2018 Summer Student Fellows

Student Fellow: Jesse Birsching, Optometry
Faculty mentor: Dr. Amy Dinardo, Optometry
Project: Not a Dry Eye in the House?: A Clinical Analysis of the Meibomian Gland Structure and Function in Patients Without Dry Eye Disease

Dry eye disease (DED) is a multifactorial disease affecting millions of people, and it is believed that a majority of these cases result from dysfunction of the Meibomian glands, secretory oil glands in the eyelids. Originally it was thought that morbidity was highest for the patient population 50 years of age and older. However, eye care professionals are beginning to notice a large increase in DED in the younger population. Since the majority of previous research has been conducted on the older population, it is of interest to investigate DED in those under 50 years old. The primary interest of this study is to investigate the relationship between Meibomian gland morphology or atrophy, cosmetic use, contact lens use, and the markers associated with DED in subjects between 18-40 years old. Participants completed the Ocular Surface Disease Index (OSDI), a survey commonly used in DED diagnosis, and a survey on cosmetic use, contact lens use, and general lifestyle. Non-invasive infrared imaging of the Meibomian glands was also conducted. This study was unable to significantly relate Meibomian gland morphology or atrophy to cosmetic use or contact lens use in this age range. It is of note, though, that both gland loss and tortuosity (twisting) between the upper/lower lids differ and are statistically significant (p=0.0157 and p<0.001 respectively). Many eye care professionals only assess the lower lids upon examination, and this study shows that it may be of benefit to assess all four lids for differing morphology and gland loss.

Student Fellow: Andrew Bobian, Pre-Pharmacy
Faculty mentor: Dr. Dan Adsmond, Arts & Sciences, Physical Science
Project: Using Amidines as Centerpieces of Ternary Cocrystal Design

A cocrystal is a crystal encompassing two or more different compounds hydrogen bonding to one another. In ternary cocrystal design, a central compound with two hydrogen bonding sites is combined with two different compounds also capable of hydrogen bonding, possibly bringing three distinct compounds into a single crystal. The goal of this project was to explore the possibility of using amidines as the central molecule in ternary cocrystal design, specifically sulfa drugs and pyrimidines. 115 experiments were run, testing the ability of each compound to form binary and ternary cocrystals. Before running experiments with our chosen amidines, I first ran solubility tests on all the compounds we would be using. Cocrystal formation is more likely to occur when the compounds have similar solubilities in the solvent used, so running solubility tests helped to determine which solvents I would use with different compounds. Experiments were initially ran on only the central amidine with the other compounds individually. This was to make sure the central amidine could form cocrystals with each respective compound before putting all three together. While no new ternary cocrystals were discovered, a possible 5 new binary cocrystals were found.

Student Fellow: Roan Fahey, Biology
Faculty mentor: Dr. Christopher Westerkamp, Arts & Sciences, Biological Sciences
Project: Effect of Chronotype and Time of Day on Exercise Performance

The purpose of this project was to study if there is an effect of chronotype (a preference to sleep and wake at different times) on the time-of-day exercise performance of humans. To determine chronotype, a morning/evening person survey was used to sort the subjects into AM, PM, and neither AM/PM groups. Subjects performed a maximal graded exercise test once in the morning and on a separate day in the evening. The test directly measured the lactate threshold (LT, blood lactate of 4-mmol·L-1) at a given velocity. Other data collected was aerobic capacity (VO2 max), heart rate, and ventilation using a heart rate monitor and a metabolic cart. From this data, we were able to determine each subjects’ efficiency of movement and their maximal aerobic capacity. We hypothesized that there was an effect of chronotype and time of day on exercise performance. We further hypothesized that the PM group and neither group will have a better performance compared to the AM group in the evening, and that the AM group will have a better performance compared to the PM group and the neither group in the morning test. We did not find and significant differences between time of day for LT, VO2 and running velocity.
Furthermore, while sample sizes are inadequate, there are likely not significant differences between chronotypes and time of exercise. These findings suggest that time of day and chronotype did not have an effect on LT, VO2, or running velocity.

