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Research in Bioengineering

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Suicide Vectors for allelic exchange in Cellulophaga Lytica

By Mehul Puri

Introduction 

C. Lytica 

The gram-negative marine-based bacteria C. lytica has a genome of 3,765,936 base pairs, including 3,303 protein-coding genes and 55 RNA genes (Pati et al., 2011). It can grow in a wide range of temperatures between 4 °C and 40 °C in an 8% NaCl concentration with optimal growth between 22 °C to 30 °C (Pati et al., 2011). In the absence of flagella and pili, C. lytica cells depend on gliding motility to transport themselves. This translocation mechanism is also used to form biofilm colonies, which consist of colonies of C. lytica cells that can grow on non- biological surfaces such as rocks and metals. This allows them to survive in hostile environments and colonize new environments with ease. (Hall-Stoodley et al., 2004). 

Additionally, biofilm colonies serve as a foundation for larvae growth due to chemical and physical cues (Unabia et al., 1999). The formation of biofilm colonies by C. lytica also produces iridescence or coloration created by light reflection on intricately organized cells resembling crystals, as shown in Figure 1 (DeSimone, 2021). Though this iridescence has not been observed in natural environments, it has been observed in colonies grown in lab environments, and the significance of this iridescence in nature has not been elucidated to date. It is one of the identifiers or markers of biofilm colonies for C. lytica cells (Kientz et al., 2016). 

Figure 1. Colonies of C. lytica grown on Black ink plates (Adapted from M. DeSimone’s thesis, DeSimone 2021) 

Goal 

This study hypothesizes that deletion of the GldB gene in the bacteria Cellulophaga Lytica is responsible for gliding motility and can disrupt the formation of colonies. The ability to disrupt the formation of uniform colonies of C. lytica can impact the biofilm formation and allow us to control the iridescence of the bacteria. 

Approach/Methodology 

Designing the Vector 

In this project, the PYT313 suicide vector (Donated by a collaborator’s lab, Dr. Yontao Zhu, Minnesota State University Mankato) was used as it works with F. johnsoniae related to C. lytica. The suicide vector, as shown in Figure 2, contains sacB and the promoter of F. johnsoniae, ompA, which is used to construct chromosomal gene deletions specific to gliding (Zhu, 2017). Additionally, PYT313 is resistant to the antibiotic ampicillin due to the presence of AmpR. 

Figure 2. The plasmid map of PYT313 donated by Dr. Yongtao Zhu indicating the presence of the sacB, erythromycin resistance (ermF), and the promoter of F. johnsoniae, ompA (Zhu et al., 2017). 

Four primers are designed to isolate the GldB (gliding motility) gene within the C. Lytica DNA and are then used to create a new suicide vector using PYT313. As shown in figure 3, primers a and d contain restriction enzyme sites on their 3’ and 5’ sites, respectively. These sites correspond to specified restriction enzyme sites on the PYT313 vector. Primers c and d are homologous 1 kb upstream and downstream of the GldB gene from the start and stop codons, respectively. Through three polymerase chain reactions (PCR), the AB fragment and CD fragment are used to create the AD fragment which contains the GldB gene with restriction enzyme sites upstream and downstream of the DNA (Francis et al.). 

 

Figure 3. Four primers are designed for Overlap PCR. Through two PCR rounds, the gene is removed from the bacteria C. Lytica and ligated onto the PYT313 suicide vector. Figure from (Francis et al.). 

Then, through double digestion, the PYT313 vector is digested at the two specified restriction enzyme sites. After running the gel purification through electrophoresis, the larger digested PYT313 DNA is extracted and ligated with the AD fragment containing the GldB gene. This creates a new vector specifically designed to replace the GldB gene within C. Lytica with an inactive copy of the gene through transformation and conjugation processes (Francis et al.). 

Transformation and Conjugation 

Bacterial transformation is the process of environmental DNA uptake by competent cells. In this project, chemically competent E. Coli S17 λ Pir cells are used to uptake the GldB gene-inclusive PYT313 suicide vector. S17 cells allow for better DNA transfer during conjugation, which is why DNA uptake during transformation is crucial for GldB gene deletion. (Chen et al.) Then, the transformed E. Coli S17 cells are conjugated with C. Lytica cells for a direct transfer of DNA. 

Bacterial conjugation directly transfers genetic material from the E. Coli S17 λ Pir cells to the C. Lytica. During the conjugation, the mutant GldB gene is introduced to the recipient C. Lytica. As shown in figure 4, a two-step homologous recombinant event occurs: first and second crossover. 

During the first crossover, C. Lytica acquires the plasmid from the S17 cells, including the ampicillin antibiotic resistance. The conjugated bacteria is isolated using antibiotics, and a second crossover event occurs using the SacB sucrose counter-selection gene. During this event, the remaining part of the vector is removed from the C. Lytica, including the ampicillin resistance, leaving behind the mutant GldB gene or a wild-type GldB gene. Colony PCR is then conducted to differentiate between the two outcomes. C. Lytica cells with mutant GldB gene are grown and tested for results (Old Reliable: Two-Step Allelic Exchange by Bitesize Bio). 

Figure 4. Two Step Homologous Recombinant Event occurs, creating two types of bacteria. First conjugated C. Lytica has a wild-type allele, and second has the desired mutant allele. Adapted from Old Reliable: Two-Step Allelic Exchange by Bitesize Bio. 

Projected Outcome 

To test if the combination of DNA transfer is successful, the conjugated C. Lytica cells are introduced to antibiotic ampicillin. If the cells survive, the conjugation was successful, otherwise the cells would deteriorate. Additionally, successful conjugation will result in the C. Lytica cells losing their ability of iridescence. 

Results 

Spring 2024 

During the Spring 2024 semester, getting results on the transformation and conjugation was emphasized rather than altering the PYT313 to create a new vector. C. Lytica and PYT313 were grown in BB2 Agar and LB Agar plates respectively, as shown in Figure 5. Afterwards, a 50 ml culture was made using the colonies from both plates. Additionally, a 50 ml culture of S17 λ pir cells was grown for transformation. Transformation protocol was conducted using the S17 cells and PYT313. Transforming the S17 λ pir cells using the PYT313 was successful as it resulted in colony growth on LB Agar plates. Colony growth on multiple plates with antibiotic present is shown in Figure 6.


Figure 5. Growth of C. Lytica and PYT313 in BB2 Agar and LB Agar plates. LB Agar plate has ampicillin antibiotic added to it which demonstrates PYT313 ampicillin resistance. 

Figure 6. LB Agar plates with PYT313 suicide vector transformed S17 λ pir cells. Growth shows successful transformation due to the presence of antibiotic ampicillin. 

However, conjugation was unsuccessful, as no growth was present in the Conjugation Plates after a week of incubating. This could be due to multiple factors such as too many antibiotics or less cell density of C. Lytica or transformed S17 cells before conjugation. Additionally, due to time constraints, multiple trials of transformation and conjugation could not be completed. 

Summer 2024 

During Summer 2024, our aim is to establish which primers to use for the PYT313 for the two- step allelic exchange and complete successful transformation and conjugation with the new vector. Because we know transformation is possible, and conjugation can also be achieved with multiple trials, we hope to achieve complete deletion/replacement of the GldB by the end of summer. Additionally, the OUR grants have greatly enhanced this project by providing funds for resources and have made my research goals possible.

References 

ChenInês, et al. “The Ins and Outs of DNA Transfer in Bacteria.” Science, vol. 310, no. 5753, 2 Dec. 2005, pp. 1456–1460, www.ncbi.nlm.nih.gov/pmc/articles/PMC3919525/, https://doi.org/10.1126/science.1114021. 

DeSimone, Mark, Development of Genetic Engineering Tools for the Iridescent Bacteria Cellulophaga lytica, A Thesis (2021) 

 Francis, Matthew S, et al. “Site-Directed Mutagenesis and Its Application in Studying the Interactions of T3S Components.” Methods in Molecular Biology, 12 Nov. 2016, pp. 11–31, pubmed.ncbi.nlm.nih.gov/27837478/, https://doi.org/10.1007/978-1-4939-6649-3_2. 

Accessed 13 June 2024. 

Hall-Stoodley, L., Costerton, J. W., & Stoodley, P. (2004). Nature Reviews Microbiology, 2(2), 95–108. https://doi.org/10.1038/nrmicro821 

Kientz, B. et al. A unique self-organization of bacterial sub-communities creates iridescence in Cellulophaga lytica colony biofilms. Sci. Rep. 6, 19906; doi: 10.1038/srep19906 (2016). 

McBride, M. J., & Zhu, Y. (2013). Gliding Motility and Por Secretion System Genes Are Widespread among Members of the Phylum Bacteroidetes. Journal of Bacteriology, 195(2), 270–278. https://doi.org/10.1128/jb.01962-12 

“Old Reliable: Two-Step Allelic Exchange.” Bitesize Bio, 17 July 2018, bitesizebio.com/41461/old-reliable-two-step-allelic- exchange/#:~:text=The%20idea%20behind%20suicide%20vectors,understandably%2C%20can’t %20replicate. 

Pati, A., Abt, B., Teshima, H., Nolan, M., Lapidus, A., Lucas, S., Hammon, N., Deshpande, S., Cheng, J.-F., Tapia, R., Han, C., Goodwin, L., Pitluck, S., Liolios, K., Pagani, I., Mavromatis, K., Ovchinikova, G., Chen, A., Palaniappan, K., & Land, M. (2011). Complete genome sequence of Cellulophaga lytica type strain (LIM-21T). Standards in Genomic Sciences, 4(2), 221–232. https://doi.org/10.4056/sigs.1774329

Unabia, C., Hadfield, M. Role of bacteria in larval settlement and metamorphosis of the polychaete Hydroides elegans. Marine Biology 133, 55–64 (1999). https://doi.org/10.1007/s002270050442

Research in Sociology

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The Impact Green Spaces Have on Mental and Physical Health: Buttonwood Park and Riverside Park

By Faith Unwin

 

Introduction

This research focuses on the economic differences between two parks in New Bedford and how they affect people’s mental and physical health. Buttonwood Park and Riverside Park are in two economically different parts of New Bedford, but both provide their populations with green space to exercise and play. According to Barton, et al., (2017, p.80), “[Green space] is an umbrella term used to describe either maintained or unmaintained environment areas, which can include nature reserves, wilderness environments, and urban parks.” The park’s communities exhibit differences in average income, race, and housing. Buttonwood Park sits on the border of New Bedford and Dartmouth, with a community of high-income earners and primarily single-family homes surrounding it. Frederick Law Olmsted created this park in 1895, and its different elements of nature and activity have been well-funded since (The Friends of Buttonwood Park). The city built Riverside Park in the early 2000s on the north end of New Bedford, along the Acushnet River. The houses surrounding Riverside Park are primarily triple-decker homes, with a large community of members in the working class. Historically, older Portuguese families lived near the park, but more recently, Central American immigrant families have settled there. Ramirez (2022) describes this area as “an area burdened by disproportionate climate and public health impacts.” People of this community may face stronger economic challenges compared to other areas. Economic differences play a huge role, not just in people’s individual lives but in communities. 

