• A Solution to Controlled Substance Abuse: Community Partnerships

      Cook, Laurie B.; Uttaro, Elizabeth; The College at Brockport (2017-04-25)
      Controlled substance abuse is a serious societal issue, particularly at the level of the actual addiction itself. Opioid analgesics are painkillers that are nationally recognized as a major cause of death by overdose. A number of policies and regulations have been put in place to curb addiction, without much success. One potential solution is to strengthen community partnerships between pharmacies, clinics, physicians and other medical care providers such that communication is improved and red flags can be addressed more quickly. These partnerships have more potential than individual parties because information is gathered from multiple different areas of the issue. Using information from multiple aspects of the issue, a solution can be created to reduce addiction at multiple points. Partnerships may be able to help prevent double filling of prescriptions, doctor shopping, and over-prescribing reducing the amount of drugs on the street. By reducing the amount of drugs available on the streets, lower amounts of addiction will occur and hopefully prevent it.
    • An Investigation into the Role of Arf6 in MCH-mediated Rearrangements in 3T3-L1 Preadipocytes

      Cook, Laurie B.; Smich, Bohdan; State University of New York College at Brockport (2020-09-16)
      Obesity has become a pandemic in our society. One potential method to alleviate this crisis is the use of pharmaceutical therapy to manage how our bodies metabolize the energy that we consume. Chronic overconsumption can lead to the activation of inflammatory responses and metabolic dysregulation, such as insulin resistance, and promote the obese condition. The pathways involved in the motility of pre-adipocyte cells are important in understanding how our bodies interpret and react to specific biochemical signals. This research is focused on a pathway that activates the expansion and migration of pre-adipocytes. Melanin-concentrating Hormone (MCH) is a neuropeptide that is known for regulating appetite and metabolism within adipocytes through the G protein-coupled receptor, MCHR1. MCHR1 is known to act through a Gq/PLC pathway to destabilize actin. Our proposed pathway is that MCHR1 activates ARNO, a guanine nucleotide exchange factor that is required in the subsequent activation of ADP-ribosylation factor 6 (Arf6). These events in succession may result in destabilized actin in murine 3T3-L1 pre-adipocytes. A pharmaceutical inhibitor of Arf6, NAV-2729, as well as the overexpression of dominant negative ARNO/Arf6 plasmids were used to determine if Arf6 was indeed a downstream signaling component in this pathway. Fluorescence microscopy was used to visualize the actin stress fibers and morphology of the cell and a scratch wound was performed to determine if the migration rate was affected. Preliminary results from our fluorescence stain show that the structure of actin was affected by NAV-2729 after MCH addition, however, additional trials and analysis are required.
    • Analyzing the Importance of Phosphorylation and Ubiquitination of Amino Acids in the Enhancer of Rudimentary Protein, ERH

      Tsubota, Stuart; Voyer, Amber L.; The College at Brockport (2018-05-04)
      The enhancer of rudimentary gene, e(r), has been found to posses certain amino acids with the potential for phosphorylation or ubiquitination. All of these amino acids are highly conserved among species. It is possible that any of these sites might play a role in Enhancer of Rudimenary Homolog protein, ERH, activation by their respective modifications. This possibility has been analyzed through the creation of transgenes with codon changes at the identified sites intended to either prevent or mimic phosphorylation or ubiquitination. Upon insertion of the e(r) transgenes into the Drosophila melanogaster genome, they have been crossed with two lethal and one low viability stock of flies. The location of the e(r) gene on the X chromosome allows basic phenotypic observations based on eye color to reveal whether any mutations exhibit wild-type ratios of males to females. It has been determined that the phosphorylation of Y19, Y22, S47, and Y92 are not necessary in the activation of ERH. Likewise, the prevention of ubiquitination at K41 and K90 does not effect protein function. A double amino acid change at T18 and S24 in the same transgene has resulted in low viability when compared to the wild-type, suggesting a decreased amount of ERH activity as a result of this mutation in regards to the Notch signaling pathway.
    • Arginine Methylation of TbLpn, a Trypanosome Lipin Homologue, by TbPRMT Enzymes