Student Fellow: Breanna Frasher, Public Health
Faculty mentor: Dr. Michael Reger, Health Professions, Public Health
Project: Phytoestrogen Intake and the Risk of Total and Advanced Colorectal Cancer

Colorectal cancer is the third most common cancer diagnosed, and the third leading cause of cancer related deaths among men and women in the U.S. It is understood that some non-modifiable risk factors such as age, race, and family history affect the development and outcome of colorectal cancer, but less exploration has taken place regarding modifiable risk factors. The dietary intake of phytoestrogens is believed to be one of these modifiable risk factors. Because of their biological similarity to human estrogens and their ability to bind to the receptors, these phytoestrogens are thought to affect the progression of colorectal cancer. The objectives of the current study are to 1) evaluate the association between dietary intake of phytoestrogens and risk of total colorectal cancer, as well as advanced colorectal cancer and 2) differentiate between risks of colorectal cancer associated separately with the intake of 6 main phytoestrogens. The data used in the study was pulled from the PLCO dataset, which hosts an intervention arm of 58,477 subjects. These subjects were further separated into quintiles based on their dietary intake of phytoestrogens. After adjusting for variables like race, age, BMI, alcohol intake, smoking status, and family history, hazard ratios were calculated for intake of separate phytoestrogens and effect on colorectal cancer. Statistically significant values were recorded for total isoflavone intake, and for the individual phytoestrogens genistein and glycitein.

Student Fellow: Zackary Hren, Chemistry
Faculty mentor: Dr. Luis Rivera, Arts & Sciences, Physical Sciences
Project: Pressure Effects on the Relaxation of an Excited Ethane Molecule in an Argon Bath

Understanding the mechanisms of vibrational energy transfer within and among molecules is fundamental to understanding many physical and chemical processes such as pyrolysis and combustion. There are two primary pathways by which a vibrationally highly-excited molecule can release its energy: reaction or relaxation. Little work has given attention to understanding vibrational energy relaxation at high pressures, where the isolated binary collision approximation breaks down. In order to study high-pressure vibrational energy relaxation, molecular dynamics simulations were run where an excited ethane molecule and 1,000 argon atoms were placed in a periodic cubic box. The system comprising the cubic box was allowed to reach thermal equilibrium, and the energy transfer from the excited molecule to the bath gas was calculated. After separating vibrational and rotational energies, it was found that the rotational energy decay is ~49 times faster than the vibrational energy decay. The decay of both vibrational and  rotational energies is characterized by poly-exponential behavior. Furthermore, molecular relaxation rates seem to increase proportionally with an increase in pressure up to quite high pressures (~250 atm) but begin to deviate from proportionality at greater pressures (> 250 atm). Non-binary collisions and bath gas clustering appear to be responsible for this deviation, which indicates a breakdown in the isolated binary collision approximation.

Student Fellow: Tayler Jones, Pharmacy
Faculty mentor: Dr. Kim Hancock, Pharmacy, Pharmaceutical Science
Project: Dissolution of Antidepressant Medications in a Vertical Sleeve Gastrectomy Dissolution Simulation

According to the National Center for Health Statistics, the prevalence of obesity in the United States was 39.8% of adults in 2016. With the rate of obesity rising, patients are searching for all available weight loss options. Along with diet and exercise, bariatric surgery may be a solution for some patients. Out of all the bariatric surgery procedures, vertical sleeve gastrectomy (VSG) is the most popular. VSG changes gastric physiology, creating a new gastric environment with a pH near 5.8 and a volume of 100 mL. Volume and pH are believed to have an effect on the dissolution of drugs; limited research has been done to look at the effects of VSG on drug dissolution. Two antidepressants, sertraline and venlafaxine, were selected as drugs of interest due to high incidence of use in bariatric patients pre and post-surgery. Dissolution tests were performed simulating both a non-surgical patient, who has low gastric pH and large stomach volume, and a VSG patient who has higher gastric pH and smaller stomach volume. For venlafaxine, volume had a significant effect on drug dissolution and pH may play a role in small volumes. For sertraline, only pH had a significant effect on drug dissolution. Both drugs saw a decrease in dissolution in the VSG dissolution model as compared to the non-surgical dissolution model; this could explain lower drug concentrations seen in the clinical setting.

