Detection, imaging and quantification of gene expression in living cells can provide essential information on basic biological issues and disease processes. To establish this technology, we need to develop molecular probes and cellular delivery methods to detect specific RNAs in live cells with potential for in vivo applications. In this thesis work, the major focus is placed on the development of molecular beacons and biochemical approaches (peptides etc.) to deliver such probes to different cellular compartments. These approaches are then employed to study the expression and localization of mRNAs, co-localization of mRNAs with cytoplasmic organelles and cytoskeleton, and co-localization of RNA molecules in the nuclei of living cells. Further along this direction, we were interested in developing a better understanding of the functional states of mRNAs and the fluorescent signal observed in optical imaging experiments. To acheive this goal, we altered the translational process and studied its effect on the detection of mRNAs in living cells. The results of these studies indicate that the translational state of mRNAs favors the hybridization of molecular beacon with its target sequence. This study has also provided the evidence that molecular beacons are reversibly bound to target mRNAs and the repression of the translational process can prevent molecular beacon from binding to its target mRNA. Further, using these approaches in combination with FRAP based biophysical analysis, the dynamics of endogenous RNA in living cells are studied. These studies revealed the possible subcellular organization of RNA molecules and their dynamics in living cells. The results also demonstrated the role of cytoskeleton and ATP in the mobility of specific mRNAs in the cytoplasm. In addition to optical probes, studies have been carried out to develop an MRI contrast agent using iron-oxide nanoparticles for deep tissue molecular imaging. Specifically, we have functionalized magnetic nanoparticles that are water-soluble, mono-dispersed, biocompatible, and easily adaptable for multifunctional bioconjugation of probes and ligands. We have successfully delivered magnetic nanoparticle bioconjugates into live cells and demonstrated their effect on relaxivity. We have further studied the role of coating thickness for optimization of contrast and further enhance the fundamental understanding of contrast mechanisms.

Size and Shape Controlled Synthesis and Superparamagnetic Properties of Spinel Ferrites Nanocrystals Qing Song 216 pages Directed by Dr. Z. John Zhang The correlationship between magnetic properties and magnetic couplings is established through the investigations of various cubic spinel ferrite nanocrystals. The results of this thesis contribute to the knowledge of size and shape controlled synthesis of various spinel ferrites and core shell architectured nanocrystals as well as the nanomagnetism in spinel ferrites by systematically investigating the effects of spin orbital coupling, magnetocrystalline anisotropy, exchange coupling, shape and surface anisotropy upon superparamagnetic properties of spinel ferrite nanocrystals. A general synthetic method is developed for size and shape control of metal oxide nanocrystals. The size and shape dependent superparamagnetic properties are discussed. The relationship between spin orbital coupling and magnetocrystalline anisotropy is studied comparatively on variable sizes of spherical CoFe2O4 and Fe3O4 nanocrystals. It also addresses the effect of exchange coupling between magnetic hard phase and soft phase upon magnetic properties in core shell structured spinel ferrite nanocrystals. The role of anisotropic shapes of nanocrystals upon self assembled orientation ordered superstructures are investigated. The effect of thermal stability of molecular precursors upon size controlled synthesis of MnFe2O4 nanocrystals and the size dependent superparamagnetic properties are described.

