This thesis presents some experimental work in the area of manipulation of microparticles. Manipulation of both magnetic and non magnetic beads as well as microorganisms are addressed. The work on magnetic bead manipulation is focused on controlled transport and release, on a micrometer level, of proteins bound to the bead surface. Experimental results for protein transport and release using a method based on magnetization/demagnetization of micron-sized magnetic elements patterned on a modified chip-surface are presented. Special attention has been placed on minimizing bead-surface interactions since sticking problems have shown to be of major importance when protein-coated beads are used. The work with non-magnetic microparticles is focused on the dielectrophoretic manipulation of microorganisms. Preliminary experimental results for trapping and spatial separation of bacteria, yeast and non-magnetic beads are presented. The overall goal was to investigate the use of dielectrophoresis for the separation of sub-populations of bacteria differing in, for example, protein content. This was, however, not possible to demonstrate using our methods.Within the non-magnetic microparticle work, a method for determining the conductivity of bacteria in bulk was also developed. The method is based on the continuous lowering of medium conductivity of a bacterialsuspension while monitoring the medium and suspension conductivities.

No description available.

Coronary artery disease and chronic pain are serious health issues that cause severe discomfort and suffering in society today. Antithrombotic agents and highly potent analgesics play a critical role in improving the recovery process for patients being treated for these diseases. This thesis focuses on the design and study of pellet-based drug dosage forms which allow diffusion-controlled delivery of drugs with the aim of achieving optimal therapeutic outcomes. A wet slurry process was used to mix the drug and the polymer and/or clay precursor excipients into a paste. The pellets were then shaped via ionotropic gelation (alginate hydrogel beads/pellets), extrusion/spheronization (halloysite clay pellets) or geopolymerization. The decrease in the drug diffusion rate in the alginate beads was affected by the drug's molecular size and charge and the characteristics (such as concentration and chemical structure) of the surrounding alginate gel. The halloysite clay pellets provided sustained release of the highly potent drug fentanyl at both gastric pH 1 and intestinal pH 6.8. As expected, crushing the pellets reduced the diffusion barrier, resulting in more rapid release (dose dumping). The use of mechanically strong geopolymer gels was investigated as a potential means of preventing dose dumping as a result of crushing of the dosage form. Variations in the synthesis composition resulted in drastic changes in the microstructure morphology, the porosity, the mechanical stability and the drug release rate. Pore network modeling and finite element simulations were employed to theoretically evaluate the effects of porosity and drug solubility in the geopolymer structure on the drug release process. Fitting the model parameters to experimental data showed that increased average pore connectivity, a greater pore size distribution, and increased drug solubility in the pellet resulted in an increased drug release rate. Furthermore, incorporation of pH-sensitive organic polymers in the geopolymer structure reduced the high drug release rate from the pellets at gastric pH. These results indicate that geopolymers have potential for use in pellet form; both the release rate of the drug and the mechanical stability of the pellets can be optimized to prevent dose dumping.

This research is directed towards developing a more sensitive and rapid electrochemical sensor for enzyme labeled immunoassays by coupling redox cycling at interdigitated electrode arrays (IDA) with the enzyme label b-galactosidase. Coplanar and comb IDA electrodes with a 2.4 mm gap were fabricated and their redox cycling currents were measured. ANSYS was used to model steady state currents for electrodes with different geometries. Comb IDA electrodes enhanced the signal about 3 times more than the coplanar IDAs, which agreed with the results of the simulation. Magnetic microbead-based enzyme assay, as a typical example of biochemical detection, was done using the comb and coplanar IDAs. The enzymes could be placed close to the sensing electrodes (~10 mm for the comb IDAs) and detection took less than 1 min with a limit of detection of 70 amole of b-galactosidase. We conclude that faster and more sensitive assays can be achieved with the comb IDA. A paramagnetic bead assay has also been demonstrated for detection of bacteriophage MS2, used as a simulant for biothreat viruses, such as small pox. The immunoassay was carried out in a microfluidic format with the IDA, reference and counter electrodes integrated on the same chip. Detection of 90 ng/mL MS2 or 1.5x1010 MS2 particles/mL was demonstrated.

