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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Suspended Polypyrrole Films Supporting Alamethicin Reconstituted Bilayer Membranes

Northcutt, Robert 03 August 2012 (has links)
This thesis presents a novel architecture for a sensing element fabricated from a conducting polymer and a bioderived membrane. The thin film device provides controlled, selective ion transport from a chemical concentration and produces measurable electrical signals, ion storage, and small scale actuation. A chemical gradient applied across a bioderived membrane generates ion flow through protein transporters in the presence of a gating signal. A conducting polymer undergoes ion ingress/egress in the presence of an electrical and chemical potential, which causes a change on the polymers conformal backbone. A ligand (or) voltage gated protein in the bioderived membrane results in ion transport through the bioderived membrane. Integrating the two electroactive materials provides a unique architecture which takes advantage of their similarities in ionic function to produce a device with controlled and selective ion transport. The chemoelectromechanical device is one that couples chemical, electrical, and mechanical potentials through number of ions, dielectric displacement, and strain. The prototype consists of a stacked thin conducting polymer film and bioderived membrane which form three aqueous chambers of varying ionic concentrations. The top chamber contains an electrolytic solution, and the bottom chamber contains deionized water adjacent to the conducting polymer. The current that passes through a conducting polymer for an applied electrical signal is based on the level of doping/undoping and therefore can be used as a method of sensing protein function in the sensing element. This architecture results in a sensing element applicable in real time chemical sensors, volatile organic compound detectors, and bioanalytical sensors. The conducting polymer layer is formed from polypyrrole (PPy) doped with sodium dodecylbenzenesulfonate (NaDBS), and the bilayer lipid membrane is formed from 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) reconstituted with the protein alamethicin. The magnitude of current required to span a 175 µm pore was empirically found to be 326.5 A/cm2 and is based on electrode condition, electrode surface area, pyrrole concentration, and electrical potential. A micron-scale pore through a silicon substrate is spanned by a thin PPy(DBS) layer, forming a bridge which supports the bioderived membrane. The bioderived membrane is reconstituted with alamethicin, a voltage-gated protein extracted from trichoderma viride. Ion transport experiments were performed to characterize the PPy(DBS) layer and the bioderived membrane and are represented as electrical equivalents for subsequent analysis. The equivalent impedance of polypyrrole was calculated to be 1.7847±0.1735Ωcm2 and capacitance was calculated to be 1.2673±0.1823µF/cm2. The equivalent impedance of a bioderived membrane was calculated to be 1.654±1.9894MΩcm2, capacitance was calculated to be 1.1221± 0.239µF/cm2, and alamethicin resistance was calculated to be 1.025± 0.7228MΩcm2. Thus, using impedance measurements in the conducting polymer layer, it is proposed that a scaled up sensing element can be fabricated using the suspended polypyrrole supported bioderived membrane.
2

Effects of carbon nanotubes on airway epithelial cells and model lipid bilayers : proteomic and biophysical studies

Li, Pin January 2014 (has links)
Indiana University-Purdue University Indianapolis (IUPUI) / Carbon nanomaterials are widely produced and used in industry, medicine and scientific research. To examine the impact of exposure to nanoparticles on human health, the human airway epithelial cell line, Calu-3, was used to evaluate changes in the cellular proteome that could account for alterations in cellular function of airway epithelia after 24 h exposure to 10 μg/mL and 100 ng/mL of two common carbon nanoparticles, singleand multi-wall carbon nanotubes (SWCNT, MWCNT). After exposure to the nanoparticles, label-free quantitative mass spectrometry (LFQMS) was used to study differential protein expression. Ingenuity Pathway Analysis (IPA) was used to conduct a bioinformatics analysis of proteins identified by LFQMS. Interestingly, after exposure to a high concentration (10 μg/mL; 0.4 μg/cm2) of MWCNT or SWCNT, only 8 and 13 proteins, respectively, exhibited changes in abundance. In contrast, the abundance of hundreds of proteins was altered in response to a low concentration (100 ng/mL; 4 ng/cm2) of either CNT. Of the 281 and 282 proteins that were significantly altered in response to MWCNT or SWCNT, respectively, 231 proteins were the same. Bioinformatic analyses found that the proteins common to both kinds of nanotubes are associated with the cellular functions of cell death and survival, cell-to-cell signaling and interaction, cellular assembly and organization, cellular growth and proliferation, infectious disease, molecular transport and protein synthesis. The decrease in expression of the majority proteins suggests a general stress response to protect cells. The STRING database was used to analyze the various functional protein networks. Interestingly, some proteins like cadherin 1 (CDH1), signal transducer and activator of transcription 1 (STAT1), junction plakoglobin (JUP), and apoptosis-associated speck-like protein containing a CARD (PYCARD), appear in several functional categories and tend to be in the center of the networks. This central positioning suggests they may play important roles in multiple cellular functions and activities that are altered in response to carbon nanotube exposure. To examine the effect of nanotubes on the plasma membrane, we investigated the interaction of short purified MWCNT with model lipid membranes using a planar bilayer workstation. Bilayer lipid membranes were synthesized using neutral 1, 2-diphytanoylsn-glycero-3-phosphocholine (DPhPC) in 1 M KCl. The ion channel model protein, Gramicidin A (gA), was incorporated into the bilayers and used to measure the effect of MWCNT on ion transport. The opening and closing of ion channels, amplitude of current, and open probability and lifetime of ion channels were measured and analyzed by Clampfit. The presence of an intermediate concentration of MWCNT (2 μg/ml) could be related to a statistically significant decrease of the open probability and lifetime of gA channels. The proteomic studies revealed changes in response to CNT exposure. An analysis of the changes using multiple databases revealed alterations in pathways, which were consistent with the physiological changes that were observed in cultured cells exposed to very low concentrations of CNT. The physiological changes included the break down of the barrier function and the inhibition of the mucocillary clearance, both of which could increase the risk of CNT’s toxicity to human health. The biophysical studies indicate MWCNTs have an effect on single channel kinetics of Gramicidin A model cation channel. These changes are consistent with the inhibitory effect of nanoparticles on hormone stimulated transepithelial ion flux, but additional experiments will be necessary to substantiate this correlation.

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