IP Peg is a member of a family of star systems known as cataclysmic variable stars, CVs. CVs are star systems in which a red dwarf star orbits a white dwarf and mass transfer is taking place from the red dwarf to the white dwarf. The mass ratio of the two component stars has been found the most difficult parameter to determine. Tools used in the analysis of mass-ratio included the differential photometric light curve of IP Peg, Doppler tomograms, and spectral data covering the HP emission-line. The mass ratio was found to lie in the range, q = 0.44 ± 0.14. The inclination lies in the range i = 81.5° ± 8.5°. Finally, the accretion disk radius was 0.342 times the distance between the stars. The analysis of the parameters of eclipsing CV systems like IP Peg provides a rare opportunity that helps in the understanding of other CV systems. / Department of Physics and Astronomy

Glaucoma is a group of optic nerve degenerative diseases, which leads to gradual and permanent vision loss. Recent developments in the field of gene therapy have proposed increasingly promising treatments for glaucoma, in the form of delivery of neuro-protective or neuro-regenerative genes to the retina. Despite these developments, there are concerns related to the biocompatibility and invasiveness of common gene delivery systems, since they are commonly mediated by viral gene carriers and invasive administration methods. Non-viral gene delivery systems offer a safe and increasingly efficient alternative to deliver therapeutic genes to the retina. An example of these systems is gemini-phospholipid nanoparticles (GL-NPs), which have been successfully used to deliver genes in similarly challenging anatomical settings, such as the skin. The objective of this thesis is to demonstrate the potential of GL-NPs, as candidate gene delivery vehicles for topically administered genes, targeted to the retina. The dicationic gemini surfactant, 12-7NH-12 was used, along with the helper lipids, 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), to prepare various types of GL-NPs, and assess their transfection efficiency in the rat retinal ganglion cell line (RGC-5). The transfection efficiency was evaluated using flow cytometry, as a function of several physical and chemical parameters of GL-NPs. These include a range of charge ratios (5:1 to 15:1 ????), helper lipid composition (several DOPE: DPPC ratios), order of assembly (plasmid-gemini + lipid versus gemini-lipid + plasmid), and manufacturing method of helper lipid vesicles (thin film versus high pressure homogenization method). Size and zeta (??) potential characterization of GL-NPs was carried out in parallel, using dynamic light scattering, to relate the physical parameters of GL-NPs to their respective transfection efficiency. A comprehensive toxicological evaluation was undertaken to assess the extent of GL-NP???s toxicity in RGC-5 cells, using the resazurin-based PrestoblueTM cell toxicity assay. Optimized GL-NPs were used to induce expression of the brain derived neurotrophic factor (BDNF) in RGC-5 cells, and were assessed in terms of their capacity to induce neurite outgrowth. Quantification of neurite outgrowth was carried out by measuring average neurite length in RGC-5 cells, by confocal microscopic imaging of immunostained neurites. Furthermore, confocal microscopic studies were carried out to assess the extent of GL-NP???s corneal permeation in a 3-D human corneal epithelial (HCE) model. A parallel toxicological evaluation was completed to ensure GL-NP???s biocompatibility with the corneal epithelial cells. Finally, GL-NP biodistribution pattern and gene transfer capacity was assessed in a mouse model, following topical and intravitreal administration. The transfection efficiency in RGC-5 cells, which ranged between 2.1 ?? 0.3% and 14.5 ?? 1.4%, was highly dependent on GL-NP???s charge ratio, helper lipid composition, order of assembly, and manufacturing technique. GL-NPs at 10:1 ???? charge ratio, assembled with homogenized DOPE (25%)-DPPC (75%) helper lipid vesicles, in the plasmid-gemini + lipid order, mediated the highest transfection efficiency in RGC-5 cells. These GL-NPs had a size of 222.8 ?? 4.2 nm and a ?? potential of +33.5??2.9 mV. Optimized GL-NPs were highly biocompatible with both RGC-5 and HCE model cells, with viability values ranging between 94.8 ?? 6 % to 100 ?? 3.4 %. Assessment of corneal permeation showed that GL-NPs were able to bind to the corneal epithelial surface and achieve a moderate permeation depth (35-40 ??m), following topical application in the HCE model. Intravitreal injection of the non-viral GL-NPs in mice has successfully led to their localization within the nerve fiber layer (NFL) of the retina. Finally, GL-NPs were non-invasively delivered to several anterior chamber tissues, including the limbus, the iris and conjunctiva, following topical administration. GL-NPs offer several advantageous features critical to topical and intravitreal ocular administration of gene carriers, including in vitro corneal binding and effective biodistribution following in vivo topical and intravitreal administration, high biocompatibility, and a highly tunable transfection efficiency. The current data presents 12-7NH-12 GL-NPs as a promising candidate for ocular gene therapy applications.

