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Size, Charge and Dose Dependent In-vitro Kinetics of Polystyrene Nanoparticles
Title
Size, Charge and Dose Dependent In-vitro Kinetics of Polystyrene Nanoparticles.
Creator
Abdellatif, Yasmine, Gesquiere, Andre, Kang, Hyeran, Rajaraman, Swaminathan, University of Central Florida
Abstract / Description
The aim of the study described herein is to quantify the in-vitro kinetics of internalization of polystyrene nanoparticles (PS NPs) by cells. We used different charges, sizes and doses of fluorescently labelled PS NPs. Nanoparticles were characterized with UV-Vis, Fluorescence emission Dynamic Light Scattering (DLS) and Zeta potential for knowing their absorption, fluorescence spectra, size, charge, respectively. Additionally, cell viability was tested to know the toxicity of PS NPs. The...
Show moreThe aim of the study described herein is to quantify the in-vitro kinetics of internalization of polystyrene nanoparticles (PS NPs) by cells. We used different charges, sizes and doses of fluorescently labelled PS NPs. Nanoparticles were characterized with UV-Vis, Fluorescence emission Dynamic Light Scattering (DLS) and Zeta potential for knowing their absorption, fluorescence spectra, size, charge, respectively. Additionally, cell viability was tested to know the toxicity of PS NPs. The quantitative uptake, the kinetics profile and rate of uptake were studied by using a new in-vitro fluorescence assay. This was achieved quantitatively and qualitatively by fluorescent plate reader and confocal imaging, respectively. It was found that the amine PS NPs are higher in cytotoxicity than the carboxy PS NPs due to the proton sponge phenomenon. It was observed that the fraction uptake of PS NPs changes by changing the physiochemical properties as charge, size (&) dose. The fraction uptake of neutral and amine PS NPs was higher than that of carboxy PS NPs. For the neutral PS NPs, the uptake depends on the macropinocytosis. For the amine PS NPs, the uptake depends on the electrostatic interaction and the rapid regeneration of new binding sites. Regarding the dose of PS NPs, for the amine PS NPs, it was found that the concentrations lower and higher than 5nM had lower fraction uptake, because the 5nM achieved the balance between the available number of binding sites and the rapid regeneration of new binding sites. For the kinetics profile of the amine and carboxy PS NPs, by comparing both of them, it was observed that the rate of uptake of applied doses lower than 5nM was different, but higher than 5nM was similar. However, for the neutral Ps NPs, they exhibit a steady state of rate of uptake in between the amine and carboxy PS NPs. Also, it was confirmed by the confocal images that as the concentration of amine PS NPs increase, the stress on the cells increase, leading to the cell death. These results were aligned with the results obtained from the cytotoxicity test.
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Date Issued
2018
Identifier
CFE0007386, ucf:52744
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0007386
Interplay of Molecular and Nanoscale Behaviors in Biological Soft Matter
Title
Interplay of Molecular and Nanoscale Behaviors in Biological Soft Matter.
Creator
Ciaffone, Nicholas, Tetard, Laurene, Kang, Hyeran, Santra, Swadeshmukul, University of Central Florida
Abstract / Description
The complexity of biological soft matter at the sub-micrometer level is fundamentally correlated to the functionalities at the larger scale. Reflecting the level of heterogeneities in the properties of systems remains challenging when probing small scales, due to the mismatch between the area surveyed with the tools offering nanoscale resolution, such as atomic force microscopy (AFM), and the scale of natural variations inherent to biology. Hence, to understand the physiological and...
