Micro and nanostructured impedance sensors for biological and biomedical applications

This thesis work covered the fabrication and characterisation of impedance sensors for biological applications aiming in particular to the cytotoxicity monitoring of cultured cells exposed to different kind of chemical compounds and drugs and to the identification of different types of biological tissue (fat, muscles, nerves) using a sensor fabricated on the tip of a commercially available needle during peripheral nerve block procedures. Gold impedance electrodes have been successfully fabricated for impedance measurement on cells cultured on the electrode surface which was modified with the fabrication of gold nanopillars. These nanostructures have a height of 60nm or 100nm and they have highly ordered layout as they are fabricated through the e-beam technique. The fabrication of the threedimensional structures on the interdigitated electrodes was supposed to improve the sensitivity of the ECIS (electric cell-substrate impedance sensing) measurement while monitoring the cytotoxicity effects of two different drugs (Antrodia Camphorata extract and Nicotine) on three different cell lines (HeLa, A549 and BALBc 3T3) cultured on the impedance devices and change the morphology of the cells growing on the nanostructured electrodes. The fabrication of the nanostructures was achieved combining techniques like UV lithography, metal lift-off, evaporation and e-beam lithography techniques. The electrodes were packaged using a pressure sensitive, medical grade adhesive double-sided tape. The electrodes were then characterised with the aid of AFM and SEM imaging which confirmed the success of the fabrication processes showing the nanopillars fabricated with the right layout and dimensions figures. The introduction of nanopillars on the impedance electrodes, however, did not improve much the sensitivity of the assay with the exception of tests carried out with Nicotine. HeLa and A549 cells appeared to grow in a different way on the two surfaces, while no differences where noticed on the BALBc 3T3 cells. Impedance measurements obtained with the dead cells on the negative control electrodes or the test electrodes with the drugs can be compared to those done on the electrodes containing just media in the tested volume (as no cells are attached and cover the electrode surface). The impedance figures recorded using these electrodes were between 1.5kΩ and 2.5 kΩ, while the figures recorded on confluent cell layers range between 4kΩ and 5.5kΩ with peaks of almost 7 kΩ if there was more than one layer of cells growing on each other. There was then a very clear separation between the values of living cell compared to the dead ones which was almost 2.5 - 3kΩ. In this way it was very easy to determine whether the drugs affected the cells normal life cycle on not. However, little or no differences were noticed in the impedance analysis carried out on the two different kinds of electrodes using cultured cells. An increase of sensitivity was noticed only in a couple of experiments carried out on A549 cells growing on the nanostructured electrodes and exposed to different concentration of a solution containing Nicotine. More experiments to achieve a higher number of statistical evidences will be needed to prove these findings with an absolute confidence. The smart needle project aimed to reduce the limitations of the Electrical Nerve Stimulation (ENS) and the Ultra Sound Guided peripheral nerve block techniques giving the clinicians an additional tool for performing correctly the peripheral nerve block. Bioimpedance, as measured at the needle tip, provides additional information on needle tip location, thereby facilitating detection of intraneural needle placement. Using the needle as a precision instrument and guidance tool may provide additional information as to needle tip location and enhance safety in regional anaesthesia. In the time analysis, with the frequency fixed at 10kHz and the samples kept at 12°C, the approximate range for muscle bioimpedance was 203 – 616 Ω, the approximate bioimpedance range for fat was 5.02 - 17.8 kΩ and the approximate range for connective tissue was 790 Ω – 1.55 kΩ. While when the samples were heated at 37°C and measured again at 10kHz, the approximate bioimpedance range for muscle was 100-175Ω. The approximate bioimpedance range of fat was 627 Ω - 3.2 kΩ and the range for connective tissue was 221-540Ω. In the experiments done on the fresh slaughtered lamb carcass, replicating a scenario close to the real application, the impedance values recorded for fat were around 17 kΩ, for muscle and lean tissue around 1.3 kΩ while the nervous structures had an impedance value of 2.9 kΩ. With the data collected during this research, it was possible to conclude that measurements of bioimpedance at the needle tip location can give valuable information to the clinicians performing a peripheral nerve block procedure as the separation (in terms of impedance figures) was very marked between the different type of tissues. It is then feasible to use an impedance electrode fabricated on the needle tip to differentiate several tissues from the nerve tissue. Currently, several different methods are being studied to fabricate an impedance electrode on the surface of a commercially available needle used for the peripheral nerve block procedure.