In recent years there has been increasing interest in novel forms of surfactants. Of particular interest are gemini surfactants, which consist of two conventional surfactants joined by a spacer at the head groups, as they exhibit lower critical micelle concentrations than can be achieved by conventional surfactants. In this work, the self-assembly behaviour of several nonionic gemini surfactants with polyethylene oxide head groups (GemnEm, where n (= 20) is the number of carbons per tail and m (= 10, 15, 20 and 30) is the number of ethylene oxides per head group) were investigated. The Critical Micelle Concentrations (CMCs) were measured using a fluorescence probe technique. The CMCs are all ~2 x 10?7 M, with almost no variation with m. The CMCs are several orders of magnitude lower than conventional C12Em nonionic surfactants. The mixing behaviour of the gemini surfactants with conventional surfactants was also studied. They obeyed ideal mixing behaviour with both ionic and nonionic surfactants. Micelle morphologies were studied using Small Angle Neutron Scattering. The gemini surfactants with the larger head groups (i.e. Gem20E20 and Gem20E30) formed spherical micelles. Gem20E15 showed strong scattering at low Q, characteristic of elongated micelles. As the temperature was increased towards the cloud point, the scattering approached the Q-1 dependence predicted for infinite, straight rods. The existence of anisotropic micelles was supported by the viscosity of Gem20E15, which increases by several orders of magnitude on heating towards its cloud point. Phase behaviour was determined using Diffusive Interfacial Transport coupled to near-infrared spectroscopy. Much of the behaviour of these systems is similar to conventional nonionic surfactants. For example, Gem20E10 forms a dilute liquid isotropic phase (W) coexisting with a concentrated lamellar phase (La) at around room temperature and forms a sponge phase at higher temperatures. This is similar to the behaviour of C12E3 and C12E4. The other surfactants studied are all quite soluble in water and form liquid isotropic and hexagonal phases from room temperature. At higher concentrations Gem20E15 formed a cubic and then a lamellar phase while Gem20E20 formed a cubic phase and then an intermediate phase. This is also comparable to the phase behaviour of conventional nonionic surfactants except the intermediate phase, which is often only observed for surfactant systems with long alkyl tails.

