Upregulation of nicotinic acetylcholine receptors (nAChRs) is a well-documented response to chronic nicotine exposure. Nicotinic acetylcholine receptors are pentameric ligand-gated ion channels consisting of alpha (α2-10) and beta (β2-4) subunits. Nicotine, an agonist of nAChRs, alters trafficking and assembly of some subtypes of nAChRs, leading to an increase in expression of high sensitivity receptors on the plasma membrane. These physiological changes in nAChRs are believed to contribute to nicotine addiction, although the mechanism of these processes has not been resolved. Recently, many studies have converged on the idea that nicotine induces upregulation by an intracellular mechanism. In this dissertation, expression levels of nAChRs were quantified upon exposure to nicotine and its primary metabolite, cotinine. A pH sensitive variant of GFP, super ecliptic pHluorin (SEP), was integrated with a nAChR subunit to study expression and trafficking of nAChRs by differentiating intracellular and plasma membrane inserted receptors. In this work, cotinine is shown to increase the number of α4β2 nAChRs within a cell. Cotinine also affects trafficking of α4β2, evident by a redistribution of intracellular receptors and an increase in single vesicle insertion events on the plasma membrane. This work shows both nicotine and cotinine alter the overall assembly of α4β2 to favor the high sensitivity (α4)2(β2)3 version. Since cotinine and nicotine induce similar physiological changes in nAChRs, the metabolite potentially plays a role in the mechanism of nicotine addiction. Although an intracellular mechanism for upregulation has been supported, a shift in assembly to the high sensitivity (α4)2(β2)3 version exclusively in the endoplasmic reticulum has not previously been detected. In order to study organelle specific changes in stoichiometry, a novel method was developed to isolate single nAChRs in nanovesicles derived from native cell membranes. Separation of nanovesicles originating from the endoplasmic reticulum and plasma membrane, encompassing isolated nAChRs, allows precise changes in stoichiometry to be monitored in subcellular regions. In this work, single molecule bleaching steps of green fluorescent protein (GFP) encoded in each alpha subunit of the pentamer are detected. The number of bleaching steps, or transitions to a nonfluorescent state upon continuous excitation, corresponds to the number of GFP-labeled alpha subunits present. Therefore, the stoichiometry can be deduced by detection of two bleaching steps, as in (α4)2(β2)3, or three bleaching steps, seen in (α4)3(β2)2. Using this method on isolated nAChRs, a shift to assembly of high sensitivity (α4)2(β2)3 receptors is detected definitively within the endoplasmic reticulum. In addition, an increase in (α4)2(β2)3 receptors located on the plasma membrane is shown when nicotine is present. This work provides convincing evidence that nicotine acts intracellularly, within the endoplasmic reticulum, to alter stoichiometry of nAChRs.

Nanopore sensing is a label-free method used to characterize water-soluble molecules. Recent work describes how Au25(SG)18 clusters improve the single molecule nanopore spectrometry (SMNS) technique when analyzing polyethylene glycol (PEG). This thesis will further study and optimize the enhancement effect resulting from a cluster’s presence. Additionally, a model describing the interaction between a cluster and PEG is developed to assist in understanding this mechanism of enhancement. This thesis will also discuss expanding the SMNS method to detect peptides, using Au25(SG)18 for enhancement, and adjusting solution conditions to improve the sensitivity of the SMNS system for peptide detection. Finally, a model describing the relationship between nanopore current blockades and molecular weight is developed to demonstrate the feasibility of using SMNS as a viable analytical technique for characterizing a wide variety of water-soluble molecules.

The effect of a magnetic field on copper electrodeposition was investigated. Copper was electrodeposited onto square copper cathodes 1 sq cm in area from an aqueous solution (0.5 M CuSO4, 0.5 M H2SO. A glass cell was placed between the pole pieces of an electromagnet, and the magnetic fields applied were in the range from 0 to 12.5 kG. The current density was in the range from 80 mA/sq cm to 880 mA/sq cm. In each of the experiments, cell current, cell voltage, and cell temperature were monitored with a microcomputer. The weight change, deposit surface and cross section morphology, and the hardness were also found. Anodes used in the experiments were studied to see the effect of various conditions on the surface finish. Copper was also electrodeposited onto copper grids in order to study how the uniformity of the deposit is affected by an applied magnetic field.

In this dissertation we discuss the electronic properties, structural configurations, and reaction mechanisms of chemically functionalized graphene and charged atomic metals. In general, we analyze fundamental atomic scale and nanoscale systems with density functional theory in order to investigate chemical reaction energetics for peroxide synthesis as well as methanol production without carbon emission. These systems were found to be tunable via the addition of cationic and anionic charges, change in transition metal type, and modification through chemical functionalization. Furthermore, transition state theory was used to predict an optimal configuration for chemically functionalized graphene, efficient use of anionic atomic gold and palladium for synthesis of water to peroxide, and clean conversion of methane to methanol without carbon dioxide emission utilizing anionic gold.

