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Biological and immunological characterization of plant-produced HIV-1 Gag/dgp41 virus-like particles
Description
Anti-retroviral drugs and AIDS prevention programs have helped to decrease the rate of new HIV-1 infections in some communities, however, a prophylactic vaccine is still needed to control the epidemic world-wide. Despite over two decades of research, a vaccine against HIV-1 remains elusive, although recent clinical trials have shown promising results. Recent successes have focused on highly conserved, mucosally-targeted antigens within HIV-1 such as the membrane proximal external region (MPER) of the envelope protein, gp41. MPER has been shown to play critical roles in the viral mucosal transmission, though this peptide is not immunogenic on its own. Gag is a structural protein configuring the enveloped virus particles, and has been suggested to constitute a target of the cellular immunity potentially controlling the viral load. It was hypothesized that HIV-1 enveloped virus-like particles (VLPs) consisting of Gag and a deconstructed form of gp41 comprising the MPER, transmembrane, and cytoplasmic domains (dgp41) could be expressed in plants. Plant-optimized HIV-1 genes were constructed and expressed in Nicotiana benthamiana by stable transformation, or transiently using a tobacco mosaic virus-based expression system or a combination of both. Results of biophysical, biochemical and electron microscopy characterization demonstrated that plant cells could support not only the formation of HIV-1 Gag VLPs, but also the accumulation of VLPs that incorporated dgp41. These particles were purified and utilized in mice immunization experiments. Prime-boost strategies combining systemic and mucosal priming with systemic boosting using two different vaccine candidates (VLPs and CTB-MPR - a fusion of MPER and the B-subunit of cholera toxin) were administered to BALB/c mice. Serum antibody responses against both the Gag and gp41 antigens could be elicited in mice systemically primed with VLPs and these responses could be recalled following systemic boosting with VLPs. In addition, mucosal priming with VLPs allowed for a robust boosting response against Gag and gp41 when boosted with either candidate. Functional assays of these antibodies are in progress to test the antibodies' effectiveness in neutralizing and preventing mucosal transmission of HIV-1. This immunogenicity of plant-based Gag/dgp41 VLPs represents an important milestone on the road towards a broadly-efficacious and inexpensive subunit vaccine against HIV-1.
ContributorsKessans, Sarah (Author) / Mor, Tsafrir S (Thesis advisor) / Matoba, Nobuyuki (Committee member) / Mason, Hugh (Committee member) / Hogue, Brenda (Committee member) / Fromme, Petra (Committee member) / Arizona State University (Publisher)
Created2011
Description
This study presents the results of one of the first attempts to characterize the pore water pressure response of soils subjected to traffic loading under saturated and unsaturated conditions. It is widely known that pore water pressure develops within the soil pores as a response to external stimulus. Also, it has been recognized that the development of pores water pressure contributes to the degradation of the resilient modulus of unbound materials. In the last decades several efforts have been directed to model the effect of air and water pore pressures upon resilient modulus. However, none of them consider dynamic variations in pressures but rather are based on equilibrium values corresponding to initial conditions. The measurement of this response is challenging especially in soils under unsaturated conditions. Models are needed not only to overcome testing limitations but also to understand the dynamic behavior of internal pore pressures that under critical conditions may even lead to failure. A testing program was conducted to characterize the pore water pressure response of a low plasticity fine clayey sand subjected to dynamic loading. The bulk stress, initial matric suction and dwelling time parameters were controlled and their effects were analyzed. The results were used to attempt models capable of predicting the accumulated excess pore pressure at any given time during the traffic loading and unloading phases. Important findings regarding the influence of the controlled variables challenge common beliefs. The accumulated excess pore water pressure was found to be higher for unsaturated soil specimens than for saturated soil specimens. The maximum pore water pressure always increased when the high bulk stress level was applied. Higher dwelling time was found to decelerate the accumulation of pore water pressure. In addition, it was found that the higher the dwelling time, the lower the maximum pore water pressure. It was concluded that upon further research, the proposed models may become a powerful tool not only to overcome testing limitations but also to enhance current design practices and to prevent soil failure due to excessive development of pore water pressure.
