l/s (Fr1 = 10,35), 42,7 l/s (Fr1 = 12,12) e 50l/s (Fr1 = 14,23), além de vinte (20) diferentes espessuras de colchão d'água. Com os dados de pressão, efetuou-se análises no domínio do tempo, da frequência e da posição. No domínio do tempo, analisou-se a variação dos parâmetros estatísticos essenciais (média, desvio padrão, assimetria e curtose) bem como a identificação de valores máximo e mínimos das pressões. No domínio da frequência, procurou-se avaliar por meio da análise espectral a identificação de frequências significativas associadas à energia macroturbulenta. No que tange à análise no domínio da posição buscou-se analisar a faixa de influência do jato junto ao fundo, a região de subpressões e o alcance das pressões dinâmicas. Outro aspecto importante abordado é a apresentação dos resultados em gráficos adimensionalizados, o que confere ao estudo um caráter geral dentro da faixa de valores ensaiados. Os resultados definem um colchão mínimo de água da ordem de 3 vezes a altura crítica do escoamento, para o amortecimento das pressões dinâmicas. A análise dos momentos estatísticos demonstra que a distribuição das pressões não segue a distribuição normal. Dentro disso, estão disponibilizados coeficientes estatísticos de probabilidade para a determinação das pressões extremas. Ainda, estão definidas quatro zonas a jusante do impacto do jato: Zona I - região do impacto, Zona II - reflexão do jato, Zona III - Acomodação do escoamento e por fim, a Zona IV - caracterizada pelo fim da influência do jato. Além disso, uma análise deste escoamento sob a perspectiva da formação de um ressalto hidráulico é realizada, indicando que apenas os parâmetros utilizados, não descrevem o escoamento como tal. Esta tese é produto da parceria entre Laboratório de Obras Hidráulicas do Instituto de Pesquisas Hidráulicas da Universidade Federal do Rio Grande do Sul (LOH/IPH/UFRGS) e o Laboratório de Hidráulica Experimental e Recursos Hídricos de Furnas Centrais Elétricas S. A. (LAHE/FURNAS). / This thesis aims to characterize the flow prompted by the incidence of a vertical jet on a flat bottom, thus describing the mean and floating characteristics of the pressures from the experimental investigation. The model used is comprised of a rectangular jet positioned perpendicularly at a distance of 0.92 m from the flat bottom of a channel. Using pressure transducers installed in the jet incidence position, the "instantaneous" pressure values were acquired at a frequency of 128 Hz for 10 minutes. Five (5) discharges were tested: 25 l/s (Fr1 = 7.12), 30 l/s (Fr1 = 8.51), 36.6 l/s (Fr1 = 10.35), 42.7 l/s (Fr1 = 12.12) and 50 1/s (Fr1 = 14.23), as well as twenty (20) different waterbed thicknesses. The pressure data were analyzed in terms of time, frequency and position domains. In the time domain, the variation of the essential statistical parameters (mean, standard deviation, asymmetry and kurtosis) was analyzed, as well as the identification of maximum and minimum pressure values. In the frequency domain, it was sought to assess, by means of the spectral analysis, the identification of significant frequencies associated with macroturbulent energy. Regarding the analysis in the position domain, this study aimed to analyze the range of influence of the jet near the bottom, the region of subpressions and the reach of the dynamic pressures. Another important aspect is the presentation of the results in dimensionless graphs, which grants the study a general character within the range of values tested. The results define a minimum waterbed of the order of 3 times the critical height of the flow for the damping of the dynamic pressures. Statistical moments analysis shows that the distribution of pressures does not follow the normal distribution. Therein, probability statistical coefficients are available for the determination of extreme pressures. Also, four zones are defined downstream of the impact of the jet: Zone I - region of impact, Zone II - reflection of the jet, Zone III - accommodation of the flow and, finally, Zone IV - characterized by the end of the influence of the jet. Furthermore, an analysis of this flow from the perspective of the formation of a hydraulic jump is conducted, indicating that the parameters used alone do not describe the flow as such. This thesis is a product of the partnership between the Hydraulic Works Laboratory of the Hydraulic Research Institute of the Federal University of Rio Grande do Sul (LOH/IPH/UFRGS) and the Laboratory of Experimental Hydraulics and Hydraulic Resources of Furnas Centrais Elétricas SA (LAHE/FURNAS).

