Engineering the transition from non-living to living matter

Re-creating and understanding the origin of life represents one of the major challenges facing the scientific community. We will never know exactly how life started on planet Earth, however, we can reconstruct the most likely chemical pathways that could have contributed to the formation of the first living systems. Traditionally, prebiotic chemistry has investigated the formation of modern life’s precursors and their self-organisation under very specific conditions thought to be ‘plausible’. So far, this approach has failed to produce a living system from the bottom-up. In the work presented herein, two different approaches are employed to explore the transition from inanimate to living matter. The development of microfluidic technology during the last decades has changed the way traditional chemical and biological experiments are performed. Microfluidics allows the handling of low volumes of reagents with very precise control. The use of micro-droplets generated within microfluidic devices is of particular interest to the field of Origins of Life and Artificial Life. Whilst many efforts have been made aiming to construct cell-like compartments from modern biological constituents, these are usually very difficult to handle. However, microdroplets can be easily generated and manipulated at kHz rates, making it suitable for high-throughput experimentation and analysis of compartmentalised chemical reactions. Therefore, we decided to develop a microfluidic device capable of manipulating microdroplets in such a way that they could be efficiently mixed, split and sorted within iterative cycles. Since no microfluidic technology had been developed before in the Cronin Group, the first chapter of this thesis describes the soft lithographic methods and techniques developed to fabricate microfluidic devices. Also, special attention is placed on the generation of water-in-oil microdroplets, and the subsequent modules required for the manipulation of the droplets such as: droplet fusers, splitters, sorters and single/multi-layer micromechanical valves. Whilst the first part of this thesis describes the development of a microfluidic platform to assist chemical evolution, finding a compatible set of chemical building blocks capable of reacting to form complex molecules with endowed replicating or catalytic activity was challenging. Abstract 10 Hence, the second part of this thesis focuses on potential chemistry that will ultimately possess the properties mentioned above. A special focus is placed on the formation of peptide bonds from unactivated amino acids, despite being one of the greatest challenges in prebiotic chemistry. As opposed to classic prebiotic experiments, in which a specific set of conditions is studied to fit a particular hypothesis, we took a different approach: we explored the effects of several parameters at once on a model polymerisation reaction, without constraints on hypotheses on the nature of optimum conditions or plausibility. This was facilitated by development of a new high-throughput automated platform, allowing the exploration of a much larger number of parameters. This led us to discover that peptide bond formation is less challenging than previously imagined. Having established the right set of conditions under which peptide bond formation was enhanced, we then explored the co-oligomerisation between different amino acids, aiming for the formation of heteropeptides with different structure or function. Finally, we studied the effect of various environmental conditions (rate of evaporation, presence of salts or minerals) in the final product distribution of our oligomeric products.

Jung and Goddess: the significance of Jungian and post-Jungian theory to the development of the Western Goddess Movement

This study is concerned with the significance of Jungian and post-Jungian theory to the development of the contemporary Western Goddess Movement, which includes the various self-identified nature-based, Pagan, Goddess Feminism, Goddess Consciousness, Goddess Spirituality, Wicca, and Goddess-centred faith traditions that have seen a combined increase in Western adherents over the past five decades and share a common goal to claim Goddess as an active part of Western consciousness and faith traditions. The Western Goddess Movement has been strongly influenced by Jung’s thought, and by feminist revisions of Jungian Theory, sometimes interpreted idiosyncratically, but presented as a route to personal and spiritual transformation. The analysis examines ways in which women encounter Goddess through a process of Jungian Individuation and traces the development of Jungian and post-Jungian theories by identifying the key thinkers and central ideas that helped to shape the development of the Western Goddess Movement. It does so through a close reading and analysis of five biographical ‘rebirth’ memoirs published between 1981 and 1998: Christine Downing’s (1981) The Goddess: Mythological Images of the Feminine; Jean Shinoda Bolen’s (1994) Crossing to Avalon: A Woman’s Midlife Pilgrimage; Sue Monk Kidd’s (1996) The Dance of the Dissident Daughter: A Woman’s Journey from Christian Tradition to the Sacred Feminine; Margaret Starbird’s (1998) The Goddess in the Gospels: Reclaiming the Sacred Feminine; and Phyllis Curott’s (1998) Book of Shadows: A Modern Woman’s Journey into the Wisdom of Witchcraft and the Magic of the Goddess. These five memoirs reflect the diversity of the faith traditions in the Western Goddess Movement. The enquiry centres upon two parallel and complementary research threads: 1) critically examining the content of the memoirs in order to determine their contribution to the development of the Goddess Movement and 2) charting and sourcing the development of the major Jungian and post-Jungian theories championed in the memoirs in order to evaluate the significance of Jungian and post-Jungian thought in the Movement. The aim of this study was to gain a better understanding of the original research question: what is the significance of Jungian and post-Jungian theory for the development of the Western Goddess Movement? Each memoir is subjected to critical review of its intended audiences, its achievements, its functions and strengths, and its theoretical frameworks. Research results offered more than the experiences of five Western women, it also provided evidence to analyse the significance of Jungian and post-Jungian theory to the development of the Western Goddess Movement. The findings demonstrate the vital contributions of the analytical psychology of Carl Jung, and post-Jungians M Esther Harding, Erich Neumann, Christine Downing, E.C. Whitmont, and Jean Shinoda Bolen; the additional contributions of Sue Monk Kidd, Margaret Starbird, and Phyllis Curott, and exhibit Jungian and post-Jungian pathways to Goddess. Through a variety of approaches to Jungian categories, these memoirs constitute a literature of Individuation for the Western Goddess Movement.