Over the past several years, some of the platforms in the Gulf of Mexico have been damaged completely, such that conventional P&A operations may not be possible. In these cases, plugging fluid needs to be pumped through an intervention well and dropped several thousand feet in water to settle above a packer and seal the well. The current P&A material of choice is cement, but cement is miscible in water, which dilutes and contaminates the cement. Therefore, alternate plugging materials need to be used for these operations. This paper discusses the development of a cost-effective Epoxy P&A method and the challenges of using Epoxy. First, the impact of seawater, oil, and pipe dope on the curing process remains unknown. Secondly, the yield strength of Epoxy with and without the contaminating chemicals must be equal to or better than cement. Finally, previous tests have shown significant losses of Epoxy to the walls of the wellbore during the 7,000-ft drop. 2 High temperature curing and compression tests were performed on contaminated epoxy samples to determine the effectiveness of the epoxy plug. To reduce material losses, an improved method for introducing the epoxy into the target zone was developed. This method takes advantage of a narrow window in the cure process where the curing process can be suspended by quenching the partially cured liquid epoxy in water at room temperature, thereby changing the liquid epoxy into solid beads. The beads can then be pumped into the wellbore, where they liquefy at wellbore temperature, 200°F, then cure into a solid plug. Seawater was found to accelerate the cure time, while all contaminants tested reduced the fracture strength by more than 25% compared to pure resin. The yield strengths of contaminant mixtures, however, remained relatively constant, with the greatest drop being only 11%. The use of solid epoxy beads was found to have a compressive strength 50% greater than Portland cements I&II. In addition, the application mentioned herein eliminates the need to prepare the plug material on site. These advantages greatly contribute to reducing the costs of an epoxy P&A operation, to potentially being USD 0.7 million cheaper than a Portland cement operation.

No description available.

The aim of this research project was to test new collection and cultivation techniques that may increase the range of cultivable diversity of soil bacteria. Fortified BioSep beads were employed in situ to capture soil bacteria, and the success of the beads was analyzed using Phylochip microarray analysis. In the cultivation phase, three different media substrates and increased incubation period were evaluated for the ability to select novel or rare bacteria. Over 700 agricultural soil bacterial isolates were classified, including a rare Gemmatimonadetes sp., a rare Verrucomicrobia sp., several Acidobacteria sp., and many novel isolates. Land management, media, and incubation period each resulted in lineage specific preferences. The yeast fortified BioSep bead cultivation collection was significantly different from the bulk soil or acyl homoserine lactone (AHL) fortified bead cultivation collections, and there were lineage specific differences in all three collection types. Phylochip analysis showed a significant difference between bulk soil and all BioSep bead (water, yeast, or AHL fortified) communities based on microarray analysis of 16S rDNA. The yeast fortified BioSep bead community was richer in operational taxonomic units (OTU) than all others. The number of phyla determined by the Phylochip analysis was much higher than that seen in the overall cultivation collection.Prophage induction assays of 21 isolates were performed, using mitomycin C (mitC) and a mixture of six AHLs, to examine soil lysogenic phage-host interactions. The fraction induced by mitC was 29%, and 10% were induced by AHL. There was no correlation between induction and land management or host growth rate. This research showed that increases in cultivable diversity can be attained by the use of BioSep beads in the collection process, varying media substrates, and by extending incubation of inoculate cultures. Phylochip analysis, however, revealed that even with altered cultivation methods, there is still a wealth of soil bacterial diversity that remains to be cultivated from this site. We also found that AHLs impact the interactions between soil bacterial hosts and prophage.

This dissertation describes the development of a new method for increasing the resolution of the current protein microarray technology, down to the single molecule detection level. By using a technique called size-dependent self-assembly, different proteins can be bound to different sized fluorescent nanostructures, and then located on a patterned silicon substrate based on the sized pattern which is closest to the size of the bead diameter.The protein nanoarray was used to detect antibody-antigen binding, specifically anti-mouse IgG binding to mouse IgG. The protein nanoarray is designed with the goal of analyzing rare proteins. However, common proteins, such as IgG, are used in the initial testing of the array functionality. Mouse IgG, representing rare proteins, is conjugated to fluorescent beads and the beads are immobilized on a patterned silicon surface. Then anti-mouse IgG binds to the mouse IgG on the immobilized beads. The binding of the antibody, anti-mouse IgG, to the antigen, mouse IgG is determined by fluorescent signal attenuation.The first objective was to bind charged nanoparticles, conjugated with proteins, to an oppositely charged silicon substrate. Binding of negatively charged gold nanoparticles (AuNP), conjugated with mouse IgG, to a positively charged silicon surface was successful.The second objective was to demonstrate the method of size-dependent self-assembly at the nanometer scale (<100 >nm). Different-sized, carboxylated, fluorescent beads and AuNP, which were conjugated with proteins, were serially added to a patterned polymethyl methacrylate (PMMA) coated silicon surface. Size-dependent self-assembly was successfully demonstrated, down to the nanometer scale.The final objective was to obtain a signal from antibody-antigen binding within the protein array. Conjugated fluorescent beads were bound to e-beam patterns and signal attenuation was measured when the antibodies bound to the conjugated beads. The size-dependent self-assembly is a valuable new method that can be used for the detection and quantification of proteins.

