Honors Projects for Biology

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    (University of Hawaii at Manoa, 2021) Natale, Jessica ; Agsalda, Melissa ; Biology
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    Effects of Hypoxia on Cardiomyocyte Chromatin
    (University of Hawaii at Manoa, 2021) Markell, Elliott ; Knutson, Andrew ; Biology
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    The Gut Microbiome of the Snails Lissachatina fulica and Parmarion martensi, Infected and Uninfected by the Rat Lungworm, Angiostrongylus cantonensis
    (University of Hawaii at Manoa, 2020) Uemura, Leina ; Cowie, Robert ; Biology
    This project focused on a parasite, the rat lungworm (Angiostrongylus cantonensis), and its intermediate snail hosts (rats are the definitive hosts in which the worms reproduce). This parasite is important because it is the main etiological agent causing human eosinophilic meningitis. The intermediate hosts are important because people are infected by the parasite when they consume infected hosts, deliberately or inadvertently. Once consumed, the parasites eventually reach the brain, where they remain, feeding and moving until they die. This can lead to a wide range of signs and symptoms resulting from the neurological damage and inflammation the worms cause; in severe cases it may lead to coma and death. The bacterial microbiota in the gut of the snail host may be important in influencing the likelihood of infection of the host by the parasite. Therefore the goal of this project was to characterize the gut microbiome of two snail species, Lissachatina fulica and Parmarion martensi, to determine whether their microbiomes differed and whether being infected by A. cantonensis led to a change in their microbiomes. The snails were collected in Heeia, island of Oahu, Hawaii. Molecular screening for A. cantonensis infection permitted selection of 20 infected and 17 uninfected L. fulica and 13 infected and 20 uninfected P. martensi for analysis. High throughput sequencing of the gut microbiomes of these snails identified four dominant bacterial phyla: Proteobacteria, Tenericutes, Firmicutes, and Bacteroidetes. However, there were significant differences between the microbiomes of the two snail species and between infected and uninfected individuals. The significance of this project is two-fold. First it has shed light on the gut microbiomes of snails and how biotic interactions may impact them. Second, by increasing current knowledge about the impact of this parasite on its host it may lead to novel methods to hinder the ability of the parasite to effectively complete its life cycle and thereby decrease the number of accidental human infections.
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    Investigating the role of altered immune cell metabolism and inflammation in type 2 diabetes
    (University of Hawaii at Manoa, 2020) Lee, Ashley Mainani ; Dye, Christian ; Biology
    Type 2 diabetes (T2D) is a worldwide epidemic, with a disproportionate affect on the Native Hawaiian population and economy in Hawaiʻi. Inflammation associated with obesity is a major risk factor in T2D and often preceeds T2D, but the role of inflammation and its link to altered immune cell metabolism and inflammation in T2D is unknown. Thus, there is a need to characterize and understand the inflammatory mechanisms contributing to metabolic abnormalities in T2D. Herein, we characterized and compared innate immune cell metabolism of lymphocytes (T cells, B cells, NK cells) and monocytes in PBMCs and phenotyped inflammatory immune CD4+ and CD8 + T cell subset populations (senescent, naïve, central, intermediate, and effector) in individuals with T2D and controls. First, to characterize and compare immune cell metabolism in individuals with T2D, we compared the metabolic profiles of T2D and healthy control peripheral blood mononuclear cells (PBMCs) by measuring glycolysis and mitochondrial respiration rates on the Agilent Seahorse XFe96 Analyzer. Individuals with T2D showed abnormal altered metabolic function of immune cells by decreasing glycolysis and increasing oxidative phosphorylation in PBMCs. Second, to phenotype and quanitfy the immune cell subset populations in individuals with T2D, we used a BD LSR Fortessa flow cytometer capable of detecting fluorochrome-conjugated antibodies specific for markers of particular immune cell subsets. Inflammatory CD8+ central memory T cells were increased and CD8+ naïve T cells were decreased in T2D compared to controls. In summary, this project helped fill a gap in knowledge on the role of altered cellular metabolism and the immune cell types that may contribute to disease. Moreover, this novel information may be relevant for improving the treatment and management of T2D.