This research aims to understand whether the differences between the communities affect who and how many people visit the parks. All community members should have the ability to go outside in a safe environment with enough space to be active. Green spaces provide “creative opportunities for open space preservation that could help connect the community and revitalize its economy and social connectivity” (EPA). However, some areas may find it more difficult to do this because of their location and resources. With some parks being in different areas, there may be differing opinions on when to use the parks and for what reasons. According to Grinspan, et al., (2020), green spaces have many benefits, including climate control, community involvement, exercise, and relaxation. Both parks seemingly provide these benefits to their areas as they are green spaces, but they are different. They are very spacious, but they do not all have the same qualities. Both parks have basketball courts, a walking trail, a playground, benches, and other green spaces where you can enjoy nature and perform other activities. In addition, Buttonwood Park has a greenhouse, zoo, senior center, pond, and numerous monuments. The only additional attraction Riverside Park has is a skateboard park, and it is located on the Acushnet River. Although parks do not need the same attractions, Buttonwood Park may have the advantage of possibly attracting more people. One also must consider whether the additional attractions at Buttonwood Park result from the park’s location and the economic differences. This project reflects ideas from researchers and their findings on green spaces as well as their effect on people’s health. It will also use researchers’ findings on why green spaces are created and will discuss the hands-on research in the two parks and how people involved with the park feel it contributes to their lives.  

Methods 

I collected ethnographic data through observations, participant surveys, and semi-structured participant interviews. I collected this data at Riverside Park and Buttonwood Park, visiting on multiple days at various times. I began with observations, then moved to participant data collections after understanding roughly how many people visit the parks and at what times. 

This project has collaborated with the New Bedford Parks, Recreation, and Beaches Department. I will share the collected data with them so they can better understand the surrounding or adjacent communities and determine how to meet their needs. They want to provide the communities with what they want and need to the best of their abilities, and this will help them. The department wants safe spaces for people living in New Bedford, and they want to make sure they are sufficiently meeting the members’ needs for physical and mental health activities and spaces. 

The methods of collecting data underwent an approval process with the Institutional Review Board (IRB) of the University of Massachusetts Dartmouth. The IRB needed to approve the survey and interview questions, consent forms for the interviews, and the cover letters for the surveys. This process was necessary since human subjects were involved, and the privacy and confidentiality of their identities must be protected. Although the interview questions do not ask for identifiable information, participants must still sign a form agreeing to be recorded for the interview.  

I made sure to visit both parks during the same time of day and under similar weather conditions. I observed who was in the park, the demographics of park users, and what they were doing. After completing observations, I began conducting surveys and interviews at both parks. There were 23 survey questions (see Appendix A) and 10 interview questions (see Appendix B). 

Data Analysis

Many people visit Buttonwood Park throughout the day. Based on observations, approximately 43 people were using the park from 7:00  to 9:00 in the morning. Based on what was observed, it seemed as though the majority (38) of people were white, 1 person was black, 1 person was Asian, and 3 other people’s race could not be determined due to hoods and distance. All appeared to be adults over 18 years old. Most people were walking or jogging the perimeter paths alone, with friends, or with dogs. Of the people utilizing the paths, 19 were men, and 17 were women. People of all ages were at the park, but many seemed to be over 60 years old. 7 people were outside of the Senior Center, likely getting ready to go inside. Due to the park’s size, I had to drive around the park multiple times to count park users accurately. 

During the afternoon, there were approximately 102 people at the park within the two-hour window. 78 were adults over 18 years old, while 27 were minors. Again, the majority (around 90) of people were white, 5 were black, 5 were Latino, and 2 were Asian. Most people were in their 30s and 40s, with the exception of minors who were accompanied by adults. Most were utilizing the walking paths, though some were using the playground, open field (for a girls’ youth soccer practice) and going in and out of the senior center. The colder weather on this day may have impacted playground usage as only a few people were using it.  

Early morning observations at Riverside Park differed from observations from Buttonwood Park. 9 adults were observed using the park. Approximately 3 people were white, 2 were black, 3 were Latino, and 1 was too far away to be seen. 5 people were walking or jogging on the path, 2 appeared to be using the park as a shortcut to work while carrying work bags, and 2 people were playing soccer. Only one person appeared to be female while everyone else was male. Additionally, 10 high school-age students were waiting for the bus. They were standing outside the park bounds. 

Riverside Park proved to be a busy park in the afternoon. There were approximately 81 people at this park between the two hours; 51 of them were adults over the age of 18, and 30 were minors. In contrast to Buttonwood Park, the majority (57) of people were Latino, while there was a much smaller number of white people (13) and black people (8), and again, there were people (3) whose race could not be determined. Approximately 14 people were by the playground, 47 were on the soccer field, 18 were using the walking path, and 2 were at the skatepark. According to these results, usage of the park was evenly distributed among the park’s amenities and area.  

Over 2 days, 20 survey responses were collected from users of Buttonwood Park. Based on results from the people willing to participate, 80% identified as white, 15% as Latino/Hispanic, and 5% as mixed races (See Graph 1). 70% were female, 25% male, and 5% non-binary. Most participants were 60 years old or older (30%), in their 30’s (30%), or in their 40’s (25%). Only 1 person was between 19 and 24 years old, 25 and 29 years old, and in their 50’s. In terms of marital status, 55% were married/partnered, 40% were single/divorced, and 5% were widowed. 75% of people lived in households with 3 or more people, 15% lived with 2 people in their household, and 10% lived alone.  

Graph 1: Percentages of respondents’ identified race at Buttonwood Park. 

Graph 1: Percentages of respondents’ identified race at Buttonwood Park. 

Regarding income, of those living alone, 5 people who responded were below the median Massachusetts personal income of $49,746. For multi-income households (14), 57.1% were above, 21.4% were around, and 21.4% were below the median Massachusetts household income of $93,550 (See graph 2). 1 person chose not to respond to either question about income. 25% had no children under 18 years old living with them, while 30% had 1 child, 25% had 2 children, and 20% had 3 or more children. 75% of people owned their homes, while 25% rented. 

Graph 2: Percentages of how much a multi-income family makes of Buttonwood Park visitors. 

Outside of demographics, there were questions focusing on park usage. One of the survey questions asked how frequently people visited the park. 45% said “often,” 35% said “sometimes,” and 20% answered “rarely” (see Graph 3). Popular times were afternoons (90%), before 9am (15%), 9am-12pm (15%), and evenings (5%). Everybody felt safe at the park, but 1 person expressed  concern about the lack of lights in the park at nighttime. 80% of people would not visit the park at night. 84.2% of people said they would not send their children or grandchildren alone to the park. Only 15% of people feel the park is not kept clean. The other 85% think it is clean and 94.7% feel the structures are well-maintained. 50% of people walk the path, 35% play a sport or watch their friends and family play a sport, 10% bird-watch, 15% sit and read, and 30% bring their kids to the playground (See Graph 4). 63.2% felt community members took care of the park more than the city while the other 36.8% felt the city took better care. Only 20% were aware of the “Friends of Buttonwood.” 

Graph 4: Activities performed by participants at Buttonwood Park. 

There was a section at the end of the survey asking for comments from the participants. Of the 5 comments, the city and community were both acknowledged for their maintenance. Participants suggested more frequent grass cutting, adding restrooms inside the park, improving park cleanliness, and upgrading the playground equipment. Overall, everyone praised the park, its location, quietness, and cleanliness. 

 

Picture 1: Collecting Survey and Interview Responses at Buttonwood Park 

Over 3 trips were made to Riverside Park to collect 20 survey responses. Respondents identified as Black (35%), White (40%), and Latino/Hispanic (40%), with some selecting multiple races (See Graph 5). 55% were female, and 45% were male. Most were in their 30s (65%) or 40s (10%), with few other age groups represented. 80% were married or partnered, and 20% were single or divorced. 90% had 3 or more people in their household. 

Graph 5: Percentages of respondents identified race at Riverside Park. 

Only two people responded for single-income households; both were below the Massachusetts median personal income. For multi-income households (18), 38.9% reported making around the median and 61.1% reported making below that income (See Graph 6). 15% had no children under the age of 18 living with them, 45% had 1 child, 35% had 2 children, and 5% had 3 or more children. 35% of people owned their homes, and 65% rented.  

Graph 6: Percentages of how much a multi-income family makes of Riverside Park visitors. 

Park usage and visitation differed slightly from Buttonwood Park. 55% visited sometimes, 25% often, and 20% rarely. Most visited in the afternoon, with fewer people visiting in the mornings (5% before 9:00 a.m., 10% between 9:00 a.m. and 12:00 p.m.), and in the evenings (10%) (See Graph 7). Everyone felt safe at the park; however, 75% of people would not visit at night and 90% would not let their children go to the park alone. 75% felt the park was clean and all felt the park structures were well-maintained. Activities included playing a sport or watching friends and family play a sport (55%), bringing their children (45%), walking alone or with someone else (25%), birdwatching (10%), and reading (20%). 55% of people felt the city provided more care than the community. 84.2% were unaware of the “friends groups,” and after learning about them, 89.5% of people said they would like a “friend group” for Riverside Park. Only one person wrote in the comment section requesting increased police presence at the park during its busiest hours.  

Graph 8: Activities performed by participants at Riverside Park. 

Picture 2: Collecting Survey and Interview Responses at Riverside Park 

Buttonwood Park’s respondents to the interview questions were alike regarding their positive feelings about the park and what they felt could be added or taken better care of. Everyone felt it was safe and had plenty of space for activities. Some people only visit this park, though a few said they have traveled to others for new experiences or convenience. Everyone said they visited the park for its space and ability to relax and have fun. It was described as a great all-season park for its numerous amenities. When asked about their opinions on green spaces, everyone said they are important, and the world needs more. One person said, “We are paving the world” and there are not as many green spaces as before. Another person acknowledged the promotion of health and safety that comes from green spaces. 