The thesis entitled “The Role of Liposomal Hybrids and Gold Nanoparticles in the Efficacious Transport of Nucleic Acids and Small Molecular Drugs for Cancer Nanomedicine” elucidates the preparation of various liposomal formulations of cationic monomeric and gemini lipids where hydrophobic domains were consisted of tocopherol, cholesterol and pseudoglyceryl backbone for the cellular transport of nucleic acids. The thesis continues while elucidating the role of various pH sensitive molecules and gold nanoparticles in liposomes to improve the delivery efficacy levels. This thesis also elucidates the role of gold nanoparticles stabilized with natural pH sensitive molecules for efficacious drug delivery applications. Additionally, the role of such pH sensitive gold nanoparticles in association with liposomes for the co-delivery of drug and gene has been discussed. The work has been divided into six chapters. Chapter 1A: Dimeric Lipids Derived from α-Tocopherol as Efficient Gene Transfection Agents. Mechanistic Insights into Lipoplex Internalization and Therapeutic Induction of Apoptotic Activity In this chapter, we present cationic dimeric (gemini) lipids for significant plasmid DNA (pDNA) delivery to different cell lines without any marked toxicity in the presence of serum. The six gemini lipids possess α-tocopherol as their hydrophobic backbone and differ from each other in terms of their spacer chain lengths. Each of these gemini lipids mixed with a helper lipid 1, 2-dioleoyl phosphatidyl ethanolamine (DOPE), was capable of forming stable aqueous suspensions. These co-liposomal systems were examined for their potential to transfect pEGFP-C3 plasmid DNA in to nine cell lines of different origins. The transfection efficacies noticed in terms of EGFP expression levels using flow cytometry were well corroborated using independent fluorescence microscopy studies. Significant EGFP expression levels were reported using the gemini co-liposomes which counted significantly better than one well known commercial formulation lipofectamine 2000 (L2K). Transfection efficacies were also analyzed in terms of the degree of intracellular delivery of labeled plasmid DNA (pDNA) using confocal microscopy which revealed an efficient internalization in the presence of serum. The cell viability assays performed using optimized formulations demonstrated no significant toxicity towards any of the cell lines used in the study. We also had a look at the lipoplex internalization pathway to profile the uptake characteristics. A caveolae/lipid raft route was attributed to their excellent gene transfection capabilities. The study was further advanced by using a therapeutic p53-EGFP-C3 plasmid and the apoptotic activity was observed using FACS and growth inhibition assay. Figure 1. The co-liposomes of tocopheryl gemini lipids and DOPE for efficient delivery of p53-EGFP-C3 plasmid DNA that induces significant apoptotic response. Chapter 1B: Efficacious Gene Silencing in Serum and Significant Apoptotic Activity Induction by Survivin Downregulation Mediated by Cationic Gemini Tocopheryl Lipids Non-viral gene delivery offers cationic liposomes as promising instruments for the delivery of double-stranded RNA (ds RNA) molecules for successful sequence-specific gene silencing (RNA interference). The efficient delivery of siRNA (small interfering RNA) to cells while avoiding the unexpected side effects is an important prerequisite for the exploitation of the power of this excellent tool. We discuss in this chapter about six tocopherol based cationic gemini lipids, which induce substantial gene knockdown without any obvious cytotoxicity. All the efficient co-liposomal formulations derived from each of these geminis and a helper lipid, dioleoyl phosphatidyl ethanolamine (DOPE) were well characterized using physical methods such as atomic force microscopy (AFM) and dynamic light scattering (DLS). Zeta potential measurements were conducted to estimate the surface charge of these formulations. Flow cytometric analysis showed that the optimized co-liposomal formulations could transfect anti-GFP siRNA efficiently in three different GFP expressing cell lines, viz. HEK 293T, HeLa and Caco-2 significantly better than a potent commercial standard Lipofectamine 2000 (L2K) both in the absence and presence of serum (FBS). Notably, the knockdown activity of co-liposomes of gemini lipids was not affected even in the presence of serum (10% and 50% FBS) while it dropped down for L2K significantly. Observations under a fluorescence microscope, RT-PCR and western blot analysis substantiated the flow cytometry results. The efficient cellular entry of labeled siRNA in GFP expressing cells as evidenced from confocal microscopy put forward these gemini lipids among the potent lipidic carriers for siRNA. The efficient transfection capabilities were also profiled in a more relevant fashion while performing siRNA transfections against survivin (an anti-apoptotic protein) which induced substantial apoptosis. Furthermore, the survivin downregulation improved the therapeutic efficacy levels of an anticancer