The human impacts of flower farm development in the Ethiopian Rift Valley region

The flower industry has a reputation for heavy usage of toxic chemicals and polluting the environment, enormous consumption of water, and poor working condition and low wage level in various parts of the world. It is unfortunate that this industry is adamant to change and repeating the same mistakes in Ethiopia. Because of this, - there is a growing concern among the general public and the international community about sustainability of the Ethiopian flower industry. Consequently, working conditions in the flower industry, impacts of wage income on the livelihoods of employees, coping strategies of low wage flower farm workers, impacts of flower farms on the livelihoods of local people and environmental pollution and conflict, were analysed. Both qualitative and quantitative research methods were employed. Four quantitative data sets: labour practice, employees’ income and expenditure, displaced household, and flower grower views survey were collected between 2010 and 2012. Robust regression to identify the determinants of wage levels, and Multinomial logit to identify the determinants of coping strategies of flower farm workers and displaced households were employed. The findings show the working conditions in flower farms are characterized by low wages, job insecurity and frequent violation of employees’ rights, and poor safety measures. To ensure survival of their family, land dispossessed households adopt a wide range of strategies including reduction in food consumption, sharing oxen, renting land, share cropping, and shifting staple food crops. Most experienced scarcity of water resources, lack of grazing areas, death of herds and reduced numbers of livestock due to water source pollution. Despite the Ethiopian government investment in attracting and creating conducive environment for investors, not much was accomplished when it comes to enforcing labour laws and environmental policies. Flower farm expansion in Ethiopia, as it is now, can be viewed as part of the global land and water grab and is not all inclusive and sustainable. Several recommendations are made to improve working conditions, maximize the benefits of flower industry to the society, and to the country at large.

An exploration of the impact of the Green-Schools programme on the development of sustainable behaviours in the home

Concern for the sustainability of our planet is widespread. The ever-increasing economic activity and large scale industralisation our consumer society requires has increased concerns among academics, politicians, and consumers alike on natural resource depletion, waste management, dangers of toxic chemicals, and climate change. Human consumption is causing major issues for the space we inhabit. Much work has been done over the past four decades to remedy human impact on our environment at corporate, policy and consumer level. But concerns on our ability to progress the sustainability agenda remain. Consumer behaviour plays a pivotal role in sustainable development. In light of this, we need to explore and understand the ways in which consumption occurs in consumers lives, with an aim to changing behaviours that do not support the natural environment. Questions on how to change consumer behaviour dominate much of the literature on sustainable consumption, but substantial behaviour change among individuals has not occurred as predicted. Some focus has shifted to look at upstream interventions, such as education. The Green-Schools Programme (known internationally as Eco-Schools) is one such intervention. The aim of this thesis was to explore consumption in the context of the Green-Schools Programme. The main research question asks: in the context of the Green-Schools, how are sustainable behaviour practices developed in the home? The findings presented in this thesis show that sustainable behaviour has developed in the home from both internal and external factors, the Green-Schools effect being one such factor; the programme does influence behaviour in the home context to some degree. One of the main findings of this research indicates that schoolchildren are imparting ‘positive pester power’ on household behaviour practices and the majority of households are passively practicing sustainable consumption. These findings contribute to knowledge on sustainable consumption in the home context.

Towards the enantioselective synthesis of Plakortide P and its application to the development of drugs for the treatment of Chagas disease

The primary focus of this thesis was the asymmetric peroxidation of α,β-unsaturated aldehydes and the development of this methodology to include the synthesis of bioactive chiral 1,2-dioxane and 1,2-dioxalane rings. In Chapter 1 a review detailing the new and improved methods for the acyclic introduction of peroxide functionality to substrates over the last decade was discussed. These include a detailed examination of metal-mediated transformations, chiral peroxidation using organocatalytic means and the improvements in methodology of well-established peroxidation pathways. The second chapter discusses the method by which peroxidation of our various substrates was attempted and the optimisation studies associated with these reactions. The method by which the enantioselectivity of our β-peroxyaldehydes was determined is also reviewed. Chapters 3 and 4 focus on improving the enantioselectivity associated with our asymmetric peroxidation reaction. A comprehensive analysis exploring the effect of solvent, concentration and temperature on enantioselectivity was examined. The effect that different catalytic systems have on enantioselectivity and reactivity was also investigated in depth. Chapter 5 details the various transformations that β-peroxyaldehydes can undergo and the manipulation of these transformations towards the establishment of several routes for the formation of chiral 1,2-dioxane and 1,2-dioxalane rings. Chapter 6 details the full experimental procedures, including spectroscopic and analytical data for the compounds prepared during this research.