Life Sciences Commons^™

Impact Of Lyophilization On Porcine Hemoglobin Properties., Mustafa Almosawi

College of Arts & Sciences Senior Honors Theses

Blood transfusion is the single most often performed lifesaving procedure in hospitals worldwide. Unfortunately, packed red blood cells (RBCs) used for transfusion can only be stored for 42 days at 4 °C before being discarded due to irreversible damage that occurs during storage. Any reduction in available RBCs for an extended period can lead to blood shortages. To increase the shelf-life of RBCs, we investigated freeze-drying (lyophilizing) in the presence of the non-toxic sugar trehalose as a method for long-term preservation. However, the oxidative stress of the lyophilization and storage processes can compromise the functionality of these cells, and the …

The Emerging Oral Pathogen, Filifactor Alocis, Modulates Antimicrobial Responses In Primed Human Neutrophils., Ian J. Snider

College of Arts & Sciences Senior Honors Theses

Periodontitis is an irreversible, chronic inflammatory, infectious disease of the oral cavity that affects approximately half of all adults 30 years or older in the USA. The oral cavity is under high immune surveillance because of its constant exposure to microbes in the environment. The primary immune cell responsible for this surveillance is the neutrophil. Pathogens associated with periodontitis possess virulence factors and have evolved strategies to evade neutrophil antimicrobial responses to survive. One such pathogen is Filifactor alocis, whose presence is associated with the progression of periodontitis. F. alocis manipulates several neutrophil antimicrobial functional responses to avoid killing, an …

Impacts Of Dietary Restriction On A Drosophila Model Of Werner Syndrome, Eileen Sember

College of Arts & Sciences Senior Honors Theses

Werner syndrome (WS) is an autosomal recessive disorder that results in premature aging and occurs in 1 in 1,000,0000 to 1 in 10,000,000 people. In humans, WS is the result of mutations that render the WRN gene, that contains a helicase and an exonuclease domain, non-functional. Currently, there is no cure for WS in humans, making dietary and lifestyle interventions attractive for increasing the quality and longevity of lives. Diet restriction (DR) has been shown to extend the lifespan of several model organisms, including Drosophila melanogaster, making it a strong candidate for WS treatment. In this thesis, mutant flies …

The Effects Of Ppal-1 In Arabidopsis Gamete Development., Amanda White

College of Arts & Sciences Senior Honors Theses

Prenylation is a type of post-translational modification in which a 15- or 20-carbon lipid is added to the carboxyl (C) terminus of the protein. Arabidopsis thaliana contains the PROTEIN PRENYLTRANSFERASE ALPHA SUBUNIT-LIKE (PPAL) gene, which encodes a protein with homology to the α-subunits of the three known prenylation enzymes, PFT, PGGT, and Rab-GGT. We previously identified two mutations in PPAL, one of which is ppal-1, which contains a T-DNA insertion in the fourth intron. We have previously observed that self-fertilizing heterozygous ppal-1 plants produce progeny in which homozygous ppal-1 is underrepresented. This project attempts to ascertain …

Phytochemical Content And Anti-Breast Cancer Activity Of Kentucky Native Plants., Nicole A. Mcgrath

College of Arts & Sciences Senior Honors Theses

Plants are a proven and rich source of chemical diversity for the development of pharmaceuticals. Plants that are native to Kentucky are an underexplored resource that could lead to advances in treating human diseases such as cancer. Kentucky native plants, especially those with highly diverse or enriched chemical compositions are possible leads for therapeutic drug development. To explore the potential of native Kentucky plants for treatment of breast cancer, a list of prospective target plants was developed based on literature reports. This list was narrowed to 10 tissues for further in-depth chemical analysis including estimation of alkaloids, terpenes, and phenolics, …

Relationships Of Protein Biomarkers Of The Urokinase Plasminogen Activator System With Expression Of Their Cognate Genes In Primary Breast Carcinomas., Seth B. Sereff

College of Arts & Sciences Senior Honors Theses

Background: Urokinase plasminogen activator (uPA), its receptor uPAR and serine protease inhibitors PAI-1 or PAI-2 play key roles in tissue membrane remodeling and invasion of basement membranes by induction of a fibrinolytic pathway. Earlier studies reported that uPA and PAI-1 protein levels in breast carcinomas assist in prediction of response to chemotherapy. Our goal is to develop molecular signatures of candidate genes and identify novel relationships with these four protein biomarkers that demonstrate clinical utility for assessment of breast carcinoma outcomes.

