Matching Items (89)
Description
Pathogenic Gram-negative bacteria employ a variety of molecular mechanisms to combat host defenses. Two-component regulatory systems (TCR systems) are the most ubiquitous signal transduction systems which regulate many genes required for virulence and survival of bacteria. In this study, I analyzed different TCR systems in two clinically-relevant Gram-negative bacteria, i.e., oral pathogen Porphyromonas gingivalis and enterobacterial Escherichia coli. P. gingivalis is a major causative agent of periodontal disease as well as systemic illnesses, like cardiovascular disease. A microarray study found that the putative PorY-PorX TCR system controls the secretion and maturation of virulence factors, as well as loci involved in the PorSS secretion system, which secretes proteinases, i.e., gingipains, responsible for periodontal disease. Proteomic analysis (SILAC) was used to improve the microarray data, reverse-transcription PCR to verify the proteomic data, and primer extension assay to determine the promoter regions of specific PorX regulated loci. I was able to characterize multiple genetic loci regulated by this TCR system, many of which play an essential role in hemagglutination and host-cell adhesion, and likely contribute to virulence in this bacterium. Enteric Gram-negative bacteria must withstand many host defenses such as digestive enzymes, low pH, and antimicrobial peptides (AMPs). The CpxR-CpxA TCR system of E. coli has been extensively characterized and shown to be required for protection against AMPs. Most recently, this TCR system has been shown to up-regulate the rfe-rff operon which encodes genes involved in the production of enterobacterial common antigen (ECA), and confers protection against a variety of AMPs. In this study, I utilized primer extension and DNase I footprinting to determine how CpxR regulates the ECA operon. My findings suggest that CpxR modulates transcription by directly binding to the rfe promoter. Multiple genetic and biochemical approaches were used to demonstrate that specific TCR systems contribute to regulation of virulence factors and resistance to host defenses in P. gingivalis and E. coli, respectively. Understanding these genetic circuits provides insight into strategies for pathogenesis and resistance to host defenses in Gram negative bacterial pathogens. Finally, these data provide compelling potential molecular targets for therapeutics to treat P. gingivalis and E. coli infections.
ContributorsLeonetti, Cori (Author) / Shi, Yixin (Thesis advisor) / Stout, Valerie (Committee member) / Nickerson, Cheryl (Committee member) / Sandrin, Todd (Committee member) / Arizona State University (Publisher)
Created2013
Description
The study of bacterial resistance to antimicrobial peptides (AMPs) is a significant area of interest as these peptides have the potential to be developed into alternative drug therapies to combat microbial pathogens. AMPs represent a class of host-mediated factors that function to prevent microbial infection of their host and serve as a first line of defense. To date, over 1,000 AMPs of various natures have been predicted or experimentally characterized. Their potent bactericidal activities and broad-based target repertoire make them a promising next-generation pharmaceutical therapy to combat bacterial pathogens. It is important to understand the molecular mechanisms, both genetic and physiological, that bacteria employ to circumvent the bactericidal activities of AMPs. These understandings will allow researchers to overcome challenges posed with the development of new drug therapies; as well as identify, at a fundamental level, how bacteria are able to adapt and survive within varied host environments. Here, results are presented from the first reported large scale, systematic screen in which the Keio collection of ~4,000 Escherichia coli deletion mutants were challenged against physiologically significant AMPs to identify genes required for resistance. Less than 3% of the total number of genes on the E. coli chromosome was determined to contribute to bacterial resistance to at least one AMP analyzed in the screen. Further, the screen implicated a single cellular component (enterobacterial common antigen, ECA) and a single transporter system (twin-arginine transporter, Tat) as being required for resistance to each AMP class. Using antimicrobial resistance as a tool to identify novel genetic mechanisms, subsequent analyses were able to identify a two-component system, CpxR/CpxA, as a global regulator in bacterial resistance to AMPs. Multiple previously characterized CpxR/A members, as well as members found in this study, were identified in the screen. Notably, CpxR/A was found to transcriptionally regulate the gene cluster responsible for the biosynthesis of the ECA. Thus, a novel genetic mechanism was uncovered that directly correlates with a physiologically significant cellular component that appears to globally contribute to bacterial resistance to AMPs.