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    Characterization of the Root-Nodulating Rhizobia Associated with Native and Non-Native Legumes in Hawai‘i and Their Potentials for Restoration of Native Wiliwili (Erythrina sandwicensis) Trees
    (University of Hawaii at Manoa, 2020) Abe, Jon ; Nguyen, Nhu H. ; Biology
    Legumes are highly nutritional, pod-bearing plants that account for more than a quarter of all agricultural crops. In addition to being a staple of global diets, they are feasible alternatives to nitrogenous fertilizers as they form a symbiotic relationship with rhizobia. Through biological nitrogen fixation, rhizobia are able to transform N 2 gas into ammonia, a readily accessible form for living organisms. Because nodulation of legumes is important for both agricultural and native plants, it is imperative to understand which species of rhizobia associates with which legumes. My objective in Part I of this project was to identify native and non-native legume- rhizobia associations by sampling the nodules of 22 legume species across the island of ‘Oahu. Following the isolation and culturing of the corresponding rhizobia within these nodules, molecular techniques were used to isolate, amplify, and sequence the DNA. Nodules from 51 different leguminous individuals, including 22 different species, were sampled, yielding 20 different rhizobia strains from four different genera. On the island of ‘Oahu, Bradyrhizobium sp. strain c020_F was found to be a generalist, forming nodules on nine different leguminous species. My objective in Part II of this project was to develop an Erythrina sandwicensis (wiliwili) restoration protocol using the two strains of Bradyrhizobium isolated from its roots in Part I. This protocol tested the feasibility and effectiveness of sowing seeds into soil inoculated by a rhizobia slurry of 10 6 rhizobia/mL. After three months, nodulated wiliwili seedlings had a significantly larger biomass than non-nodulated seedlings. Both strains of Bradyrhizobium had a nodulation rate of 80%; however, there were no significant differences in biomass between the two strains. Results of this experiment suggest that soil inoculation of wiliwili seedlings yields larger, healthier individuals and should be considered in future restoration protocols.
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    Bacterial Communities of Invasive Aphids Co-occurring on Novel Host Plants on Oahu
    (University of Hawaii at Manoa, 2020) Ota, Elissa ; Chong, Rebecca ; Biology
    Aphids have had invasive success in Hawaii that has been facilitated by rapid asexual reproduction. Another contributor to the invasive success of aphids in Hawaii involves the symbiotic interactions between aphids and their obligate and facultative endosymbionts, which are intracellular bacteria that dwell within hosts and constitute microbial communities. The obligate endosymbiont of aphids, Buchnera aphidicola, provides nutrients needed for the aphid’s survival and reproduction, while facultative endosymbionts may confer novel traits to the aphid. As both obligate and facultative endosymbionts affect aphid fitness, the composition and diversity of the aphid bacterial community may be important to the invasive success of aphids. An examination of the bacterial communities of two invasive aphid species across two novel host plants on the Island of Oahu was conducted to resolve the differences between aphid bacterial communities across aphid species, host plants, and sampling localities. The diversity within each aphid’s bacterial community did not significantly vary across aphid species, host plants, nor sampling locality. When assessing the diversity across the bacterial communities of multiple aphid individuals, bacterial communities differed significantly across aphid species as predicted since both obligate and facultative endosymbionts undergo vertical transmission from mother to clonal offspring. B. aphidicola was present in every sample, but the abundance of B. aphidicola did not vary across host plants. Additionally, facultative endosymbionts including Pseudomonas, Chryseobacterium, Wolbachia, Rickettsia, and Sphingomonas were discovered in some of the aphids sampled. Overall, this study concludes that aphid species is the most prominent factor that shapes the aphid bacterial community.