      Pelletier, Michel; Flint, Alexandra; The College at Brockport (2018-05-04)
      Phospholipids biosynthesis, particularly phosphatidylcholine (PC) and phosphatidylethanolamine (PE) plays a major role in the survival of T.brucei. Of great importance is the fact that, as opposed to other parasitic organisms, trypanosomes synthesize phospholipids de novo. Although the pathways for phospholipids biosynthesis have not been very well characterized, recent data have helped to better understand how trypanosomes are able to assemble phospholipids. Previous work in our lab has shown that a protein, termed TbLpn, is a phosphatidic acid phosphatase potentially involved in phospholipid biosynthesis in T.brucei. In addition, TbLpn contains methylated arginine residues and interacts with T.brucei major Protein Arginine Methyltransferases, TbPRMT7. The major focus of my project is to identify the effect of TbLpn methylation by TbPRMT7 on its enzymatic activity and cellular localization.
    • Building Profiles for miRNA Target Prediction

      Shen, Rongkun; Galbier, Lucas; The College at Brockport (2017-05-09)
      MicroRNAs are very short non-coding RNAs. Since microRNAs play important roles in many biological process, the research of microRNAs is a burgeoning field with much promise. Due to the high cost of experimental approaches, many computational techniques and algorithms have been implemented to study microRNAs. However, current methods for determining the targets for miRNAs are far from accurate. To address this issue, we developed algorithms that produced profiles of miRNA recognition elements and features such binding energy threshold and conservation score. These profiles will be used to train a machine learning algorithm for miRNA target prediction.
    • Characterization and Identification of Bacterial Flora from Infected Equine Hooves

      Pelletier, Michel; Draper, Janna; The College at Brockport (2013-05-10)
      The aim of this research was to characterize and identify the bacterial flora associated with a specific hoof infection in a population of horses in the Rochester NY area. Samples from horses showing symptoms of infection were first grown on Brain-Heart Infusion (BHI) solid medium, a rich medium that allow growth of all bacteria. Forty different bacterial colonies were obtained and characterized microscopically and biochemically. We have identified normal inhabitants of the skin such as Staphylococcus epidermidis, Staphylococcus saprophyticus, Micrococcus varians, and Micrococcus luteus. Bacteria commonly found in soil such as Bacillus megaterium, Bacillus azotoformans, Bacillus insolitus, and Bacillus popilliae were also identified along with Enterococcus faceium, which is found in the gastrointestinal tract of mammals (1, 2, 3). Interestingly bacteria such as Corynebacterium xerosis, known to cause animal diseases were also isolated from infected horses (4). Several of these bacteria were confirmed by sequencing 16S ribosomal DNA. Surprisingly Enterococcus gallinarum, Bacillus subtilis, and Macrococcus were also identified. Bacillus subtilis is commonly found in soil; therefore its presence is understandable. Enterococcus gallinarum is very rare and is found in the intestinal tract of mammals. One species of Macrococcus, Macrococcus epuipercicus, has been found on horse’s skin (5). The bacteria responsible for the hoof infection have yet to be identified, although it is hypothesized that the bacteria are anaerobic and prefer moist environments.
    • Determination of the roles of the Trypanosoma brucei lipin homologue TbLpn

      Pelletier, Michel; Mousso, Thomas (2020-05-01)
      African Sleeping Sickness is caused by Trypanosoma brucei. T.b is a parasitic protozoan that affects humans and other animals alike in sub-Saharan Africa. Epidemics have occurred throughout the 20th century, including Uganda and Kenya in 1900 and Alego and Kenya from 1976 to 1983. Due to its antigenic variation, which helps it evade the immune system, there have only been five drugs approved for the treatment of this disease and they tend to be toxic. Therefore, in order to better understand how T.b functions, a lipin homologue termed TbLpn was examined. Lipins help catalyze the formation of diacylglycerol (DAG), which is used to form triacylglycerol (TAG). TAG is then used to form phosphatidylethanolamine (PE) and phosphatidylcholine (PC), which are major lipids that constitute the membrane of T.b. In order to better understand how TbLpn functions, we checked if it was phosphorylated in vivo by protein methyl transferases termed (TbPRMT1 and TbPRMT7) in procyclic T.b forms. TbLpn was immunoprecipitated from whole cell extracts using anti-TbLpn. Then, TbLpn treated with CIAP (dephosphorylates proteins) was compared to non-CIAP treated TbLpn using an SDS/Western blot. The cellular localization of TbLpn was also determined by first preparing both cytosolic and nuclear extracts. TbPRMT1 and 7 were downregulated by RNA interference in both the cytosolic and nuclear extracts and compared to normal TbPRMT1 and 7 using SDS/Western blot. The results show that whether or not TbPRMT1 and 7 are downregulated or not, TbLpn is primarily found in the cytosol. Thus, it can be concluded that TbLpn is primarily found in the cytosol, however its location within the cytosol, due to phosphorylation, is unknown. The results also show that TbLpn is in fact phosphorylated in vivo, due to the difference in the mobility shift between the treated and non-treated CIAP samples. The CIAP treated sample should have been higher because in general, if a protein is dephosphorylated, its shift should be lower than usual. However, its mobility shift was higher than that of the non-CIAP treated. It is unknown why that is, but phosphorylation could be linked to another process that affects its mobility shift in some way. Learning that TbLpn is phosphorylated is of great importance, since it could play a key factor in which TbLpn catalyzes the beginning stages of the formation of PE and PC, which are found in high abundance on the plasma membrane.
    • Disruption of Caveolae Lipid Rafts and the Effects on Melanin-concentrating Hormone Receptor-1 Localization: A Pharmacological Study

