Adoption of no-till (NT) and reduced tillage management is widespread on the Canadian prairies and together form the basic platform of soil management upon which most crop production is based. Elimination of tillage in cropping systems changes the physical and chemical characteristics of the soil profile and can affect crop growth and ultimately yield. As such, understanding how soil biota, as drivers of nutrient turnover, adapt to NT is important for maximizing crop productivity and mitigating environmental damage in agroecosystems. This work aims to achieve a greater understanding of microbial community structure and function in long-term NT versus conventionally tilled (CT) soils. Community phospholipid and DNA fingerprinting did not reveal any consistent tillage-induced shifts in microbial community structure, but demonstrated a clear influence of depth within the soil profile. While tillage did not result in broad changes in the community structure, total, bacterial and fungal biomass was consistently greater near the surface of NT soils. Further examination at one site near Swift Current, SK revealed differences in microbial biomass and community structure in NT and CT in field-formed aggregate size fractions. Measurement of mineralization and nitrification at the same site indicated that differences in the early-season turnover of N may be related to physical rather than microbial differences in NT and CT soils. Potential nitrification was higher prior to seeding than mid-season, was not affected by tillage and was correlated with ammonia oxidizer population size of archaea, but not bacteria. This work indicates that edaphic soil properties and spatial distribution of resources in the soil profile, rather than tillage management, are the primary factors driving microbial community structure in these soils.

Phospholipid vesicles are biocompatible, and have potential for intracellular applications, but minimal membrane integrity limits their use in membrane-rich environments. Stabilized membranes overcome this limitation while maintaining biocompatible surface structures. Additionally, the modularity of phospholipid bilayer makes them ideal components when designing biologically inspired sensors. Membrane composition can be tailored to specific applications, transmembrane proteins can provide added functionalities, and the isolated interior can prevent cytotoxic and interfering detection chemistries from altering the cellular environment. This work has focused on expanding the capabilities of stabilized phospholipid membranes, and determining which formulations hold promise in developing stabilized phospholipid vesicle nanosensors.Current membrane stabilization methods suffer from either incomplete stabilization, or irreversible stabilization limiting the applications of vesicle nanosensors. Therefore, a facile method to prepare robust phospholipid vesicles using commonly available phospholipids stabilized via the formation of an interpenetrating, acid-labile, cross-linked polymer network that imparts controlled polymer destabilization and subsequent vesicle degradation was developed. Upon exposure to acidic conditions, the highly cross-linked polymer network was converted to linear polymers, substantially reducing vesicle stability upon exposure to chemical and physical insults. The resultant transiently stabilized vesicles have potential for enhanced drug delivery and chemical sensing applications requiring minimal membrane defects, and allow for improved physiological clearance.Some vesicle nanosensor schemes may require the passive diffusion of low molecular weight species across the membrane in addition to controllable degradation. Therefore, the acid-degradable, polymer-stabilized, phospholipid vesicle production method was extended to bis-SorbPC membranes by simultaneously polymerizing the vesicle with an acetal-containing cross-linker. The vesicles display prolonged stability under physiological conditions, and significant additional stability compared to vesicles composed of naturally occurring phospholipids. The vesicles demonstrated potential utility for sensing and therapeutic applications.Phospholipid vesicles can also serve as labels to observe movement in macromolecular biological assemblies, but a dearth of caged fluorescent labels limits design and function. Therefore, the first caged fluorescent thiol was synthesized, shown to label amines rapidly, and demonstrated the required photolytic properties. The caged fluorescent thiol has potential as a label in observing the movement of macromolecular biological assemblies and as a fluorescent probe for observing endosomal trafficking and release.

