Oklahoma Louis Stokes Alliance for Minority Participation (OK-LSAMP) Program

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Recent Submissions

  • Publication
    Synthesizing Tetraphenylporphyrins and Metalloporphyrins using microwave and ultrasonic reactors
    (Oklahoma State University, 2022-10-01) Moro, Nathalie
    Tetraphenylporphyrin (TPP) is a macrocyclic molecule found in biological compounds. TPP's UV-Vis spectrum ranges from 380-500 nanometers. Metalloporphyrins (MeP) are considered “bioinspired oxidation catalysts” and are synthesized from TPP and metal anhydrous salts. Therefore, hypothesizing that the UV-Vis spectrum of the MeP will be altered by the type of benzaldehyde (electron-donating and electron-withdrawing groups attached to the benzene ring) which are added to the TPP ligand during the MeP synthesis reaction. In this project investigated solvent-free conditions, first with a silica gel support and then without it. The TPPs were produced in a Chemical Explorer Microwave (CEM) Reactor with a commercial 950W/1000W microwave oven. The conventional reflux method and an ultrasonic reactor were used to develop the MePs. TPPs were synthesized with substituted and unsubstituted benzaldehydes to investigate the effect of electron-donating and electron-withdrawing groups on the complex synthesis and the UV shift. The UV-Vis spectra for each MeP confirmed the formation of complexes with transition and p-block metals. The data displayed that the Tin MeP showed a red-shift UV-Vis spectrum (446 nm) compared to the initial TPP (417 nm). Complexation of Tin with the 4-chloroTPP displayed a maximum peak at 473 nm. This reaffirms that the electron-withdrawing benzaldehydes worked better than the electron-donating when added to the TPP. Thus, causing the UV-Vis spectra to have a red shift compared to the initial product.
  • Publication
    Understanding awareness of food security resources available to university students
    (Oklahoma State University, 2022-04-19) Posey, Isabelle H.; McClanahan, Kristen E.; Blevins, Brian
    Food insecurity is a prevalent issue in college campuses across the country. The Hunger on Campus study found that 48% of college students face food insecurity. Oklahoma State University (OSU) also experiences high rates of food insecurity. A recent study found that approximately 42% of OSU Stillwater students were food insecure and sophomores, upperclassmen, and minority students were at the highest risk. The 2020 study found the food insecure students at OSU were not utilizing the food security resources available to them. The purpose of this mixed methods study was to understand why students are not utilizing the available food security resources. The first phase was qualitative and utilized interviews with sophomores, juniors, and senior-level OSU students to identify obstacles preventing resource utilization. Randomized email lists were used for recruitment, and participation was encouraged in ethnically diverse campus organizations. Interviews were transcribed and analyzed for emerging themes. The quantitative phase was informed by the interview results, and included a social media campaign promoted throughout campus using A-Frames. This intervention utilized the social media platform Instagram to promote available resources to students. Student engagement in the intervention was measured by the number of QR scans and Instagram analytics. To measure the effectiveness of increasing resource awareness, a survey was administered to Instagram followers through a direct message. The results of the qualitative phase demonstrated a major limitation in student resource usage is a lack of awareness, and the best way to distribute information on campus was social media and A-Frames. Overall, students at OSU Stillwater are facing food insecurity but are not aware of the resources available; the use of A-frames and social media demonstrate potential to increase awareness. The long-term implication of this study is a reduced prevalence of food insecurity at OSU Stillwater's campus.
  • Publication
    Microplastic extraction from sediments with a continuous flow elutriation process
    (Oklahoma State University, 2021-10-09) Egermeier, Mason; Forsythe, Kyle
  • Publication
    Analysis of heme-binding proteins from Listeria monocytogenes using differential scanning calorimetry
    (Oklahoma State University, 2021-10-09) Craig, Kaylee
    Listeria monocytogenes is a gram-positive bacterium that can cause severe infection in immunocompromised individuals. It is reliant on the acquisition of iron from its host to continuously spread throughout the body. Here we describe the thermal denaturation points of heme binding proteins Hbp1 and Hbp2, two iron scavenging proteins, using Differential Scanning Calorimetry. DSC is a useful technique used to characterize the thermal denaturation point of protein folding and unfolding events. As a result, the enthalpy and entropy of the protein unfolding event can be calculated. Conventional DSC results showed a reproducible denaturation point of 59.6 +/- 0.3 °C for Hbp1 and a denaturation point of 65.6 +/- 0.2 °C for Hbp2. Further investigation was completed to determine the role of bound ligands, such as heme, on overall protein stability. These thermodynamic values for protein denaturation can be applied to pharmaceutical studies to understand the role of these proteins as virulence factors for disease and their potential use as therapeutic targets.