Show moreThe complexity of biological soft matter at the sub-micrometer level is fundamentally correlated to the functionalities at the larger scale. Reflecting the level of heterogeneities in the properties of systems remains challenging when probing small scales, due to the mismatch between the area surveyed with the tools offering nanoscale resolution, such as atomic force microscopy (AFM), and the scale of natural variations inherent to biology. Hence, to understand the physiological and mechanical alterations that occur within a single cell relative to a cell population, a multiscale approach is necessary. In this work we show that it is possible to observe molecular, chemical and physical alterations in both plant and human cells with a multiscale approach. Biophysical and biochemical traits of cell populations are studied with Fourier Transform infrared spectroscopy (FTIR) and in turn, guide higher resolution discovery with Raman spectroscopy and nanoscale infrared spectroscopy using AFM (NanoIR) to access finer details. We illustrate this with three examples of biological soft matter systems: 1) a preliminary study of cellular interactions with naturally occurring vehicles applicable to human health, 2) a qualitative examination of antibiotics and new pesticide treatments in food crop systems, and 3) a fundamental investigation of the deconstruction mechanisms of plant cells during pre-treatments in preparation for biofuel production.
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Date Issued
2018
Identifier
CFE0007395, ucf:52058
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0007395
A Multisystem Approach for the Characterization of Bacteria for Sustainable Agriculture
Title
A Multisystem Approach for the Characterization of Bacteria for Sustainable Agriculture.
Creator
Lee, Briana, Tetard, Laurene, Kang, Hyeran, Mason, Chase, University of Central Florida
Abstract / Description
The chemical, physical, and biological properties of bacteria developing resistance have been explored in animal based bacteria while plant bacteria have been largely neglected. Thus, the ability to probe changes in stiffness, adhesion, binding interactions and molecular traits of bacteria causing plant diseases is of great interest to develop a new generation of more potent, yet sustainable, pesticides. Our study aims to investigate the physical and chemical properties of bacterial systems,...
Show moreThe chemical, physical, and biological properties of bacteria developing resistance have been explored in animal based bacteria while plant bacteria have been largely neglected. Thus, the ability to probe changes in stiffness, adhesion, binding interactions and molecular traits of bacteria causing plant diseases is of great interest to develop a new generation of more potent, yet sustainable, pesticides. Our study aims to investigate the physical and chemical properties of bacterial systems, in particular their cell walls. Building upon this fundamental understanding of the cells, we also investigate the physicochemical responses associated to multivalent nanoparticle-based bactericide treatments on bacterial systems identified as pathogens in plant diseases. Here our efforts focus on developing a protocol for the fundamental understanding of Xanthomonas perforans, a strain known for causing bacterial spot in tomatoes and causing close to 50% losses in production. To support the design and accelerate the development of pesticides and treatments against this disease, we evaluate the changes bacteria undergo in the presence of the treatment. Using a silica nanoparticle-based treatment designed with a shell containing multivalent copper and quaternary ammonium, we compare bacteria pre- and post-treatment with infrared spectroscopy, atomic force microscopy (AFM)-based techniques, and TIRF microscopy. Statistical data analysis enables the identification of attributes that can potentially serve as markers to track the bacterial responses to the treatment in the future. Finally, we will discuss the exciting implications of this work, such as potential clues for the development of more potent treatments for resistant bacteria.
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Date Issued
2018
Identifier
CFE0007038, ucf:52005
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0007038
Mechanism of Actin Bundle Assembly, Mechanics and Structure by Ion Interaction
Title
Mechanism of Actin Bundle Assembly, Mechanics and Structure by Ion Interaction.
Creator
Castaneda, Nicholas, Kang, Hyeran, Huo, Qun, Gesquiere, Andre, University of Central Florida
Abstract / Description
The assembly of actin filaments into bundles plays an essential role in mechanical strength and dynamic reorganization of cytoskeleton. Divalent counterions at high concentrations promote bundle formation through electrostatic attraction between charged filaments. Although it has been hypothesized that specific cation interactions may contribute to salt-induced bundling, molecular mechanisms of how salt modulates bundle assembly and mechanics are not well established. Here we determine the...