Mining activities cause severe disturbance to the soil environment in terms of soil quality and productivity and are of serious concern worldwide. Under South African legislation, developers are required to ecologically rehabilitate damaged environments. The application of agronomic approaches for the rehabilitation of coal discard sites has failed dismally in the arid areas of southern Africa. It is obvious that compliance with mitigation and rehabilitation requirements cannot be enforced without a thorough understanding of the ecological principles that ensure ecological stability and subsequent sustainability of soil ecosystems. Soil micro organisms are crucial role-players in the processes that make energy and nutrients available for recycling in the soil ecosystem. Poor management practices and other negative impacts on soil ecosystems affect both the physical and chemical properties of soil, as well as the functional and structural properties of soil microbial communities. Disturbances of soil ecosystems that impact on the normal functioning of microbial communities are potentially detrimental to soil formation, energy transfers, nutrient cycling, plant reestablishment and long-term stability. In this regard, an extensive overview of soil properties and processes indicated that the use of microbiological and biochemical soil properties, such as microbial biomass, enzymatic activity and the analysis of microbial community structure by the quantification of specific signature lipid biomarkers are useful as indicators of soil ecological stress or restoration properties because they are more responsive to small changes than physical and chemical characteristics. In this study, the relationship between the physical and chemical characteristics and different biological indicators of soil quality in the topsoil covers of seven coal discard sites under rehabilitation in South Africa, as well as three reference sites was investigated. Through the assimilation of basic quantitative data and the assessment of certain physical, chemical and biological properties of the topsoil covers obtained from the various coal discard sites as well as the reference sites, the relative success or progress of rehabilitation and the possible correlation between the biological indicators of soil quality and the establishment of self sustaining vegetation covers was determined. Results from soil physical and chemical analyses and percentage vegetation cover were correlated with the results obtained for the functional and structural diversity of microbial communities at the various sites. All results were investigated through statistical and multivariate analysis and the most prominent physical and chemical parameters that influence the biological and biochemical properties of the soil and possibly the establishment of self-sustainable vegetation cover on these mine-tailing sites were identified. Results obtained from this study indicated no significant difference (p>0.05) between the various discard sites based on conventional microbiological enumeration techniques. However, significant differences (p<0.05) could be observed between the three reference sites. All enzymatic activities assayed for the rehabilitation sites, with the exception of urease and alkaline phosphatase displayed a strong, positive association with the organic carbon content (%C). Ammonium concentration had a weak association with all the enzymes studied and pH only showed a negative association with acid phosphatase activity. A positive association was observed between the viable microbial biomass, vegetation cover and the organic carbon content, ammonium, nitrate and phosphorus concentrations of the soil. The various rehabilitation and reference sites could be differentiated based on the microbial community structure as determined by phospholipid fatty acid (PLFA) analysis. It is hypothesised that the microbial community structure of the Hendrina site is not sustainable when classified along an r-K gradient and that the high percentage of vegetation cover and high levels of estimated viable microbial biomass are an artificial reflection of the current management practices being employed at this site. Results obtained during this study, suggest that an absence or low percentage of vegetation cover and associated lower organic matter content of the soil have a significant negative impact on soil biochemical properties (enzymatic activity) as well as microbial population size. Furthermore, prevailing environmental physico-chemical and management characteristics significantly influences the vegetation cover and subsequently the microbial community structure. The results indicate that the microbial ecosystems in the coal discard sites could become more stable and ecologically self-regulating, provided effective management to enhance the organic carbon content of the soil. This could enhance nutrient cycling, resulting in changes of soil structure and eventually an improved soil quality which could facilitate the establishment of self sustaining vegetation cover. Results obtained during this study suggest that a polyphasic assessment of physical and chemical properties; microbial activities by enzymatic analysis; the characterisation of microbial community structure by analysis of phospholipid fatty acids; and the multifactorial analysis of the data obtained can be used as complementary assessment criteria for the evaluation of the trend of rehabilitation of mine tailings and discard sites. Strategic management criteria are recommended based on the soil quality environmental sustainability indices to facilitate the establishment of self sustainable vegetation covers. The contribution of this research to soil ecology is significant with regards to the intensive investigation and explanation of characteristics and processes that drive ecological rehabilitation and determine the quality of the soil environment. The multidisciplinary approach that is proposed could, furthermore, assist in the successful rehabilitation and establishment of self-sustaining vegetation covers at industrially disturbed areas, as well as assist in improving degraded soil quality associated with both intensive and informal agriculture. Additionally, this approach could negate the negative social and environmental impacts frequently associated with these activities. / Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2004.