  • Publication
    Tagging endogenous mitochondrial proteins using CRISPR/Cas9 technology
    (Oklahoma State University, 2021-10-09) Mejia, Laura
  • Publication
    Synthesis, purification, and characterization of guest molecules for inclusion in cucurbit[n]urils
    (Oklahoma State University, 2021-10-09) Webb, Ryan
    Cucurbit[n]uril (CB[n]) is a unique macrocycle that can bind small molecules with promising potential applications in drug delivery, molecular machines, and smart materials. This work focuses on the supramolecular equilibrium binding modes and the equilibrium binding constants of CB[n] with a variety of viologens, pyridinium species which have not previously been studied. The synthesis, purification and characterization of these guest species will be presented. The knowledge gained from the study of physical properties of these host guest systems which will aid in the future development of more complex systems.
  • Publication
    Antifungal activity of novel compound EIPE-1 against Cryptococcus neoformans
    (Oklahoma State University, 2021-10-09) Chatman, Priscilla; Conn, Brittney; Maritz, Emma; Nelson, Toby L.
    Cryptococcus neoformans is an opportunistic fungal pathogen of the respiratory tract, which is responsible for over 200,000 deaths annually. Antifungal drugs have been used to treat fungal infections for many decades; however, due to similarities between fungal and mammalian cells, these drugs are often toxic. In these last few decades, the fungi have also become resistant to the antifungal drugs. EIPE-1 was synthesized from vanillin, and was shown to have activity against methicillin resistant S. aureus (MRSA), and other gram-positive bacterial pathogens. We hypothesized that EIPE-1 could be used to kill fungal pathogens. For this study, we tested EIPE-1 against C. neoformans using a minimum inhibitory concentration (MIC) assay and an in vitro model of intracellular macrophage growth using RAW macrophages. EIPE-1 has antifungal activity in our MIC assay, with an MIC value of 1.749 ug/ml. In addition, after incubation of C. neoformans with RAW macrophages and EIPE-1, treatment with EIPE-1 had significant antifungal effects on C. neoformans compared to C. neoformans alone and compared to C. neoformans with RAW macrophages. In further studies, we will examine the mechanism of EIPE-1 anti-fungal activity, and we will also test EIPE-1 against other fungal pathogens including Candida albicans.
  • Publication
    Preliminary exploration of molecular mechanism of the anticancer activity of a novel lead lactate dehydrogenase inhibitor against pancreatic cancer
    (Oklahoma State University, 2021-10-09) Urquiza, Uzziah; Sharma, Horrick
    Pancreatic ductal adenocarcinoma (PDAC) is a leading cause of cancer-related death in the United States, with a long-term survival rate of just under 10%. Chemotherapy is one of the standard treatment options for pancreatic cancer; however, the rapid development of resistance to chemotherapeutic agents represents a significant bottleneck. Therefore, identifying new biological drug targets and exploring biomolecular mechanisms that lead to chemoresistance in pancreatic cancer is urgently required. Cancer cells rewire their metabolic pathways to support their growth and proliferation. Unlike normal cells, cancer cells prefer aerobic fermentation, a phenomenon known as the Warburg effect, where they convert glucose to lactic acid even in the presence of oxygen. Warburg effect in cancer cells is regulated by lactate dehydrogenase-A (LDHA) that is overexpressed in many tumor types. Here, we report our preliminary studies on the molecular mechanisms underlying the anticancer activity of a novel LDHA inhibitor, compound 23, in pancreatic cancer cell lines.