      Cook, Laurie B.; King, Colin; The College at Brockport (2014-05-15)
      Melanin-concentrating hormone (MCH) is integral to the regulation of human appetite. MCH targets G protein-coupled receptors in the brain and peripheral tissues. When MCH receptor 1 binds MCH on the surface of cells, it activates multiple signaling pathways, then desensitizes. Internalization of MCH-bound MCHR1 is only thought to be partially responsible for the loss of MCH signaling capacity of cells. Previous research has shown that MCH receptors are enriched in caveolae and specifically complex with caveolin-1. Caveolin-1 is a key structural component of caveolae, which are cholesterol-based lipid rafts that are known for concentrating signaling molecules and clathrin-independent endocytosis. This study aims to investigate how MCH signaling would be affected if MCH receptors weren’t enriched in these regions. Pharmacological treatments were used to achieve this goal as caveolae formation was disrupted with the antibiotic nystatin, a cholesterol inhibitor. It has been shown that sodium carbonate-based extraction procedure followed by flotation on sucrose density centrifugation isolates caveolae from other cell contents. Caveolae-isolation procedures to detect caveloin-1 were undertaken to indicate that untreated BHK-570 cells contained caveolin-1 in fractions 4 and 5 of the sucrose gradients while a gradient shift of caveolin-1 to fractions 7-10 in nystatin-treated cells occurred. Such shift confirmed that there was a partial disruption of caveolae within the treated cells. Future experiments will test whether other pharmacological inhibitors such as filipin and methyl-?-cyclodextrin as well as caveolin-1 RNAi are better able to deplete caveolae from cells as well as their impact on MCH signaling.
    • Disruption of TbLpn Expression in T. brucei by RNA Interference

      Pelletier, Michel; Serbonich, Matthew; The College at Brockport (2018-05-17)
      Trypanosoma brucei, the causative agent of human African trypanosomiasis, also known as African sleeping sickness, expresses a protein known as TbLpn which plays a critical role in arginine methylation and phospholipid synthesis within T. brucei. In order to determine the effect and potential medical benefits of downregulated TbLpn expression in T. brucei, RNA interference was used by transfecting T. brucei with plasmid DNA through electroporation. After conducting this experimental protocol, T. brucei was unable to be grown successfully in media following transfection and the effect of downregulated TbLpn in T. brucei requires further investigation in order to determine its potential for developing new drug targets and treatment methods for human African trypanosomiasis. Experimental protocol changes and repetition and verification of the validity of current protocols could lead the way for successful growth of T. brucei with downregulated TbLpn.
    • Effect of Dietary Magnesium Manipulation on the Gastrointestinal Microbiome of a Mouse Model of Ulcerative Colitis

      Ortega, Bernardo; Carlson, Christopher; The College at Brockport (2018-12-17)
      Ulcerative colitis (UC) is a disease characterized by inflammation of the GI tract, which disturbs the mucosal lining and hinders magnesium (Mg2+) absorption. Research has shown that increasing the dietary intake of Mg2+ decreases the severity of the colitis symptoms, but there is no data on the effect this has on the microbiota of the GI tract or the blood. We found that, in DSS-treated mice, the amount of bacteria in the colon increases with a decrease in dietary Mg2+, and that the concentration of bacteria in the spleen does not correlate to symptom severity or to colonic bacterial amounts. Mg2+ could be used as a supplement for UC patients, treating both hypomagnesemia and lowering colonic bacteria closer to healthy levels.
    • Effects of Magnesium Deprivation on the Flora of the Gastrointestinal Tract