L’auto-assemblatge de molècules en nano- i micro-estructures és una àrea de gran interès, sent els lípids particularment atractius en la formació de diverses estructures incloent els liposomes. Hi ha un gran número de mètodes reportats en la literatura per a la preparació de liposomes, però els inconvenients que limiten l’ús generalitzat dels liposomes són; els passos de preparació que requereixen de molt de temps donant lloc a poblacions heterogènies de liposomes de mida incontrolable, l’ús de solvents orgànics i la necessitat de passos per a reduir la mida dels liposomes. Per tant; l’objectiu d’aquest doctorat és la optimització d’un mètode ultra-ràpid per a la preparació de liposomes en un sol pas i lliure de dissolvents orgànics. Anomenat “Curvature tuned preparation method” ha estat implementat en diverses formulacions lipídiques per a la formació de liposomes i d’altres superestructures de lípids. Aquestes estructures s’han emprat en diverses aplicacions, com ara en nanoreactors i plantilles per a la síntesis a mida de nanopartícules d’or, liposomes per encapsular enzims com a potenciadors de senyal en el desenvolupament de immunosensors i finalment, com a vehicles per l’alliberament controlat de fàrmacs. / The self-assembly of molecules into nano- or microstructures is an area of intense interest, with lipids being particularly attractive in the formation several structures including liposomes. There are numerous methods reported for the preparation of liposomes, however, time-consuming preparative steps resulting in heterogeneous liposome populations of incontrollable size, the use of organic solvents and the need of further size-reducing steps are the drawbacks limiting wide-spread use of liposomes. Therefore; the main concern of this PhD thesis is optimization of a one-step, organic solvent-free, ultra rapid method for the preparation of liposomes. So called “Curvature tuned preparation method” was later implemented in several lipid formulations which resulted in liposomes and other lipid superstructures. Those structures were further used in several applications, such as nanoreactors and templates for tailored synthesis of gold nanoparticles, enzyme encapsulating liposomes as signal enhancers in immunosensor development, and finally as carriers for controlled release of drugs

Cancer is the leading cause of death worldwide and one third of its burden can be decreased with early detection. Surface enhanced Raman spectroscopic (SERS) based imaging is a promising new technique for non-invasive detection of tumours due to its ultra-sensitivity and multiplexing capabilities. For in vivo SERS molecular imaging, a biocompatible, robust and targeted nanoparticle is required to attain high sensitivity and specificity. In this thesis, a SERS capable gold nanoparticle was rationally designed by encapsulation with a phospholipid bilayer which conferred biocompatibility, colloidal stability and versatility to changing surface chemistry. Moreover, validation of this SERS probe with a specific targeting ligand for carcinoma cells was studied through the targeting of a commonly overexpressed cancer receptor, epidermal growth factor receptor. Using this phospholipid design, optimizations with differing chemistries, targeting ligand or modifications for additional functionalities can be achieved for further development as a viable in vivo molecular imaging tool.

Cancer is the leading cause of death worldwide and one third of its burden can be decreased with early detection. Surface enhanced Raman spectroscopic (SERS) based imaging is a promising new technique for non-invasive detection of tumours due to its ultra-sensitivity and multiplexing capabilities. For in vivo SERS molecular imaging, a biocompatible, robust and targeted nanoparticle is required to attain high sensitivity and specificity. In this thesis, a SERS capable gold nanoparticle was rationally designed by encapsulation with a phospholipid bilayer which conferred biocompatibility, colloidal stability and versatility to changing surface chemistry. Moreover, validation of this SERS probe with a specific targeting ligand for carcinoma cells was studied through the targeting of a commonly overexpressed cancer receptor, epidermal growth factor receptor. Using this phospholipid design, optimizations with differing chemistries, targeting ligand or modifications for additional functionalities can be achieved for further development as a viable in vivo molecular imaging tool.

The phospholipid membrane is a self-assembled, dynamic molecular system that may exist alone in association with only water, or in complex systems comprised of multiple lipid types and proteins. In this dissertation the intra- and inter-molecular forces responsible for the atomistic, molecular and collective equilibrium structure and dynamics are studied by nuclear magnetic resonance spectroscopy (NMR). The multinuclear NMR measurements and various experimental techniques are able to provide data that enable the characterization of the hierarchical spatio-temporal organization of the phospholipid membrane. The experimental and theoretical studies conducted target membrane interactions ranging from model systems composed of only water and lipids, to multiple component domain forming membranes that are in association with peripheral and trans-membrane proteins. These measurements consisit of frequency spectrum lineshapes and nuclear-spin relaxation rates obtained using 2 H NMR, 13 C NMR, 31 P NMR and 1 H NMR. The changes of these experimental observables are interpreted within a statistical thermodynamic framework that allows the membrane structure, activation energies, and correlation times of motion to be determined. The cases presented demonstrate how fundamental principles of NMR spectroscopy may be applied to a host of membranes, leading to the biophysical characterization of membrane structure and dynamics.

