Thesis (M. Sc.)--York University, 2002. Graduate Programme in Kinesiology and Health Science. / Typescript. Includes bibliographical references (leaves 74-82). Also available on the Internet. MODE OF ACCESS via web browser by entering the following URL: http://gateway.proquest.com/openurl?url_ver=Z39.88-2004 & res_dat=xri:pqdiss & rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation & rft_dat=xri:pqdiss:MQ71602.

Thesis (M.S.)--Worcester Polytechnic Institute. / Keywords: implant; medical; atomic force microscopy; fungi; bacteria. Includes bibliographical references (p. 82-100).

Thesis (M.S.)--Mechanical Engineering, Georgia Institute of Technology, 2004. / Dr. David Ku, Committee Chair ; Dr. Raymond Vito, Committee Member ; Dr. Alexander Rachev, Committee Member. Includes bibliographical references (leaves 103-106).

Thesis (Ph.D.)--Brown University, 2008. / Vita. Advisor : L. Ben Freund. Includes bibliographical references (leaves 89-97).

Thesis (Ph. D.)--Institute of Paper Chemistry, 1971. / Includes bibliographical references (p. 86-91).

Bacterial resistance to antibiotics is increasing at a high rate in both developing and developed countries. To circumvent the problem of drug-resistant bacterial pathogens, we need to develop new effective methods, substances, and materials that can disarm and prevent them from causing infections. However, to do this we first need to find new possible targets in bacteria to approach and novel strategies to apply.Escherichia coli (E. coli) bacteria is a normal member of the intestinal microflora of humans and mammals, but frequently cause diverse intestinal and external diseases by means of virulence factors, which leads to hundreds of million sick people each year with a high mortality rate. An E. coli bacterial infection starts with adhesion to a host cell using cell surface expressed adhesion polymers, called adhesion pili. Depending on the local environment different types of pili are expressed by the bacteria. For example, bacteria found in the gastrointestinal tract commonly express different pili in comparison to those found in the urinary tract and respiratory tract. These pili, which are vital for bacterial adhesion, thereby serve as a new possible approach in the fight against bacterial infections by targeting and disabling these structures using novel chemicals. However, in order to develop such chemicals, better understanding of these pili is needed.Optical tweezers (OT) can measure and apply forces up to a few hundred pN with sub-pN force resolution and have shown to be an excellent tool for investigating mechanical properties of adhesion pili. It has been found that pili expressed by E. coli have a unique and complex force-extension response that is assumed to be important for the ability of bacteria to initiate and maintain attachment to the host cells. However, their mechanical functions and the advantage of specific mechanical functions, especially in the initial attachment process, have not yet been fully understood.In this work, a detailed description of the pili mechanics and their role during cell adhesion is presented. By using results from optical tweezers force spectroscopy experiments in combination with physical modeling and numerical simulations, we investigated how pili can act as “shock absorbers” through uncoiling and thereby lower the fluid force acting on a bacterium. Our result demonstrate that the dynamic uncoiling capability of the helical part of the adhesion pili modulate the force to fit the optimal lifetime of its adhesin (the protein that binds to the receptor on the host cell), ensuring a high survival probability of the bond.iiiSince the attachment process is in proximity of a surface we also investigated the influence of tether properties and the importance of different surface corrections and additional force components to the Stokes drag force during simulations. The investigation showed that the surface corrections to the Stokes drag force and the Basset force cannot be neglected when simulating survival probability of a bond, since that can overestimate the probability by more than an order of magnitude.Finally, a theoretical and experimental framework for two separate methods was developed. The first method can detect the presence of pili on single cells using optical tweezers. We verified the method using silica microspheres coated with a polymer brush and E. coli bacteria expressing; no pili, P pili, and type 1 pili, respectively. The second method was based on digital holography microscopy. Using the diffraction of semi-transparent object such as red blood cells, we showed that this method can reconstruct the axial position and detect morphological changes of cells.