  • Publication
    Discovery and characterization of novel fungal diversity in the equine alimentary tract
    (Oklahoma State University, 2021-10-09) Arreola, Alex X.; Meili, Casey H.; Youssef, Noha H.
    Fungi provide crucial ecosystem services in multiple ecosystems. Most fungi thrive as free-living organisms, but many forge symbiotic, predatory, pathogenic, and commensal relationships with algae, plants, and animals. One of the most peculiar groups of fungi are the anaerobic gut fungi (phylum Neocallimastigomycota) that reside in the alimentary tract of herbivores. Little is known regarding the scope of diversity of these elusive, anoxic microorganisms. Our work aims to characterize the diversity of AF’s in the herbivorous gut on a global scale using culture independent approaches. We have collected >1,000 samples, from >50 type of animals, across 5 continents. My focus in this broader project is to examine patterns and determinants of the diversity of AF in equine alimentary tract. To this end, I extracted DNA from >100 fecal horse samples, and used polymerase chain reaction (PCR) to amplify a specific marker gene (D1/D2 LSU). High throughput sequencing was conducted, and analysis of the sequence data is currently underway. My preliminary analysis revealed a high level of AF diversity within the equine alimentary tract, as evident by the detection of > 30 different cultured and yet-uncultured AF genera in these samples. Moreover, a fraction of the sequenced obtained belong to multiple novel, hitherto undiscovered lineages, clearly indicated that the scope of AF diversity in the equine alimentary tract is much broader than previously suggested. Currently, I am attempting to identify and quantify the impact of various factors (animal feed, age, sex, location) in shaping the AF community in horses via implementing a wide range of statistical and phylogenetic approaches.
  • Publication
    Bacterial two hybrid analysis of Chlamydia trachomatis proteins
    (Oklahoma State University, 2021-10-09) Nail, Kayli; Holcomb, Christian
    Chlamydia trachomatis is an obligate intracellular bacterial pathogen which poses severe health problems throughout the world. There are over 90 million new cases annually, making it the most common sexually transmitted disease in the world. Chlamydia can pose significant problems during and after infection. It is imperative to understand how Chlamydia manipulates the host cell and potentially develop future treatments. The mechanisms by which C. trachomatis alters immune response is not well understood, but recent work has identified an interaction between the chlamydial inclusion membrane protein, CT226, and the potential interacting host proteins, Flightless homologue II (FLII), Leucine Rich-Repeat Flightless-Interacting Proteins 1&2 (LRRFIP1 and LRRFIP2) and TMOD3. FLII, LRRFIP1 and LRRFIP2 are known to interact as a complex and are upstream regulators of the inflammasome. Currently, it is unknown if CT226 interacts with one or all of the interacting partners and needs the actual interaction needs investigation. My hypothesis is that CT226 will directly interact with one of the 3 potential interacting partners (LRRFIP1, LRRFIP2, and FLII) and that we will be able to detect this interaction in the bacterial two hybrid system. Current efforts have focused on cloning CT226 into PUT18 and the three host proteins (FLII, LRRFIP1 and LRRFIP2) individually into PKNT25. PUT18 and PKNT25 are the bait and prey plasmids that have been adapted for use in the bacterial two hybrid system. Once each pair of plasmids are properly transformed (each E. coli strain will have two plasmids, one carrying CT226 and one carrying the potential interacting partner), they will be screened using traditional B-galactosidase assays or cAMP assays to determine the specific interactions.
  • Publication
    Using fluorescent UPEC as a model to study interactions with human bladder cells
    (Oklahoma State University, 2021-10-09) Eberhard, Alissa D.