Group A streptococcal disorders can result from autoantibodies generated against M proteins. These autoantibodies cross react with the basal ganglia resulting in movement disorders. Previously, we demonstrated binding of streptococcal mAb10F5, with CPu and phospholipids. To determine if mAb10F5 binding to basal ganglia and phospholipids is due to virulence of the antibody or antibody subtype, rats were injected with control IgG2 antibodies and euthanized after 24, 48, or 72 hours. Brains were harvested and immunofluorescence was used to analyze brain slices. Control IgG2 rats showed significantly less fluorescence in the CPu than mAb10F5 injected rats at every time point. These findings reaffirm 10F5 is an anti-basal ganglia antibody. To evaluate mechanism of antibody entry, mAb10F5 was examined for anti-phospholipid activity. MAb10F5 displayed greater affinity to phospholipids when compared to IgG2 controls. Our findings support mAb10F5 is an anti-basal ganglia and anti-phospholipid antibody due to its own virulence. / Access to thesis permanently restricted to Ball State community only / Department of Physiology and Health Science

Background: Dietary fat intake particularly of omega-3 and omega-6 fatty acids play an important role in growth and development and influence the risk of nutrition related non communicable diseases. These dietary intakes are reflected in the red blood cell (RBC) fatty acid profile. Aim: The aim of this study was to assess the fatty acid profile (%) of red blood cell membrane phospholipids in relation to the dietary intake of South African children aged 2 to 5 years, and of their mothers/caregivers from three communities, each with distinct dietary patterns. Method: In this cross-sectional observational study, approximately 105 children, aged: 2-5 years and their mother/caregivers were selected from three different geographical areas. These included; the urban community of De Aar (n=105), the urban coastal community Ocean View (n=93) and the rural community of Sekhukhune District (n=104). The red blood cell membrane total phospholipid fatty acid profile was determined by gas chromatography. A 24-hour dietary recall was done for each child and mother/caregiver as well as a socio-demographic questionnaire answered by each mother/caregiver. The mean and standard deviations of the RBC fatty acids were determined and compared through an analysis of variance (ANOVA) test followed by a Bonferroni post hoc test. Age and gender were controlled for in the children and age was controlled for in the mothers/caregivers. The median dietary intake (quartile range) was compared between communities by a Kruskal-Wallis test. The relationship between RBC membrane total phospholipid fatty acid profile and dietary fatty acid intake was done by stratifying the data for the three groups combined into tertiles according to RBC fatty acid profile and comparing the median (quartile range) of the dietary fatty acid intake in the different strata. Results: In the children, the total dietary fat, SFA and PUFA and omega-3 intake of De Aar (34.2%, 11.9%, 5.9% and 0.2% of energy, respectively) and Ocean View (33.0%, 11.2%, 7.4% and 0.2% of energy, respectively) was significantly different to Sekhukhune (19.9%, 6.5%, 3.0% and 0.1% of energy, respectively). Eicosaipentanoic (EPA) and docosahexaenoic (DHA) and α-linolenic acid (ALA) mean intake in children in all three sites was lower than recommended. In children from De Aar the RBC membrane total phospholipids contained significantly higher SFA and trans-fat percentages, while children in Sekhukhune District had significantly higher PUFA, omega-6 and omega-3 percentages. The linoleic acid (LA) profile in children from Ocean View was significantly higher than in those from De Aar and Sekhukhune District. The mother/caregivers’ dietary fat intake of total fat, SFA, PUFA and trans-fat was significantly higher in De Aar (31.7%, 10.5%, 6.3% and 0.2% of energy respectively) and Ocean View (37.4%, 12.1%, 8.5% and 0.59% of energy respectively) in comparison to Sekhukhune District (15.7%, 3.0%, 3.2% and 0.02% energy respectively). PUFA intakes were significantly higher in Ocean View (8.5% of energy). EPA, DHA and ALA dietary intakes were lower than recommended. The mother/caregiver’s RBC membrane total phospholipid SFA percentage was significantly higher in mothers/caregivers from De Aar and Ocean View whereas those from Sekhukhune District had significantly higher PUFA and omega-3 percentage. Conclusion: Differences particularly between the urban areas of De Aar and Ocean View and the rural area of Sekhukhune District were observed in dietary fat intake which was reflected in the red blood cell membrane total phospholipid fatty acid profile for children and mother/caregivers. Dietary omega-3 fatty acid intake was low in both children and mother/caregivers from all three study sites and is of concern. / MSc (Nutrition), North-West University, Potchefstroom Campus, 2014