Show moreThe assembly of actin filaments into bundles plays an essential role in mechanical strength and dynamic reorganization of cytoskeleton. Divalent counterions at high concentrations promote bundle formation through electrostatic attraction between charged filaments. Although it has been hypothesized that specific cation interactions may contribute to salt-induced bundling, molecular mechanisms of how salt modulates bundle assembly and mechanics are not well established. Here we determine the mechanical and dynamic properties of actin bundles with physiologically relevant cations. Using total internal reflection fluorescence (TIRF) microscopy, we measure the bending stiffness of actin bundles determined by persistence length analysis. We characterize real-time formation of bundles by dynamic light scattering intensity and direct visualization using TIRF microscopy. Our results show that divalent cations modulate bundle stiffness as well as time-dependent average bundle size. Furthermore, molecular dynamic simulations propose specificity for cation binding on actin filaments to form bundles. The work suggests that cation interactions serve a regulatory function in bundle assembly dynamics, mechanics, and structure.
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Date Issued
2017
Identifier
CFE0006572, ucf:51307
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0006572
Tunable Effect of Metal Ions on Polyelectrolyte Mechanics
Title
Tunable Effect of Metal Ions on Polyelectrolyte Mechanics.
Creator
Diaz, Angie, Kang, Hyeran, Zhai, Lei, Tetard, Laurene, University of Central Florida
Abstract / Description
Polyelectrolyte based hydrogel fibers can mimic extracellular matrix and have applications such as drug delivery and tissue scaffolding. Metal ions play a critical role in hydrogel fiber stability via electrostatic interactions, but knowledge of how they modulate mechanical properties of individual polyelectrolyte polymers is lacking. In this study, electrospun polyacrylic acid with chitosan is used as a model system to evaluate ferric ion effect on nanofiber mechanics. Using dark field...
Show morePolyelectrolyte based hydrogel fibers can mimic extracellular matrix and have applications such as drug delivery and tissue scaffolding. Metal ions play a critical role in hydrogel fiber stability via electrostatic interactions, but knowledge of how they modulate mechanical properties of individual polyelectrolyte polymers is lacking. In this study, electrospun polyacrylic acid with chitosan is used as a model system to evaluate ferric ion effect on nanofiber mechanics. Using dark field microscopy imaging and persistence length analysis, we demonstrate that ferric ions modulate the bending stiffness of nanofibers. Young's modulus of individual nanofibers is estimated at values of a few kilopascals, suggesting that electrospun nanofibers possibly exist in a hydrated state. Furthermore, Fourier Transform Infrared (FTIR) spectra indicate the effect of ferric ions on polyacrylic acid molecular bonds. Our results suggest that metal ions can regulate single nanofiber stiffness, thereby providing designs to fabricate hydrogels in a tunable fashion.
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Date Issued
2018
Identifier
CFE0006993, ucf:51625
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0006993
Light Scattering Property of Gold Nanoparticles with Applications to Biomolecule Detection and Analysis
Title
Light Scattering Property of Gold Nanoparticles with Applications to Biomolecule Detection and Analysis.
Creator
Zheng, Tianyu, Huo, Qun, Zou, Shengli, Gesquiere, Andre, Kang, Hyeran, Zhai, Lei, University of Central Florida
Abstract / Description
Gold nanoparticles (AuNPs) have unique optical and chemical properties. Dynamic light scattering (DLS) is an analytical tool used routinely for nanoparticle size measurement. The combined use of AuNPs and DLS has led to a novel analytical assay technology called D2Dx (from diameter to diagnostics). Herein, my dissertation highlights the extended use of D2Dx for biomolecule detection and analysis. Under this general theme, Chapter 1 provides some background information of AuNPs, DLS, the...