Controlled release formulations offer many advantages over conventional dosage forms. These include reduced plasma fluctuations and improved patient comp1i:nce. Complex controlled release formulations such as those with enteric release properties, often require additional steps in the production phase. The costs and economic impact associated with these complex controlled release dosage formulations often outweigh the immediate benefits. Thus the development of an economic method to produce controlled release particles is of great importance especially in third world countries. In controlled release formulations, the drug is generally dispersed throughout a polymer matrix. The rate of drug release is often determined by the viscosity or complexity of the polymer matrix through which the drug needs to diffuse in order to be released. With enteric release the polymer coating, insoluble in an acidic environment is often applied in the final phase of production. Chitosan is a versatile polymer of natural origin with many favourable characteristics. These include its safety, biocompatibility, and biodegradability. Simple methods can be applied and modified to produce controlled release particles form chitosan. The effect of modern controlled release polymers such as Aqoat AS-HF, Eudragit SlOO and Kollidon SR was investigated. Chitosan beads and chitosan-polymer beads, as well as chitosan granules and chitosan-polymer granules, were prepared and investigated as possible controlled release formulations. Ketoprofen was chosen as the model drug. Chitosan beads and chitosan-polymer beads were prepared by inotropic gelation in tripolyphosphate. Chitosan granules and chitosan-polymer matrix granules were prepared by binding chitosan with an acetic acid solution as a granulating system. The beads and granules appeared differed in appearance as well as in the results obtained from various experiments. Granules prepared in the study did not appear to be effective with regards to enteric and controlled release. Beads prepared form Kollidon SR appeared to be effective with regards to enteric and controlled release, with Kollidon 1% and 5% w/v chitosan beads achieving good drug loading of up to 73.13% and releasing less than 15 % of the total drug content in 0.1 M HCI after 60 minutes. Drug release continued steadily for up to 360 minutes in pH 7.2. It was concluded that Kollidon SR loaded chitosan beads nay be a viable controlled release dosage form with enteric release properties, and that future experiments, possibly with lower polymer concentrations, are worthwhile / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2006.

Biphasic systems such as two-phase partitioning bioreactors (TPPBs) have been used to alleviate biological inhibition by sequestering inhibitory compounds within an immiscible phase. The use of solid polymer beads as this auxiliary phase provides a fully biocompatible alternative to commonly used yet potentially toxic organic solvents. This work focused on the application of solid-liquid TPPBs to the bioproduction of the pharmaceutical precursor L-phenylacetylcarbinol (PAC), a biotransformation which suffers from substrate (benzaldehyde), product (PAC), and by-product (benzyl alcohol) inhibition, and simple strategies to improve TPPB performance in general. A wide range of commercially available, biocompatible, and non-bioavailable polymers were screened for their affinity for benzaldehyde, PAC, and benzyl alcohol. Hytrel G3548L demonstrated the highest affinity for all three target compounds and was subsequently used in solid-liquid TPPBs for PAC production. Using 15% v/v polymer beads, PAC concentration was increased by 104% and benzyl alcohol concentration decreased by 38% over the single phase control. The delivery of benzaldehyde from polymer beads demonstrated only a 6-8% reduction in mass productivity with improved operational simplicity and reduced operator intervention. The final objective of this work was to independently investigate various aspects of the aqueous phase composition and determine how each factor affects the partition coefficient of benzaldehyde in Hytrel G3548L. Temperature and pH were observed to have no significant effect on partitioning. Salt and glucose additions increased the partition coefficient by 173% and 30% respectively compared to RO water, while ethanol was found to decrease the partition coefficient from 44 (±1.6) to 1 (±0.3). These findings may be applied to solid-liquid TPPBs to increase or decrease partitioning as required, leading to improved bioreactor performance. This work has successfully shown that with careful polymer selection, solid-liquid TPPBs can be used to increase the productivity of a biotransformation without the associated biocompatibility problems that have sometimes been observed with organic solvents. The delivery of inhibitory substrate from the polymer phase was successfully accomplished, which is a novel demonstration in the field of solid-liquid TPPBs for biocatalysis. Finally this work contributes a range of simple strategies to improve the partitioning behavior of solid-liquid TPPBs using the aqueous phase composition. / Thesis (Master, Chemical Engineering) -- Queen's University, 2010-08-26 10:53:38.569