No one would allow their children or grandchildren to visit the park alone. Two people attributed this to the day and age we live in. One person reflected on when they grew up, saying if it was around the 1970’s, then they would send them alone. Three people said it is due to their young age and fear of them getting stolen or hurt. These concerns are not specific to the park but general feelings. None of the participants would visit the park at night because of possible danger. One long-time resident of the area mentioned the park previously not having a good reputation at nighttime and attributed this to why they won’t visit at night. Another concern is the lighting around the area and it being too dark. In terms of safety concerns, other than suggesting a fenced-in playground, there were no major concerns. 

Not everyone could speak on the funding of the park, but some felt there must be more funding for this park due to its location and how well-maintained it is. One person said they believe there could always be more funding for green spaces and for Buttonwood Park, and they would like to see public restrooms. They understood the possible concern of homeless people camping in there or vandalism and suggested an electronic lock with a phone number to access the bathroom. This person also suggested more maintenance work on the playground. Overall, everyone enjoys the park and believes it is well taken care of by both the city and the “Friends of Buttonwood.”  

Additionally, everyone talked about how the park benefits their physical and mental health, as well as their children and grandchildren. Exercise, relaxation, and positive feelings come from being at the park and are what they enjoy so much. All respondents see the impact it has on their children and grandchildren. Mentioned activities included walking, playing on the playground, and summer evening yoga classes. People find the overall park positively affects their well-being and their children or grandchildren.  

Similar to Buttonwood Park, participants from Riverside Park had the same positive feelings regarding it. Only one respondent said they have traveled to other parks for experience, but prefer Riverside’s location, stating they would still visit if it was smaller. Another said they came here for the soccer field since none existed near their home in Dartmouth. All respondents stated they come to the park for convenience, experience, and the amenities it offers. Every participant felt safe when visiting and felt the park had a great amount of space. They all believe in the importance of green spaces and the necessity of more for people’s health benefits and the environment. 

When asked about allowing their children to go alone to the park, only one person said they would send their children alone, but not at night. Another said if their children were older, they could go alone. Safety concerns of the world today were also a factor for some people. Also, the frequent sightings of emergency vehicles, homeless people, and drug users were reasons cited for not sending their children alone. Participants had no concerns about allowing their children to visit the park at night if they were together. Without bringing the children, no one had concerns about visiting by themselves at night, although one person did say they would be more vigilant than if they went to a park in Dartmouth.  

Two participants think Riverside Park receives more funding because of the enclosed turf soccer field. Another person believes all New Bedford parks receive the same amount of funding, which is not enough in their opinion. They would like to see more maintenance work, especially on the water fountains that do not work when the splash pads are on in the summer. This person would also like to see a “friends group” started for the park. More than one person mentioned wanting the park cleaner, with less broken glass, and for homeless people to stop using the splash pads to shower and leave a mess. 

Similar to Buttonwood Park, almost everyone felt Riverside Park positively impacts their own and their children’s mental and physical health. This park is seen as a great available space by everyone. Only one person did not see an impact on their own mental health, but they do believe it impacts their child’s mental health since the child uses the park more.

Discussion 

Buttonwood Park and Riverside Park are both very lively, especially in the afternoon; however, there were differences in the demographics of visitors, particularly in race and gender. There were more white people at Buttonwood Park while there were more Latinos at Riverside Park. There was an even number of men and women observed at Buttonwood, while a majority of Riverside’s visitors were men. Results also show that activity at Buttonwood was geared towards the walking path, whereas the soccer field saw the most usage at Riverside. Walking paths at Buttonwood were primarily used for walking with some riding bikes. Riverside’s path was used for walking, bike riding, and skateboarding. Lastly, there was also a big difference in the ages of visitors between the two parks. The afternoon hours at Buttonwood saw more older adults (30s/40s) while Riverside saw more minors with more use of the playground.  

There were notable differences in survey responses between the two parks. The first notable difference is collecting responses at Riverside Park took one extra day. Riverside Park had more racial diversity and a more even split between females and males, while responses from Buttonwood Park were skewed heavily toward white women. There was an approximate 30-year age gap between responses, with respondents from Buttonwood being older on average. There were more divorced or single people at Buttonwood Park, ] one being a widow, whereas more people at Riverside Park were married or partnered. All single people from both parks said they make below the median Massachusetts personal income of $49,746. For married persons, no one from Riverside Park said they make above the median Massachusetts household income of $93,550; however, there were a few responses stating ‘above’ from Buttonwood Park. Child household percentages were similar. Housing status differed; most people from Buttonwood Park owned homes, while most people from Riverside Park rented. The afternoon was the busiest time for both parks, though some respondents visited at other times. Everyone felt safe at the parks and there was a mutual feeling of not wanting to visit at night. Most people from both parks would not send their children alone. People felt structures were well-maintained at both parks. Only a few more people at Buttonwood Park said they feel that the community takes more care of the park. Most participants from both parks had never heard about the “friends groups,” but most were interested in it. Many things are similar in this study, with the exception of the demographics involved. 

When reviewing the interview responses from both parks, there were both similarities and differences. Everyone felt safe and comfortable at the parks with positive feelings about the amenities and space provided. Both parks were described as wonderful green spaces that provided health, relaxation, and helped the environment. Most respondents said they would not send their children alone to the park and though reasons for this varied, general safety concerns were the number one reason. Everyone also recognized the mental and physical benefits the parks provide to them, especially their children and grandchildren.  

Differences primarily included park use, beliefs about funding, and desired improvements. Many people at Buttonwood Park had general safety concerns that did not pertain specifically to the park but to today’s world, while people at Riverside Park had concerns regarding homeless people, drug use, and uneasy feelings from emergency vehicle sightings. This is not to say there are no homeless people at Buttonwood Park. A person outside of the research collected shared that they had seen tents in the wooded area of Buttonwood Park and on a side street. Though people are unaware, this is important to note because it shows that homeless people can camp at any kind of park in any area.  

People seemed to have different beliefs about funding. Some people felt Riverside Park receives more funding because of the turf soccer field, while others said they believed Buttonwood receives more funding because of its location. In terms of improvements, people at Buttonwood mentioned playground upgrades and restrooms being added to the park. Riverside Park respondents expressed wanting better maintenance of the splash pad, more frequent cleanup of the park, and a “friends group.” Overall, the parks are valued by their visitors and people are not negatively impacted by them.  

Conclusion 

Green spaces provide people with numerous physical and emotional benefits while benefiting the environment. This study has shown that Buttonwood Park and Riverside Park positively impact community members in different areas of New Bedford. Although the parks are in different neighborhoods and include different amenities, data results have shown that visitors enjoy the parks and what they have to offer. It is evident that people with different socioeconomic statuses are separated in the city. Based on the collected data, people with lower incomes tended to live near Riverside Park and those with higher incomes tended to live near Buttonwood Park. However, this is not to say that people do not travel to each park from other areas. Visitors value both parks for their fields, playgrounds, and walking paths. They provide joy to their visitors and although some may have general safety concerns about the world, the parks instill feelings of relaxation and enjoyment. No one is overly concerned about the maintenance of either park and people expressed being happy when they visit. Some people believe that city planners create parks to separate people; however, this study shows the greater impact parks have when located in different neighborhoods. A park in a lower-income neighborhood may not attract many high-income earners, but that does not mean the park itself deters people. It may be difficult to look at Buttonwood Park the same way because of its history, but the important fact learned from this study is the importance of green spaces and their impact on the community they serve. Ultimately, this research highlights the importance of green spaces and their influence on community members’ mental and physical health. People largely view these parks as being welcoming to diverse backgrounds, as well as fostering activity and community involvement.

References 

Amano, T., Butt, I., & Peh, K. S. ‐H. (2018). The importance of green spaces to public health: A multi‐continental analysis. Ecological Applications, 28(6), 1473–1480. https://doi.org/10.1002/eap.1748 

Barton, J., & Rogerson, M. (2017). The importance of greenspace for mental health. BJPsych International, 14(4), 79–81. https://doi.org/10.1192/s2056474000002051 

Browning, M. H. E. M., & Rigolon, A. (2019). Could nature help children rise out of poverty? green space and future earnings from a cohort in ten U.S. cities. Environmental Research, 176. https://doi.org/10.1016/j.envres.2019.04.016 

Cole, H. V., Lamarca, M., Connolly, J. J. T., & Anguelovski, I. (2017). Are green cities healthy and equitable? Unpacking the relationship between health, green space and gentrification. Journal of Epidemiology and Community Health (1979-), 71(11), 1118–1121. https://www.jstor.org/stable/26383998 

Environmental Protection Agency. (n.d.). Green Streets and Community Open Space. EPA. Retrieved April 25, 2023, from https://www.epa.gov/G3/green-streets-and-community-open-space 

The Friends of Buttonwood Park. The Friends of Buttonwood Park. (2022, February 2). https://buttonwoodpark.org/ 

Graeber, D., & Wengrow, D. (2021). Free People, the Origin of Cultures, and the Advent of Private Property. In The Dawn of Everything (pp. 112–155). 

Grinspan, D., Pool, J.-R., Trivedi, A., Anderson, J., & Bouyé, M. (2020, September 29). Green space: An underestimated tool to create more equal cities. World Resources Institute. Retrieved from https://www.wri.org/insights/green-space-underestimated-tool-create-more-equal-cities#:~:text=Green%20space%20can%20help%20make,air%20quality%20and%20mitigating%20flooding. 

Low, S., Scheld, S., & Taplin, D. (2005). Rethinking Urban Parks: Public Space & Cultural Diversity (1st ed.). University of Texas Press. 

Ramirez, R. (2023, May 10). Building community, one urban garden at a time. Groundwork USA. https://groundworkusa.org/building-community-one-urban-garden-at-a-time/ 

Rigolon, A., Yañez, E., Aboelata, M. J., & Bennett, R. (2022). “A park is not just a park”: Toward counter-narratives to advance equitable green space policy in the United States. Cities, 128. https://doi.org/10.1016/j.cities.2022.103792 

Rosenzweig, R., & Blackmar, E. (1992). The Park and the People: A History of Central Park. Cornell University Press. 

Scott, James C. (1998). Seeing Like a State, Yale University Press. 