ContributorsCary, Carlos (Author) / Zapata, Claudia E (Thesis advisor) / Wiczak, Matthew W (Thesis advisor) / Kaloush, Kamil (Committee member) / Sandra, Houston (Committee member) / Arizona State University (Publisher)
Created2011
Description
A recent joint study by Arizona State University and the Arizona Department of Transportation (ADOT) was conducted to evaluate certain Warm Mix Asphalt (WMA) properties in the laboratory. WMA material was taken from an actual ADOT project that involved two WMA sections. The first section used a foamed-based WMA admixture, and the second section used a chemical-based WMA admixture. The rest of the project included control hot mix asphalt (HMA) mixture. The evaluation included testing of field-core specimens and laboratory compacted specimens. The laboratory specimens were compacted at two different temperatures; 270 °F (132 °C) and 310 °F (154 °C). The experimental plan included four laboratory tests: the dynamic modulus (E*), indirect tensile strength (IDT), moisture damage evaluation using AASHTO T-283 test, and the Hamburg Wheel-track Test. The dynamic modulus E* results of the field cores at 70 °F showed similar E* values for control HMA and foaming-based WMA mixtures; the E* values of the chemical-based WMA mixture were relatively higher. IDT test results of the field cores had comparable finding as the E* results. For the laboratory compacted specimens, both E* and IDT results indicated that decreasing the compaction temperatures from 310 °F to 270 °F did not have any negative effect on the material strength for both WMA mixtures; while the control HMA strength was affected to some extent. It was noticed that E* and IDT results of the chemical-based WMA field cores were high; however, the laboratory compacted specimens results didn't show the same tendency. The moisture sensitivity findings from TSR test disagreed with those of Hamburg test; while TSR results indicated relatively low values of about 60% for all three mixtures, Hamburg test results were quite excellent. In general, the results of this study indicated that both WMA mixes can be best evaluated through field compacted mixes/cores; the results of the laboratory compacted specimens were helpful to a certain extent. The dynamic moduli for the field-core specimens were higher than for those compacted in the laboratory. The moisture damage findings indicated that more investigations are needed to evaluate moisture damage susceptibility in field.
ContributorsAlossta, Abdulaziz (Author) / Kaloush, Kamil (Thesis advisor) / Witczak, Matthew W. (Committee member) / Mamlouk, Michael S. (Committee member) / Arizona State University (Publisher)
Created2011
Description
The cyanobacterium Synechocystis sp. PCC 6803 performs oxygenic photosynthesis. Light energy conversion in photosynthesis takes place in photosystem I (PSI) and photosystem II (PSII) that contain chlorophyll, which absorbs light energy that is utilized as a driving force for photosynthesis. However, excess light energy may lead to formation of reactive oxygen species that cause damage to photosynthetic complexes, which subsequently need repair or replacement. To gain insight in the degradation/biogenesis dynamics of the photosystems, the lifetimes of photosynthetic proteins and chlorophyll were determined by a combined stable-isotope (15N) and mass spectrometry method. The lifetimes of PSII and PSI proteins ranged from 1-33 and 30-75 hours, respectively. Interestingly, chlorophyll had longer lifetimes than the chlorophyll-binding proteins in these photosystems. Therefore, photosynthetic proteins turn over and are replaced independently from each other, and chlorophyll is recycled from the damaged chlorophyll-binding proteins. In Synechocystis, there are five small Cab-like proteins (SCPs: ScpA-E) that share chlorophyll a/b-binding motifs with LHC proteins in plants. SCPs appear to transiently bind chlorophyll and to regulate chlorophyll biosynthesis. In this study, the association of ScpB, ScpC, and ScpD with damaged and repaired PSII was demonstrated. Moreover, in a mutant lacking SCPs, most PSII protein lifetimes were unaffected but the lifetime of chlorophyll was decreased, and one of the nascent PSII complexes was missing. SCPs appear to bind PSII chlorophyll while PSII is repaired, and SCPs stabilize nascent PSII complexes. Furthermore, aminolevulinic acid biosynthesis, an early step of chlorophyll biosynthesis, was impaired in the absence of SCPs, so that the amount of chlorophyll in the cells was reduced. Finally, a deletion mutation was introduced into the sll1906 gene, encoding a member of the putative bacteriochlorophyll delivery (BCD) protein family. The Sll1906 sequence contains possible chlorophyll-binding sites, and its homolog in purple bacteria functions in proper assembly of light-harvesting complexes. However, the sll1906 deletion did not affect chlorophyll degradation/biosynthesis and photosystem assembly. Other (parallel) pathways may exist that may fully compensate for the lack of Sll1906. This study has highlighted the dynamics of photosynthetic complexes in their biogenesis and turnover and the coordination between synthesis of chlorophyll and photosynthetic proteins.