Methods: This retrospective study used de-identified biomarker results and clinical outcomes from primary breast cancers that were stored in …

A Bug Eat Bug World : Does Urbanization Decrease Survivorship Of Pollinators., Amy L. Cherry

College of Arts & Sciences Senior Honors Theses

Human impact on the environment is responsible for the sixth mass extinction on Earth as well as rapid population declines in many taxa. Land use change, via increasing urbanization and intensity of agricultural practices, is the most important aspect of human impact on the environment, and it is associated with population decline and extinction in several taxa, but especially in pollinator species. Pollinators, which are essential for crop production, are experiencing declines as a result of the loss of habitat and host plants associated with urbanization. Urbanization negatively affects pollinators, but in the wake of lost naturalized habitat, some researchers …

Pparγ And High Molecular Weight Adiponectin In White Adipose Tissue In An Obesity Model., Katelyn M. Mcdowell

College of Arts & Sciences Senior Honors Theses

Adiponectin is a protein secreted from adipose (fat) tissue. It is secreted into the blood as either a trimer (low molecular weight), hexamer (middle molecular weight) or an 18-mer (high molecular weight). Adiponectin, particularly the high molecular weight (HMW) isoform, has been shown to increase insulin sensitivity, which is important for maintaining healthy blood glucose levels. Adiponectin levels have been shown to be decreased in individuals who are obese, and this may play a role in the diabetes which often accompanies obesity. PPARγ is a protein that is highly expressed in white adipose tissue and is responsible for regulating portions …

Indole Sensitivity In Various Insect Herbivores., Rakhi Patel

College of Arts & Sciences Senior Honors Theses

When plants in nature are attacked by herbivores, they frequently release airborne volatiles. These herbivore-induced plant volatiles (HIPVs) help plants defend themselves against herbivore attack by calling in natural enemies, but HIPVs may also be directly toxic to herbivores. Indole (C₈H₇N) is a common HIPV that is toxic to caterpillars, but its relative toxicity to different caterpillars has not been explored. I tested the toxicity of indole on six caterpillar species (Spodoptera frugiperda, Trichoplusia ni, Spodoptera exigua, Anticarsia gemmatalis, Heliothis virescens, and Helicoverpa zea). Results show that indole is …

Dsba-L Protein Levels In White Adipose Tissue In An Obesity Model., Madison E. Kerley

College of Arts & Sciences Senior Honors Theses

Adiponectin is an adipose tissue-secreted protein found in three isoforms. Disulfide bond A oxidoreductase-like protein (DsbA-L) is thought to assist with protomeric disulfide bonding to generate the HMW (high molecular weight) isoform, which regulates insulin sensitivity. Both adiponectin and DsbA-L were previously found to negatively correlate with obesity.

Immunoblots were conducted on adipose tissue samples from male mice fed a high or low-fat diet for 6, 10, or 16 weeks. Immunoblots from high-fat diet-fed mice revealed double bands for DsbA-L. High and low molecular weight bands were analyzed together, revealing significantly higher relative band densities with 10 and 16-week high-fat …

Bipolar Cell Dendritic Morphology Analysis In Mice With Model Retinitis Pigmentosa., Simra Ahmed

College of Arts & Sciences Senior Honors Theses

No abstract provided.