ContributorsWeatherspoon-Griffin, Natasha (Author) / Shi, Yixin (Thesis advisor) / Clark-Curtiss, Josephine (Committee member) / Misra, Rajeev (Committee member) / Nickerson, Cheryl (Committee member) / Stout, Valerie (Committee member) / Arizona State University (Publisher)
Created2013
Description
For over a century, researchers have been investigating collective cognition, in which a group of individuals together process information and act as a single cognitive unit. However, I still know little about circumstances under which groups achieve better (or worse) decisions than individuals. My dissertation research directly addressed this longstanding question, using the house-hunting ant Temnothorax rugatulus as a model system. Here I applied concepts and methods developed in psychology not only to individuals but also to colonies in order to investigate differences of their cognitive abilities. This approach is inspired by the superorganism concept, which sees a tightly integrated insect society as the analog of a single organism. I combined experimental manipulations and models to elucidate the emergent processes of collective cognition. My studies show that groups can achieve superior cognition by sharing the burden of option assessment among members and by integrating information from members using positive feedback. However, the same positive feedback can lock the group into a suboptimal choice in certain circumstances. Although ants are obligately social, my results show that they can be isolated and individually tested on cognitive tasks. In the future, this novel approach will help the field of animal behavior move towards better understanding of collective cognition.
ContributorsSasaki, Takao (Author) / Pratt, Stephen C (Thesis advisor) / Amazeen, Polemnia (Committee member) / Liebig, Jürgen (Committee member) / Janssen, Marco (Committee member) / Fewell, Jennifer (Committee member) / Hölldobler, Bert (Committee member) / Arizona State University (Publisher)
Created2013
Description
The spread of invasive species may be greatly affected by human responses to prior species spread, but models and estimation methods seldom explicitly consider human responses. I investigate the effects of management responses on estimates of invasive species spread rates. To do this, I create an agent-based simulation model of an insect invasion across a county-level citrus landscape. My model provides an approximation of a complex spatial environment while allowing the "truth" to be known. The modeled environment consists of citrus orchards with insect pests dispersing among them. Insects move across the simulation environment infesting orchards, while orchard managers respond by administering insecticide according to analyst-selected behavior profiles and management responses may depend on prior invasion states. Dispersal data is generated in each simulation and used to calculate spread rate via a set of estimators selected for their predominance in the empirical literature. Spread rate is a mechanistic, emergent phenomenon measured at the population level caused by a suite of latent biological, environmental, and anthropogenic. I test the effectiveness of orchard behavior profiles on invasion suppression and evaluate the robustness of the estimators given orchard responses. I find that allowing growers to use future expectations of spread in management decisions leads to reduced spread rates. Acting in a preventative manner by applying insecticide before insects are actually present, orchards are able to lower spread rates more than by reactive behavior alone. Spread rates are highly sensitive to spatial configuration. Spatial configuration is hardly a random process, consisting of many latent factors often not accounted for in spread rate estimation. Not considering these factors may lead to an omitted variables bias and skew estimation results. The ability of spread rate estimators to predict future spread varies considerably between estimators, and with spatial configuration, invader biological parameters, and orchard behavior profile. The model suggests that understanding the latent factors inherent to dispersal is important for selecting phenomenological models of spread and interpreting estimation results. This indicates a need for caution when evaluating spread. Although standard practice, current empirical estimators may both over- and underestimate spread rate in the simulation.
ContributorsShanafelt, David William (Author) / Fenichel, Eli P (Thesis advisor) / Richards, Timothy (Committee member) / Janssen, Marco (Committee member) / Arizona State University (Publisher)
Created2012
Description
Hepatitis C virus (HCV) is a globally prevalent infection which is a main contributor to the global burden of liver disease. Due to its ability to establish a chronic infection, and the lack of usefulness of traditional neutralizing antibody vaccine design in producing a protective immune response, a preventative vaccine has been notoriously difficult to produce. To overcome this, a vaccine using non-structural protein 3 (NS3) as a target to elicit a T cell specific immune response is thought to be a possible strategy for eliciting a protective immune response against hepatitis C infection. In this paper, a recombinant strain of measles virus (MV) that expresses HCV NS3 protein was analyzed. The replication fitness of this recombinant virus also indicates that this construct replicates at a higher rate than parental measles strain. It is also demonstrated through western blot analysis of protein expression and immunofluorescence that this recombinant virus expresses both the inserted HCV NS3 protein, as well as native measles proteins.
ContributorsWoell, Dana Marie (Author) / Reyes del Valle, Jorge (Thesis director) / Nickerson, Cheryl (Committee member) / Julik, Emily (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Human Evolution and Social Change (Contributor)
Created2015-05
Description
Clean water for drinking, food preparation, and bathing is essential for astronaut health and safety during long duration habitation of the International Space Station (ISS), including future missions to Mars. Despite stringent water treatment and recycling efforts on the ISS, it is impossible to completely prevent microbial contamination of onboard water supplies. In this work, we used a spaceflight analogue culture system to better understand how the microgravity environment can influence the pathogenesis-related characteristics of Burkholderia cepacia complex (Bcc), an opportunistic pathogen previously recovered from the ISS water system. The results of the present study suggest that there may be important differences in how this pathogen can respond and adapt to spaceflight and other low fluid shear environments encountered during their natural life cycles. Future studies are aimed at understanding the underlying mechanisms responsible for these phenotypes.