ContributorsYao, Cheng I Daniel (Author) / Vermaas, Wim (Thesis advisor) / Fromme, Petra (Committee member) / Roberson, Robert (Committee member) / Webber, Andrew (Committee member) / Arizona State University (Publisher)
Created2011
Description
Non-native saltcedar (Tamarix spp.) has invaded many riparian communities and is the third most abundant tree in Southwestern riparian areas. I evaluated lizard populations and microhabitat selection during 2009 and 2010 along the Virgin River in Nevada and Arizona to determine the impact of saltcedar. Along the riparian corridor, I observed common side-blotched lizards (Uta stansburiana) within two vegetation types: monotypic non-native saltcedar stands or mixed stands of cottonwood (Populus fremontii), willow (Salix spp.), mesquite (Prosopis spp.) and saltcedar. I predicted that population parameters such as body condition, adult to hatchling ratio, abundance, and persistence would vary among vegetation types. Also, I predicted the presence of saltcedar influences how lizards utilize available habitat. Lizard population parameters were obtained from a mark-recapture study in which I captured 233 individual lizards. I examined habitat selection and habitat availability using visual encounter surveys (VES) for lizards and recorded 11 microhabitat variables where 16 lizards were found. I found no significant difference in population parameters between mixed and non-native saltcedar communities. However, population parameters were negatively correlated with canopy cover. I found that lizards selected habitat with low understory and canopy cover regardless of vegetation type. My results indicate that lizards utilize similar structural characteristics in both mixed and non-native vegetation. Understanding impacts of saltcedar on native fauna is important for managers who are tasked with control and management of this non-native species.
ContributorsNielsen, Danny (Author) / Bateman, Heather L. (Thesis advisor) / Miller, William H. (Committee member) / Sullivan, Brian K. (Committee member) / Arizona State University (Publisher)
Created2011
Description
There is increasing evidence that ovarian status influcences behavioral phenotype in workers of the honey bee Apis mellifera. Honey bee workers demonstrate a complex division of labor. Young workers perform in-hive tasks (e.g. brood care), while older bees perform outside tasks (e.g. foraging for food). This age correlated division of labor is known as temporal polyethism. Foragers demonstrate further division of labor with some bees biasing collection towards protein (pollen) and others towards carbohydrates (nectar). The Reproductive Ground-plan Hypothesis proposes that the ovary plays a regulatory role in foraging division of labor. European honey bee workers that have been selectively bred to store larger amounts of pollen (High strain) also have a higher number of ovarioles per ovary than workers from strains bred to store less pollen (Low strain). High strain bees also initiate foraging earlier than Low strain bees. The relationship between ovariole number and foraging behavior is also observed in wild-type Apis mellifera and Apis cerana: pollen-biased foragers have more ovarioles than nectar-biased foragers. In my first study, I investigated the pre-foraging behavioral patterns of the High and Low strain bees. I found that High strain bees progress through the temporal polyethism at a faster rate than Low strain bees. To ensure that the observed relationship between the ovary and foraging bias is not due to associated separate genes for ovary size and foraging behavior, I investigated foraging behavior of African-European backcross bees. The backcross breeding program was designed to break potential gene associations. The results from this study demonstrated the relationship between the ovary and foraging behavior, supporting the proposed causal linkage between reproductive development and behavioral phenotype. The final study was designed to elucidate a regulatory mechanism that links ovariole number with sucrose sensitivity, and loading decisions. I measured ovariole number, sucrose sensitivity and sucrose solution load size using a rate-controlled sucrose delivery system. I found an interaction effect between ovariole number and sucrose sensitivity for sucrose solution load size. This suggests that the ovary impacts carbohydrate collection through modulation of sucrose sensitivity. Because nectar and pollen collection are not independent, this would also impact protein collection.