CNR Web (2022, April 20). How green spaces can improve your health. College of Natural Resources News. Retrieved from https://cnr.ncsu.edu/news/2022/04/parks-green-spaces-improve-health/ 

Appendix A 

Survey Questions 

Research in Nursing

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Perceived Barriers to Medication Adherence in Latinos with Hypertension Using the World Health Organization Multidimensional Adherence Model

By Chelsea Martinez

 

Project Description 

Recent data indicates that hypertensive Latino adults reported the lowest adherence to their medications (67%) when compared with Black and White adults (77% for both groups) (Schoenthaler et al., 2019). As a result, a lack of medication adherence (MA) among Latinos/Hispanics with hypertension (HTN) leads to higher HTN-related mortality at a faster rate in Latinos than in other ethnic groups (Schoenthaler et al., 2019). Suboptimal MA is a main contributor to chronic conditions in this community, including HTN. For context, Hispanics/Latinos are an underrepresented population in research (Dreyfus et al., 2023). The current research on barriers to MA in the hypertensive population is often not tailored to Latinos, the largest racial and ethnic minority in The United States (US Census Bureau, 2023). Currently, Hispanics have a prevalence of HTN of 43.7 percent (Ostchega et al., 2020). With a rapidly growing population and significant HTN prevalence, it is imperative that effective interventions to increase MA are developed. However, without research that aims to identify the primary barriers to MA within the population, effective and efficient interventions cannot be tailored. 

       Therefore, the objective of this study is to identify the primary barriers to MA within the hypertensive Latino population in the Greater New Bedford area by utilizing the World Health Organization’s Multidimensional Adherence Model (WHOs MAM) (Sabaté, 2003). The barriers within the MAM include personal, socioeconomic, therapy, health care, and condition-related factors. Using quantitative data, this project aims to find the positive and negative associations between certain dimensions delineated by the MAM and self-reported MA. Participants have been chosen based on a convenience sample of 20 Hispanics/Latinos with HTN from the health clinic at the Community and Economic Development Center (CEDC) in New Bedford, as facilitated by the director, Corinne Williams. Participants have also been recruited from Southcoast Health’s Cardiology office, facilitated by Vicki Saint-Paine, a nurse practitioner at the clinic. Materials include instruments that measure MA and personal beliefs about medication and healthcare. An iPad is being used to show educational videos that compose the brief teaching intervention on MA. Participants have only spoken English and Spanish so far. As a native Spanish speaker, I have translated the surveys into Spanish and have distributed the translations as needed. 

       Ultimately, the purpose of this research is to build on the information known about MA and the barriers that affect the Latino population. This will be done with the hope of discovering the prevalent personal, condition, therapy, socioeconomic, and healthcare-related barriers to MA to understand and discover specific aspects of adherence that can influence the establishment of effective interventions. 

Progress 

As of June 28, 2024, I have 20 enrolled participants for the study. This is the total number of participants needed for the study. Only thirteen participants have completed the study; the remaining seven participants are scheduled to participate in the study within the next week. This also signifies that thirteen of the ten-dollar Market Basket gift cards have been distributed, with seven left. Thirteen out of 20 participants have been recruited from the CEDC. The paper surveys are used and then entered into Qualtrics. My goal is to finish surveying participants by the second week of July. I am grateful to the OUR and its committee once again for giving me this opportunity and allowing me to provide participants with an incentive.

 

References 

Dreyfus, B., Kuri, L., Ferri, M., Doykos, P., Fazeli, M. S., Hofer, K., Andonova, A., Ferri, L. (2023). Understanding Hispanic/Latino Participation in Clinical Trials and Observational Studies, and Strategies to Increase Participation: A Targeted Literature Review. Journal of Health Care for the Poor & Underserved, 34(1), 399–424. https://doi- org.libproxy.umassd.edu/10.1353/hpu.2023.0026 

Kvarnström, K., Westerholm, A., Airaksinen, M., & Liira, H. (2021). Factors Contributing to Medication Adherence in Patients with a Chronic Condition: A Scoping Review of Qualitative Research. Pharmaceutics, 13(7), 1100. https://doi.org/10.3390/pharmaceutics13071100 

Ostchega, Y., Fryar, C. D., Nwankwo, T., & Nguyen, D. T. (2020, April 24). Hypertension Prevalence Among Adults Aged 18 and Over: United States, 2017–2018. Centers for Disease Control and Prevention. https://www.cdc.gov/nchs/products/databriefs/db364.htm#:~:text=In%20survey%20perio d%202017%E2%80%932018,%25%20(60%20and%20over). 

Sabaté, E. (2003). Adherence to long-term therapies: Evidence for Action. World Health Organization 

Schoenthaler, A., de la Calle, F., Pitaro, M., Lum, A., Chaplin, W., Mogavero, J., & Rosal, M. C. (2020). A Systems-Level Approach to Improving Medication Adherence in Hypertensive Latinos: a Randomized Control Trial. Journal of general internal medicine, 35(1), 182–189. https://doi.org/10.1007/s11606-019-05419-3 

US Census Bureau. (2023, September 28). Hispanic Heritage Month: 2023. Census.gov. https://www.census.gov/newsroom/facts-for-features/2023/hispanic-heritage-month.html 

Research in Bioengineering

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Examining the Effect of Vitamin D on Melanoma

 By Mary Goodrow

 

Introduction 

Over the summer and during the spring semester, I was introduced to research and working with live animals. I also learned to keep a lab notebook and record lab activities during this introductory process. The first thing I did was separate fertilized fish eggs from unfertilized ones. First, it was hard to distinguish fertilized eggs from unfertilized ones. However, it was cool to see under the microscope. Later, after the semester had ended, I started learning how to make agarose plates. Agarose plates are needed in the process of producing Zebrafish with melanoma, which is done via recombinant DNA technology. It was a fun learning process being introduced to the lab equipment and procedures for using it. One of the graduate students was nice enough to offer some help along the way. I made some mistakes, especially since I am not used to the sterile technique. I made the mistake of placing the lids of the plates on the counter. It’s now ingrained in me to open the top of the lid only slightly when working with Petri dishes.  

Methods 

In my lab notebook, I kept track of all the instructions given to me by Dr. Ferreira, mainly the ones directly related to the Agarose plates. Each page was formatted based on the date and name of the procedure we performed that day. I initially listed the procedure for my main experiment, following space set aside for a table of contents.  

Results 

Despite the premature discontinuation of the experiment, I am proud to share that my agar plates were a success. Though new to me, the process of changing the tanks and feeding the Zebrafish was also carried out successfully. This first-time experience, without any prior knowledge, was a testament to the effectiveness of our approach, even in the face of unexpected circumstances. 

Discussion/Turn of Events 

In a significant turn of events, I have made the difficult decision to continue my academic journey at Vanderbilt University as a Biomedical Engineering student. This decision was not easily made, and it brings with it the uncertainty of being able to complete my experiment before my departure. However, I am certain that the lessons I have learned will undoubtedly shape my future endeavors.  

Research in Biology

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Invasive Species and Their Effect on Relative Abundance of Native Species in the Wild

By Brent Mello

My research is intertwined with the research of a graduate student, Eleanor DiNuzzo. Through the last two summers, she has surveyed the same three rocky intertidal sites in Massachusetts for four species: Hemigrapsus sanguineus (Asian shore crab), Carcinus maenas (green crab), Nucella lapillus (Atlantic dogwhelk), and Mytilus edulis (blue mussel). I first learned about these four intertidal species last year when I started to help with Eleanor’s intertidal surveys. I was taught how to identify the different species, as well as the sex of the crabs. 

In early April, the sites were visited for the first time for the season. We were making sure the sites were still prepped for our surveys. At each tide height, 0m, 0.5m, and 1m, we located the rocks that had been tagged with “dog tags.” If any rock had lost the tag applied from the previous years, we had to apply a new tag. Only two new tags had to be applied this year. We utilized marine epoxy that we had in the lab. I have included a picture below of me using epoxy to bond our tag to the rock. I have also included a picture of the tag bonded to the rock two months later during our June surveys.  

In my original grant proposal, I had planned for the possibility that most of the tags had fallen off over the winter. Since this was not the case, the money for the drill, masonry bits, and anchor kit was not needed. I did not want to spend the granted money on unnecessary tools. 

Our next visits to the sites were planned for May 5th-7th, but we encountered external issues that made us have to drop these survey dates from our data.  

The June surveys were a success and were conducted on June 6th-8th. During our surveys, we would get to the site about a half hour prior to the A.M. low tide. This would give us time to prep for our surveys. Once we got to the sites, we located the 0m marker as quickly as possible. In some locations, the tide could chase us out, so getting to the mark as early as possible allotted more time for our surveys. After the 0m mark was found, a 30-yard transect was set up. Five random numbers were generated per tide height between 1 and 30, which were used to place down the quadrat for surveying. I have included a picture of the 1m X 1m quadrat we used in our surveys.  

Once each quadrat was placed, surveying began. One person was set as the data recorder, while the other 2 or 3 surveyors were tasked with searching each quadrat for the four targeted species. Each quadrat followed the same procedure, which I will give step by step. Firstly, we began by moving seaweed out of the way while also searching through it for any of our species of interest. We tried to be minimally invasive and would remove seaweed as little as possible, more so trying to rearrange it. The next step was to search the rocks for and collect Nucella, Mytilus, Carcinus, and Hemigrapsus. We would collect a max of 10 mussels (Mytilus) and as many of the other species present in the quadrat. Crabs (Hemigrapsus or Carcinus) smaller than 10mm were not collected and were called out to the recorder as small, followed by species type. Once the visible species were collected without disturbance, we proceeded to turn over rocks and collect every individual we could, placing them in a bucket. After all rocks were overturned and returned to their initial locations, we began measuring the individuals collected while simultaneously sexing the crabs. Their sizes were measured using battery-powered calipers. The calipers from the previous year were not damaged by the moisture, and we were able to use them. For this reason, I did not use the original plan to buy the waterproof calipers yet. We may need to order more calipers if the current ones get damaged during our future surveys. The research will proceed through the summer with three survey days each month ending in September. After each month’s surveys, data will be transferred from the survey sheets to an Excel spreadsheet. I have included a picture of the survey sheet below from one of our June surveys. 