      Ortega, Bernardo; Cooke, Tricia; The College at Brockport (2018-05-15)
      Crohn’s disease, a pathological condition characterized by gastrointestinal (GI) inflammation and mucosal changes is often associated with hypomagnesemia resulting from changes in the GI mucosa and an increased GI transit rate. Furthermore, certain types of bacteria are also implicated in the formation of GI neoplasia and carcinogenesis. A variety of physiologic changes such as an increase in the systemic stress response, severity of the inflammatory response, or an exaggerated immune response is known to occur in Magnesium (Mg2+) deficient mice and rats. To understand to what extent Mg2+ deficiency could contribute to the aggravation of such diseases by inducing specific changes in the GI microbiome, this study will focus on the effect of a Mg2+ deficient diet on the GI flora of Black-6 mice. Mice were fed diets containing known concentrations of Mg2+ for 1 week before being placed in a metabolic cage for 24 hours. After 24 hours the mice were sacrificed, blood, feces, and urine samples were collected, and total food consumption, urine excretion and feces excretion was determined. Bacterial analysis of feces, collected via dissection of the colon, revealed that a decrease in the amount of dietary Mg2+ consumed is associated with an increase in the number of Lactic Acid Bacteria (LAB) and Bifidobacterium Colony-Forming Units (CFU) found per gram of feces. Based on the average amount of Mg2+ excreted in the feces and urine of the mice on each diet (Regular, 2% Mg oxide, 2% Mg citrate, 1% Mg citrate, 0.5% Mg citrate, or Mg free) it was also determined that the Mg2+ in the 2% Mg citrate diet is absorbed into systemic circulation more efficiently than the Mg2+ in the other diets tested. Further research is still required to determine if the changes in GI flora associated with a lower dietary Mg2+ consumption are due to systemic hypomagnesemia or due to decreased luminal availability of Mg2+ in the GI tract, but based on the results of this study, it can be concluded that decreased dietary Magnesium consumption has a notable effect on the flora of the GI tract.
    • Human Genetic Enhancement: Is it Cheating?

      Long, Joseph; Hull, Lucas; State University of New York College at Brockport (2020-09-28)
      Genetically modifying organisms has been a very useful technology in the development of ways that we can solve many agricultural problems. This technology, which has been around since the 1990s, is starting to be used on humans in an effort to combat many genetic diseases. But does human genetic enhancement (HGE) cross a moral line? Many consider HGE to be a form of cheating since people who have been enhanced would have many advantages over those who have not been enhanced. To address this issue, I first distinguish between modifications and enhancements. Then, in light of Ken Kirkwood’s analysis of cheating, I describe four ways in which someone can be said to cheat. I conclude that, whatever other moral lines HGE might cross, HGE is not a form of cheating.
    • Nuclear Encoded Proteins Important in Mitochondrial Genome Stability

      Sia, Rey; Krembs, Luke; The College at Brockport (2012-05-04)
      The mitochondrion is widely known to be the site of cellular respiration and the factory of cellular energy. Similar to the nucleus, mitochondria house genetic material (mtDNA), which is responsible for the production of proteins essential to mechanisms required for cellular respiration. Furthermore, if there is a mutation or deletion in the mtDNA there can be ramifications in terms of energy production, which will hinder cell viability. Additionally, mutations in the mtDNA are associated with certain neuromuscular diseases as well as contributing to the aging process. The focus of this research is to identify genes that contribute to the maintenance of the mtDNA. Our data from genetic assays indicate that loss of the Clu1p protein exhibits an increase respiration loss as well as increase spontaneous point mutations. In addition, loss of Clu1p alters mitochondrial morphology.
    • Significance of the Nuclear Gene RAD54 in Mitochondrial Genome Stability of Saccharomyces cerevisiae