ContributorsSiegel, Adam J (Author) / Page, Jr., Robert E (Thesis advisor) / Hamilton, Andrew L. (Committee member) / Brent, Colin S (Committee member) / Amdam, Gro V (Committee member) / McGraw, Kevin J. (Committee member) / Arizona State University (Publisher)
Created2011
Description
The discipline of rhetoric and composition established the Writing Program Administrators Outcomes Statement (WPA OS) to fulfill a general expectation about the skills and knowledge students should be able to demonstrate by the end of first-year composition. Regardless of pedagogy used, academic preparation of the teacher, or preference of particular topics or types of assignments, the WPA OS is versatile. This dissertation employs a problem-solution argument showcasing methods to improve assignments through intentional use of the WPA OS for a fluid conversation throughout first-year composition and a more clear articulation of course goals. This dissertation includes summation, analysis, and synthesis of documents that inform first-year composition curriculum from foundational organizations within the field, including National Council of Teachers of English, Council of Writing Program Administrators, National Writing Project, and Conference on College Composition and Communication. This study uses the WPA OS as a lens to examine and revise writing assignments that aid in students' comprehension of the WPA OS with particular focus on the areas of rhetorical knowledge and critical thinking, reading, and writing. Framing assignment design with theoretically grounded content and the use of common topics throughout first-year composition is one way to operationalize the WPA OS. Using common topics throughout course content presents opportunities for teachers to include detailed scaffolding in assignments that expand students' literate practices and engage students as critical thinkers and writers. This study explores how using the topic of family, a common topic to all students, provides a rich bank of social, historical, and cultural elements for research and writing. The topic of family seamlessly employs multimodal composition, which presents students with opportunities for developing rhetorical knowledge and expanding students' literacies. This dissertation displays evidence of praxis of the WPA OS from assignment development to presentation of student samples. This study recommends the use of common topics and intentional application of the WPA OS to construct assignments that clearly articulate learning goals in first-year composition.
ContributorsRankins-Robertson, Sherry, 1976- (Author) / Roen, Duane H. (Thesis advisor) / Miller, Keith (Committee member) / Lauer, Claire (Committee member) / Arizona State University (Publisher)
Created2011
Description
The infrastructure is built in Unsaturated Soils. However, the geotechnical practitioners insist in designing the structures based on Saturated Soil Mechanics. The design of structures based on unsaturated soil mechanics is desirable because it reduces cost and it is by far a more sustainable approach. The research community has identified the Soil-Water Characteristic Curve as the most important soil property when dealing with unsaturated conditions. This soil property is unpopular among practitioners because the laboratory testing takes an appreciable amount of time. Several authors have attempted predicting the Soil-Water Characteristic Curve; however, most of the published predictions are based on a very limited soil database. The National Resources Conservation Service has a vast database of engineering soil properties with more than 36,000 soils, which includes water content measurements at different levels of suctions. This database was used in this study to validate two existing models that based the Soil-Water Characteristic Curve prediction on statistical analysis. It was found that although the predictions are acceptable for some ranges of suctions; they did not performed that well for others. It was found that the first model validated was accurate for fine-grained soils, while the second model was best for granular soils. For these reasons, two models to estimate the Soil-Water Characteristic Curve are proposed. The first model estimates the fitting parameters of the Fredlund and Xing (1994) function separately and then, the predicted parameters are fitted to the Fredlund and Xing function for an overall estimate of the degree of saturation. Results show an overall improvement on the predicted values when compared to existing models. The second model is based on the relationship between the Soil-Water Characteristic Curve and the Pore-Size Distribution of the soils. The process allows for the prediction of the entire Soil-Water Characteristic Curve function and proved to be a better approximation than that used in the first attempt. Both models constitute important tools in the implementation of unsaturated soil mechanics into engineering practice due to the link of the prediction with simple and well known engineering soil properties.