drug, doxorubicin significantly. In short, the new tocopherol based gemini lipids appear to be highly promising for achieving siRNA mediated gene knockdown in various cell lines. Figure 2. The co-liposomes of tocopheryl gemini lipids and DOPE for efficient delivery of siRNA against survivin that induces significant apoptotic response. Chapter 2: Efficacious in Vitro EGFP Expression and Silencing in Serum by Cationic Pseudoglyceryl Gemini Lipids To elicit the desirable efficacy levels in cationic liposome mediated nucleic acid therapeutics has been part of extensive scientific efforts. This chapter describes three cationic gemini lipids and application of their co-liposomes with DOPE as potent pDNA (plasmid DNA) and siRNA (small interfering RNA) cytofectins for remarkably advanced efficacy levels in numerous cell lines in the presence of serum. The hydrophobic structural lineament of cationic gemini lipids is made up of pseudoglyceryl backbone linked to the hydrocarbon chains via oligo-oxyethylene units. The stable aqueous co-liposomal suspensions of gemini lipids showed an efficient binding to pDNA or siRNA and their significant intracellular delivery in various cell lines. The transfection capabilities of different co-liposomal formulations were profiled based on EGFP expression (pEGFP-C3 pDNA transfection) and EGFP knockdown (anti-GFP siRNA transfections) in EGFP expressing cell lines. The cellular EGFP expression levels and intracellular delivery of labeled nucleic acids were thoroughly studied using flow cytometry (FACS), fluorescence and confocal microscopy. The MTT based cell viability assay revealed no loss in cell viabilities for all of the transfection optimized lipoplexes of siRNA or pDNA. The transfection profile of gemini co-liposomes was noted to be significantly much better than a commercial lipofection reagent, Lipofectamine 2000 used for pDNA and siRNA applications in each of the cell lines studied. The co-liposomes and their transfection optimized lipoplexes were physiochemically characterized extensively by means of zeta potential, dynamic light scattering (DLS) and atomic force microscopy (AFM). In brief, these new gemini co-liposomal formulations seem to offer a great opportunity for successful nucleic acid (DNA and siRNA) delivery in a practical scenario. Figure 3. Efficacious EGFP expression (pDNA transfection) and EGFP silencing (anti GFP siRNA transfection) mediated by co-liposomes of pseudoglyceryl gemini lipids and DOPE. Chapter 3: Efficient Elicitation of Liposomal Nucleic acid delivery through the Eminence of Gold Nanoparticles Stabilized with pH Responsive Short Tripeptide Derived from Tyrosine Kinase NGF Receptors The prerequisite in the area of gene therapy today is to serve transfection efficient formulations nullifying the enduring key issues. To this end, we discuss in this chapter, the role of hybrid liposomal formulations derived from structurally distinct cationic lipids, a neutral lipid (DOPE) and pH responsive short tripeptide (KFG, Lys-Phe-Gly) capped gold nanoparticles (PAuNPs). The hybrid liposomes are presented to be efficient enough to transfect pDNA leading to remarkably high gene expression levels in various cell lines of different origins in the presence of serum (FBS). Hybrid liposomes could deliver pDNA more effectively than the native liposomes and commercial standard lipofectamine 2000 (L2K) across the entire range of N/P ratios studied under the influence of intracellular pH response and gold nanoparticles prominence. The gene transfection capabilities are profiled based on transfections performed using two different plasmids (pGL3, luciferase activity and p-EGFP-C3, green fluorescent protein expression). pDNA cellular internalization and subsequent gene expression levels are studied using flow cytometry, fluorescence microscopy and confocal microscopic studies. The extensive physiochemical characterization of hybrid liposomal formulation and their complexes with pDNA in comparison with respective native liposomes was performed using AFM, TEM, Zeta, DLS, gel retardation assay, U.V. and fluorescence emission measurements. The hybrid liposomes are shown to possess significantly higher fusion activity at lowered pH of intracellular compartments. These hybrid liposomes are fairly biocompatible across the concentration range used in transfection experiments. Precisely, introduction of these pH responsive tripeptide capped gold nanoparticles in to liposomal formulations straightforwardly must be more advantageous for a practical application in biomedical scenario to achieve therapeutic levels. Figure 4. The hybrid of liposomes and tri-peptide capped gold nanoparticles for significantly improved gene expression levels. Chapter 4: RNA Aptamer Decorated pH Sensitive Liposomes for Active Transport of Nucleic Acids in Specific Cancer Cells This chapter describes the target specific transport of pH sensitive liposomes loaded with a RNA aptamer for promising nucleic acid therapeutics. The pH sensitive liposomes are constructed from a cationic cholesteryl gemini lipid (CGL), neutral helper lipid (DOPE) and gemini analog of a pH sensitive lipid, palmitoyl homocysteine (GPHC). The liposomes are shown to be significantly fusogenic that deliver the cargoes upon lowerin the pH (6.0). The fusogenic behaviour of the liposomes was thoroughly studied by means of dynamic light scattering (DLS), zeta potential, lipid mixing, calcein dequenching and atomic force microscopy (AFM). The facile integration of cholesterol conjugated RNA aptamer in liposomes derived from cholesteryl gemini lipids was exploited for their delivery to specific cancer cells. The RNA aptamer specifically binds to epithelial cell adhesion molecule (EpCAM) with high affinity which is a cell surface marker in various solid cancers such as colorectal and breast carcinoma. These aptamer decorated pH sensitive liposomes could efficiently enter the EpCAM expressing COLO-205, Caco-2, MCF-7 and MDA-MB-231 cell lines while no such noticeable liposome transport was observed in EpCAM negative HEK 293T cells as evidenced by flow cytometry and confocal microscopy. Additionally, the liposomes are shown to be actively transported inside the cells, i.e., receptor mediated endocytosis. These liposomes could complex the nucleic acids (pDNA) in an efficient manner. The MTT based cell viability assay accounted no noticeable loss in cell viabilities for liposome treatments. Concisely, we have formulated RNA aptamer loaded pH sensitive liposomes that would certainly be promising tool in target based cancer nanomedicine. Figure 5. (A) Cellular internalization of DY-647 labeled aptamer loaded pH sensitive liposomes. (B) The liposomes were actively internalized through receptor mediated endocytosis. Each panel (A and B) represents (from left to right) bright field image, aptamer fluorescence, DAPI stained nuclei and merge of previous three impressions. Chapter 5: Natural Tri-peptide Capped Gold Nanoparticles for Efficacious Doxorubicin Delivery in Vitro and in Vivo Nanotechnology has gained ever increasing interest for the successful implementation of chemotherapy based treatment of cancer. This chapter describes the role of gold nanoparticles (AuNPs) capped with a natural pH responsive short tri-peptide (Lys-Phe–Gly or KFG) for significant intracellular delivery of an anti-cancer drug, doxorubicin (DOX). A significantly increased apoptotic response was noted for DOX treatments mediated by KFG-AuNPs in comparison with drug alone treatments in various cell lines (BT-474, HeLa, HEK 293T and U251) in vitro. Furthermore, KFG-AuNPs mediated DOX treatment significantly decreased cell proliferation and tumor growth in BT-474 cell xenograft model in nude mice. In addition, KFG-AuNPs showed efficacious drug delivery in DOX-resistant HeLa cells (HeLa-DOXR) in comparison with drug alone treatments. Figure 6. Representative images of excised tumors after doxorubicin treatment mediated by pH responsive tri-peptide capped gold nanoparticles (DOX-KFG-AuNPs) (C) in comparison with doxorubicin alone treatments (B) and untreated tumors (A). Extensive cell death as observed under Hematoxylin/eosin (H&E) (D) and TUNEL (E) staining of DOX-KFG-AuNPs treated tumor sections. Chapter 6: Significant Apoptotic Activity Induction by Efficacious Co-delivery of p53 Gene and Doxorubicin Mediated by the Combination of Co-liposomes of Cationic Gemini lipid and pH Responsive Tri-peptide Combining chemotherapy with gene therapy has appeared as an efficient tool to treat complex biological disorder like cancer. Herein, we show efficient co-delivery of DNA and an anti-cancer drug, doxorubicin (DOX) by means of gemini cationic liposome (GCL) based lipoplex nanoaggregates that are coated with DOX encapsulated pH responsive tripeptide nanovesicles. The lipoplex, tripeptide vesicles and their association was thoroughly studied using dynamic light scattering (DLS), zeta potential, atomic force microscopy (AFM). Flow cytometry, fluorescence and confocal microscopic analysis revealed that the GCL-tripeptide association could significantly co-deliver the p53 expression plasmid (p53-EGFP-C3) and DOX in HeLa and HEK 293T cells in the presence of serum. A synergistic increase in gene expression level and DOX internalization was observed in co-delivery which was even substantially higher than individual lipoplex transfection and DOX treatment. The apoptosis induced due to p53 expression and DOX was profiled with the help of annexin-V positivity analysis under flow cytometry and nuclear damage analysis by DAPI nuclei counterstaining under confocal microscopy which noted to be significantly higher in cells during co-delivery. The MTT based cell viability assay revealed a significantly increased loss in cell viability counts for co-delivery treatments. Such a system delivering synergistically increased significant efficacy levels in combinatorial drug and nucleic acid therapeutics would be certainly advantageous for practical biomedical applications. Figure 7. The co-delivery of pDNA and drug (doxorubicin) mediated by GCL-tripeptide association as observed under (A) confocal microscopy (pDNA; green and doxorubicin; red) and (B) flow cytometry.