Exploring coordination-driven self-assembly with autonomous chemical robots

The work presented herein focused on the automation of coordination-driven self assembly, exploring methods that allow syntheses to be followed more closely while forming new ligands, as part of the fundamental study of the digitization of chemical synthesis and discovery. Whilst the control and understanding of the principle of pre-organization and self-sorting under non-equilibrium conditions remains a key goal, a clear gap has been identified in the absence of approaches that can permit fast screening and real-time observation of the reaction process under different conditions. A firm emphasis was thus placed on the realization of an autonomous chemical robot, which can not only monitor and manipulate coordination chemistry in real-time, but can also allow the exploration of a large chemical parameter space defined by the ligand building blocks and the metal to coordinate. The self-assembly of imine ligands with copper and nickel cations has been studied in a multi-step approach using a self-built flow system capable of automatically controlling the liquid-handling and collecting data in real-time using a benchtop MS and NMR spectrometer. This study led to the identification of a transient Cu(I) species in situ which allows for the formation of dimeric and trimeric carbonato bridged Cu(II) assemblies. Furthermore, new Ni(II) complexes and more remarkably also a new binuclear Cu(I) complex, which usually requires long and laborious inert conditions, could be isolated. The study was then expanded to the autonomous optimization of the ligand synthesis by enabling feedback control on the chemical system via benchtop NMR. The synthesis of new polydentate ligands has emerged as a result of the study aiming to enhance the complexity of the chemical system to accelerate the discovery of new complexes. This type of ligand consists of 1-pyridinyl-4-imino-1,2,3-triazole units, which can coordinate with different metal salts. The studies to test for the CuAAC synthesis via microwave lead to the discovery of four new Cu complexes, one of them being a coordination polymer obtained from a solvent dependent crystallization technique. With the goal of easier integration into an automated system, copper tubing has been exploited as the chemical reactor for the synthesis of this ligand, as it efficiently enhances the rate of the triazole formation and consequently promotes the formation of the full ligand in high yields within two hours. Lastly, the digitization of coordination-driven self-assembly has been realized for the first time using an in-house autonomous chemical robot, herein named the ‘Finder’. The chemical parameter space to explore was defined by the selection of six variables, which consist of the ligand precursors necessary to form complex ligands (aldehydes, alkineamines and azides), of the metal salt solutions and of other reaction parameters – duration, temperature and reagent volumes. The platform was assembled using rounded bottom flasks, flow syringe pumps, copper tubing, as an active reactor, and in-line analytics – a pH meter probe, a UV-vis flow cell and a benchtop MS. The control over the system was then obtained with an algorithm capable of autonomously focusing the experiments on the most reactive region (by avoiding areas of low interest) of the chemical parameter space to explore. This study led to interesting observations, such as metal exchange phenomena, and also to the autonomous discovery of self assembled structures in solution and solid state – such as 1-pyridinyl-4-imino-1,2,3-triazole based Fe complexes and two helicates based on the same ligand coordination motif.