ContributorsTorres Hernández, Gustavo (Author) / Zapata, Claudia (Thesis advisor) / Houston, Sandra (Committee member) / Witczak, Matthew (Committee member) / Arizona State University (Publisher)
Created2011
Description
Social insect colonies exhibit striking diversity in social organization. Included in this overwhelming variation in structure are differences in colony queen number. The number of queens per colony varies both intra- and interspecifically and has major impacts on the social dynamics of a colony and the fitness of its members. To understand the evolutionary transition from single to multi-queen colonies, I examined a species which exhibits variation both in mode of colony founding and in the queen number of mature colonies. The California harvester ant Pogonomyrmex californicus exhibits both variation in the number of queens that begin a colony (metrosis) and in the number of queens in adult colonies (gyny). Throughout most of its range, colonies begin with one queen (haplometrosis) but in some populations multiple queens cooperate to initiate colonies (pleometrosis). I present results that confirm co-foundresses are unrelated. I also map the geographic occurrence of pleometrotic populations and show that the phenomenon appears to be localized in southern California and Northern Baja California. Additionally, I provide genetic evidence that pleometrosis leads to primary polygyny (polygyny developing from pleometrosis) a phenomenon which has received little attention and is poorly understood. Phylogenetic and haplotype analyses utilizing mitochondrial markers reveal that populations of both behavioral types in California are closely related and have low mitochondrial diversity. Nuclear markers however, indicate strong barriers to gene flow between focal populations. I also show that intrinsic differences in queen behavior lead to the two types of populations observed. Even though populations exhibit strong tendencies on average toward haplo- or pleometrosis, within population variation exists among queens for behaviors relevant to metrosis and gyny. These results are important in understanding the dynamics and evolutionary history of a distinct form of cooperation among unrelated social insects. They also help to understand the dynamics of intraspecific variation and the conflicting forces of local adaptation and gene flow.
ContributorsOverson, Rick P (Author) / Gadau, Jürgen (Thesis advisor) / Fewell, Jennifer H (Committee member) / Hölldobler, Bert (Committee member) / Johnson, Robert A. (Committee member) / Liebig, Jürgen (Committee member) / Arizona State University (Publisher)
Created2011
Description
The focus of this study is statistical characterization of the significant duration of strong ground motion time histories. The significant duration is defined as the time needed to build up between five and seventy five (SD575) and ninety five percent (SD595) of the energy of a strong motion record. Energy is measured as the integral of the square of the acceleration time history and can be used to capture the potential destructiveness of an earthquake. Correlations of the geometric means of the two significant duration measures (SD575 and SD595) with source, path, and near surface site parameters have been investigated using the geometric mean of 2,690 pairs of recorded horizontal strong ground motion data from 129 earthquakes in active plate margins. These time histories correspond to moment magnitudes between 4.8 and 7.9, site to source distances up to 200 km, and near surface shear wave velocity ranging from 120 to 2250 m/s. Empirical relationships have been developed based upon the simple functional forms, and observed correlations. The coefficients of the independent variables in these empirical relationships have been determined through nonlinear regression analysis using a random effects model. It is found that significant duration measures correlate well with magnitude, site to source distance, and near surface shear wave velocity. The influence of the depth to top of rupture, depth to the shear wave velocity of 1000 m/s and the style of faulting were not found to be statistically significant. Comparison of the empirical relationship developed in this study with existing empirical relationships for the significant duration shows good agreement at intermediate magnitudes (M 6.5). However, at larger and smaller magnitude, the differences between the correlations developed in this study and those from previous studies are significant.
ContributorsGhanat, Simon T (Author) / Kavazanjian, Jr., Edward (Thesis advisor) / Houston, Sandra (Committee member) / Arrowsmith, Ramon (Committee member) / Arizona State University (Publisher)
Created2011