After the last survey in September, statistical analyses will begin. The statistical tests will be run in R, which I learned the basics of in Bio 430 last semester. I very much look forward to applying the use of R to real-life biological studies, as well as becoming more familiar with other features of the system. The goal is to use non-metric multidimensional scaling (nMDS) in R to compare community composition at different tide heights and locations. I will be using the last three years of survey data for these statistical comparisons aiming to find a significance difference in community composition to see the effects of the presence of the invasive Hemigrapsus in New England coastal rocky intertidal zones. By December, I plan on completing the statistical analyses and having a manuscript ready for submission. 

I would like to thank the OUR department for the grant approval back in January, as well as the support this provides not just for me, but for all undergraduate researchers. Since there may be unexpected expenses that arise over the summer, I would like to request an extension on the grant money. I plan to use it wisely for tools or programs that may be necessary or will benefit the research as a whole.  

Extra pictures from the surveys: 

Picture of me holding a gravid (pregnant) Carcinus maenas 

 

         Hemigrapsus and how they are measured and sexed 

 

         Nucella 

 

Every survey is accomplished with a team effort, so I have included a team picture from one of our June surveys.  

RESEARCH IN PSYCHOLOGY

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Impostor Syndrome and Diversity: Race, Ethnicity, and Generation
By Alejandra M. Ponce

 

INTRODUCTION

While high-achieving students are admirable in their efforts and accomplishments, their personal well-being is cause for concern. While one would assume that hard-working academic students acknowledge themselves for the work they accomplish, sometimes students will trudge through their scholastic journeys and accept their accolades with shame, guilt, and/or fear—even after being awarded for their achievements. This phenomenon can be attributed to impostor syndrome, which involves chronic self-doubt and fear of being exposed as a fraud in connection to feelings of incompetence, inadequacy, and an inability to feel self-pride or internalize accomplishments (Bravata et al., 2019; Cokley et al., 2017). Previous research has linked impostor syndrome to emotional and physical exhaustion, generalized anxiety, diminished workplace performance, and even decreased diversity in institutional leadership positions (Bravata et al., 2019; Cokley et al., 2017; Moyer et al., 2021; Rivera et al., 2021). Although research findings recognized that impostor syndrome is common as well as personally and socially detrimental to the individuals experiencing it, there seems to be conflicting evidence about how exactly it affects people, specifically students in universities.

Other factors that have been previously implicated in research on impostor syndrome include general mental health (Bernard et al., 2017; Cokley et al., 2013; Cokley et al., 2017; Cusack et al., 2013), perfectionism (Cusack et al., 2013, Pannhausen et al., 2020; Rohrmann et al., 2016), and self-esteem (Cusack et al., 2013; Ibrahim et al., 2020). Moreover, previous research has suggested complex associations between impostor syndrome and race and ethnicity (Ayesiga, 2022; Bernard et al., 2017; Cokley et al., 2013; Cokley et al., 2017; Jackson et al., 2022) while leaving first-generation student status largely unexplored. Therefore, this study aimed to explore the possible differences in experiences of impostor syndrome in ethnically and racially underrepresented and first-generation students in comparison to represented and continuing-generation students, respectively. In this mixed methods study, potential links between diversity-related variables such as race, ethnicity, and first-generation student status relative to impostor syndrome and other personal variables were investigated.

METHODS

Quantitative Study

Participants

Upon approval from the Institutional Review Board (IRB) at the University of Massachusetts Dartmouth, participants were recruited through physical flyers (see Appendix A) posted around academic buildings on campus, where a quick response (QR) code directed them to the survey on Qualtrics. Data was collected from a sample of 39 participants. 7 of the survey responses were removed from the data analysis due to incomplete responses. Demographic information of the sample (n = 32) is summarized in Table 1. The sample mostly consisted of female (81.3%), White (81.3%) upperclassmen in the College of Arts & Sciences (71.9%) at the University of Massachusetts Dartmouth. Participants were given $10 gift cards upon participation.

Instruments

Demographics. Participants were asked to report demographic information such as their gender assigned at birth, gender identity, age, race, ethnicity, current cumulative grade point average (GPA), major, current year of study, whether they identified as a first-generation student, and the educational levels of their parents or legal guardians. All these variables were suspected, or have shown, to be associated with impostor syndrome.

Impostor Syndrome. Feelings of impostor syndrome among participants were measured using the Clance Impostor Phenomenon Scale (CIPS; Clance & Imes, 1978). The CIPS is a 20-item scale rated on a 5-point Likert scale ranging from 1 (not at all true) to 5 (very true). Sample items include “I can give the impression that I’m more competent than I really am” and “I’m afraid people important to me may find out that I’m not as capable as they think I am.” Total scores on the instrument are summed with final scores between 20-100. Higher scores indicate greater feelings of impostor syndrome. Holmes and colleagues (1993) reported high internal consistency reliability of the scale ( = .96). The scale had an internal consistency of = .93.

General Mental Health. To account for general state of mental well-being, participants completed the Mental Health Inventory-5 (MHI-5; Berwick et al., 1991). A 6-point scale ranging from 1 (all of the time) to 6 (none of the time) is used to measure the 5-item scale. Sample items include “During the past month, how much of the time were you a happy person?” and “How much of the time, during the past month, have you been a very nervous person?” The scale is scored from 0 to 100 (items 1 and 2 are reverse-coded) with a score of 100 indicating excellent mental health. A Cronbach’s alpha of .84 was previously found for the scale (Nearchou et al., 2019), but had a poor internal consistency of = .25.

Perfectionism. The Frost Multidimensional Perfectionism Scale (FMPS; Frost et al., 1990) is a 35-item scale that measures perfectionism on four subscales: concern over mistakes and doubts about actions, concern with parental expectations, high personal standards, and concern with organization. However, a total score was used in this study. Sample items include “My parents set very high standards for me” and “Organization is very important to me.” A 5-point Likert scale ranging from 1 (strongly disagree) to 5 (strongly agree) is used to measure responses, with higher scores indicating higher perfectionistic tendencies. A Cronbach’s alpha of .93 was found for the total scale (Pannhausen et al., 2020). The scale had an internal consistency of = .93.

Self-Esteem. As for self-esteem, the Rosenberg Self-Esteem Scale (RSES; Rosenberg, 1965) was used. The RSES contains 10 items and measures responses with a 4-point Likert scale ranging from 1 (strongly disagree) to 4 (strongly agree). Sample items include “On the whole, I am satisfied with myself” as well as “At times I think I am no good at all.” Scores on the scale are summed and total scores range from 4 to 16 with higher scores indicating higher self-esteem. A Cronbach’s alpha of .84 was previously found for the scale (von Collani & Herzberg, 2003), but had a very poor internal consistency of = .09.

Diversity-Related Stressors. To measure stressors that are generally experienced by the student population as well as those uniquely experienced by students that are underrepresented, the Minority Status Stress Scale (MSSS; Smedley et al., 1993) was used. The MSSS is a 33-item scale utilizing a 6-point Likert scale ranging from 0 (does not apply) to 5 (extremely stressful). There are six subscales: achievement stressors, environmental stressors, race-related stressors, intrapersonal stressors, interpersonal stressors among ethnic minority groups, and interpersonal stressors with White people. Sample items include “The university is an unfriendly place” as well as “Being treated rudely or unfairly because of my race.” Subscale and total scores may be used, but for the purposes of this study, only total scores were used. Total scores on the scale are summed and range from 0 to 185 with higher scores suggesting more feelings of diversity-related stress. Internal consistency of the scale has been reported with a Cronbach’s alpha of .92 (Greer & Chwalisz, 2007). The scale had an internal consistency of = .96.

Qualitative Study

Participants

Upon approval from the IRB at the University of Massachusetts Dartmouth, participants were asked at the end of the Qualtrics survey whether they were willing to take part in the second phase of the research. Participants were given the option to input their school email address as an indication that they were interested in being contacted to schedule an in-person interview. Participants were made aware that agreeing to participate and inputting their school emails would allow the student and primary investigators access to their quantitative survey data, which was used to identify whether the participant met the inclusion criteria for the in-person interviews. Data was collected from a sample of 4 participants, all female, between the ages of 20 and 22. All the participants fit the inclusion criteria, which included identifying as a non-White race, as part of an ethnic group, or as a first-generation student. Each participant was also required to have experienced moderate to intense impostor feelings, according to their CIPS scores (Clance & Imes, 1978). Participants were given $20 gift cards upon participation.

Instruments

In-Person Interviews. A qualitative descriptive design, as explained by Sandelowski (2000) was used for the method. An in-person, responsive interview technique following a conversational guide (Rubin & Rubin, 2012) was conducted. Interviews were 15-30 minutes in duration and were conducted on campus in quiet, private locations. The student researcher completed the interviews. All interviews were audio recorded using Zoom, so than an electronic transcript could be prepared later on. Only voices were heard, and no images were recorded of the student researcher or conversational partner. Questions on the interview guide included general, open-ended inquiries about impostor syndrome as experienced by the individual (see Appendix B).

RESULTS

Quantitative Study

Spearman’s rho correlations were conducted to analyze potential relationships between study variables, as illustrated in Table 2. No correlations were found relating to race. As for ethnicity, no significant correlations were found with regards to the personal variables analyzed such as impostor syndrome, mental health, perfectionism, self-esteem, and diversity-related stressors. However, significant correlations were found for impostor syndrome and low self-esteem (r = -.36, p = .045), and impostor syndrome and perfectionism (r = .65, p < .001). No other significant correlations among the personal variables were observed.

A hierarchical multiple regression analysis showed that model 1: first-generation student status and ethnicity (step 1) explained 4% of the variance but was nonsignificant. Entry of personal variables, such as perfectionism, mental health, self-esteem, and diversity-related stress added a significant amount of variance to the model, explaining 55% of the variance, More specifically, perfectionism was the only significant factor ( = .60, p  < .001). The hierarchical multiple regression analysis for the variables of interest is displayed in Table 3.

 

Table 1: Demographic information of the sample (n = 32).