Filifactor Alocis, A Newly Appreciated Oral Pathogen, Fails To Induce The Respiratory Burst Response Of Human Neutrophils., Jacob S. Edmisson

College of Arts & Sciences Senior Honors Theses

Almost 50% of adult Americans suffer from periodontitis which is a bacterially induced inflammation of the tissue that surround and support the tooth. The accumulation of neutrophils, a critical cell component of the innate immune system, in the gingival crevice contributes to tissue damage. Filifactor alocis is a newly appreciated pathogen present in oral biofilms at periodontal disease sites. Studying the interactions between neutrophils and F. alocis will provide valuable information for delineating the role of this bacterium in periodontal disease and enhance our understanding of bacterial strategies to evade leukocytes’ antimicrobial mechanisms. The hypothesis that F. alocis modulates human …

An Investigation Into The Adiponectin Receptor Levels In Cardiac, Liver And Skeletal Muscle Of Mature Rats., Jacqueline Sippel

College of Arts & Sciences Senior Honors Theses

Both obesity and diabetes are domestic and international health problems. Adiponectin is a protein secreted from adipocytes and other tissues such as muscle which influences carbohydrate and lipid metabolism. For this reason, Adiponectin is a primary target for therapeutic treatment of metabolic diseases. Adiponectin interacts with receptors termed AdipoR1 and AdipoR2, which are structurally similar to G protein-coupled receptors except with a reverse membrane topology. The aim of this study is to determine the levels of AdipoR1 and AdipoR2 within cardiac, skeletal, and liver tissue samples of mature non-obese rats. The objective of determining potential differences in the tissue levels …

Antibiotic Resistant Bacteria In Natural Environments Within Kentucky., Steve Tran

College of Arts & Sciences Senior Honors Theses

Rates of antibiotic resistance in bacteria are rapidly rising; this is, in part, due to overuse of antibiotics resulting in a great burden on the U.S. healthcare system. With the rise of resistant bacteria, a large-scale outbreak of antibiotic resistant bacterial infections may also occur. Research on developing antibiotics has also decreased. Thus healthcare is at a great disadvantage in the arms race against bacteria. The environmental reservoirs of antibiotic resistance are currently being explored in Kentucky, in which bacteria have been collected from a broad spectrum of natural environments to analyze their antibiotic resistant capabilities and their interactions with …

Movement Patterns Of The Cave Salamander (Eurycea Lucifuga) In Sauerkraut Cave, Kentucky., Paige N. Wilson

College of Arts & Sciences Senior Honors Theses

The Cave Salamander, Eurycea lucifuga, is a common but poorly understood species of salamander that inhabits limestone features and karst systems across the eastern and midwest United States. As a species that can bridge the gap between subterranean and epigean environments, E. lucifuga may behave as a top predator and act as a significant facilitator in the flow of energy from the outside environment into the dark zone. The goal of this study was to document the seasonal movements of a local population of cave salamanders in Louisville, Kentucky, and the relationship between those movements and abiotic factors. Secondarily, …

Habitat Effects On Blood Adiponectin Isoforms In Black Bears., James-Dean Wood

College of Arts & Sciences Senior Honors Theses

No abstract provided.

Syndecan-1 Tagged Liposomes As A Theranostic Nanoparticle For Pancreatic Adenocarcinoma., Wenyuan Yin

College of Arts & Sciences Senior Honors Theses

Theranostic nanoparticles are emerging as a novel mechanism for detecting and treating cancer. Due to the difficulties in detection and treatment of pancreatic cancer, these particles could serve within this unique niche. In this study, a Syndecan-1 ligand was utilized to increase tumor specificity of fluorescent dye encapsulated liposomes which were evaluated as a potential theranostic nanoparticle for pancreatic adenocarcinoma. Their diagnostic capabilities and specificity to pancreatic adenocarcinoma were determined in vitro using immunocytochemistry and in vivo using multi-spectral optoacoustic tomography (MSOT). Immunocytochemistry showed that liposomes preferentially bound and released their contents into cells expressing high levels of Insulin-Like Growth …