    Escherichia coli (E. coli) are opportunistic bacteria that reside in the intestines of humans and contribute to gastrointestinal health. However, there are some strains that can cause a variety of diseases including urinary tract infections (UTIs). UTIs caused by E. coli are the most common type of bacterial infections seen in women and are a significant public health concern. Uropathogenic E. coli (UPEC) have acquired specific virulence factors including adhesions and fimbriae, which lead to increased adherence and invasion into urinary tract cells in the host. The pathogenic mechanisms employed by UPEC that promote adhesion and invasion have yet to be fully elucidated. We propose to study the mechanisms of adherence and invasion of UPEC to host cells by generating UPEC expressing the green fluorescent protein (GFP). we hypothesize that the GFP-expressing UPEC will assist in studying host-pathogen interactions. To test this hypothesis, we transformed UPEC with a GFP encoding plasmid and successfully generated fluorescent UPEC. These fluorescent UPEC were used to infect human bladder epithelial cells (5637) at increasing multiplicities of infection (MOI) to study adherence and invasion. We successfully detected and quantified adherence and invasion of the fluorescent UPEC by different methods that include fluorescent microscopy, flow cytometry, and gentamicin-based invasion assays. Thus, with the assistance of GFP-expressing UPEC, we can efficiently gain more insight on host proteins that mediate adherence and invasion of UPEC. These findings will shed more light on the different mechanisms utilized by UPEC in establishing UTIs, which will in turn lead to the development of more effective therapies for the prevention and treatment of UTIs.
  • Publication
    Studying the effects of antibiotics on mitochondrial inhibitors
    (Oklahoma State University, 2021-10-09) Saleh, Maryam
    The usage of ivermectin (ivm) to treat COVID-19 is controversial. According to a study, ivermectin helped treat COVID-19, but clinical trials showed that ivermectin with the tetracycline-class antibiotic doxycycline overcame the effects of ivermectin to a certain degree2. The goal of this project is to understand why this occurs. Cell cultures were used as a model. Rather than overcoming the effects of ivermectin, tetracycline seems to prevent ivermectin from mitochondrial inhibition. This effect seems to occur when tetracycline is administered first, with ivermectin administered after one hour or more; the effect is prevented when ivermectin is administered, left for two hours, and then tetracycline is added and left for one hour.
  • Publication
    Screening Warrior Hops for inhibitors of cancer cell metabolism
    (Oklahoma State University, 2021-10-09) Okolie, Star
    The purpose of this project is to examine the effects of components of Warrior Hops on cancer cell metabolism to better understand the mechanism of infection for SARSCoV-2. In h293 cells (human embryonic kidney cells that metabolically mimic cancer cells), several fractions have been obtained that showed mitochondrial inhibition and the inhibition of ATP production in cells similarly to hydroxychloroquine, which acts as the positive control. This project is still in progress and will provide a better understanding of the mechanism of infection for COVID-19 on the cellular level.
  • Publication
    Enzymatic assessment of cells with distinct TP53 mutations
    (Oklahoma State University, 2021-10-09) Kouplen, Kate; Hermann, Evan
    The p53 transcription factor, encoded by the human gene TP53, is the most commonly mutated gene in human cancers. Once the TP53 gene is mutated, p53 proteins not only lose their tumor suppression abilities but can also contribute to tumor growth. One-way mutant p53 can do this is by increasing iron uptake and availability by altering the regulation of iron-sulfur (Fe-S) cluster containing proteins. Understanding such alterations are important because Fe-S cluster proteins play a role in both energy metabolism and DNA repair enzymes, which can suppress the progression and expansion of cancerous tumors. This study’s main focus was to examine how TP53 mutation status influences Fe-S cluster protein regulation, and subsequently iron homeostasis in tumor cells. First, we assessed the activity of the Fe-S containing enzymes cytosolic and mitochondrial aconitase in cell lines expressing a variety of TP53 mutations. Specifically, we will use cell lines that express the six most commonly observed TP53 mutations, which represent nearly 25% of all TP53 mutations in human cancers. We found that while aconitase activity decreased as expected following the removal of iron in cells expressing WTP53, the influence of iron chelation on aconitase activity in cells expressing mutant TP53 varied depending on the mutation type. This data strongly suggests that TP53 mutations significantly influence how the activity of Fe-S cluster enzymes is regulated. Thus, our ongoing studies are focused on investigating how TP53 mutation status influences the regulation of genes involved with Fe-S cluster biogenesis and consequently, Fe-S cluster containing protein activity.
