Bacterial Growth and Search for Antibiotic Producers at Springfield College
Subject: Science (
Biology & Chemistry)
Presenters: Alyssa Paquette & Mackenzie Gingras
Faculty Sponsor:
Dr. Sally Chamberland
Discovery for new antibiotics is ever growing in need as many bacterial species evolve antibiotic resistance. A goal of this research was to identify an antibiotic producer sourced from Springfield College’s campus in the hopes of discovering a microbe that generates antibiotics against one of the known ESKAPE pathogens. A soil sample was collected by Watershops Pond in Springfield, MA. The soil sample was then cultured using luria broth agar. Throughout the testing process, LBA was used and bacteria were grown at room temperature conditions. Testing against ESKAPE pathogens using safe relatives was conducted. After identification of an antibiotic producer, isolation of the bacteria was completed. One isolate (number 18) had shown a zone of inhibition when plated with B. subtilis. Identification of the bacteria was attempted through characterization. Diagnostic tests provided insight into the antibiotic producer; however, the isolate was never identified. The results from this project may contribute to the scientific community as the search for new antibiotics continues.
Sequence Analysis of CG15365 as a Potential LZTS1 Ortholog
Subject: Science (Biology & Chemistry)
Presenters: Julianna Faillace
Faculty Sponsor:
Christopher Abdullah
Hypermobile Ehlers-Danlos Syndrome (hEDS) is a connective tissue disease with unknown etiology characterized by generalized joint hypermobility and tissue fragility. Several questions as to the etiology behind this condition remain. Mutation of the human gene LZTS1 has recently been proposed to have a role in the development of hEDS. We aimed to determine whether an orthologous gene to LZTS1 is present in Drosophila melanogaster as this is an inexpensive model and no genetic models of hEDS currently exist. Identification of such a gene in Drosophila would be significant as it would allow for the characterization of a genetic model of an LZTS1 mutation to be analyzed for hEDS-related phenotypes. We performed several computational analyses to determine if an orthologous gene exists in Drosophila. Initial analyses revealed CG15365 to be a candidate ortholog and several experiments were conducted to investigate possible similarity. Amino acid sequences were aligned to compare protein similarity between LZTS1 and CG15365. We examined 3-D protein structures and conserved domains of LZTS1 and CG15365 proteins to analyze both predicted structural and functional similarities. Our results suggest that CG15365 may not be orthologous to LZTS1 and raises questions about the proposed relationship between the two genes. While CG15365 and LZTS1 share some similarities, further research is needed to determine any functional relevance of CG15365 to LZTS1 and whether LZTS1 plays a role in hEDS pathology.
Identification and Characterization of Antibiotic-Producing Bacteria from Soil Samples
Subject: Science (Biology)
Presenters: Felix Kriedemann
Faculty Sponsor:
Dr. Sally Chamberland
The increase of antibiotic resistant bacteria poses an urgent challenge to modern medicine. The aim of this research is to investigate the potential of bacteria isolated from soil collected from the shore of Watersheds Pond as a source of new antibiotic compounds. This research investigates how the proximity to freshwater and more consistent temperatures of the collection site favors a diverse bacterial population with potential antibiotic activity. Systematic research was conducted where soil samples were collected and plated on media with different pH values to promote diverse bacterial growth. These bacterial isolates were then tested against ESKAPE safe relatives, and were bacteria noted for having antimicrobial properties. These isolates were then characterized using a number of different biochemical tests. The results show a higher abundance of antibiotic-producing isolates when grown at a pH 5 compared to pH 8, suggesting that acidic conditions may promote the production of antibiotic compounds. The results of this research suggest that environmental manipulation, such as adjusting the pH of growth media, may be a viable strategy to increase the detection of antibiotic-producing bacteria from soil samples.