The interactive wireless sensor detects many hazardous gases such as Hexane, Propane, Carbon monoxide and Hydrogen. These gases are highly toxic and used in different kinds of manufacturing industries, domestic purpose and so on. So, building a sensor that can detect this kind of gases can save the environment; prevent the potential for explosion, and endangering human life. In long term, interactive wireless sensor can also prevent the financial losses that might occur due to the hazardous incident that might occur due to these toxic gases. Hexane is a colorless, strong gas which inhaled in significant amounts by a person then he may suffer with hexane poisoning and suffocation. It also causes skin burns when exposed in high concentrations. Propane, carbon monoxide and hydrogen can easily freeze in room temperature, if in contact with eye, it could permanently damage eye or cause blindness. The advantage of this wireless sensor is the use of artificial olfactory system (electronic nose) that can be taught to detect these hazardous gases. This sensor has a unique molecular combination of analysts, impurities and background that corresponds to a gas leak. It consists of a chemiresistor, such as an array of conductometric sensors, and a mechanism analyzing the data in real time. A smell-print is composed of many molecules which reaches receptor in the human nose. When a specific receptor receives a molecule, it sends a signal to the brain where the smell is identified and associated with that particular molecule. Similar manner, albeit substituting sensors for the receptors, and transmitting the signal to a machine learning algorithm for processing, rather than to the brain. This wireless gas leak sensing consists of microchip Pic 32, integrated electronic nose, automated data analysis unit, power supply, and communications. The communication channel will use the ZigBee link, or the cellular links, or other specific frequency wireless link. The time-stamped and position-stamped sensor measurement data are transmitted to the central computer in predetermined periods of time. The data will be stored in the computer database for possible future analysis of the gas leak development process.

Methylation is involved in the regulation of varied biological processes, from cancer to embryonic development. In B-DNA, methylation can alter the frequency with which a dinucleotide step samples one of two substates: BI and BII. Changing the BI-BII equilibrium can in turn affect the ability of proteins to bind to DNA, which may ultimately alter the transcription of genes. Using MD simulations, we evaluate the effects of cytosine methylation on the BI-BII equilibrium and the RMS atomic fluctuations of the backbone of two sequences: (GC)5 and the Dickerson dodecamer (DD). Methylation in (GC)5 stabilizes the BII state in CpG steps and stabilizes the BI state in GpC steps, but the BII state is always less favored than the BI state. The activation energy between the BI/BII states in (GC)5 increases in GpC steps and decreases in CpG steps upon methylation, indicating that backbone dynamics are affected by methylation in a step-dependent manner. The DD simulations suggest that methylation stabilizes the BII state in both CpG and GpC steps, although more sampling is needed to determine the significance of these results. Methylation has little effect on the atomic fluctuations of the backbone in (GC)5 or DD simulations. Thus, in these sequences, methylation does not uniformly stabilize one state, nor does methylation stabilize the state that is favored in the native sequence.

Using iron super-paramagnetic and ferromagnetic nanoparticles composed of Fe3O4 molecules, scientists analyze the effectiveness and practicality of this new treatment theory, hyperthermia. The problems of magnetic particle density, isothermal barriers/cellular cooling thresholds, and nanoparticle specific targeting are addressed in this review. Iron magnetic nanoparticles were chosen due to their relatively low biological reactivates and lack of subsequent cellular toxicity. However, there are significant heating problems associated with these magnetic nanoparticles due to their relative size and short thermal time constants or thermal half-lives. Effectively, these aforementioned issues create a phenomenon where cancerous cells, surrounded by unheated healthy tissue, exhibit properties similar to those of an isothermal barrier. As a result, target cells experience limited gross heating, which is localized to the area directly surrounding the active magnetic nanoparticle within the cytoplasm. The effects of isothermal barriers and HSP up regulation on particle-based hypothermia are profound and prevent therapeutic temperatures from being achieved in single cell heating limiting the applications for Fe3O4 magnetic nanoparticle hyperthermia applications. It has been shown that reaching a certain magnetic nanoparticle density within the cell can result in a larger heating capacity, though this effect is also dependent on the particle dispersion pattern within cytoplasm. It has yet to be concluded whether ferromagnetic particles or super-paramagnetic particles are superior or more practical for hyperthermic treatments as they each have distinct benefits, and further study is needed. Finally, the popular targeting mechanism associated with magnetic nanoparticle research, monoclonal antibodies, require that they have an organic coating (such as starch) as a means of both providing an organic binding point and as camouflage for avoiding host filtration pathways. Forgoing this organic coating could lead to increased particle density within the cell and the adoption of a more specific targeting mechanism such as virus like particles (VLPs) altered to target HSP’s could lead to an increase in yield. Furthermore the up regulation of HSPs in response to therapeutic temperature is problematic for the therapies practically.