Student Fellow: Samuel Joseph, Pre-Pharmacy
Faculty Mentor:  Dr Luis A. Rivera, Arts & Sciences, Physical Sciences
Project: A Molecular Dynamics Study of an Order-Disorder Transition in a Small Confined System

Molecular Dynamics simulations were used to investigate an order-disorder transition in a small confined system. Using the LAMMPS program, particles were placed in a cuboid box with confining walls perpendicular to X (located at X = 0 and X = 6.25) and periodic boundary conditions in the Y and Z directions (Y = Z = 20). Simulations were performed for 2,000 particles at a density of 0.8, and various values of temperature (T = 0.3 to 10). After thermal equilibration, the particles settled into layers parallel to the confining walls. Then the heat capacity was calculated to determine a baseline for when a quasi-phase transition would take place. The transition temperature is indicated by a spike in the heat capacity vs. temperature curve. Once the heat capacity was calculated, the X-density profile and the 2-D Radial Distribution Function were calculated to form a clearer picture of the behavior of the particles in the layers parallel to the wall. A quasi-phase transition was not observed for simulations of 8,000 particles at density of 0.8.

Student Fellow: Micaela Rice, Biology/Pre-Med
Faculty Mentor: Dr. Dan Adsmond, Arts & Sciences, Physical Sciences
Project: Investigation of Amide Coformets for Sulfa Drug Cocrystals

Cocrystallization is a process that can occur when hydrogen bonds form between an acceptor atom of one compound and a donor atom on a different compound. Cocrystal formation, which brings different types of molecules together in the same crystal, causes changes in the physical properties of the reactant compounds, which makes them of great interest to the pharmaceutical industry. In this project, eleven different amides and four different sulfa drugs were combined in 1:1 and 2:1 molar ratios in order to test each amide’s ability to form cocrystals with sulfa drugs. Each compound was measured out separately, dissolved in warm solvent (acetonitrile), filtered into a third beaker, and covered with aluminum foil. Holes in the aluminum foil allowed the solvent to evaporate at a constant rate, resulting in crystals precipitating out of the solution. These crystals were analyzed by NMR (nuclear magnetic resonance) which showed the ratio of components in the cocrystal and IR (infrared) spectroscopy, which showed changes in hydrogen bonding in the crystal. Crystals that contained both compounds in simple whole number ratios by NMR and also showed significant movement in the N-H peaks in both the amide and sulfa drug in the IR were likely cocrystals. In the end, out of 11 combinations, sulfamethazine formed 9 cocrystals, sulfamethoxypyridazine showed 5, sulfameter formed 2, and sulfathiazole showed none.

Student Fellow: Gregory Shaffer, Pharmacy
Faculty mentor: Dr. Jennifer Lamberts, Pharmacy, Pharmaceutical Sciences
Project: Understanding the Role of Neuroinflammation in Parkinson’s Disease

It is estimated that 7-10 million people worldwide, including 1 million Americans, have Parkinson’s disease (PD). Symptoms including tremors, unstable posture, stiff or weakened muscles, and difficulty with fine motor skills severely affect patients’ quality of life. While the cause of PD is unknown, it is believed that neuroinflammation (inflammation of the brain) plays a role in the progression and severity of the disease. While limited inflammation can be helpful in fighting off infections, chronic inflammation is harmful to the brain and can contribute to worsening of the disease. In PD, chronic neuroinflammation starts with activation of microglial cells, the resident immune cells of the brain. Once they become activated, they release inflammatory factors, which have damaging effects on neurons. The goal of this project was to create a model of neuroinflammation in the lab to further understand this process. To do this, we separately cultured both microglial cells and neuronal cells and exposed the cells to inflammatory/toxic substances. We then measured the effect of these substances on cell health. Rotenone, a neurotoxin, killed more neurons as exposure time and toxin concentration increased. Treatment of microglial cells with LPS, a bacterial endotoxin, caused an increase in the release of inflammatory factors with increased concentration and time. Lastly, we tested the effect of naltrexone, a drug with anti-inflammatory properties, in these models. Ultimately, we hope to discover potential drug targets that can stop or reduce this process of neuroinflammation in PD. 