Background: Dietary fat intake particularly of omega-3 and omega-6 fatty acids play an important role in growth and development and influence the risk of nutrition related non communicable diseases. These dietary intakes are reflected in the red blood cell (RBC) fatty acid profile. Aim: The aim of this study was to assess the fatty acid profile (%) of red blood cell membrane phospholipids in relation to the dietary intake of South African children aged 2 to 5 years, and of their mothers/caregivers from three communities, each with distinct dietary patterns. Method: In this cross-sectional observational study, approximately 105 children, aged: 2-5 years and their mother/caregivers were selected from three different geographical areas. These included; the urban community of De Aar (n=105), the urban coastal community Ocean View (n=93) and the rural community of Sekhukhune District (n=104). The red blood cell membrane total phospholipid fatty acid profile was determined by gas chromatography. A 24-hour dietary recall was done for each child and mother/caregiver as well as a socio-demographic questionnaire answered by each mother/caregiver. The mean and standard deviations of the RBC fatty acids were determined and compared through an analysis of variance (ANOVA) test followed by a Bonferroni post hoc test. Age and gender were controlled for in the children and age was controlled for in the mothers/caregivers. The median dietary intake (quartile range) was compared between communities by a Kruskal-Wallis test. The relationship between RBC membrane total phospholipid fatty acid profile and dietary fatty acid intake was done by stratifying the data for the three groups combined into tertiles according to RBC fatty acid profile and comparing the median (quartile range) of the dietary fatty acid intake in the different strata. Results: In the children, the total dietary fat, SFA and PUFA and omega-3 intake of De Aar (34.2%, 11.9%, 5.9% and 0.2% of energy, respectively) and Ocean View (33.0%, 11.2%, 7.4% and 0.2% of energy, respectively) was significantly different to Sekhukhune (19.9%, 6.5%, 3.0% and 0.1% of energy, respectively). Eicosaipentanoic (EPA) and docosahexaenoic (DHA) and α-linolenic acid (ALA) mean intake in children in all three sites was lower than recommended. In children from De Aar the RBC membrane total phospholipids contained significantly higher SFA and trans-fat percentages, while children in Sekhukhune District had significantly higher PUFA, omega-6 and omega-3 percentages. The linoleic acid (LA) profile in children from Ocean View was significantly higher than in those from De Aar and Sekhukhune District. The mother/caregivers’ dietary fat intake of total fat, SFA, PUFA and trans-fat was significantly higher in De Aar (31.7%, 10.5%, 6.3% and 0.2% of energy respectively) and Ocean View (37.4%, 12.1%, 8.5% and 0.59% of energy respectively) in comparison to Sekhukhune District (15.7%, 3.0%, 3.2% and 0.02% energy respectively). PUFA intakes were significantly higher in Ocean View (8.5% of energy). EPA, DHA and ALA dietary intakes were lower than recommended. The mother/caregiver’s RBC membrane total phospholipid SFA percentage was significantly higher in mothers/caregivers from De Aar and Ocean View whereas those from Sekhukhune District had significantly higher PUFA and omega-3 percentage. Conclusion: Differences particularly between the urban areas of De Aar and Ocean View and the rural area of Sekhukhune District were observed in dietary fat intake which was reflected in the red blood cell membrane total phospholipid fatty acid profile for children and mother/caregivers. Dietary omega-3 fatty acid intake was low in both children and mother/caregivers from all three study sites and is of concern. / MSc (Nutrition), North-West University, Potchefstroom Campus, 2014