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Tumor associated microglia-and-macrophages (TAMS) may constitute up to 30% of the composition of glioblastoma. Through mechanisms not well understood, TAMS are thought to aid the progression and invasiveness of glioblastoma. In an effort to investigate properties of TAMS in the context of glioblastoma, I utilized data from a PDGF-driven rat

Tumor associated microglia-and-macrophages (TAMS) may constitute up to 30% of the composition of glioblastoma. Through mechanisms not well understood, TAMS are thought to aid the progression and invasiveness of glioblastoma. In an effort to investigate properties of TAMS in the context of glioblastoma, I utilized data from a PDGF-driven rat model of glioma that highly resembles human glioblastoma. Data was collected from time-lapse microscopy of slice cultures that differentially labels glioma cells and also microglia cells within and outside the tumor microenvironment. Here I show that microglia localize in the tumor and move with greater speed and migration than microglia outside the tumor environment. Following previous studies that show microglia can be characterized by certain movement distributions based on environmental influences, in this study, the majority of microglia movement was characterized by a power law distribution with a characteristic power law exponent lower than outside the tumor region. This indicates that microglia travel at greater distances within the tumor region than outside of it.
ContributorsJuliano, Joseph Dominic (Author) / Kostelich, Eric (Thesis director) / Nagy, John (Committee member) / Swanson, Kristin (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2013-12
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Description
Protein AMPylation is a recently discovered and relatively unstudied post-translational modification (PTM). AMPylation has previously been shown to play an important role in metabolic regulation and host pathogenesis in bacteria, but the recent identification of potential AMPylators across many species in every domain of life has supported the possibility that

Protein AMPylation is a recently discovered and relatively unstudied post-translational modification (PTM). AMPylation has previously been shown to play an important role in metabolic regulation and host pathogenesis in bacteria, but the recent identification of potential AMPylators across many species in every domain of life has supported the possibility that AMPylation could be a more fundamental and physiologically significant regulatory PTM. For the first time, we characterized the auto-AMPylation capability of the human protein SOS1 through in vitro AMPylation experiments using full-length protein and whole-domain truncation mutants. We found that SOS1 can become AMPylated at a tyrosine residue possibly within the Cdc25 domain of the protein, the Dbl homology domain is vital for efficient auto-AMPylation activity, and the C-terminal proline-rich domain exhibits a complex regulatory function. The proline-rich domain alone also appears to be capable of catalyzing a separate, unidentified covalent self-modification using a fluorescent ATP analogue. Finally, SOS1 was shown to be capable of catalyzing the AMPylation of two endogenous human protein substrates: a ubiquitous, unidentified protein of ~49kDa and another breast-cancer specific, unidentified protein of ~28kDa.
ContributorsOber-Reynolds, Benjamin John (Author) / LaBaer, Joshua (Thesis director) / Borges, Chad (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Life Sciences (Contributor)
Created2014-05
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Description
Using weather data from the Weather Research and Forecasting model (WRF), we analyze the transport of inertial particles in Hurricane Katrina in order to identify coherent patterns of motion. For our analysis, we choose a Lagrangian approach instead of an Eulerian approach because the Lagrangian approach is objective and frame-independent,

Using weather data from the Weather Research and Forecasting model (WRF), we analyze the transport of inertial particles in Hurricane Katrina in order to identify coherent patterns of motion. For our analysis, we choose a Lagrangian approach instead of an Eulerian approach because the Lagrangian approach is objective and frame-independent, and gives results which are better defined. In particular, we locate Lagrangian Coherent Structures (LCS), which are smooth sets of fluid particles which are locally most hyperbolic (either attracting or repelling). We implement a variational method for locating LCS and compare the results to previous computation of LCS using Finite-Time Lyapunov Exponents (FTLE) to identify regions of high stretching in the fluid flow.
ContributorsDeibel, Angelica Rae (Author) / Tang, Wenbo (Thesis director) / Moustaoui, Mohamed (Committee member) / Kostelich, Eric (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Department of Physics (Contributor)
Created2013-05
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Description
In this research we consider stochastic models of Glioblastoma Multiforme brain tumors. We first look at a model by K. Swanson et al., which describes the dynamics as random diffusion plus deterministic logistic growth. We introduce a stochastic component in the logistic growth in the form of a random growth

In this research we consider stochastic models of Glioblastoma Multiforme brain tumors. We first look at a model by K. Swanson et al., which describes the dynamics as random diffusion plus deterministic logistic growth. We introduce a stochastic component in the logistic growth in the form of a random growth rate defined by a Poisson process. We show that this stochastic logistic growth model leads to a more accurate evaluation of the tumor growth compared its deterministic counterpart. We also discuss future plans to incorporate individual patient geometry, extend the model to three dimensions and to incorporate effects of different treatments into our model, in collaboration with a local hospital.
ContributorsManning, Michael Clare (Author) / Kostelich, Eric (Thesis director) / Kuang, Yang (Committee member) / Gardner, Carl (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / School of Letters and Sciences (Contributor) / School of Human Evolution and Social Change (Contributor)
Created2013-12
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Description
In this thesis, glycan nodes, the basic subunits of complex biological sugars, were studied to determine the reproducibility of gas chromatography-mass spectrometry (GC/MS) based methylation analysis of whole blood plasma by normalization using an internal standard of heavy permethylated glycans. Glycans are complex biological sugars that have a variety of