Ovarian cancer is the most lethal gynecological cancer. This is mainly due to widespread peritoneal dissemination and malignant ascites, in which angiogenesis, the formation of new blood vessels, is critical to both ascites development and its metastasis. Loss of E-cadherin is a well-established marker that characterizes the progression of metastatic tumors, including ovarian cancer. The release of a soluble form of E-cadherin (sE-cad) has been frequently associated with a rapid reduction of functional E-cadherin at the cell surface. Importantly, sE-cad is significantly present in ascites from women with stage III/IV ovarian cancer when compared to women with benign ovarian cysts. However, despite the clinical significance, most studies have focused on its role in weakening cell-cell adhesion, whether sE-cad itself has any biological function is not fully understood. Here it is shown for the first time a potent angiogenic role for sE-cad released from ovarian carcinoma. Soluble form of E-cadherin promoted the migration, permeability, and tubulogenesis of endothelial cells. These activities were also observed with a sE-cad/Fc chimera, and targeted inhibition using E-cadherin blocking antibodies completely prevented the sE-cad mediated effects. In addition, it was further revealed that sE-cad could be released from ovarian cancer cells in form of exosomes, a form of extracellular vesicles that play an important role in distant intercellular communication. sE-cad-positive exosomes were able to stimulate the angiogenic phenotype in vitro and functional neovascularization in a Matrigel implant model in vivo. The use of E-cadherin blocking antibodies resulted in diminished angiogenesis, confirming that the effect was sE-cad-positive exosomes specific. In search of the underlying mechanism by which sE-cad-positive exosomes promoted angiogenesis in endothelial cells which lacked E-cadherin, sE-cad was found to heterodimerize with VE-cadherin. This effect was associated with constitutive activation of phosphatidylinositol 3-kinase (PI3K)/Akt and its effector β-catenin, but not p120 catenin. Similarly, the angiogenic phenotype could be reversed by inhibition of VE-cadherin, PI3K/Akt and β-catenin. A mass spectrometric proteomic analysis of the isolated exosomes revealed distinct membrane-bound proteases, especially disintegrin and metalloproteinase 10 (ADAM10) and matrix metalloproteinase 25 (MMP25) commonly associated with ovarian cancer progression, are implicated in sE-cad production. Small interfering RNA-mediated down-regulation of ADAM10 and MMP25 significantly inhibited sE-cad production. Moreover, hepatocyte growth factor, a multifaceted cytokine which is frequently elevated in ovarian cancer ascites, was shown to increase the expression of ADAM10 and MMP25 concomitant with an elevated level of sE-cad. Together, these results uncover a novel angiogenic role of sE-cad and a new mechanism of the action of sE-cad in tumor progression. / published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy

Cell adhesion to a substrate or another cell plays an important role in the activities of the cell, such as cell growth, cell migration and cell signaling and communication with extracellular environment or other cells. The adhesion of the cell to the extracellular matrix also plays a vital role in life, as it involves in healing process of a wound and formation of the blood clot inside a vessel. The spread of cancer metastasis tumors inside the body is mostly dependent on the mechanisms of the cell adhesion. The current work is devoted to studying deformation and adhesion of the cell membrane mediated by receptors and ligands in order to enhance the existing models. In fact phospholipid molecules as the constructive units of the cell membrane grant sufficient in-plane continuity and fluidity to the cell membrane that it can be acceptably modeled as a continuum fluid medium. Therefore a two dimensional isotropic continuum fluid model is proposed in here for cell under implementation of membrane theory. In accordance to lack of sufficient study on direct effect of presence of receptors on membrane dilation, the developed model engages the intensity of presence of receptors with membrane deformation and adhesion. This influence is considered through introduction of spontaneous areal dilation. Another innovation is introduced regarding conception of receptor-ligand bonds formation such that a nonlinear constitutive relation is developed for binding force based on charge-induced dipole interaction, which is physically admissible. This relation becomes also enriched by considering one-to-one shielding phenomenon. Diffusion of the receptors is formulated along the membrane under the influence of receptor-receptor and receptor-ligand interactions. Then the presented models in this work are implemented to an axisymmetric configuration of a cell to study the deformation and adhesion of its membrane. Another target of this work is to clarify the impacts of variety of material, binding and diffusion constitutive factors on membrane deformation and adhesion and to declare a rational comparison among them. / Graduate / 0548 / 0346 / golestan@uvic.ca

This thesis presents an experimental study of the effect that particle size and shape have on nanoparticle magination and adhesion propensity in micro-capillaries. With the use of half elliptical cross-section microfluidic channels that were fabricated using photolithography as well as wet and dry etching techniques and geometrically mimetic of human microcirculation, particles ranging from 93 to 970 nm were flown and imaged individually adhering to the channel walls. The results show a significant increase in particle adhesion below 200 nm as well as the emergence of a critical particle diameter above which no particle adherence was observed. The volume delivery efficiency was also shown to increase below 200 nm, providing insight for the rational design of nanocarriers for targeted cancer therapeutics. / text

published_or_final_version / abstract / toc / Surgery / Master / Master of Philosophy