Although the association between social inequality and poor adult health is well established, the mechanisms by which inequality is translated into poor adult health are less clear. Increasingly, evidence suggests that many adult health problems and health disparities have their origins in early life; the developmental origins of health and disease (DOHaD) hypothesis provides an explanatory mechanism linking adverse early life conditions with permanent structural or functional changes that increase the risk for disease. This hypothesis is consistent with bioarchaeological research noting reduced lifespan among individuals exhibiting signs of childhood stress. The principal aim of this dissertation is to contribute a bioarchaeological perspective to health disparities research by investigating how health disparities can be measured and understood in the past. This study focuses on early life conditions as a source of adult health disparity by examining a skeletal sample for the association between childhood stress and adult longevity; the relationship between childhood stress and the presence of adult health conditions; and sex, ancestry, and regional differences in these relationships. The study sampled 830 age-documented, U.S. born African American males and females and Euro-American males from the Terry and the Hamann-Todd anatomical collections, representing socially-marginalized individuals from the late 19th- to early 20th centuries. Enamel hypoplasia, femoral length, and vertebral neural canal diameters represented childhood stress; skeletal fractures, tibial periostosis, and the diseased, missing, and filled tooth index represented adult health. Longevity was modeled with Kaplan-Meier survival curves and adult health relationships were modeled with logistic regression. Additionally, cause of death data from historic health department publications and the study sample morgue records were examined for disparity in the epidemiological transition from infectious to degenerative cause of death. The study found mixed results for all analyses. There was no reduction in longevity for the presence of enamel hypoplasia, short femoral length, or reduced thoracic neural canal diameter. African American males had statistically significant reduced longevity for small lumbar vertebral neural canal diameters. African American males from the Hamann-Todd Collection and Euro-American males from both collections had significant relationships between vertebral neural canal diameters and adult conditions; these relationships varied among the groups but in most cases demonstrated reduced odds for having the adult condition for individuals with smaller canal diameters. African American females had no differential survival or relationships between variables over the lifecourse. All groups except for the Terry Collection Euro-American males continued to have more infectious disease deaths than degenerative disease deaths. The study results contribute to disparities research by demonstrating that the consequences of childhood stress varied by sex and ancestry and by demonstrating within-population variation in timing of the epidemiological transition. Additionally, the study results support the contention of greater male sensitivity to environmental conditions and contributes evidence supporting the DOHaD hypothesis.

This study explores the relationship between tourism, the nutrition transition, and prenatal dietary practices in the Monteverde Zone, Costa Rica. This rural tourism community, located in the central highlands of Costa Rica, has seen rapid growth and development since the tourism boom in the early 1990s, leading to changes in the local food system and increased food insecurity. This investigation added to this work by identifying the ways that prenatal dietary practices have shifted over time in the context of increased tourism and the concomitant nutrition transition, and by describing the relationship between food insecurity and nutritional status among pregnant women. In applying a critical biocultural approach, this study drew on both quantitative and qualitative methods. Pregnant women were recruited to participant in twenty-four hour diet recalls (n=21), the Household Food Insecurity and Access Scales (n=20), and semi-structured interviews (n=22). Women who had older children were also recruited for semi-structured interviews (n=20) to explore prenatal dietary practices and decision-making over time. Focus groups (N=2, n=15) and surveys with a free listing component (n=52) were administered to better understand the cultural construction of nutrition in this region, and how tourism and the nutrition transition have interacted with the local dietary norms. Overall this study found that there was a relationship between tourism, the nutrition transition, and diet, although findings suggest that pregnant women may be buffered from these effects by cultural factors. Food insecurity was present in the sample (n=7) and was associated with numerous variables, including saturated fat and zinc intake.

Amyloid fibril aggregation is associated with several horrific diseases such as Alzheimer’s, Creutzfeld-Jacob, diabetes, Parkinson’s and others. The process of amyloid aggregation involves forming myriad different metastable intermediate aggregates. Amyloid fibrils are composed of proteins that originate in an innocuous α-helix or random-coil structure. The α-helices convert their structure to β-strands that aggregate into β-sheets, and then into protofibrils, and ultimately into fully formed amyloid fibrils. On the basis of experimental data, I have developed a mathematical model for the kinetics of the reaction pathways and determined rate parameters for peptide secondary structural conversion and aggregation during the entire fibrillogenesis process from random coil to fibrils, including the molecular species that accelerate the conversions. The specific steps of the model and the rate constants that are determined by fitting to experimental data provide insight on the molecular species involved in the fibril formation process. To better understand the molecular basis of the protein structural transitions and aggregation, I report on molecular dynamics (MD) computational studies on the formation of amyloid protofibrillar structures in the small model protein ccβ, which undergoes many of the structural transitions of the larger, naturally occurring amyloid forming proteins. Two different structural transition processes involving hydrogen bonds are observed for aggregation into fibrils: the breaking of intrachain hydrogen bonds to allow β-hairpin proteins to straighten, and the subsequent formation of interchain hydrogen bonds during aggregation into amyloid fibrils. For my MD simulations, I found that the temperature dependence of these two different structural transition processes results in the existence of a temperature window that the ccβ protein experiences during the process of forming protofibrillar structures. Both the mathematical modeling of the kinetics and the MD simulations show that molecular structural heterogeneity is a major factor in the process. The MD simulations also show that intrachain and interchain hydrogen bonds breaking and forming is strongly correlated to the process of amyloid formation.