      Sia, Rey; Stoj, Melissa; The College at Brockport (2017-05-04)
      Mitochondria are essential organelles in eukaryotic organisms that synthesize the energy-providing molecule, ATP, through the process of oxidative phosphorylation. As explained by the endosymbiotic theory, mitochondria contain mitochondrial DNA (mtDNA), distinct from nuclear DNA (nDNA). When mitochondrial function is impaired, and mtDNA stability is compromised, detrimental neuromuscular and neurodegenerative disorders such as Mitochondrial Encephalomyopathy, Lactic acidosis and Stroke-like episodes (MELAS) and Leber’s Hereditary Optic Neuropathy (LHON) have the potential to occur. The purpose of this study was to determine the role of the nuclear gene RAD54 in maintaining mtDNA stability in the budding yeast, Saccharomyces cerevisiae. Although the role of Rad54p in maintaining nDNA stability is understood, its impact on mtDNA stability is relatively unknown. RAD54 is a member of the RAD52 epistasis group, coding for a protein vital to the initial steps of homologous recombination and double-stranded break (DSB) repair. Given that members of the RAD52 epistasis group have been shown to contribute to homologous recombination and DSB repair in mtDNA of S. cerevisiae, we hypothesized that loss-of-function RAD54 would decrease the rate at which homologous recombination in mtDNA occurred (Stein, Kalifa & Sia, 2015). A phenotypic respiration loss assay was performed in a rad54? strain to determine the frequency of spontaneous mutations in mtDNA that blocked the oxidative phosphorylation process. The mutant strain demonstrated a 1.56-fold decrease in spontaneous respiration loss when compared to wild type (p-value = 0.0574). Interestingly, previous research has demonstrated that the nature of these spontaneous mutations is due to large deletions in the mtDNA. To investigate the role of Rad54p in preventing these deletions from occurring, a direct repeat-mediated deletion (DRMD) assay was performed. The DRMD assay demonstrated a significant 3.23-fold increase in nDNA homologous recombination events (p-value = 0.0158) and a statistically insignificant 1.08-fold increase in mtDNA homologous recombination events (p-value = 0.8741) between rad54? and wild type strains. Given the present findings of this study, it appears the nuclear gene RAD54 does not play a significant role in maintaining mtDNA stability in respiration loss or DRMD assays.
    • The Role of RAD55 in Mitochondrial DNA Stability in Saccharomyces cerevisiae

      Sia, Rey; McAtee, Kyle J.; The College at Brockport (2016-04-06)
      Mitochondria are organelles present in eukaryotic cells. Through the process of cellular respiration mitochondria produce ATP; a vital molecule for the completion of many cellular processes. Mitochondria are unique in that they contain their own DNA separate from the DNA within the nucleus. Mutations in mitochondrial DNA have notable connections to several human pathologies such as various neuromuscular and neurodegenerative disorders. The focus of this study was to determine the role of the nuclear gene RAD55 in maintaining mitochondrial DNA in budding yeast, Saccharomyces cerevisiae. The gene product of RAD55 cooperates with other proteins to bring about the repair of double-stranded DNA breaks in the nucleus. Specifically, RAD55 is a member of the RAD52 epistasis group whose gene product functions as a heterodimer with Rad57p. The Rad55p/57p heterodimer promotes Rad51p filament assembly on single-stranded DNA. Once assembled, Rad51p filaments displace Replication Protein A from single-stranded DNA and its recombinase activity is initiated. To determine the effect loss of Rad55p had on the stability of mitochondrial DNA, two genetic assays were performed. The first assay measured the frequency of spontaneous respiration loss in rad55? mutants. The lab observed rad55? mutants did not show a significant increase in spontaneous respiration loss compared to that of the wild type. An additional direct repeat-mediated deletion assay was performed to determine if Rad55p played a role in stabilizing the mitochondrial genome from mutations caused by recombination events. It was discovered that the rate of direct repeat-mediated deletions for rad55? in the nuclear genome increased 5.8-fold compared to that of the wild type. Surprisingly, the lab found the rate of direct repeat-mediated deletions for rad55? in the mitochondrial genome decreased by 1.5-fold compared to that of the wild type.
    • The Role of the Aryl Hydrocarbon Receptor in Tumor Growth and Chemoresistance