Variable Mean SD n %
Age (years) 22.1 6.48
GPA 3.30 0.782
Gender Assigned at Birth

Female

Male

  

 

  

28

4

  

87.5

12.5
Gender Identity

Female

Male

Prefer not to say

  

26

5

1

  

81.3

15.6

3.1
Race

American Indian or Alaska Native

Asian

Black or African American

Native Hawaiian or Pacific Islander

White or Caucasian

Other

  

 

  

1

1

5

1

26

1

  

3.1

3.1

15.6

3.1

81.3

3.1
Ethnicity

Hispanic or Latino or Spanish Origin

Not Hispanic or Latino or Spanish Origin

  

4

28

  

12.5

87.5
First-Generation Student

Yes

No

  

11

21

  

34.4

65.6
Parent or Legal Guardian Education Level

Less than College

Some College or More

  

25

39

  

39.1

60.9
Major

Charlton College of Business

College of Arts and Sciences

College of Engineering

College of Nursing & Health Sciences

College of Visual & Performing Arts

  

2

23

3

3

1

  

6.2

71.9

9.3

9.3

3.1
Year

Freshman (1st year)

Sophomore (2nd year)

Junior (3rd year)

Senior (4th year)

  

2

8

13

9

  

6.3

25.0

40.6

28.1

 

Table 2: Correlations between race, ethnicity, personal variables, and diversity-related stressors (n = 32).

Variables 1 2 3 4 5 6 7
1. Race –
2. Ethnicity – –
3. Impostor Syndrome – .18 –
4.Perfectionism – -.31 .65** –
5. Self-Esteem – -.22 -.36* -.21 –
6.Diversity-Related Stressors – -.25 -.05 .11 -.21 –
7.Mental Health – -.13 -.29 -.16 .53 .30 –
Key: ** p < .001, * p < .05

 

Table 3: Hierarchical multiple regression analysis for demographic and personal variables influencing impostor syndrome (n = 32).

Step and Predictor Variable B SE B Beta R2 R2
Step 1:

Demographic Variables

 .04
First-Generation Studenta 2.48 6.17 .08
Ethnicityb 7.63 8.89 .16
Step 2:

Personal Variables

 .55 .52
Perfectionism .44 .10 .60***
Mental Health -1.54 .91 -.27
Self-Esteem -.71 .87 -.13
Diversity-Related Stressors .02 .09 .04
Note. *** p < .001

a Coded: 0 = yes, 1 = no

b Coded: 0 = Hispanic, Latino, Spanish origin, 1 = not Hispanic, Latino, Spanish origin

Qualitative Study

Transcripts underwent inductive content analysis. Individual transcripts were reviewed for relevant segments that were coded based on study aims. Coded segments were entered into an excel file, collapsed into categories, and finally into representative themes. Any quotations from participants used in the results and discussion were presented anonymously.

The participants reported experiencing impostor syndrome mostly at school, and in some cases, at work, particularly when put into leadership roles. Participants described impostor syndrome related to academic pursuits as feeling like they are “not good enough.” They also reported “feeling like [they] didn’t belong there,” especially in regard to science, technology, engineering, and mathematics (STEM) majors and the Honors College. One participant reported changing majors, and another reported dropping out of the Honors program altogether due to severe feelings of impostor syndrome. They both described that the feelings of impostor syndrome “didn’t allow [them] to continue” in either major or program.

Moreover, in both student and work roles, participants described having chronic, significant self-doubt and engaged in negative self-talk. In most cases, the participants’ family members, supervisors, and professors believed in the ability of the participant more than the participants believed in their own abilities. Some contributing factors to feelings of impostor syndrome, as reported by the participants, included being a first-generation college student, experiencing parental pressure to achieve, and competition with siblings. These contributing factors appeared to promote more negative self-talk, including statements like “I feel like I am not as smart as my siblings.”

When receiving awards and accolades, all participants reported discomfort and claimed feeling like “[they] didn’t deserve it” and “didn’t want to sound like [they] have a big ego.” One participant reported that she hides her awards under her bed rather than displaying them.

All participants described themselves as being kind, caring, and giving individuals with more introverted personality types and perfectionistic tendencies. Interestingly, despite their feelings of impostor syndrome, all participants also reported moderate to moderately high self-esteem.

Lastly, when asked whether their feelings of impostor syndrome progressed over the years, participants reported decreased feelings of impostor syndrome. They attributed their lessened feelings of impostor syndrome with increased self-assurance facilitated by faculty, therapists, and friends. Religious and spiritual faith also seemed to be a protective factor against feelings of impostor syndrome.

Discussion

Research investigating impostor syndrome and its manifestation and influence in underrepresented groups are often complex and inconclusive (Cokley et al., 2013; Cokley et al., 2017), especially when also considering more personal variables, such as perfectionism and self-esteem (Cusack et al., 2013; Ibrahim et al., 2020; Pannhausen et al., 2020; Rohrmann et al., 2016). This study investigated the potential relationship between impostor syndrome and diverse social identifiers such as race, ethnicity, and first-generation status, and how it manifests in the experience of impostor feelings. The study was also concerned with determining predictors of impostor syndrome. It was hypothesized that underrepresented and first-generation students would experience impostor syndrome more severely than majority and continuing-generation students as a product of interpersonal and environmental stressors that may come with identifying as a non-White race, being part of an ethnic group, or being a first-generation student. It was also hypothesized that those variables would serve as predictors of impostor syndrome. It was observed, however, that students experienced impostor syndrome similarly across the board, with no notable difference in the manner in which underrepresented and first-generation students experienced impostor syndrome. It was also observed that identifying as a non-White race, as part of an ethnic group, or as a first-generation student were not effective predictors of impostor syndrome. Instead, a significant positive relationship was found for impostor syndrome and perfectionism, and a significant negative relationship was found for impostor syndrome and self-esteem. As for predictors, only perfectionism was shown to be a significant predictor of impostor syndrome.

Correspondence with Prior Research

Aligned with prior research investigating the variables of perfectionism and self-esteem in relation to impostor syndrome, this study also found evidence of a significant positive relationship between impostor syndrome and perfectionism (Cusack et al., 2013; Pannhausen et al., 2020; Rohrmann et al., 2016). As for the variable of self-esteem, this study was able to find a significant negative correlation for impostor syndrome and self-esteem, which supports findings in some prior research (Pannhausen et al., 2020; Rohrmann et al., 2016), but opposes other studies that did not find a significant relationship between impostor syndrome and self-esteem (Cusack et al., 2013), or only found a significant relationship between self-esteem and a specific aspect of impostor syndrome rather than with the concept as a whole (Ibrahim et al., 2020).

As for the variables that yielded nonsignificant results, this study was unable to find a significant correlation between impostor syndrome and mental health that has previously been found in other studies (Cokley et al., 2013; Cusack et al., 2013). While other studies have revealed correlations between minority status stress and impostor syndrome (Cokley et al., 2013), as well as evidence of influences of impostor syndrome on ethnic minority students (Cokley et al., 2017), none of the findings in this study yielded significant correlations between impostor syndrome and race or impostor syndrome and ethnicity.

Considering the resulting themes of the qualitative interviews, the findings of this study resembled findings of prior qualitative studies (Ayesiga, 2022; Jackson et al., 2022). Primarily, feelings of invalidation and not belonging were echoed in the interviews in this study. However, themes involving race or ethnicity were not overarching themes. If mentioned, they were only mentioned briefly.

Potential Mechanisms

Although not a major focus of this study, perfectionism was found to be significantly, positively correlated with impostor syndrome. As the concept of impostor syndrome heavily focuses on aspects of inadequacy and incompetence, it is logical that perfectionism would be related to impostor syndrome. Whether as a means of compensating or as a means of disguising oneself, it is possible that people experiencing impostor syndrome will develop perfectionistic tendencies as a way of managing their impostor feelings. The opposite may be true as well. It may be that people who develop excessive perfectionistic tendencies succumb to feelings of impostor syndrome as a result of always needing everything to be perfect and the impossibility of a perfect outcome. Regardless, perfectionism is often one of the factors contributing to student success because of their self-discipline, attention to detail, and organizational qualities. Thus, perfectionism may perpetuate a cycle with impostor syndrome, where the higher the perfectionistic tendencies are, the more likely people are to become wary of their success, and the more fearful people are of being discovered as a fraud, the more they become perfectionists to presumably avoid being exposed.

Furthermore, perfectionism was also found to be an independent predictor of impostor syndrome. This suggests that higher perfectionistic tendencies may become maladaptive to the individual, and thus create significant dissatisfaction with the self, followed by feelings of unworthiness and insufficiency—both of which are significant aspects of impostor syndrome. As mentioned before, as perfectionism and perfectionistic tendencies are often contributors to student success, this also suggests that there may be an optimal level of perfectionism, and any higher than the optimal level may contribute to feelings of impostor syndrome and become detrimental to the individual.

Moreover, as impostor syndrome is conducive to negative feelings of the self, including feelings of unworthiness, it is almost intuitive that self-esteem would be negatively impacted, despite prior research not finding consistently significant correlations between impostor syndrome and self-esteem. However, this significant finding needs to be taken with a grain of salt. The scale used to measure the general mental health of the participants yielded a poor value of internal consistency in the study. As such, the validity of the significant finding is compromised and requires further evaluation.

Unexpected Results

It was expected that significant correlations between impostor syndrome and race, ethnicity, and first-generation status would occur, as well as some evidence that these variables can act as predictors of impostor syndrome. However, that was not the case. None of these variables yielded significant results. This may be for several reasons. Primarily, it is possible that the demographic composition of the sample was not diverse enough to detect any differences in impostor syndrome between races or ethnicities. As the sample of the quantitative data was 81.3% White (Table 1), it is highly likely that the sample lacked the representation it needed in order to find significant findings for factors of race and ethnicity. Not to mention, only 34.4% of the participants were first-generation students, and thus may have also affected the results of the study.

However, the participants in the qualitative study, all of whom were either or both Black and Hispanic, also placed little importance on the role of their race or ethnicity in its contribution to their feelings of impostor syndrome. As such, it is possible that the demographic composition of the University of Massachusetts Dartmouth affected the environmental stressors that come with identifying as a non-White race, as part of an ethnic group, or as a first-generation student, which then affected the results of the study. Prior research on these variables has been conducted with samples of students in predominately White universities with very little diversity, whereas the University of Massachusetts Dartmouth often prides itself on its diverse student body (UMass Dartmouth, 2023). As such, it is possible that the environment on campus does not produce feelings of hypervisibility in terms of race (Jackson et al., 2022) or perhaps students on campus do not face as many instances of discrimination relative to other students in predominately White universities (Cokley et al., 2013), which may neutralize factors of race, ethnicity, or first-generation status when considering variables related to impostor syndrome.

Furthermore, although general mental health was not a variable that was heavily focused on, it was expected that it would yield a significant, negative correlation with impostor syndrome. However, a nonsignificant correlation was found (Table 2), opposing previous research (Cokley et al., 2013; Cusack et al., 2013). This may be due to the poor internal consistency of the scale used ( = .25.) and the small sample size employed for the statistical analyses.