Mammalian Cellular Model of Fibrodysplasia Ossificans Progressiva (FOP)
Subject: Science (Biology & Chemistry)
Presenters: Leismi Guerrero & Rasika Adhikari
Faculty Sponsor:
Hai Kinal
Fibrodysplasia Ossificans Progressiva (FOP) is a rare genetic disorder characterized by abnormal bone formation in soft tissues, caused by a specific mutation in the ACVR1 gene. With an estimated worldwide prevalence of approximately 900 cases, FOP presents as an excruciating and disabling condition, typically manifesting before the age of 6 and leading to lifelong confinement to a wheelchair. The rarity of FOP poses significant challenges in comprehending its underlying biology, primarily due to limited research efforts. In response, our research team is dedicated to developing a cellular model to facilitate a deeper understanding of FOP pathogenesis and ultimately contribute to the development of novel therapeutic interventions. To create a cellular model of FOP, we are using CRISPR cas9 genome editing technology to perform site-directed mutagenesis in the ACVR1 gene. This involves changing the non-mutated codon CGC (coding for Arginine) to mutated codon CAC (coding for Histidine). Once a cellular model of FOP is available, molecular targets may be determined and used to halt the progression of abnormal bone formation. This is especially important because current treatments only ease the suffering of FOP patients, and any surgical attempts to remove the malformed bone tissue only exacerbate the problem.
Understanding Bacterial Susceptibility to Ferulic Acid Derivatives
Subject: Science (Sports Biology)
Presenters: Abigail Taylor
Faculty Sponsor:
Dr. Chamberland and Dr. Blauvelt
Ferulic Acid (FA), is a component found in the cell walls of plants. Research has shown that phenolic acids like FA have many biological effects such as antibacterial, antifungal, anticarcinogenic, and anti-inflammatory. This research works to determine if derivatives of FA have any antibacterial properties as well. Many common bacterial pathogens are developing antibiotic resistance and are no longer as effective. It is imperative for the medical field to find alternative options. Here two different FA derivatives were made in the organic chemistry lab, methyl ferulate and butyl ferulate. These derivatives were then tested for antibiotic activity using the common disc diffusion assay. Three different bacterial species were tested, Escherichia coli, Staphylococcus epidermidis and Enterobacter aerogenes. Butyl ferulate showed broad antibacterial activity against both gram-negative bacteria, E. coli and E. aerogenes, as well as gram-negative bacteria S. epidermidis. Whereas methyl ferulate only showed antibacterial activity to the gram-negative bacteria.
The Comparison of Red Russian and Lacinato Kale Varieties Grown Using Aquaponics VS. Organic Soil
Subject: Science (Health Science: Pre-PA)
Presenters: Julianna Torres
Faculty Sponsor:
Gemma Bartha
Aquaponics is a process which utilizes both hydroponics and aquaculture together in order to grow a variety of agriculture. The aquaculture utilizes fish, and in this study tilapia, as a source of nutrients through their waste. The waste acts as nourishment for the plants by producing necessary nitrate and nitrite compounds aiding in plant growth by use of the nitrogen cycle. Through the use of this minimal system, plants have the potential to be grown in a number of locations that have minimal resources. This research study aims to analyze how aquaponics grown plants differ from those grown in soil. The overall objective was to provide statistical data showing a difference in the two methods, explaining why such differences were present. Due to this system's inexpensive nature and ease with which it was run, it proposes the question which emphasizes how third world countries and food deserts may find use in it. It would have the capacity to grow medicinal plants with minimal resources helping to slowly pave the way for community grown agriculture.