Non-common path aberrations (NCPA) in an adaptive optics system are static aberrations that appear due to the difference in optical path between light arriving at the wavefront sensor (WFS) and at the science detector. If the adaptive optics are calibrated to output an unaberrated wavefront, then any optics outside the path of the light arriving at the WFS inherently introduce aberrations to this corrected wavefront. NCPA corrections calibrate the adaptive optics system such that it outputs a wavefront that is inverse in phase to the aberrations introduced by these non-common path optics, and therefore arrives unaberrated at the science detector, rather than at the output of the corrective elements. Focal plane sharpening (FPS) is one technique used to calibrate for NCPA in adaptive optics systems. Small changes in shape to the deformable element(s) are implemented and images are taken and analyzed for image quality (IQ) on the science detector. This process is iterated until the image quality is maximized and hence the NCPA are corrected. The work carried out as described in this paper employs two FPS techniques at Gemini North to attempt to mitigate up to 33% of the adaptive optics performance and image quality degradations currently under investigation. Changes in the NCPA correction are made by varying the Zernike polynomial coefficients in the closed-loop correction file for Altair (the facility adaptive optics system). As these coefficients are varied during closed-loop operation, a calibration point-source at the focal plane of the telescope is imaged through Altair and NIRI (the facility near-infrared imager) at f/32 in K-prime (2.12 μm). These images are analyzed to determine the Strehl ratio, and a parabolic fit is used to determine the appropriate coefficient correction that maximizes the Strehl ratio. Historic calibrations of the NCPA file in Altair's control loop were done at night on a celestial point source, and used a separate, high-resolution WFS (with its own inherent aberrations not common to either NIRI nor Altair) to measure phase corrections directly. In this paper it is shown that using FPS on a calibration source negates both the need to use costly time on the night sky and the use of separate optical systems (which introduce their own NCPA) for analysis. An increase of 6% in Strehl ratio is achieved (an improvement over current NCPA corrections of 11%), and discussions of future improvements and extensions of the technique is presented. Furthermore, a potentially unknown problem is uncovered in the form of high spatial frequency degradation in the PSF of the calibration source.

No description available.

Monte Carlo simulations have been used in the past to investigate a variety of surfactant systems; however, there is little published literature for double-headed and gemini surfactants with asymmetric tails. We perform Larson-type Monte Carlo simulations of double-headed and gemini surfactant systems with asymmetric tails in two- and three-dimensions. The model predicts that the addition of a second head group to form a double-headed surfactant results in an increase in the critical micelle concentration (CMC) compared to a single-headed surfactant, in agreement with experiment. It also indicates that the placement of the second head group has an impact on the final CMC value. We study a series of gemini surfactants with asymmetric tails and find no change in the value of the CMC as the ratio of the lengths of the two tails increases. This is contrary to the only experimental study that found there was a slight decrease in the CMC as the ratio of the lengths of the two tails increases. We examine this difference in terms of the relatively small effect surfactant asymmetry has on value of the CMC and the fact that the model is capable of qualitatively reproducing the known dependence of the CMC on other architectural properties. This initial probe into systems of double-headed and gemini surfactants with asymmetric tails confirms many of the previously published findings and provides avenues for possible future research using Monte Carlo simulations.