Limitations

As the current study was conducted with very small sample sizes, the samples are likely not representative of the larger population, and thus bring to question the results that would come from larger sample sizes. In addition to sample size, it is important to consider that the sample in the quantitative study was predominately White (81.3%; Table 1), which may have also skewed the results, as other races and ethnicities may not have been adequately represented. Moreover, the quantitative study was conducted via a survey online and the qualitative survey was conducted in person with the interviews recorded on Zoom. It is possible that the results of the study were skewed by self-report bias, as participants may be unwilling to report or be unaware of the true values of the variables the study was measuring. Correspondingly, the results may also be impacted by social desirability bias, in which the participants enhance the portrayal of themselves. Especially considering most of the sample in this study had moderate to intense feelings of impostor syndrome, it is not unlikely that they would skew their responses in order to keep up with appearances and not expose themselves as frauds, as is crucial to the experience of impostor syndrome.

Implications and Future Directions

The results of the current study suggest that perfectionism is both correlated and is an independent predictor of impostor syndrome, while other variables related to diversity, such as race, ethnicity, and first-generation status are not. Future studies should attempt to replicate this study using larger and more diverse sample sizes to reconsider the potential links between diversity-related variables and impostor syndrome. In addition, more research should be conducted on this almost unanimous correlation found between impostor syndrome and perfectionism, as perfectionism may be a viable path to understanding the mechanism of impostor syndrome and can even potentially be used to screen people for impostor syndrome. These studies may also be used to develop interventions that will decrease harmful perfectionistic tendencies and, in turn, hopefully decrease impostor feelings. Additionally, future researchers should also explore the long-term implications of these elevated levels of perfectionism and impostor syndrome.

 

References

Ayesiga, Y. T. (2022). “Do I even belong here?: An Examination of Impostor Phenomenon among First-generation College Seniors and the Perceived Factors that Contribute to Resilience and Persistence on the Journey to Graduation” (dissertation). ProQuest Dissertations Publishing, Ann Arbor, MI.

Bernard, D. L., Lige, Q. M., Willis, H. A., Sosoo, E. E., & Neblett, E. W. (2017). “Impostor Phenomenon and Mental Health: The influence of racial discrimination and gender.” Journal of Counseling Psychology, 64(2), 155–166. https://doi.org/10.1037/cou0000197

Berwick, D. M., Murphy, J. M., Goldman, P. A., Ware, J. E., Barsky, A. J., & Weinstein, M. C. (1991). “Performance of a Five-item Mental Health Screening Test.” Medical Care, 29(2), 169–176. https://doi.org/10.1097/00005650-199102000-00008

Bravata, D. M., Watts, S. A., Keefer, A. L., Madhusudhan, D. K., Taylor, K. T., Clark, D. M., Nelson, R. S., Cokley, K. O., & Hagg, H. K. (2019). “Prevalence, Predictors, and Treatment of Impostor Syndrome: A Systematic Review.” Journal of General Internal Medicine, 35(4), 1252–1275. https://doi.org/10.1007/s11606-019-05364-1

Clance, P. R., & Imes, S. A. (1978). “The Imposter Phenomenon in High Achieving Women: Dynamics and Therapeutic Intervention.” Psychotherapy: Theory, Research & Practice, 15(3), 241–247. https://doi.org/10.1037/h0086006

Cokley, K., McClain, S., Enciso, A., & Martinez, M. (2013). “An Examination of the Impact of Minority Status Stress and Impostor Feelings on the Mental Health of Diverse Ethnic Minority College Students.” Journal of Multicultural Counseling and Development, 41(2), 82–95. https://doi.org/10.1002/j.2161-1912.2013.00029.x

Cokley, K., Smith, L., Bernard, D., Hurst, A., Jackson, S., Stone, S., Awosogba, O., Saucer, C., Bailey, M., & Roberts, D. (2017). “Impostor Deelings as a Moderator and Mediator of the Relationship between Perceived Discrimination and Mental Health among Racial/ethnic Minority College Students.” Journal of Counseling Psychology, 64(2), 141–154. https://doi.org/10.1037/cou0000198

Cusack, C. E., Hughes, J. L., & Nuhu, N. (2013). “Connecting Gender and Mental Health to Imposter Phenomenon Feelings.” Psi Chi Journal of Psychological Research, 18(2), 74–81. https://doi.org/10.24839/2164-8204.jn18.2.74

Dartmouth, University of Massachusetts. (2023). Fast facts. UMass Dartmouth. https://www.umassd.edu/about/facts/

Frost, R. O., Marten, P., Lahart, C., & Rosenblate, R. (1990). “The Dimensions of Perfectionism.” Cognitive Therapy and Research, 14(5), 449–468. https://doi.org/10.1007/bf01172967

Greer, T. M., & Chwalisz, K. (2007). “Minority-related Stressors and Coping Processes among African American College Students.” Journal of College Student Development, 48(4), 388–404. https://doi.org/10.1353/csd.2007.0037

Holmes, S. W., Kertay, L., Adamson, L. B., Holland, C. L., & Clance, P. R. (1993). “Measuring the Impostor Phenomenon: A Comparison of Clance’s IP Scale and Harvey’s I-P Scale.” Journal of Personality Assessment, 60(1), 48–59. https://doi.org/10.1207/s15327752jpa6001_3

Ibrahim, F., Münscher, J.-C., & Herzberg, P. Y. (2020). “The Facets of an Impostor – Development and Validation of the Impostor-profile (IPP31) for Measuring Impostor Phenomenon.” Current Psychology, 41(6), 3916–3927. https://doi.org/10.1007/s12144-020-00895-x

Jackson, A., Colson-Fearon, B., & Versey, H. S. (2022). “Managing Intersectional Invisibility and Hypervisibility during the Transition to College among First-generation Women of Color.” Psychology of Women Quarterly, 46(3), 354–371. https://doi.org/10.1177/03616843221106087

Moyer, L. P., Szmer, J., Haire, S., & Christensen, R. K. (2021). “‘All eyes are on you’: Gender, Race, and Opinion Writing on the US Courts of Appeals.” Law & Society Review, 55(3), 452–472. https://doi.org/10.1111/lasr.12559

Nearchou, F., D’Alton, P., Donnelly, A., O’Driscoll, L., O’Flanagan, S., & Kirby, B. (2019). “Validation and Psychometric Evaluation of a Brief Screening Questionnaire for Psychological Distress in Patients with Psoriasis.” Journal of the European Academy of Dermatology and Venereology, 33(7), 1325–1330. https://doi.org/10.1111/jdv.15612

Pannhausen, S., Klug, K., & Rohrmann, S. (2020). “Never Good Enough: The Relation between the Impostor Phenomenon and Multidimensional Perfectionism.” Current Psychology, 41(2), 888–901. https://doi.org/10.1007/s12144-020-00613-7

Rivera, N., Feldman, E. A., Augustin, D. A., Caceres, W., Gans, H. A., & Blankenburg, R. (2021). “Do I belong Here? Confronting Imposter Syndrome at an Individual, Peer, and Institutional Level in Health Professionals.” MedEdPORTAL. https://doi.org/10.15766/mep_2374-8265.11166

Rohrmann, S., Bechtoldt, M. N., & Leonhardt, M. (2016). “Validation of the Impostor Phenomenon Among Managers.” Frontiers in Psychology, 7. https://doi.org/10.3389/fpsyg.2016.00821

Rosenberg, M. (1965). Society and the Adolescent Self-Image. https://doi.org/10.1515/9781400876136

Rubin, H. J., & Rubin, I. S. (2012). Qualitative interviewing: The Art of Hearing Data (3rd ed.). SAGE Publications.

Sandelowski, M. (2000). “Whatever Happened to Qualitative Description?” Research in Nursing & Health, 23(4), 334–340. https://doi.org/10.1002/1098-240x(200008)23:4&lt;334::aid-nur9>3.0.co;2-g

Smedley, B. D., Myers, H. F., & Harrell, S. P. (1993). “Minority-status Stresses and the College Adjustment of Ethnic Minority Freshmen.” The Journal of Higher Education, 64(4), 434. https://doi.org/10.2307/2960051

von Collani, G., & Herzberg, P. Y. (2003). “Eine revidierte Fassung der deutschsprachigen skala zum selbstwertgefühl von rosenberg.” Zeitschrift Für Differentielle Und Diagnostische Psychologie, 24(1), 3–7. https://doi.org/10.1024//0170-1789.24.1.3

 

Appendix A

 

 

Appendix B

RESEARCH IN CHEMISTRY

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Inhibition Effects of Blueberries on α-Glucosidase

By Joshua Bernadin

 

Abstract

Type-2 diabetes is an epidemic. Blueberries could hold the key to a new natural form of prevention and possible treatment. The antioxidants in blueberries could inhibit the enzyme α-glucosidase, which can lower blood sugar spikes. An in vitro assay was performed to study the inhibition effects of blueberries on α-glucosidase. The IC50 was found to be 1.0 mg/mL.

Introduction

Type-2 diabetes is an epidemic level disease. Of the 38 million Americans who have diabetes, 90% of them have type-2 (1, 2). Type-2 diabetes is characterized by a resistance to a hormone called insulin which aids in the transfer of sugar from the blood into cells for usage and storage. A resistance to insulin causes blood sugar spikes that can lead to loss of eyesight, nerve damage, and many other problems. Better lifestyle choices like diet and exercise have been proven to help prevent type-2 diabetes. However, due to genetic predisposition to contracting type-2 diabetes and the number of people who have it already, further prevention measures and possible treatments are needed.

There are many treatments available in today’s market for type-2 diabetes (1, 2). They all have their benefits and disadvantages, but they are all effective. Insulin injections introduce more insulin into the body to go past the resistance in type-2 diabetics. These injections are taken daily and are painful due to the injection happening over the pancreas. Metformin is a drug that lowers the glucose production in the liver. If not taken with food, it can cause GI issues. DPP-4 inhibitors stop the breakdown of GLP-1 and GIP hormones. These hormones regulate glucose levels in the body. There are different agonists for GLP-1 and GIP receptors, but these cause weight loss. People started using these receptor agonists as weight loss supplements leading to a shortage. Although all of these treatments are effective, most of them have side effects and are very expensive. In 2022, over $400 billion was spent on these treatments creating a need for a cheaper alternative.