Desde sua descoberta em meados do século XIX, os reagentes organometálicos têm sido amplamente utilizados na síntese orgânica, sendo bastante eficientes na formação de novas ligações carbono-carbono. A escolha do reagente organometálico ideal para determinada reação envolve conhecimentos relacionados à sua natureza química e à própria reatividade do substrato. O núcleo quinoxalínico apresenta um grande potencial para a química medicinal, estando presente em diversos produtos naturais e sintéticos que apresentam atividade biológica. Apesar disso, existem poucos estudos na literatura sobre a aplicação de reagentes organometálicos na funcionalização de quinoxalinas. A reação de metalação dirigida usando bases organometálicas é uma poderosa ferramenta para funcionalização de substratos aromáticos. Contudo, exemplos da aplicação desta estratégia na funcionalização de quinoxalinas são bastante raros. Assim, este trabalho teve como principal objetivo investigar a reatividade dos reagentes organometálicos derivados de zinco, magnésio e lítio em reações de metalação de quinoxalinas substituídas com grupos funcionais nas posições C-2 e C-6. Embora as quinoxalinas substituídas com grupos éster, amida e fenil na posição C-2 não tenham apresentado a reatividade esperada frente aos amidetos mistos de zinco e magnésio (TMPZnCl?MgCl2?LiCl, TMP2Zn?2MgCl2?2LiCl, TMPMgCl?LiCl e TMP2Mg?2LiCl), uma nova metodologia foi desenvolvida utilizando TMPLi como base, na presença de ZnCl2, que permitiu a preparação de vários derivados difuncionalizados em bons rendimentos, muitos dos quais de estrutura inédita. Quinoxalinas não substituídas nas posições C-2 e C-3 possuem reconhecida intolerância à presença de reagentes organometálicos, rendendo preferencialmente produtos de dimerização. Desta forma, uma contribuição importante deste trabalho foi o desenvolvimento de uma metodologia de funcionalização da quinoxalina de estrutura mais simples em micro-ondas, mediada pelo sistema TMPMgCl?LiCl/ZnCl2, que permitiu a preparação de diversos derivados arilados em rendimentos que variaram de razoáveis a bons (25 a 94%). Além disso, a metodologia mostrou-se bastante eficiente para síntese de ligantes bidentados de interesse para a área de catálise / Since its discovery in the mid-nineteenth century, organometallic reagents have been widely used in organic synthesis, being very effective in the formation of new carbon-carbon bonds. The choice of the ideal organometallic reagent for certain reaction involves knowledge related to their chemical nature and reactivity of the substrate itself. The quinoxaline core has great potential for medicinal chemistry and is present in many natural and synthetic products presenting biological activity. Nevertheless, there are few studies on the application of organometallic reagents in the functionalization of quinoxalines. The directed metalation reaction using metal bases is powerful synthetic tool for functionalization of aromatic substrates. However, examples on the application of such strategy to the functionalization of of quinoxalines are very rare. This work mainly aimed to investigate the reactivity of organometallic reagents of zinc, magnesium and lithium in the metalation of quinoxalines substituted with functional groups at the C-2 and C-6 positions. Although C-2 substituted quinoxalines with ester and amide groups have not shown the expected reactivity with mixed amides of zinc, magnesium and lithium (TMPZnCl?MgCl2?LiCl, TMP2Zn?2MgCl2?2LiCl, TMPMgCl?LiCl and TMP2Mg?2LiCl), a new methodology was developed using TMPLi as a base in the presence of ZnCl2, allowing the preparation of various difuncionalized derivatives in good yields, many of them new compounds. Quinoxalines unsubstituted at the C-2 and C-3 positions have recognized intolerance to the presence of organometallic reagents, preferably yielding dimerization products. Thus, an important contribution of this work was the development of a functionalization methodology of the simplest quinoxaline in microwave mediated by the TMPMgCl?LiCl/ZnCl2, allowing the preparation of various arylated derivatives in yields ranging from reasonable to good (25 to 94%). Furthermore, the methodology could be applied to the synthesis of bidentate ligands of interest for catalysis

Directed graphs are an intuitive and versatile representation of natural language meaning because they can capture relationships between instances of events and entities, including cases where entities play multiple roles. Yet, there are few approaches in natural language processing that use graph manipulation techniques for semantic parsing. This dissertation studies graph-based representations of natural language meaning, discusses a formal-grammar based approach to the semantic construction of graph representations, and develops methods for open-domain semantic parsing into such representations. To perform string-to-graph translation I use synchronous hyperedge replacement grammars (SHRG). The thesis studies this grammar formalism from a formal, linguistic, and algorithmic perspective. It proposes a new lexicalized variant of this formalism (LSHRG), which is inspired by tree insertion grammar and provides a clean syntax/semantics interface. The thesis develops a new method for automatically extracting SHRG and LSHRG grammars from annotated “graph banks”, which uses existing syntactic derivations to structure the extracted grammar. It also discusses a new method for semantic parsing with large, automatically extracted grammars, that translates syntactic derivations into derivations of the synchronous grammar, as well as initial work on parse reranking and selection using a graph model. I evaluate this work on the Abstract Meaning Representation (AMR) dataset. The results show that the grammar-based approach to semantic analysis shows promise as a technique for semantic parsing and that string-to-graph grammars can be induced efficiently. Taken together, the thesis lays the foundation for future work on graph methods in natural language semantics.