In this thesis, glycan nodes, the basic subunits of complex biological sugars, were studied to determine the reproducibility of gas chromatography-mass spectrometry (GC/MS) based methylation analysis of whole blood plasma by normalization using an internal standard of heavy permethylated glycans. Glycans are complex biological sugars that have a variety of applications in the human body and will display aberrant compositions when produced by cancerous cells. Thus an assay to determine their composition can be used as a diagnostic tool. It was shown that the assay may have potential use, but needs further refinement to become an improvement over current methods by analyzing the results of ratio-determination and replicate experiments.
ContributorsMiyasaki, Tyler Takeo (Author) / Borges, Chad (Thesis director) / Van Horn, Wade (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / Chemical Engineering Program (Contributor)
Created2015-05
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Description
Western diets, high in dietary fat and red meat, are associated with hyperglycemia and weight gain, symptoms that promote insulin resistance and diabetes. Previous studies have shown that elevated glucose promotes glycation of circulating proteins such as albumin, which is thought to lead to hyperglycemia complications. It was hypothesized that

Western diets, high in dietary fat and red meat, are associated with hyperglycemia and weight gain, symptoms that promote insulin resistance and diabetes. Previous studies have shown that elevated glucose promotes glycation of circulating proteins such as albumin, which is thought to lead to hyperglycemia complications. It was hypothesized that diets with no meat consumption (pesco-vegetarian and lacto-vegetarian) would reduce protein glycation, in comparison to a diet with meat. Forty six healthy adult omnivorous subjects were randomized into one of three groups and instructed to either consume red meat (i.e. meat) or poultry twice per day (control), eliminate meat and increase fish consumption (pesco-vegetarian), or adopt a vegetarian diet devoid of fish, meat or poultry (lacto-vegetarian) for four weeks. Fasting plasma samples were collected from participants at baseline and after 4 weeks of the dietary intervention. Plasma glucose concentrations were measured using a commercially available kit. Percent glycated albumin was measured on a separate aliquot of plasma by mass spectrometry. Plasma glucose concentrations were significantly increased following 4-weeks of pesco-vegetarian diet (P=0.002, paired t-test). Neither the lacto-vegetarian (P=0.898) or the control diet (P=0.233) affected plasma glucose concentrations. Despite the significant increase in plasma glucose following a pesco-vegetarian diet, no change in percent glycated albumin was observed (P>0.50, ANOVA). These findings may indicate a protective effect of the pesco-vegetarian diet on protein glycation in the presence of elevated plasma glucose and suggest the need for additional studies to examine the link between increased fish consumption and glucose regulation.
ContributorsRaad, Noor (Author) / Sweazea, Karen (Thesis director, Committee member) / Borges, Chad (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2015-05
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Description
Lactate and methanol have been the most commonly used electron donors in the Krajmalnik-Brown laboratory for efficient microbial dechlorination of trichloroethene (TCE). Our goal was to assess the technical and economic feasibility of molasses and ethanol, two alternative electron donors by evaluating their costs and ability support complete TCE dechlorination

Lactate and methanol have been the most commonly used electron donors in the Krajmalnik-Brown laboratory for efficient microbial dechlorination of trichloroethene (TCE). Our goal was to assess the technical and economic feasibility of molasses and ethanol, two alternative electron donors by evaluating their costs and ability support complete TCE dechlorination to ethene. First, ethanol and molasses, with and without methanol, were evaluated for their abilities to support complete dechlorination in batch serum bottles. Molasses, the cheapest alternative, supported a similar dechlorination performance to lactate in batch experiments, so we then used it in an upflow anaerobic bioreactor (UABR) to test its ability to support rapid dechlorination in this continuous system. Molasses supported 88% TCE conversion to ethene at a hydraulic retention time (HRT) of 13 hours after 80 days of operation in continuous mode. Compared to the UABR operated previously using lactate and methanol, molasses led to a reduction of TCE conversion to ethene, and a possible increase in time required to produce culture. Additionally, when molasses was used as the electron donor, we encountered new difficulties in the operation of the UABR, such as drastic pH changes. Therefore, I conclude that the savings from using molasses is outweighed by the costs associated with the reduction in dechlorination performance and increase in reactor maintenance. I recommend that lactate and methanol continue to be used as the electron donors in the Krajmalnik- Brown dechlorination lab to support fast-rate and cost-effective production of dechlorinating culture in an UABR. Because molasses supported fast rates of dechlorination in the batch experiment, however, it is potentially a better option than lactate and methanol for batch production of culture or for biostimulation, where the aquifer resembles a batch system. I recommend that further studies be done to reach a general conclusion about the feasibility of molasses as an electron donor for other enhanced bioremediation projects.
ContributorsBondank, Emily Nicole (Author) / Krajmalnik-Brown, Rosa (Thesis director) / Delgado, Anca (Committee member) / Torres, Cesar (Committee member) / Civil, Environmental and Sustainable Engineering Programs (Contributor) / Barrett, The Honors College (Contributor)
Created2014-12
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Description