Analysis of drosophila locomotor function due to mutations in CG33090
Subject: Science (Biology & Chemistry)
Presenters: David Handron
Faculty Sponsor:
Chris Abdullah
CG33090 is an uncharacterized gene in D. melanogaster. Performing a comparative sequence alignment of the CG33090 amino acid sequence to the entire human genome predicts that CG33090 is homologous to GBA2 in H. sapiens. Compared to D. melanogaster the GBA2 protein is 62% similar to the human protein. GBA2 catalyzes the hydrolysis reaction that cleaves glucosylceramide into glucose and ceramide however similar to its paralog GBA1, GBA2 functions in the cytoplasmic side of the cell membrane while GBA1 functions in the lysosome of the cell membrane. DNA coding for green fluorescent protein (GFP) located on a plasmid is integrated into CG33090. This creates an additional region of the protein that can be visualized once the DNA is transcripted and translated to a protein. Imaging shows GFP highlighting the neuronal cell membranes, the same place that GBA2 is found in humans. Both point mutations and truncations found in human patients lead to the production of a protein that is unable to convert glucosylceramide to glucose and ceramide. A non-functional GBA2 protein leads to an accumulation of glucosylceramide in the cell, as it is never converted to glucose and ceramide. Without this recycling step, glucosylceramide accumulates. Mutations in M. musculus protein found in the same pathway as GBA2 also lead to an accumulation of membrane lipid precursors and show cerebellar atrophy This cerebellar atrophy is also seen in human patients. In D. rerio the mutation of GBA2 leads to abnormal neuronal outgrowth at the larval stage. D. melanogaster with CG33090 mutations are expected to show a deficit in their ability to crawl compared to wild-type subjects. Crawling assays with D. melanogaster larvae will be used to measure the impact of various genotypes for different CG33090 mutated flies. Flies with wild-type CG33090, a second chromosome deficiency, and an insertion in CG33090 will be crossed to create different genotypes and the crawling phenotype will be measured. We hypothesize that flies with mutations on both copies of the second chromosome will have a more severe impairment of their crawling ability. Currently, there are no known GBA2-specific drugs, and if GBA2 mutations in humans are modeled in flies there will be a model organism to cheaply and effectively screen for potential drug efficacy and specificity.
Climbing Assays Used to Assess the Motor Function of Drosophila with Mutations of Genes CG33090 & CG3376
Subject: Science (Biology & Chemistry)
Presenters: Hannah Hibbert, Lilly Vey & Abby Taylor
Faculty Sponsor:
Chris Abdullah
Drosophila share about three-fourths of the same genes that are seen in humans to cause disease. Scientists often test human disease/defects in Drosophila. We structured the experiment to analyze the effects of two genes, one common with the entire lab and one specific to each group. The common gene is CG33090 in Drosophila, a predicted ortholog of the human gene GBA2, and research shows that mutation of this gene can impact motor abilities. Our gene was CG3376, which its human ortholog is associated with Niemann-Pick disease. We tested a hypothesis about these genes by running trials of climbing assays, gathering the data of how many flies were able to reach the measured distance in the allotted time, and then analyzing it to determine if there are any motor function defects of the flies with the mutated genes. Based on the idea that both genes contributed to motor function, creating an experiment about the motor abilities of Drosophila seemed like the right direction. At the beginning of the course there was freedom to experiment with larvae versus adult flies. After these experimental days, it was concluded that assessing motor function defects in adult flies would be best fit for assessing motor function. Therefore, we hypothesized that the homozygous mutant CG3376 & CG33090 will have an additive effect in Drosophila and have the least ability to climb the six centimeter line in the allotted time trial.
Analysis of Mutations in CG33090 and CG12034 to Evaluate Resultant Locomotion Phenotype Differences in Drosophila
Subject: Science
Presenters: Morgan Stafford, Alyssa Paquette & Mackenzie Doyle
Faculty Sponsor:
Chris Abdullah
Human enzymes glucocerebrosidase beta 2 (GBA2) and sphingomyelin phosphodiesterase 2 (SMPD2) are both involved in ceramide production and sphingolipid biosynthesis; however, they work in different organelles (GBA2 in the plasma membrane and SMPD2 in the golgi apparatus). Although mutations in both human GBA2 and SMPD2 are known to cause locomotor dysfunction such as Parkinson’s Disease and Lewy Body Dementia, we hypothesize that a mutation in SMPD2 is non-additive to GBA2, and that having a mutation in one will not exacerbate the other. To test this, we proposed the use of a Drosophila model using the genes CG33090 (the predicted Drosophila version of human GBA2) and CG12034 (the known Drosophila version of human SMPD2). Seven different genotypes were analyzed to determine the number of Drosophila that reached the Line of Achievement through the use of a timed modified climbing assay. Current data suggests that there are no statistically significant differences in velocity between the genotypes. Further exploration and testing of these genotypes is planned to evaluate locomotion phenotypes due to mutations in CG33090 and CG12034.