      Sia, Rey; Lindsay, Elizabeth L.; The College at Brockport (2016-05-08)
      The aryl hydrocarbon receptor (AHR) has been shown to play a role in cancer initiation and progression in oral squamous cell carcinomas (OSCC), and other cancers. The AHR is activated by environmental toxins, including polycyclic aromatic hydrocarbons, which are commonly found in cigarette smoke. It is hypothesized that activation of the AHR by these environmental toxins can contribute to the growth and chemoresistance of OSCCs. Nude mice tongues were injected with a human OSCCs cell line, SCC2s, and treated with an AHR antagonist at 25mg/kg daily via oral gavage. Primary tumor growth was measured via calipers and IVIS imaging. RT-qPCR analysis of the harvested tongue tumors and livers was used to examine the activity of the AHR by quantifying the expression levels of Cyp1b1 and Cyp1a1. Based on the results of the in vivo experiments, continued testing was conducted to examine the role of AHR inhibition in chemoresistance. Using MTT cell viability assays coupled with dosing of commonly used chemotherapeutics, the effects of the AHR on the chemo-resistance of SCC2s was tested. Three commonly used chemotherapeutics were tested at various dose ranges: Cisplatin (0-10uM), doxorubicin (0-1uM), and 5-Fluorouracil (0-10uM). In addition, cells were co-treated with an AHR antagonist (5uM CH223191) and the chemotherapeutic to determine if decreasing AHR activity increased chemotherapeutic efficiency. ANOVAs were used to evaluate the significance of AHR activity on the effectiveness of the chemotherapeutics. It was determined that AHR antagonism with CB7993113 significantly affected OSCC primary tumor growth in vivo. Additionally, it was found that both Cyp1a1 and Cyp1b1 expression decreased after treatment with CB7993113 when compared to vehicle alone in the tongue. In the liver, it was found that both Cyp1a1 and Cyp1b1 expression also decreased after treatment with CB7993113 when compared to vehicle alone. Interestingly, we also found that decreasing AHR activity with an AHR antagonist CH223191 in addition to treatment with a chemotherapeutic lead to a significant increase in cell death when compared to treatment with the chemotherapeutic alone. This phenomenon was observed in three different frontline OSCC therapeutics. These novel findings implicate the AHR in OSCC initiation and growth, also supporting the development of AHR modulators as potential chemotherapeutics. Overall, these findings support the hypothesis that the activation of the AHR is linked to tumor growth of oral squamous cell carcinomas as well as contributing to the potential chemoresistance of these cells.
    • The Significance of the Nuclear Gene KU80 in Mitochondrial Genome Stability of Saccharomyces cerevisiae

      Sia, Rey; Scott, Brooke; State University of New York College at Brockport (2020-09-10)
      Mitochondria are essential organelles in eukaryotes. They are often referred to as the powerhouse of the cell because mitochondria manufacture ATP, which is required for the successful completion of many cellular processes. Mitochondria have individual genomes, separate from the nuclear DNA, which encode proteins required for respiration. In humans, mutations in the mitochondrial DNA (mtDNA) result in the loss of mitochondrial function which leads to neuromuscular and neurodegenerative disorders. The focus of this study is to determine the role of the nuclear gene KU80 in maintaining mtDNA stability in the budding yeast, Saccharomyces cerevisiae. The product of the KU80 gene is the protein, Ku80p. Ku80p, in humans, is encoded by the XRCC4 gene. Ku80p along with Ku70p forms a heterodimeric protein complex, which binds to DNA double-strand break ends and is required for the non-homologous end joining (NHEJ) pathway of DNA repair. The goal of this research is to determine whether loss of the KU80 gene plays a role in mitochondrial genome stability. Mitochondrial genome instability can arise via spontaneous point mutations or deletion events. Assays were performed to measure the spontaneous respiration loss rate between wild type and ku80-? mutant strains. The respiration loss assay showed a 1.90-fold increase (p=0.001256) in spontaneous respiration loss compared to the wild type strain. Strains were constructed to determine the role of KU80 in spontaneous direct-repeat mediated deletion (DRMD) events within the mitochondrial genome as well as the nuclear genome. The rate of DRMD events in the mitochondrial and nuclear genomes showed a 1.94-fold decrease (p=0.08711) in spontaneous mutation rates in mitochondrial DNA and a 5.87-fold decrease (p=0.000786) in nuclear DNA compared to the wild type. These results suggests that Ku80p plays a role in maintaining the integrity of the mitochondrial genome in budding yeast.
    • The Significance of the Nuclear Gene, SGS1, in Mitochondrial Genome Stability in Saccharomyces cerevisiae

