Biochemistry
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Item Observing the Effects of the Protein HE4 on Ovarian Cancer Progression in Danio rerio Embryos(2023) Eller, MadelineOvarian cancer is a leading cause of cancer-related death for females in the United States. Epithelial ovarian carcinomas, the most common type of ovarian cancer, are difficult to diagnose in the early stages of the disease. One of the tests used to diagnose ovarian cancer, the HE4 test, measures the amount of the human epididymis protein 4 (HE4) present. HE4 is a secretory protein shown to be overexpressed in patients with ovarian cancer and has been shown to potentially be involved in promoting cancer proliferation. This study attempted to test the following hypothesis: if HE4 promotes cancer progression, then cells that express higher levels of HE4 should cause increased tumor growth in and lower relative survival rate of zebrafish embryos because HE4 is upregulated in ovarian cancer cells. Zebrafish embryos were injected with either OVCAR8 HE4-overexpressing GFP cells or OVCAR8 wild-type cells, followed by imaging of injected embryos 2 and 3 days-post-injections and measurement of survival rate through 6 days-post-fertilization. Over the course of experimentation, however, this study turned into a methods paper due to the amount of troubleshooting that needed to be done. This paper thus details the method that should be used in the future to test this hypothesis.Item The Risk of SARS-CoV-2 Reinfection and the Limits of Inherent Immunity(Wheaton College. (Norton, Mass), 2021-05-15) Lannery, KimSARS-CoV-2 is the virus that causes the coronavirus disease 2019 (COVID-19) and is the cause of a global pandemic that has infected 155 million people worldwide as of May 2021.1 SARS-CoV-2 is an enveloped and spherical particle that contains a single-stranded positive-sense RNA genome. It is a beta-coronavirus (��CoV) belonging to the Coronavirdiae family. It originated in mammals, with the earliest documented cases being traced to bats.2 COVID-19 can spread through multiple forms of transmission but primarily spreads through respiratory droplets.3 SARS-CoV-2 enters the body and binds to the host’s ACE2 receptors. Once bound, TMPRSS2 cleaves and activates the viral spike protein. When someone is infected by SARS-CoV-2 their immune system works hard to fight the invader. The immune response produces antibodies and virus-specific memory T cells, such as CD4+ and CD8+.4 The antibodies that are produced during this response are believed to provide a certain level of immunity against future infection. The extent to which a person is immune is unknown and the duration for how long the SARS-CoV-2 specific antibodies are present is unclear. This meta-analysis aims to study the reinfection cases of SARS-CoV-2 and to identify the likelihood of a reinfection event occurring, which is a step towards understanding immunity in SARS-CoV-2.Item The Relationship Between ACE2 and the ABO Blood Groups in SARS-CoV-2 Infection Rates(Wheaton College. (Norton, Mass.), 2021-05-16) Kelly, NicholasThe COVID-19 pandemic is one of the most brutal pandemics mankind has had to endure. The novel coronavirus has taken the world by storm, with millions of deaths in just a year. Further research has indicated that the coronavirus responsible, SARS-CoV-2, may attribute to outlasting health defects, such as cardiovascular or pulmonary problems. The survival rate for people infected with this virus is grim, depending on where you are in the world, if there are any comorbidities present, or if variants are present in the area. Emerging literature on ABO blood groups suggest that there may be a correlation between a certain blood group and SARS-CoV-2 susceptibility. The objective of this study was to investigate the rate of infection according to ABO blood groups. In this study, data was compiled and analyzed in a meta-analysis to calculate odds-ratios. Our results show that blood group A individuals may be more susceptible to SARS-COV-2 infection while blood group O individuals may have a protective effect against the virus. This study may provide further insight into how blood groups may affect SARS-CoV-2 susceptibility.Item Investigation of ciliated cell types and ciliary gene expression during heart regeneration in zebrafish (Danio rerio)(Wheaton College. (Norton, Mass.), 2021-05-16) Mantineo, Helene X.Unlike the adult mammalian heart, the adult zebrafish (Danio rerio) heart can regenerate after injury. In a series of regenerative processes, the zebrafish can regenerate their hearts after up to 20% of their ventricle apex is resected. The process of regeneration is directed via highly coordinated cell signaling pathways. Primary cilia act as directors of cell signaling pathways. Important ciliary signaling pathways have been implicated in zebrafish heart regeneration, but the direct link between regeneration and cilia has not been fully elucidated. Previous studies have found cilia present in the outer region of the regenerating zone during zebrafish heart regeneration. This led to the first hypothesis that cilia were present in the epicardium in zebrafish heart regeneration because the epicardium is believed to cover the outer region of the ventricle apex during regeneration. To test this hypothesis, the present study used immunofluorescence of transgenic zebrafish to label the different cell types of the regenerating heart. It was revealed that cilia were present in the outer region regenerating zone but not on the myocardium. There was indirect evidence that the cilia are not on the epicardium. Previous studies also found a change in abundance and length of cilia during zebrafish heart regeneration. This led to the second hypothesis that ciliary genes were differentially expressed during zebrafish heart regeneration. To test this hypothesis, the present study used DESeq2 in R to determine differential gene expression of ciliary genes. A total of 196 ciliary transcripts were found to be differentially expressed in at least one time point in zebrafish heart regeneration. Overall, the ciliated cell types and ciliary gene expression may provide further evidence of the importance of cilia in zebrafish heart regeneration. Continuing to gain insight the mechanisms of zebrafish heart regeneration may lead to clinical applications for human cardiac injury.Item Structural modification of a natural angiotensin converting enzyme inhibitor and the effect on activity.(Wheaton College (MA), 2019) Swanson, Amanda ClaireHypertension is a leading risk factor for mortality that affects nearly a third of adult US residents, and it is primarily combatted through the use of angiotensin converting enzyme inhibitors (ACEIs). In addition to ACEIs currently approved as anti- hypertension drugs, there are a number of natural ACEIs that have not been studied, some of them more effective than others. This study sought to alter the structure of alanine- valine-phenylalanine (AVF), a natural, relatively ineffective ACEI, to create modified peptides that would hopefully act as more effective inhibitors. After a literature analysis, phenylalanine-valine-alanine (FVA) was determined to be more likely to act as an effective inhibitor, so four novel peptides were synthesized. One novel peptide was designed to contain unmodified phenylalanine while the other three contained one of the following para-substituted, N-protected, L-phenylalanine amino acids: L-Phenylalanine, N-benzoyl-4-methyl, L-Phenylalanine, N-benzoyl-4-methoxy-, L-Phenylalanine, N- benzoyl-4-fluoro-. In implementing these peptide modifications, first the respective modified phenylalanine amino acids were synthesized, then the FVA peptides were synthesized, with naturally occurring phenylalanine as well as the three modified phenylalanine amino acids. NMRs confirmed the identities of all products and HPLCdata confirmed the stereochemistry of the chiral amino acid intermediates.