Student Fellow: Heidi Stultz-Griffey, Pharmacy
Faculty mentor: Dr. Qian Ding, Pharmacy, Pharmaceutical Sciences
Project: Prevalence of Antipsychotic Medication Prescribing Errors for Elderly Patients Based on Medicare Claims Data

The population of the U.S. is getting older.  By the year 2060, it is projected that 25% of the population will be over the age of 65.  This increase is expected to bring with it an increase in the frequency of neurological disorders that have the ability to cause behavioral disturbances.  While antipsychotics have been shown to aid with the suppression of behavioral disturbances in these patients, their use is not recommended by the BEERS criteria due to a risk of increased mortality.  The prevalence of inappropriate use of antipsychotics in elderly patients was evaluated via data from the Medicare Current Beneficiaries Survey (MCBS).  A secondary objective of the study was to identify whether general practitioners or specialists were responsible for a greater fraction of inappropriate prescribing cases.  Patient encounters where at least one antipsychotic medication was prescribed were extracted, and data regarding patient diagnosis was analyzed via chi-squared test and two-sample t-test.  It was found that only 0.53% of evaluated claims had a diagnosis consistent with appropriate use, while 99.47% of cases were considered to be inappropriate.  Patients prescribed antipsychotics inappropriately were statistically significant more likely to be white (89.62% vs. 80.24%, p=0.000) and female (70.39% vs. 53.89%, p=0.000).  In addition, general practitioners were found to be responsible for 71.51% of claims in at least one antipsychotic was prescribed, while specialists were responsible for 4.22% of claims.

Student Fellow: Megan Van Baren, Pharmacy
Faculty mentor: Dr. Felix Amissah, Pharmacy, Pharmaceutical Sciences
Project: Impact of Cyclooxygenase Inhibitor/Polyunsaturated Fatty Acid Combination on Rho GTPase Function in Lung Cancer Cells

Lung cancer is the second most common cancer and leading cause of cancer-related death in the US. Although cessation of tobacco use decrease lung cancer incidence, after smoking cessation former smokers are still at considerable risk of developing lung cancer. This population could greatly benefit from chemoprevention. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), are polyunsaturated fatty acids (PUFAs) found in fish oils are known to possess an anticancer effect. Cyclooxygenase (COX) inhibitors, aspirin and diclofenac, have also been shown to exhibit anticancer effects. We examined whether combining a COX inhibitors with PUFAs would enhance their anticancer activity.  The key proteins involved in abnormal signaling in lung cancer are the Ras superfamily of GTPases, which include Rho family of GTPases (Cdc42, Rac1 and RhoA). These GTPases are involved in regulation of cell proliferation, survival and metastasis. In this study, we treated A549 lung cancer cells with either aspirin or diclofenac in combination with PUFAs and conducted cell proliferation assay, viability assay, western blot, and immunofluorescence. Our results indicate that the combination of a COX inhibitor and PUFA were more effective in preventing cancer cell proliferation and migration than PUFA or COX inhibitor alone. Such treatments also reduced the expression and altered the intracellular distribution of Rho GTPases.

Student Fellow: Jacklyn Zvonar, Biology/Pre-Medicine
Faculty mentor: Dr. Changqi Zhu, Arts & Sciences, Biological Sciences
Project: Mechanistic Study of the Role of Atg Genes in Fruit Fly Lifespan Regulation through Adult Muscle Tissues

Autophagy-related genes (ATG) are known to regulate autophagosomes in fruit fly tissues. ATG genes are well conserved evolutionarily from yeast, to fruit flies, and to humans. Our lab previously performed aging studies in which we found that over-expression of wild-type ATG1 gene in adult fruit fly muscle tissues can significantly shorten the lifespan, while knocking-down the levels of the transcripts of individual ATG1, ATG8a, and ATG18 genes in adult fruit fly muscle tissues has prolonged the lifespan of these fruit flies. To explore the molecular and cellular mechanisms behind these results, we dissected control and experimental adult fruit fly muscle tissue and performed three different stainings: FK2, anti-LAMP1, and an antibody against Caspase-3.  FK2 staining results indicate that autophagy was indeed increased with ATG1 expression. However, Caspase-3 staining results indicate that apoptosis does not seem to be involved in the early death of the ATG1-expressing flies as these experimental flies showed no significant difference from the control flies.