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    A Retrospective Case Study of Surf Injuries on the Hawaiian Islands
    (University of Hawaii at Manoa, 2019) Guzman, Adam ; Johnson, Sidney ; Biology
    Surfing is a popular leisure activity that comes with a high risk of injury that may participants fail to mitigate. The following research examines surfing injuries within the Hawaiian Islands and explores ways that these injuries can be prevented. It is hypothesized that surfing on a reef or sand break will be predictive of severe injuries to the head and face region and producing an innovative surf helmet would reduce these injuries. A retrospective review of the trauma registry from the main trauma center in Honolulu, Hawaii was conducted on patients admitted between January 1, 2014 and December 31, 2018. 143 patients were included and analyzed. The majority were adult (89.5%), male (85.3%), non- residents (53.2% = 37.8% U.S. Non-residents + 15.4% International). Spine (35.1%), face (17.5%), and head (12.3%) were the most common regions injured. The types of injuries included fractures (59.4%), lacerations (28.7%), contusions (22.4%), concussions (4.9%) and incomplete and complete quadriplegia (7.00%). Sustaining an injury on Maui was predictive of an overall severe injury, and particularly, severe spinal injury, as was being a patient from the mainland United States. Those who sustained severe spinal injuries were likely to remain in the hospital longer than most patients. Those patients who were severely injured were more likely to be discharged to a rehabilitation facility. Community awareness can be addressed to take precautionary measures with surfers in understanding the risks before partaking in the sport, especially for those visiting the Hawaiian islands.
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    The Impact of the Updated Cervical Cancer Screening Guidelines in Women >65 Years of Age: A 10-Year Retrospective Analysis
    (University of Hawai'i at Manoa, 2019) Sutton, Cori ; Tauchi-Nishi, Pamela ; Carlie, Robert ; Biology
    Cervical cytology screening is no longer recommended in women 65 years or older, based on the joint guidelines established by the American Cancer Society (ASC), the American Society for Colposcopy and Cervical Pathology (ASCCP), and the American Society for Clinical Pathology (ASCP) in 2012. This was reaffirmed by the United States Preventive Task Force guidelines in 2018. We examined the effect of implementing these guidelines in this elder population, in order to determine the number of abnormal Pap tests, precancerous lesions, and cancers that would have been missed if screening is discontinued. This retrospective study examined all Pap tests at the Queens Medical Center/Hawaii Pathologists Laboratory between January 2002 and December 2011, in order to determine the prevalence of abnormal Pap tests, their correlation with histologic diagnoses, and positive predictive values (PPV) for cervical intraepithelial neoplasia (CIN) 2 or higher. Chi-square testing was performed in order to detect statistically significant differences between findings in patients 65 years and older, compared to the general population (GP). Cases of malignancy in the >65 year older age group were further analyzed to determine whether the patients exhibited criteria for exclusion from cervical cancer screening, i.e. three consecutive negative Pap tests or two consecutive negative co-tests within the past ten years, with the most recent test having been performed within the past five years; no history of CIN 2 or higher; and no symptoms of abnormal bleeding at the time of Pap testing. From 2002-2011, a total of 1,026,470 Pap tests were examined. 92,247 (10%) were from the >65 year old age group. There were statistically significantly more cancers and glandular lesions found in the Pap tests of >65 year old patients compared to the GP. In contrast, more squamous intraepithelial and atypical squamous lesions were found in the GP compared to the older population. The PPVs for CIN 2 or higher were statistically significantly higher in the >65 year old age group compared to the GP for glandular abnormalities, including for atypical glandular cells of undetermined significance (AGUS) and adenocarcinoma. In contrast, there were no statistically significant differences between the two populations in the PPVs for CIN 2 or higher among any of the squamous lesions. Of the 92,247 patients aged 65 years or older, a total of 135 histologically confirmed cases of precancerous or cancerous lesions were found. If the new 2012 guidelines had been employed, 11 (8%) of these cases would have been missed, including 5 (45%) high grade squamous intraepithelial lesions (HSIL) and 6 (55%) endometrial carcinomas. Although Pap testing is not considered an effective modality for the detection of endometrial cancer, the cessation of cervical cancer screening in patients 65 years or older may lead to the delayed detection of this malignancy.