Role Of Dietary Fatty Acids In Liver Injury Caused By Vinyl Chloride Metabolites In Mice., Heegook Yeo

College of Arts & Sciences Senior Honors Theses

Background. Vinyl chloride (VC) is an environmental pollutant found in many industrial sites and it ranks 4th on the 2013 Hazardous Substances Priority List published by Agency for Toxic Substances and Disease Registry’s (ATSDR). We have previously reported increased hepatocellular necrosis with high occupational VC exposure in human and in in vitro models; however, the effect of subtoxic VC exposure on liver with underlying conditions is unknown. The purpose of the current study is to investigate the hepatic injury caused by chloroethanol (CE; VC metabolite) and inflammatory response in two experimental models of high-fat diet (HFD) induced obesity. Methods. Mice …

The Effect Of Interspecies Signaling On Stenotrophomonas Maltophilia Growth., Zachary Ethan Dickinson

College of Arts & Sciences Senior Honors Theses

Cystic fibrosis (CF) is a genetic disorder that effects over 70,000 people worldwide, and is particularly prevalent in those of Caucasian descent. CF increases susceptibility to chronic infections of the lungs and GI tract, often leading to decreased quality of life and early death to afflicted individuals. No cure is available, and so treatment is limited to treating the frequent infections that afflict CF patients. Several species of bacteria are commonly implicated in these infections, including Stenotrophomonas maltophilia. Infections of this species is of particular interest as it has been shown to be highly antibiotic resistant. This is a concern …

Swimming Behavior In Temperate Forest Ants., Sarah Frances Handlon

College of Arts & Sciences Senior Honors Theses

Most terrestrial arthropods are helpless in water, and falling from a tree into a flooded forest understory should be especially problematic for small, cursorial organisms like ants. Whereas many species of tropical arboreal ants can tread across the water surface (i.e., swim), less is known of this behavior in temperate forest ants. I tested for swimming ability in various ant species collected from tree trunks in Kentucky. Results show that Camponotus pennsylvanicus, and C. nearcticus, are strong swimmers (operationally defined as directed motion at speeds > 3 body lengths per sec.), while Crematogaster ashmeadi, and Monomorium minimum tend to struggle and …

The Effect Of Predator Positioning And Related Angles On The Flight-Initiation Distance And Refuge Choice Of The Eastern Gray Squirrel., Lindsay Nason

College of Arts & Sciences Senior Honors Theses

The purpose of this study was to determine the effect that the relative positions of squirrel, predator, and refuges, as well as the predator’s SPT angle (See Figure 1), had on a squirrel’s choice of refuge and the length of its flight initiation distance. A running person was used to simulate an attacking predator, and the relative positions of squirrel, person, and refuges were measured using both distance and compass angle. Data were collected in September and October 2014 at several urban green spaces in Louisville, Kentucky. The SPT angle measurement was not found to be particularly useful in predicting …

Identification And Expression Analysis Of Sugar Transporters In Microbotryum Violaceum., Jared Morris Andrews

College of Arts & Sciences Senior Honors Theses

No abstract provided.

Epidemiology And Virulence Of Clostridium Difficile : An Evolutionary Perspective., Lindsey E. Hastings

College of Arts & Sciences Senior Honors Theses

The emergence of virulent, antibiotic-resistant Clostridium difficile ribotypes over the past thirty years has greatly concerned both the medical and research communities. As of 2010, C. difficile has surpassed methicillin-resistant Staphylococcus aureus (MRSA) as a cause of nosocomial infections and is now found in both healthcare settings and outside communities in the United States. Attendant-borne transmission in hospitals has been associated with the evolution of increased virulence in other nosocomial pathogens, but evolutionary theory has yet to be applied to the epidemiology and virulence of C. difficile. Differences between nosocomial C. difficile infections (CDI) and community-associated CDI (CA-CDI) mortality and …