  • Publication
    Effects of light intensity on the morphology, growth rate and phenolic content of Zygnema
    (Oklahoma State University, 2021-10-09) Long, C. Ethan
    Zygnema is a freshwater alga that forms thick mats that float at the water surface. Cells located at the surface of mats are exposed to very high visible light and UVB compared to cells deep within the mats. Previous studies have shown that Zygnema grown at high visible light and UVA levels shows increased tolerance to UVB exposure that coincided with an increase in UVB absorbance suggesting the photo-induced production of UVB protective compounds. We investigated whether visible light or UVA exposure induces changes in UVB absorbance and whether increased production of phenolic compounds is responsible for the absorbance changes. We exposed two Zygnema isolates to different light treatments: high light with UVA, high light without UVA, low light with UVA, and low light without UVA. After 7 days of exposure, cells were photographed and cell lengths measured. Weighed samples were extracted in methanol. The extracts were scanned with a UV spectrophotometer to determine UVB absorbance and quantitatively tested for total phenolic compounds. Zygnema isolates exposed to high light exhibited a 296% increase in UVB absorbance which coincided with a 255% increase in total phenolic content compared to samples exposed to low light. Removing UVA at high visible light exposure resulted in decreased phenolic content for only one isolate. Cell length decreased 51% in high light treatments. Removing UVA resulted in significantly larger cells only at low visible light.
  • Publication
    Construction of a reusable Gel-Eluted Liquid Fraction Entrapment Electrophoresis (GELFrEE) using 3D printing
    (Oklahoma State University, 2021-10-09) Galie, Walter; Guo, Yanting; Cupp-Sutton, Kellye A.
    Gel-Eluted Liquid Fraction Entrapment (GELFrEE) is commonly utilized in proteomics research for size-based separation and fractionation of proteins. Commercial GELFrEE cartridges are wasteful due to their single-use application and, furthermore, are being discontinued for leaving researchers that utilize the technique without needed resources. We have designed a 3D-printed, reusable GELFrEE cartridge that can be utilized with existing GELFrEE instruments. This cartridge allows the user to clean and repeatedly refill the chambers with new gels eliminating unnecessary plastic waste. Additionally, we have optimized the construction of the cartridge with respect to the type of plastic as well as coating materials and sealing glues to avoid interaction with buffers, solvents, and proteins. The 3D-printed cartridge will further allow us to optimize the composition of the separating gel according to the experimental purposes. Lastly, the cartridge has been refined to successfully separate complex E. coli lysate utilizing twelve and fifteen percent acrylamide gel.
  • Publication
    Investigation of the mechanical, collagen microstructural, and morphological properties of human intracranial aneurysms
    (Oklahoma State University, 2021-10-09) Cabello, Eleana
    Intracranial aneurysms (ICAs) are focal dilations of cerebral arteries caused by the weakening of the arterial wall. The resulting changed hemodynamics can further lead to stress and growth of the site until it eventually ruptures. The morbidity and mortality of a ruptured aneurysm are high, even after treatment, with many dying within months of the occurrence or suffering from lifelong disability [1]. Previous studies have separately examined the hemodynamics, mechanical properties, or morphology of human aneurysm tissues. The present work provides quantitative data on a resected human cerebral aneurysm's mechanical, collagen fiber microstructural, and morphological characteristics. The mechanical properties of the tissue were characterized using biaxial tension and stress relaxation tests. The tissue's collagen fiber architecture and its load-dependent changes were then examined using a polarized spatial frequency domain imaging system. The microstructural components of the tissue were quantified using histological procedures. This investigation extends on our previously developed characterization framework [2] and provides additional quantitative information on human cerebral artery aneurysms. Such investigations of these properties can provide essential insight into the evolution of aneurysms and their associated rupture risk, which can ultimately improve our fundamental understanding of aneurysm growth critical for the future development of aneurysm therapeutics with improved outcomes.
