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Browsing by Author "Wiseman, Brittany"

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    9-(2-Phosphonyl-methoxyethyl)-adenine promotes erythrocytic differentiation and disrupts cell replication in chronic myelogenous leukemia K562 cells
    (2021) Wiseman, Brittany
    Disruption during cellular differentiation can cause hematopoietic stem cells to proliferate uncontrollably, resulting in the development of cancer. Differentiation therapies are being investigated as a type of cancer treatment which involve inducing agents that promote the differentiation of cancer cells into those with similar properties to normal blood cells. These cells can then undergo apoptosis at an accelerated and controlled rate compared to cancer cells, making this a potential therapeutic technique. In this study, the ability of human chronic myelogenous leukemia K562 cells to undergo cellular differentiation in response to the inducing agent 9-(2-Phosphonyl-methoxy ethyl)-adenine (PMEA) is investigated. PMEA has previously been shown to disrupt cell replication, and promote erythrocytic differentiation in K562 cells. In order to further test the effectiveness of this inducer, cell proliferation was measured with a cell growth curve, hemoglobin presence was measured with benzidine staining, and gamma-globin expression (a protein subunit of fetal hemoglobin) was measured in both induced and uninduced K562 cell cultures via RT-qPCR and western blotting. The results indicate that PMEA slows cell replication, and promotes hemoglobin (and subsequently gamma-globin) expression in treated cells. In summary, the findings support the conclusion that PMEA is able to promote erythrocytic differentiation in K562 cells, and provides information that supports differentiation therapies as a method for cancer treatment.
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    9-(2-Phosphonyl-methoxyethyl)-adenine promotes erythrocytic differentiation and disrupts cell replication in chronic myelogenous leukemia K562 cells
    (2021) Wiseman, Brittany; Harcombe, Kimberley; Bernstein, Nina
    Disruption during cellular differentiation can cause hematopoietic stem cells to proliferate uncontrollably, resulting in the development of cancer. Differentiation therapies are being investigated as a type of cancer treatment which involve inducing agents that promote the differentiation of cancer cells into those with similar properties to normal blood cells. These cells can then undergo apoptosis at an accelerated and controlled rate compared to cancer cells, making this a potential therapeutic technique. In this study, the ability of human chronic myelogenous leukemia K562 cells to undergo cellular differentiation in response to the inducing agent 9-(2-Phosphonyl-methoxy ethyl)-adenine (PMEA) is investigated. PMEA has previously been shown to disrupt cell replication, and promote erythrocytic differentiation in K562 cells. In order to further test the effectiveness of this inducer, cell proliferation was measured with a cell growth curve, hemoglobin presence was measured with benzidine staining, and gamma-globin expression (a protein subunit of fetal hemoglobin) was measured in both induced and uninduced K562 cell cultures via RT-qPCR and western blotting. The results indicate that PMEA slows cell replication, and promotes hemoglobin (and subsequently gamma-globin) expression in treated cells. In summary, the findings support the conclusion that PMEA is able to promote erythrocytic differentiation in K562 cells, and provides information that supports differentiation therapies as a method for cancer treatment.
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    Ethics in research: An overview of universal ethics and the perpetuation of inequality in academia
    (2021) Wiseman, Brittany; Biittner, Katie; Davis, Monica
    Ethics govern how research is conducted by Western institutions, though there are limitations in how effective codes of conduct can be in ensuring that research practice is truly ethical in all situations. Though practices have improved, there are several considerations that must still be met to ensure that research is both beneficial and respectful to all involved. The historical lack of repercussions that have accompanied Western research practice has functioned to further disadvantage Indigenous People, People of Color, and women. This has allowed for sexism, harassment, racism, and discrimination to continue. Existing ethical protocols are limited based on the inherent subjectivity in how ethics are perceived, where supplementary protocols should be created on a case by case basis that actively include and empower voices from local community members and researchers. Additionally, recognition of the past and present inequalities faced by marginalized groups is necessary to rectify the issues that these people face while they establish themselves in academic disciplines. This research project addresses the problems associated with “universal” ethical protocols, the disconnect that exists in the construction and ideological view of ethics between disciplines, and the ways that Western research practice has been shaped, including how the hierarchy present in academic institutions continues to marginalize and disadvantage certain groups of people, with particular emphasis on the perpetuation of racism and sexism.
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    Identifying the nuclear localization signal of the Arabidopsis thaliana MADS box transcription factor Flowering Locus C
    (2021) Wiseman, Brittany; Hills, Melissa
    The goal of this project is to design a research strategy to identify the Nuclear Localization Signal (NLS) of the Arabidopsis thaliana MADS box transcription factor Flowering Locus C (FLC), and to make plant transformation components that are necessary for the creation of controls for this objective. An NLS is required for the nuclear localization of nuclear proteins, and though previous work on Arabidopsis has established conserved structural-functional domains within FLC, the NLS has not yet been characterized. To complete the objectives of this project, primers were designed and used to modify FLC to be compatible with the GreenGate cloning system. FLC was cloned, sequenced, and glycerol stocks were created and stored. Future projects will use this FLC to create an FLC:GFP fusion to serve as a control where protein localization is expressed in the nucleus. Electrocompetent Agrobacterium tumefaciens were made and a pGFPGUSPLUS plasmid was introduced. Glycerol stocks were stored to be used in Agrobacterium mediated transformation in the future, which will establish another control line of Arabidopsis expressing GFP proteins that are not nuclearly localized. Finally, a literature review was completed to explore the existing understanding of NLS structure in general and in MADS box transcription factor proteins, to propose mutations that might disrupt the NLS sequence within FLC. This work aims to fill a knowledge gap in regards to the mechanism of nuclear localization of FLC and will ultimately contribute to our understanding of nuclear localization in plants and MADS box proteins in general.