There is no abstract available for this thesis. / Department of Physics and Astronomy

Research in the field of non-viral gene delivery has demonstrated that a deeper understanding of the fundamental processes of nanoparticle assembly is required in order to improve their efficacy. While gemini nanoparticles (gemini NPs) and other non-viral delivery systems have been vigorously characterized using several techniques, our knowledge is still incomplete. The first objective of this study was the development of new methodology using fluorescence correlation spectroscopy (FCS) to investigate the stages of gemini NPs assembly. It was demonstrated that by labeling the plasmid, different stages of gemini NP assembly from the gemini-plasmid pre-complex (GP) to the final gemini nanoparticle (or gemini-plasmid-lipid complex; GPL), could be studied. Based on diffusion coefficients and particle numbers extrapolated from the autocorrelation function (ACF), FCS was able to determine that each phase of assembly had distinct characteristics. The FCS study using 12-3-12 gemini surfactant showed that both the diffusion coefficient and particle number of GPs (0.98??0.31 x 10-12 m2/s) was significantly lower than the final GPL (3.11??0.41 x 10-12 m2/s). Based on the Stokes-Einstein equation the particle size was calculated to be 300-500 nm for GP and 200-300 nm for GPLs. The raw intensity histograms showed that both GPs and GPLs are composed of multiple plasmids. Furthermore the study showed that the final GPLs contain fewer plasmids compared to the intermediate GP. FCS results were validated by using existing characterization methods including dynamic light scattering (DLS), zeta potential and dye exclusion assays. The second objective involved the detailed characterization of gemini NP. Nine different gemini surfactants and two different phospholipids were used in a systematic study to assess the effect of gemini surfactant and lipid structure on the final morphology of gemini NP. The study revealed that gemini surfactant structure had a strong effect on structure of GP intermediates, but addition of phospholipids resulted in the formation of uniform gemini NPs. Based on the results of this study a new model for GP and GPL assembly is proposed based on the formation of supramolecular aggregates of gemini-plasmids, governed by gemini surfactant chemical structure, and dispersed by phospholipids to form GPLs.

There are many observational methods for studying debris disks because of constraints imposed on observing their predominately infrared wavelength emission close to the host star. Two methods which are discussed here are ground-based high contrast imaging and space-based far-IR emission. The Gemini Planet Imager (GPI) is a high contrast near-IR instrument designed to directly image planets and debris disks around other stars by suppressing star light to bring out faint sources nearby. Because debris disks are intrinsically polarized, polarimetry offers a useful way to enhance the scattered light from them while suppressing the diffracted, unpolarized noise. I discuss the characterization of GPI's microlens point spread function (PSF) in polarization mode to try to improve the quality of the processed data cubes. I also develop an improved flux extraction method which takes advantage of an empirically derived high-resolution PSF for both spectral and polarization modes. To address the instrumental effects of flexure, which affect data quality, I develop methods to counteract the effect by using the science images themselves without having to take additional calibrations. By reducing the number of calibrations, the Gemini Planet Imager Exoplanet Survey (GPIES) can stand to gain ~66 hours of additional on-sky time, which can lead to the discovery of more exoplanetary systems. The Herschel Space Observatory offers another method for observing debris disks which is ideally suited to measure the peak dust emission in the far-IR. Through a careful analysis, we look at 100/160 μm excess emission around λ Boo stars, to differentiate whether the emission is from a debris disk or a bowshock with the interstellar medium. It has been proposed that the stars' unusual surface abundances are due to external accretion of gas from those sources. We find that the 3/8 stars observed are well resolved debris disks and the remaining 5/8 were inconsistent with bowshocks. To provide a causal explanation of the phenomenon based on what we now know of their debris disks, I explore Poynting-Robertson (PR) drag as a mechanism for secondary accretion via a debris disk. However, I find that the accretion rates are too low to cause the surface abundance anomaly. Further study into the debris disks in relation to stellar abundances and surfaces are required to rule out or explain the λ Boo phenomenon through external accretion. / Graduate / 0606 / zhd@uvic.ca