Show moreGold nanoparticles (AuNPs) have unique optical and chemical properties. Dynamic light scattering (DLS) is an analytical tool used routinely for nanoparticle size measurement. The combined use of AuNPs and DLS has led to a novel analytical assay technology called D2Dx (from diameter to diagnostics). Herein, my dissertation highlights the extended use of D2Dx for biomolecule detection and analysis. Under this general theme, Chapter 1 provides some background information of AuNPs, DLS, the principle of D2Dx technique and its potential applications. Chapter 2 summarizes a study on the effect of AuNP concentrations and laser power on the hydrodynamic size measurement of AuNPs by DLS. This study demonstrated the multiple scattering effect on DLS analysis, and how to use the exceptionally high sensitivity of DLS in AuNP aggregate detection for bioassay design and development. Chapter 3 explores a cooperative interaction between AuNP and certain proteins in blood serum that are key to the immune system, leading to a novel diagnostic tool that can conveniently monitor the humoral immunity development from neonates to adults and detect active infections in animals. Chapter 4 reports an application of D2Dx technique for acute viral infection detection based on the active immune responses elicited from mouse models infected with influenza virus. Chapter 5 describes another application of D2Dx for prostate cancer detection. The D2Dx assay identifies prostate cancer patients from non-cancer controls with improved specificity and sensitivity than PSA test. Chapter 6 demonstrates the use of AuNPs and DLS for hydrodynamic size measurement of protein disulfide isomerase with two different conformations. Chapter 7 investigates the concentration-dependent self-assembling behavior of ribostamycin through its interaction with AuNPs in aqueous solution. Overall, this dissertation established several lines of applications of using AuNPs and DLS for biomolecular research and in vitro diagnostics.
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Date Issued
2018
Identifier
CFE0007385, ucf:52056
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0007385
Advanced Nanoscale Characterization of Plants and Plant-derived Materials for Sustainable Agriculture and Renewable Energy
Title
Advanced Nanoscale Characterization of Plants and Plant-derived Materials for Sustainable Agriculture and Renewable Energy.
Creator
Soliman, Mikhael, Tetard, Laurene, Vaidyanathan, Raj, Kang, Hyeran, Santra, Swadeshmukul, Zhai, Lei, Chumbimuni Torres, Karin, University of Central Florida
Abstract / Description
The need for nanoscale, non-invasive functional characterization has become more significant with advances in nano-biotechnology and related fields. Exploring the ultrastructure of plant cell walls and plant-derived materials is necessary to access a more profound understanding of the molecular interactions in the systems, in view of a rational design for sustainable applications. This, in turn, relates to the pressing requirements for food, energy and water sustainability experienced...
Show moreThe need for nanoscale, non-invasive functional characterization has become more significant with advances in nano-biotechnology and related fields. Exploring the ultrastructure of plant cell walls and plant-derived materials is necessary to access a more profound understanding of the molecular interactions in the systems, in view of a rational design for sustainable applications. This, in turn, relates to the pressing requirements for food, energy and water sustainability experienced worldwide.Here we will present our advanced characterization approach to study the effects of external stresses on plants, and resulting opportunities for biomass valorization with an impact on the food-energy-water nexus.First, the adaption of plants to the pressure imposed by gravity in poplar reaction wood will be discussed. We will show that a multiscale characterization approach is necessary to reach a better understanding of the chemical and physical properties of cell walls across a transverse section of poplar stem. Our Raman spectroscopy and statistical analysis reveals intricate variations in the cellulose and lignin properties. Further, we will present evidence that advanced atomic force microscopy can reveal nanoscale variations within the individual cell wall layers, not attainable with common analytical tools. Next, chemical stresses, in particular the effect of Zinc-based pesticides on citrus plants, will be considered. We will show how multiscale characterization can support the development of new disease management methods for systemic bacterial diseases, such as citrus greening, of great importance for sustainable agriculture. In particular, we will focus on the study of new formulations, their uptake and translocation in the plants following different application methods. Lastly, we will consider how plant reactions to mechanical and chemical stresses can be controlled to engineer biomass for valorization applications. We will present our characterization of two examples: the production of carbon films derived from woody lignocellulosic biomass and the development of nanoscale growth promoters for food crop. A perspective of the work and discussion of the broader impact will conclude the presentation.
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Date Issued
2018
Identifier
CFE0007415, ucf:52717
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0007415