The human gut microbiome is a complex community of microorganisms. These microbes play an important role in host health by contributing essential compounds and acting as a barrier against pathogens. However, these communities and associated functions can be impacted by factors like disease and diet. In particular, microbial fermentation of

The human gut microbiome is a complex community of microorganisms. These microbes play an important role in host health by contributing essential compounds and acting as a barrier against pathogens. However, these communities and associated functions can be impacted by factors like disease and diet. In particular, microbial fermentation of dietary components like polysaccharides, proteins, and fats that reach the gut are being examined to better understand how these biopolymers are utilized and affect community structure. Thus, evaluating the accuracy of methods used to quantify specific macromolecules is crucial to gaining a precise understanding of how gut microbes hydrolyze those substrates. This study presents findings on the accuracy of the Megazyme RS kit (Rapid) modified for high performance liquid chromatography (HPLC) readings and the DC Protein Assay when performed on samples from complex gut media with potato starch treatments and bovine serum albumin (BSA) treatments. Overall, our data indicates that the megazyme RS kit needs further modification to detect expected starch content with the HPLC and that the DC Protein Assay is not suitable for specific protein analysis.

ContributorsKlein, Rachel Marie (Author) / Krajmalnik-Brown, Rosa (Thesis director) / Marcus, Andrew (Committee member) / School of Life Sciences (Contributor) / School of International Letters and Cultures (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Description

Five immunocompetent C57BL/6-cBrd/cBrd/Cr (albino C57BL/6) mice were injected with GL261-luc2 cells, a cell line sharing characteristics of human glioblastoma multiforme (GBM). The mice were imaged using magnetic resonance (MR) at five separate time points to characterize growth and development of the tumor. After 25 days, the final tumor volumes of

Five immunocompetent C57BL/6-cBrd/cBrd/Cr (albino C57BL/6) mice were injected with GL261-luc2 cells, a cell line sharing characteristics of human glioblastoma multiforme (GBM). The mice were imaged using magnetic resonance (MR) at five separate time points to characterize growth and development of the tumor. After 25 days, the final tumor volumes of the mice varied from 12 mm3 to 62 mm3, even though mice were inoculated from the same tumor cell line under carefully controlled conditions. We generated hypotheses to explore large variances in final tumor size and tested them with our simple reaction-diffusion model in both a 3-dimensional (3D) finite difference method and a 2-dimensional (2D) level set method. The parameters obtained from a best-fit procedure, designed to yield simulated tumors as close as possible to the observed ones, vary by an order of magnitude between the three mice analyzed in detail. These differences may reflect morphological and biological variability in tumor growth, as well as errors in the mathematical model, perhaps from an oversimplification of the tumor dynamics or nonidentifiability of parameters. Our results generate parameters that match other experimental in vitro and in vivo measurements. Additionally, we calculate wave speed, which matches with other rat and human measurements.

ContributorsRutter, Erica (Author) / Stepien, Tracy (Author) / Anderies, Barrett (Author) / Plasencia, Jonathan (Author) / Woolf, Eric C. (Author) / Scheck, Adrienne C. (Author) / Turner, Gregory H. (Author) / Liu, Qingwei (Author) / Frakes, David (Author) / Kodibagkar, Vikram (Author) / Kuang, Yang (Author) / Preul, Mark C. (Author) / Kostelich, Eric (Author) / College of Liberal Arts and Sciences (Contributor)
Created2017-05-31
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Description

Recent studies suggest a role for the microbiota in autism spectrum disorders (ASD), potentially arising from their role in modulating the immune system and gastrointestinal (GI) function or from gut–brain interactions dependent or independent from the immune system. GI problems such as chronic constipation and/or diarrhea are common in children

Recent studies suggest a role for the microbiota in autism spectrum disorders (ASD), potentially arising from their role in modulating the immune system and gastrointestinal (GI) function or from gut–brain interactions dependent or independent from the immune system. GI problems such as chronic constipation and/or diarrhea are common in children with ASD, and significantly worsen their behavior and their quality of life. Here we first summarize previously published data supporting that GI dysfunction is common in individuals with ASD and the role of the microbiota in ASD. Second, by comparing with other publically available microbiome datasets, we provide some evidence that the shifted microbiota can be a result of westernization and that this shift could also be framing an altered immune system. Third, we explore the possibility that gut–brain interactions could also be a direct result of microbially produced metabolites.

ContributorsKrajmalnik-Brown, Rosa (Author) / Lozupone, Catherine (Author) / Kang, Dae Wook (Author) / Adams, James (Author) / Biodesign Institute (Contributor)
Created2015-03-12