Interactions Between Human Neutrophils And Mycobacterium Smegmatis : A Comparative General Analysis., Irina Miralda

College of Arts & Sciences Senior Honors Theses

Mycobacteria smegmatis is an abundant soil and water inhabitant with which humans are continuously in contact. Despite the fact that it is typically considered non-­‐pathogenic, a few rare cases of M. smegmatis-­‐caused infections have been reported and it has been shown that M. smegmatis is able to modulate inflammatory responses in macrophages. Neutrophils are the innate immune system’s first line of centralized defense against invading microbes, especially the frequently encountered M. smegmatis. However, very little information is known of the mechanism by which neutrophils eliminate environmental bacteria. In this study, the hypothesis that M. smegmatis regulates neutrophil functional responses, and …

The Human Immune Response To The Flagellins Of Burkholderia Dolosa., Molly Mcdonald

College of Arts & Sciences Senior Honors Theses

Cystic Fibrosis (CF) is the most commonly inherited lethal disorder in Caucasian populations. Chronic bacterial infection both decreases lung function and lowers the quality of the CF patient’s life. In one of the most recent CF epidemics, Burkholderia dolosa, an organism previously uncharacterized as an epidemic strain, infected over forty patients and led to the death of over seven patients at Children’s Hospital Boston. Preliminary research revealed that this strain of B. dolosa possesses genes for two distinct types of flagella, which is uncommon in bacterial pathogens. In this study we examine the interaction of human cells and B. dolosa, …

Mutant Drought Phenotype Investigation : A Look Into Effects Of Protein Prenylation On Water Utilization In Conditions Of Varying Soil Saturation And Gene Knockout., Casey Rasmussen

College of Arts & Sciences Senior Honors Theses

Plants respond to environmental stress, such as drought, with increases in levels of Abscisic Acid (ABA). This stress response allows greater survival rates but results in the plants producing lower seed yield and biomass. Certain mutant strains of the Arabidopsis plant have been identified with mutations in coding regions involved in the formation of the α and β subunits of PFT and PGGT-I genes involved in protein prenylation. We explored the possibility of these mutant strains to better preserve seed yield and plant biomass under drought stressing conditions in comparison to the wild type (Columbia) strain, based on the mutants’ …

Sinusoidal Endothelial Cell-Derived Extracellular Matrix Regulates Basal And Stimulated Macrophage Activation., Lauren G. Poole

College of Arts & Sciences Senior Honors Theses

Fatty liver disease, be it from alcohol (ALD), obesity (NAFLD) or other sources (e.g. viral infection), share similar mechanisms of disease progression. One potential mechanism is that altered sinusoidal endothelial cell (SEC) extracellular matrix (ECM) favors a more inflammatory phenotype in resident macrophages (i.e., Kupffer cells). Here, the hypothesis was tested that SEC-derived matrices directly affect the inflammatory response of macrophages. Methods. Transformed sinusoidal endothelial cells (TSECs) were cultured for 24 or 72 hours. Culture plates were then washed with a solution that selectively removed the cells, but preserved the ECM. Cultured macrophages (RAW 264.7 cells) were then seeded on …

Targeting The Anaphase Promoting Complex/Cyclosome To Induce Mitotic Arrest In Ovarian Cancer., Douglas John Saforo

College of Arts & Sciences Senior Honors Theses

Taxanes are a class of chemotherapeutic drug that act to disrupt microtubule function and cause mitotic arrest and cell death. These drugs are commonly used for cancer treatment, however their effectiveness is dependent on the presence of a functioning spindle assembly checkpoint (SAC). As a result, it is possible for cancer cells to become resistant to microtubule disrupting drugs by inactivating the SAC. The anaphase promoting complex/cyclosome (APC/C) is an E3 ubiquitin ligase that acts as the master regulator of cell cycle progression. Inhibition of the APC/C should result in disruption of the cell cycle, resulting in arrest during mitosis …