ContributorsKang, Bianca Younseon (Author) / Nickerson, Cheryl (Thesis director) / Barrila, Jennifer (Committee member) / Ott, Mark (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
Division of labor among task specialists is a key feature of the organization of insect societies. Foraging and emigration are two distinct colony tasks that nonetheless depend on very similar behaviors, including searching outside the nest, evaluating discoveries, and recruiting nestmates. These subtasks are crucial to collective decisions about forager allocation and nest site selection. It remains unclear, however, whether the same ants are responsible for similar behavior in both contexts, and to what degree they show finer specializations among common subtasks. We are investigating these issues in the ant Temnothorax rugatulus, by making detailed behavioral descriptions of individually marked colonies as they forage and emigrate. There exists considerable heterogeneity among nest-movers, with a small proportion consistently responsible for a large share of recruitment. We found a similar pattern of heterogeneity amongst ants retrieving food during foraging, but had inconclusive results when examining recruitment to the food. It also appears that the ants that complete tasks during foraging are different from the ants that complete similar tasks during emigrations. These findings will shed light on the organization of division of labor and how it contributes to collective decision-making.
ContributorsSchaper, Gage (Contributor) / Pratt, Stephen (Thesis director) / Fewell, Jennifer (Committee member) / Shaffer, Zachary (Committee member) / Barrett, The Honors College (Contributor)
Created2013-05
Description
In large part, the great success of eusocial insects is due to efficient division of labor (Duarte et al. 2011; Dornhaus 2008). Within ant colonies, the process of dividing labor is not clearly defined, but it may be key to understanding the productivity and success of these colonies. This study analyzed data from an experiment that was conducted with the goal of examining how finely division of labor is organized in ant colonies. The experiment considered the actions of all ants from three Temnothorax rugatulus colonies. The colonies were each carefully recorded during five distinct emigrations per colony. The experiment produced such a large quantity of data that it was challenging to analyze the results, a major obstacle for many studies of collective behavior. Therefore, I designed a computer program that successfully sorted all of the data and prepared it for an initial statistical analysis that was performed in R. The preliminary results suggest that while most of the ants perform little to no work, there is an overall pattern of elitism; it seems that division of labor in ants is not more finely divided than previously shown. Future studies should provide further analysis of the data and will be useful in forming a more complete understanding of the division of labor within the emigrations of Temnothorax rugatulus colonies.
ContributorsJones, Samantha (Author) / Pratt, Stephen (Thesis director) / Jones, Donald (Committee member) / Shaffer, Zachary (Committee member) / Barrett, The Honors College (Contributor) / W. P. Carey School of Business (Contributor)
Created2012-12
DescriptionThis paper provides an analysis of the differences in impacts made by companies that promote their sustainability efforts. A comparison of companies reveals that the ones with greater supply chain influence and larger consumer bases can make more concrete progress in terms of accomplishment for the sustainability realm.
ContributorsBeaubien, Courtney Lynn (Author) / Anderies, John (Thesis director) / Allenby, Brad (Committee member) / Janssen, Marco (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2013-05
Description
Temnothorax ants are a model species for studying collective decision-making. When presented with multiple nest sites, they are able to collectively select the best one and move the colony there. When a scout encounters a nest site, she will spend some time exploring it. In theory she should explore the site for long enough to determine both its quality and an estimate of the number of ants there. This ensures that she selects a good nest site and that there are enough scouts who know about the new nest site to aid her in relocating the colony. It also helps to ensure that the colony reaches a consensus rather than dividing between nest sites. When a nest site reaches a certain threshold of ants, a quorum has been reached and the colony is committed to that nest site. If a scout visits a good nest site where a quorum has not been reached, she will lead a tandem run to bring another scout there so that they can learn the way and later aid in recruitment. At a site where a quorum has been reached, scouts will instead perform transports to carry ants and brood there from the old nest. One piece that is missing in all of this is the mechanism. How is a quorum sensed? One hypothesis is that the encounter rate (average number of encounters with nest mates per second) that an ant experiences at a nest site allows her to estimate the population at that site and determine whether a quorum has been reached. In this study, encounter rate and entrance time were both shown to play a role in whether an ant decided to lead a tandem run or perform a transport. Encounter rate was shown to have a significant impact on how much time an ant spent at a nest site before making her decision, and encounter rates significantly increased as migrations progressed. It was also shown to individual ants did not differ from each other in their encounter rates, visit lengths, or entrance times preceding their first transports or tandem runs, studied across four different migrations. Ants were found to spend longer on certain types of encounters, but excluding certain types of encounters from the encounter rate was not found to change the correlations that were observed. It was also found that as the colony performed more migrations, it became significantly faster at moving to the new nest.
ContributorsJohnson, Christal Marie (Author) / Pratt, Stephen (Thesis director) / Pavlic, Theodore (Committee member) / Shaffer, Zachary (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2013-05