  • Publication
    Comparison study of 1999-2019 suicide rate per 100,000 population between Native American and other races in the United States of America
    (Oklahoma State University, 2021-10-09) Quintana, Emma
    The suicide rate in the USA has been on the rise, which affects specific age groups, genders, and race/ethnicity. Native American (NA) has been known to be on a higher level than the other races. This study focuses on differences in suicide rates between NA and the other races at particular age groups and different genders. Twenty years (1999-2019) of suicide data were retrieved from the CDC database. Microsoft Excel and t-test were employed for data processing and statistical analysis. The results showed that the suicide rate per 100,000 population increased 38.1% nationally with male increased 34.5% and female increased 55%. Male was 281.42% higher than female. NA was 44.48% higher than total nation in age group 10-39. In age group 40-84, US total was 81.46% higher than NA. Crossing race comparison among NA, African American (AA), Hispanic (His), Asian or Pacific Islander (AP), and White (W) showed NA and W were much higher than other races. NA showed 43.31% higher in the age group 10-34 than W. W showed 86.63% higher in the age group 35-84 than NA. NA showed 99.09% and 89.62% higher in the age group 10-64 than His and AP, respectively. AP showed 31.68% higher than NA in the age group 65-84. There was no significant difference between NA and His in the age group 65-84. NA was 94.07% higher than AA through all age groups. In summary, the suicide rate increased by about 38.10%. The suicide rates ranked as W>NA>AP>His and AA. NA was the highest one in the age group 10-34, which should bring more attention to the community for further investigation. This study provides information for suicide prevention and emphases the necessity of psychological/psychiatric services and family education for those affected age groups nationally so reduction in suicide may positively impact the populations.
  • Publication
    Viral oncoprotein Tax and regulation of the HTLV promoter
    (Oklahoma State University, 2021-10-09) Porter, Savannah
    Tax, an oncoprotein virally expressed from Human T-cell Leukemia Virus (HTLV), is a transcriptional regulator with incompletely understood functions in host cells. HTLV is a retrovirus of clade deltraretroviradae with two known sub-types (HTLV Type-1 and HTLV Type-2). HTLV-1 is associated with a subset of patients in development of an extremely aggressive blood cancer called Adult T-cell Leukemia/Lymphoma (ATLL) but HTLV-2 is not. It is known that Tax recruits host activator and co-activator proteins to binds to the integrated viral promoter region of HTLV and rapidly activate viral gene expression. Tax coding regions vary in HTLV-1 and type-2 in the predicted transcriptional activation domain. Exploring interaction surfaces of Tax with various host proteins involved may help us understand the basic transcriptional activation mechanisms that are manipulated by the HTLV-1. To investigate this, we accumulated a library of Tax expression plasmids with the Tax wild-type and mutant coding sequences from HTLV-1. The mutants collected have been established to impair transcriptions or cytoplasmic activities of Tax and include in contrast the Tax coding sequence from HTLV-2. Once the mutants were obtained, we established a stock that can be used in the future for downstream expression, purification, and activity binding assays.
  • Publication
    Ossetra caviar microbiome
    (Oklahoma State University, 2021-10-09) Hansen, Meadow; Faizi, Irene
    Ossetra caviar from Russian sturgeon Acipenser gueldenstaedtii is one of the most sought-after caviars in the world. Wild Russian sturgeon populations have been vastly depleted in their native locations, such as the Caspian Sea, due to overexploitation and habitat degradation. In fact, the importation of caviar from their native areas has become illegal in many countries. Russian sturgeon are now raised in aquaculture to overcome this challenge. As a result, aquaculture has become more popular as it farms sturgeon, and thus caviar, without putting them at risk. The purpose of this study was to investigate the microbiome of caviar and identify key differences among prokaryotic taxa to determine whether particular organisms correspond to better flavor profiles and overall product quality. Fifteen caviars of varying prices ($60 to $595/oz.) were purchased from online vendors. Genomic DNA was extracted from the brine of each tin of caviar and prepared for sequencing. Sequences were then analyzed using the QIIME 2 microbiome bioinformatics platform. A diverse array of microorganisms were found to be present across all caviar samples. Although no differences in diversity were discovered when comparing the grade, osmolarity, salt content, pH, presence of sodium tetraborate (chemical preservative), container type, or diameter or moisture content of the roe, there were differences in the microbial composition of the caviars. This model may be used to infer the impact of microorganisms on the quality of caviar and explain how they may be used to improve the value and appeal of caviars produced in aquaculture.