M. Tech. (Chemistry Department, Faculty of Applied and Computer Sciences) Vaal University of Technology. / Benzene, toluene and xylene (BTX) are water pollutants that appear very often in chemical and petrochemical wastewaters due to gasoline leakage from storage tanks and pipelines. These BTX compounds can cause adverse health effects on humans even at very low concentrations. Amongst the available pollutant removal methods from wastewater, adsorption has been used due to its ease of operation, simplicity and cost-effectiveness. Different adsorbents have been used for BTX removal, however the use of Magnetite-organic acid composites as an adsorbent seems to offer a much cheaper alternative. This work seeks to develop a one-step microwave synthesis and optimization of magnetite-oleic (MNP-OA) and magnetite-palmitic (MNP-PA) acid) composites. Response surface methodology was used to optimize the magnetite-organic acid composites. The optimum conditions estimated for MNP-OA acid composite were 78.3 % Fe content, 1561.9 S/cm conductivity, 82.2, 84.1, 85.3 mg/g for BTX adsorption capacity. The MNP-PA composite were 75.6 % Fe content, 1325.66 S/cm conductivity, 60.55, 64.47, 63.06 mg/g for BTX adsorption capacity. The materials were characterized, and the adsorption process was optimized for BTX removal from aqueous solution. X-ray analysis confirmed the formation of magnetite by the presence of both ferric and ferrous ion states on the surface. It was noted that after modification, the magnetite-organic acids characteristics peaks became broad and the height of the peaks decreased indicating that surface modification with organic acid controls the crystallinity of the material. The average cystalline size of MNP, MNP-OA, and MNP-PA composites were 19.7, 17.1 and 17.9 nm. FTIR analysis confirmed the target materials were produced and also to determine if the organic acids were imobilised on the surface of the magnetite. TEM images presented that the MNP, MNP-OA, and MNP-PA composites were spherical in shape with particle average sizes of 18.4 ± 0.5, 15.6 ± 0.5 and 16.5 ± 0.5 nm. The magnetite-organic acids show the particles with better isolated as compared to that of the MNP. The BET isotherms of the materials were described by a type IV characteristic related to uniform mesoporous materials. The magnetic saturation value for MNP, MNP-OA, and MNP-PA composites were 62.9, 59.0 and 51.0 emu/g. The decrease in magnetization was explained by the presence of the non-magnetic layer on magnetite surface. The pHpzc of MNP, MNP-OA, and MNP-PA composites were 6.9, 6.4 and 6.1. The decrease in pHpzc aftern modification was due to the charging acid-base interaction mechanism of metal oxide nanoparticles. The optimum pH for the adsorption of BTX onto MNP, MNP-OA, and MNP-PA composites was determined to be pH 7 for benzene, pH 8 for toluene and xylene. Among the three pollutants, xylene had the highest adsorption capacity followed by toluene and benzene. The optimum adsorbent dose for the adsorbents for the adsorption process was 0.1 g/dm3. The effect of time on the uptake of BTX onto MNP, MNP-OA, and MNP-PA composites show that initial adsorption of BTX occured between 0 and 3 min of contact time. The effect of initial concentration results shows the initial concentration of BTX increases from 100 to 350 mg/dm3 with an increase in adsorption capacity. The results suggest that the adsorption process is controlled by concentration driving force. The experimental data was fitted to the pseudo-first and pseudo-second-order kinetic models for all adsorbents and all pollutants. The pseudo-second-order models showed good correlation as compared to the first-pseudo model. Desorption studies for benzene, toluene and xylene using the pure magnetite, magnetite-palmitic and magnetite oleic acid composites indicate adsorption mrchanism can be explained in relation to acid–base chemistry. Electron donation from the phenyl ring of each benzene, toluene and xylene compound to surface iron atoms of magnetite has been suggested. The CH3OH and H2O desorbing agents were used and regeneration using five cycles show that the percentage desorption decreses from Benzene < Toluene < Xylene. The reduction in adsorption capacity after the cycles are attributed to decomposition of the adsorbents active sites and mass loss of the sample.

Most developing countries are faced with drinking water problems, with conditions becoming more severe due to water pollution. Meeting the growing demands for clean water in most countries, there are difficult challenges as the availability and supply of drinkable water are diminishing. Due to economic and environmental concerns, development of additional physical means for the removal of organic compounds from wastewater using permanent magnets, electromagnetic coils, electrodes and ultrasonic pretreatment is desirable. Improving the adsorption and separation process, magnetic field exposure method has progressively drawn consideration. Magnetic field exposure method has demonstrated its capacity for increasing the adsorptive elimination of contaminants from water as static magnetization is suitable, simple and cost-effective. The polypyrrole magnetic nanocomposite use for adsorption experiments influenced by exter-nal magnetic field was prepared using the in-situ polymerization method, which was charac-terized using TEM, SEM, EDX, XRD, BET, FTIR, VSM, and ESR spectrophotometers. The magnetic nanocomposite (PPy/Fe3O4) was observed to have an average particle size of 10 nm with the elementary composition of carbon, oxygen, nitrogen, chloride and iron. The magnetic nanocomposite had a crystalline structure of face-centred cubic lattice of Fe3O4, an adsorption-desorption isotherm shape indicating a typical type-IV mesoporous material with a surface area of 28.77 m2/g. Characteristic peaks of Fe3O4 and PPy were also observed using FTIR spectro-photometer. From the VSM and ESR characterization, the synthesized superparamagnetic ma-terial was shown to have a saturation magnetization of 23 emu/g and an effective g-value of 2.25 g which was attributed to Fe3+ spin interaction. An enhanced removal of Cr(VI), fluoride and congo red dye were observed under the influence of magnetic field, with parameters like pH, adsorbent dosage, the initial concentration of ad-sorbate, magnetic field and magnetic exposure time been varied. The enhanced adsorption of contaminants using magnetic field is attributed to the increase in the magnetic field induced on the particles over a magnetic exposure time, resulting in the rotating particles forming aggre-gates due to the increased magnetic force and torque on the particles from the PSV results. This leads to increase in the chain collision and area of particle interaction with the aqueous solution of hexavalent chromium, fluoride and congo red dye. / Physics / Ph. D. (Physics)