An Analysis of Drosophila Melanogaster Gene CG31148 and Motor Function in Adult Fruit Flies
Subject: Science (
Biology)
Presenters: Damian Gilrein, Oliver Kroon & Giancarlo Martinez
Faculty Sponsor:
Chris Abdullah
Fruit fly (Drosophila melanogaster) gene CG31148, which encodes the glucocerebrosidase-ɑ (GBA1ɑ) enzyme, has not undergone extensive research, despite its relation to the human GBA1 enzyme. The GBA1 enzyme in humans has been associated with neurological disorders like Gaucher’s and Parkinson's disease and in fruit flies is associated with a number of motor dysfunctions as well as parkinsonian signs (Maor et al., 2016). The enzyme catalyzes the hydrolysis of glucocerebroside into glucose and a ceramide. The goal of this research project is to analyze the relationship between mutations in genes CG31148 and CG33090 and motor function of adult fruit flies through the use of climbing assays. We hypothesized that the mutations related to the GBA1 enzyme would have a negative effect on locomotion in the groups of flies with hetero- and/or homozygous mutations in the CG31148 and CG33090 gene sequences when compared to the wild-type control flies. Further research of this gene is valuable because the phenotypes of these mutations have not previously been studied in adult fruit flies. Previous data on gene expression have shown that peak expression occurs during late larval stages, this should not undermine the effects GBA mutations have on later life stages. Our data suggests that expression of the GBA1ɑ gene occurs even during the flies adult life, with fruit flies in the experimental groups performing worse than the control group.
Game On! A Comparative Analysis of External Load in Division III Soccer Players Across Gender and Position
Subject: Science (Applied Exercise Science)
Presenters: Ashima Fillbach
Faculty Sponsor:
Bianca deLucia and Mary Kate Feit
OBJECTIVE: This study aimed to contribute to the limited research that examines gender differences in external load throughout a collegiate soccer season through the use of GPS.
PURPOSE: To compare game workloads across collegiate men (MSOC) and women (WSOC) soccer players. Division III MSOC and WSOC players wore GPS devices during games throughout a 15-week season.
METHODS: Metrics analyzed from GPS data were: total distance (TD), sprint distance (SD), number of deceleration efforts (decel efforts), number of acceleration efforts (accel efforts), sprint efforts, top speed (TS), player load (PL), player load per minute (PL/Min), and distances in five-speed zones. Only high-minute players were included in the analysis. An independent samples MANOVA was conducted as well as 12 univariate analyses of variance (ANOVAs).
RESULTS: Results of a 2 (Gender: men, women) by 3 (Position: defender, midfielder, forward) independent samples MANOVA showed a significant multivariate interaction effect and significant multivariate main effects of gender and position. Significant univariate interaction effects were found for 11 out of 12 dependent variables. Significant univariate main effects of gender were found for 11 out of 12 dependent variables. Means were significantly greater for men than women for TD, decel efforts, accel efforts, TS, PL/min, distance in SZ1, and distance in SZ2. Means were significantly greater for WSCO than MSCO for SD, number of sprints, and distance in SZ3 and SZ5. Significant univariate main effects of position were found for 11 out of 12 dependent variables.
DISCUSSION: Notable gender differences were found between game external load metrics, gender and position, and within-gender position across external load metrics. Further, results can be used as a framework to develop gender-specific programs to prepare athletes for the demands of their competitive seasons.