      Sia, Rey; Wershing, Kathryn C.; The College at Brockport (2016-04-26)
      The homologs that humans share with Saccharomyces cerevisiae render yeast an ideal model organism to investigate the potential importance of genes in humans. SGS1 is a nuclear gene for a RecQ helicase in yeast, known to play a role in homologous recombination in nuclear genome repair. The research in question is intended to evaluate if SGS1 has a similar importance in mitochondrial genome repair. These conclusions can be employed to better understand the disease phenotypes that humans present as a result from mitochondrial malfunction. A respiration loss assay showed that SGS1 knockout strains have a ~2.2 fold increase in spontaneous respiration loss frequency, indicating that SGS1 plays a role in mitochondrial genome stability. A direct-repeat mediated deletion assay proves SGS1 is involved in homologous recombination in mitochondria due to an ~1.6 fold decrease in rate of homologous recombination in SGS1 knockout strains. With a p value of 0.66, no significant difference was observed in an induced direct-repeat mediated deletion assay between wild type and sgs1? strains, implying that SGS1 does not play a fundamental role in double strand break repair. Future experimentation could include additional knockout strains testing other genes known to be involved in nuclear homologous recombination repair and double knockout strains to assess the relative order of active proteins involved in genetic repair mechanisms.
    • Three Gastrointestinal Assays

      Rich, Adam; Majtyka, Bailey; State University of New York College at Brockport (2020-09-16)
      Gastrointestinal (GI) functionality relies on the spontaneous, rhythmic and coordinated propagation of muscular contractions in the GI tract, or GI motility. Without these coordinated motor patterns, digestion falters, and results in problems with digestion. Disrupted or un-coordinated motor patterns are associated with altered GI transit times. GI transit is the amount of time necessary for intestinal contents to move through the GI tract. GI transit is measured in patients complaining about abdominal discomfort to determine if discomfort results from a true dysmotility or from idiopathic symptoms. GI transit assays help to determine appropriate treatments but idiopathic symptoms, or pain from an unknown cause, is very common. The zebrafish is an attractive model system for human GI motility because the entire GI tract can be observed in intact zebrafish larva. In current methods, larvae are fed food with a marker substance and movement through the intestine is viewed using a microscope and recorded using a digital camera. However, GI transit time is highly variable. It is possible that this variability is completely normal and results from variable GI physiology. Alternatively, it is possible that the variability is due to the assay. Three distinct GI transit assays have been published. The overall objective for this work is to determine the reliability for each assay and to better understand which assay is most appropriate for future work. The assays will be described and compared, and results comparing the assays will be presented.
    • What You Need to Know to Apply to Veterinary School and Career Options Available After Obtaining Your Doctorate in Veterinary Medicine

      Sia, Rey; Walker, Jacqueline; The College at Brockport (2010-05-01)
      I am a senior undergraduate student at the College at Brockport. I will be attending Cornell University of Veterinary Medicine this coming academic year. I am writing this manuscript for my thesis project which is a requirement to graduate from the Honor’s Program at the College at Brockport. For my project, I wanted to do something that would increase my knowledge on a subject that was important to me: becoming a veterinarian. Also, as an undergraduate it is hard to decide what you want to do with the rest of your life, and my hope is that this manuscript can give students interested in becoming a veterinarian a better idea of exactly what it is they want to do or if they even want to become a veterinarian at all. No one sits you down and tells you what you are getting into when you choose a course of study that interests you; it is expected that you figure out exactly what you need to do to either obtain the job you want or complete the requirements needed to go on to graduate or professional school. It is difficult figuring everything out, and sometimes mistakes are made along the way. Once someone figures everything out that is needed to complete a course of study, I believe it is in their best interest to share that information with others so they can avoid making similar mistakes. Applying for vet school and figuring out the timing of everything that I needed to have done to apply was on my shoulders: it was my future and I had to take charge. Now that I have almost completed my undergraduate degree and have been accepted to vet school, I want to share my knowledge and experience in the hopes that other undergraduate students may learn something from the path that I took to get there. The way I did things is not the only way, but I hope that my experience and mistakes can be applied to the lives of other undergraduates so that they will have a more successful future.