α-Glucosidase inhibition was used for type-2 diabetes treatment. α-Glucosidase is an enzyme that resides in the small intestine that breaks down complex carbohydrates into glucose through hydrolysis (3). In type-2 diabetics, this leads to sharper blood sugar spikes due to the absorption of glucose. Drugs, like acarbose with an IC50 of 2 µg/mL, completely inhibits the activity of α-glucosidase. This has the consequence of making these complex carbohydrates act as fiber, leading to GI issues. In order to use α-glucosidase inhibition for treatment and/or management of type-2 diabetes, partial inhibition of α-glucosidase is key.

Blueberries may hold the key to this partial inhibition. Blueberries are rich in polyphenolic antioxidants like flavonoids and anthocyanins. Previous experimentation has shown that these antioxidants have neuronal protective properties that could aid in the prevention of Alzheimer’s disease (4-7). This antioxidant activity could be used on α-glucosidase as preventative care or a possible treatment for type-2 diabetes.

Experimental

Blueberry Extract

The antioxidants in blueberries were extracted using a solvent mix of 40:40:19:1 of acetone, methanol, water, and formic acid. 500 g of fresh blueberries yielded 778 mg of the extract. This extraction created a dark violet powder (6, 7).

α-Glucosidase Inhibition Assay

The inhibition of α-glucosidase was determined in a 6.8 pH phosphate buffer in the presence and absence of the blueberry extract at 1, 0.5, 0.25, and 0.125 mg/mL concentrations in a final volume of 100 µL. Acarbose was used as a positive control. α-Glucosidase was incubated with the blueberry extract for 30 minutes in a dark area. Para-nitrophenol-α-D-glucoside (pNPG) was added to measure the activity of α-glucosidase. These samples were placed in a 96 well microplate. As soon as pNPG was added, the samples were placed in a spectrophotometer at 410 nm measuring the kinetic activity for an hour using SpectraMax M5 plate reader (Molecular Device, Sunnyvale, CA). Measurements were taken every minute. Using the average rates of absorption, the inhibition % was calculated using the following equation:

The inhibition assay was used to calculate the IC50 of the blueberry extract. All measurements had multiple runs with n= 5-8.

Results

The α-glucosidase activity assay is based on the hydrolysis of pNPG, which release the glucose and p-Nitrophenol (yellow color) with absorbance at 410 nm. As shown in Figure 1, α-glucosidase showed strong activity of hydrolysis of pNPG, and acarbose, a known inhibitor of α-glucosidase completely inhibits the activity of α-glucosidase. The assay was used to examine the inhibition effect of the blueberry extract on α-glucosidase.
Figure 1: Kinetics of α-glucosidase activity assay using pNPG as the substrate with monitoring absorbance at 410 nm.
Figure 2: These figures shows the experimental samples before (left) and after (right) analysis in the spectrophometer. From top to bottom, the rows are enzyme control and enzyme with respective contractions of blueberry extract.

As shown in Figure 2, in samples containing the blueberry extract, little color change was reported. Through the rate of absorption given by the spectrophotometer, inhibition of α-glucosidase by blueberry extract was shown in a concentration dependent manner (Table 1 and Figure 3).

Table 1: Percentage inhibition of α-glucosidase by different concentration of blueberry extract

Figure 3: This graph shows how the inhibition % changes as concentration changes.

The IC50 of blueberry extract was calculated through the concentration dependent inhibition curve (Figure 3), and was estimated to be 1 mg/mL of the extract, which is equivalent to 0.64 g/mL of fresh blueberries.

Conclusion

The inhibition of α-glucosidase was seen at each concentration tested. The IC50 was measured to be 1 mg/mL of the blueberry extract, equivalent to 0.64 g/mL of fresh blueberries. This shows that the blueberry extract, although not as potent as acarbose, can effectively inhibit α-glucosidase partially. This partial inhibition can be the key to future preventative care and treatment for type-2 diabetes.

If given the chance, future studies would measure inhibition kinetics and mechanisms to test the fidelity of the blueberry extract and which antioxidants in the extract cause the α-glucosidase inhibition. This would lead to in vivo studies to test if blood sugar spikes are lessened with the active antioxidant. Afterwards, the blueberry extract’s inhibition effects would be tested on other enzymes and oxidative species that cause other issues. In conclusion, type-2 diabetes may find a new preventative measure or treatment through blueberries. For now, a few blueberries after a carb heavy meal may go a long way for preventing type-2 diabetes.

 

References

American Diabetes Association. “What Are My Options for Type 2 Diabetes Medications? ADA.” Diabetes.org, American Diabetes Association, diabetes.org/health-wellness/medication/oral-other-injectable-diabetes-medications.

Centers for Disease Control and Prevention. “Type 2 Diabetes.” Centers for Disease Control and Prevention, 18 Apr. 2023, www.cdc.gov/diabetes/basics/type2.html.

Daou, Mariane, et al. “In Vitro α-Glucosidase Inhibitory Activity of Tamarix Nilotica Shoot Extracts and Fractions.” PLOS ONE, vol. 17, no. 3, 14 Mar. 2022, p. e0264969, https://doi.org/10.1371/journal.pone.0264969.

Costa, Sophia (2022). “Neuronal Protective Effects of Blueberries against Oxidative Stress on Human Neuroblastoma Cells and Anti-Amyloidogenic Properties.” Thesis, Chemistry and Biochemistry, University of Massachusetts Dartmouth. umassd.primo.exlibrisgroup.com/discovery/delivery/01MA_DM_INST: umassd_library/12133216540001301.

Roderick, Chelsea (2022), Phytochemical profiling of blueberries and their neuronal protection through the inhibition of tyrosinase and acetylcholinesterase: a thesis in Chemistry and Biochemistry, University of Massachusetts Dartmouth.

Samani, Pari (2022). “Anti-inflammatory Properties and Neuroprotective Effects of Blueberries – an implication for the prevention of Alzheimer’s disease.” Dissertation in Chemistry and Biochemistry, University of Massachusetts Dartmouth.

Samani, P.; S. Costa; S. Cai (2023). “Neuroprotective Effects of Blueberries through Inhibition on Cholinesterase, Tyrosinase, Cyclooxygenase-2, and Amyloidogenesis.” Nutraceuticals 3, 39-57. https://doi.org/10.3390/nutraceuticals3010004.

RESEARCH IN MECHANICAL ENGINEERING

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Fluid-Structure Interaction of Flexible Thin Sheet

By Josiah Cassidy

 

Introduction

The environmental effect and limited supplies of fossil fuel energies have prompted extensive study into the creation of innovative and diversified methods to produce electrical energy. Parallel to this, systems capable of cheaply producing limited amounts of energy to power remote or isolated devices, for which connection to the standard electrical network is unfeasible due to cost or technical complexity, have also received special attention. These two factors have heightened interest in methods capable of producing self-sustaining vibrations of a solid or flexible substrate and converting the accompanying mechanical energy into electrical power. The conversion of kinetic energy from geophysical movements such as tidal currents, winds, and river flows into electricity is particularly interesting due to this energy source’s widespread availability and low environmental impact. Several mechanisms, such as vortex-induced vibration, flutter, and coupled- mode flutter instability of the flexible plate in a steady flow, have been identified through research on fluid–structure interactions (FSI) [1]. In this scenario, it is well known that the flat equilibrium state of the plate becomes unstable above a certain flow velocity, at which point significant amplitude dynamic vibrations can form on the structure. The goal of the proposed project is to investigate the capacity to generate electrical power from the self-sustained oscillations of a flexible plate caused by this fluttering instability by converting mechanical strain into electric potential using multiple embedded piezo electronic sensors.

Methods

Through a set of water tunnel experiments, we investigated the onset of flexible plate’s dynamic instability and the occurrence of possible limit cycle oscillations as a function of flow velocity and geometrical dimensions. We studied the effect of the geometrical parameters, such as the ratio between the length, width, and thickness of the plate, on the onset of the instabilities. The flexible plate was fabricated by pouring silicon rubber into 3D-printed molds to ensure both shape accuracy and structural flexibility. During this process, multiple piezoelectric sensors were embedded at various locations across the plate. The experiments for the dynamic response measurement were conducted in a re-circulating water tunnel facility at FSI LAB at UMass Dartmouth. To be able to conduct the experiment in our water tunnel, we have also designed a setup that will secure the flexible plate in the water tunnel shown in Figure 1.
Figure 1: Model of Flexible Plate Setup

The structural response of the system was measured using the embedded piezoelectric sensors purchased with OUR grant funds. Each piezoelectric sensor was connected using full bridge rectifiers to separate inputs of the Arduino in order to collect each sensor’s data separately but simultaneously. As the flexible plate starts to oscillate, it causes a strain in the sensor which transfers data to the Arduino in the form of voltage measurements based on the amount of strain experienced. Using this setup, we performed experiments with various sensors locations along the flexible plate. This method allowed us to find how the strain in the sensor changes based on not only the fluid flow and the flexible plate’s aspect ratio but also the geometric location of the piezoelectric sensor. Throughout this spring semester, many changes needed to be made in order for this setup to operate accurately and efficiently. First, new molds need to be designed and 3D printed so that we can do testing with many different aspect ratio flexible plates. We also needed to adjust the mounting setup for the flexible plate so that it would be fixed securely in the water tunnel. We made these adjustments by redesigning the holder in SOLIDWORKS to better support the flexible plate when oriented vertically. Other modifications were added like screws to secure the flexible plate in place during testing. Next, we adjusted our Arduino setup in order to accommodate 4 data inputs. This allows us to read data from multiple piezos on a single flexible plate during testing at the same time. A picture of the flexible plate during testing in the water tunnel is shown below in Figure 2.
Figure 2: Flexible Plate in Water Tunnel

Results

Figure 3 shows one of our flexible plates and an example of piezo placement in the plate. Some preliminary results are shown in Figure 4.
Figure 3: Placement of piezoelectric sensors in flexible plate
Figure 4: Graph comparing voltage output to water tunnel speed

The y-axis shows average voltage which was found from data collection through the use of the Arduino. This data was streamed to an excel file where it was averaged for each water tunnel speed test. Across the x-axis is the speed of the water flow in the water tunnel in meters per second. From this preliminary testing, we observed a higher voltage output from piezos placed toward the front of the flexible plate when compared to the rear. This could be due to the greater amount of change in displacement that occurs at these water tunnel speeds. A sample time series of the sensor measurements is shown below in Figure 5 in which you can see the changes in voltage output as the flexible plate experiences displacement.
Figure 5: “Sample Time Series of Sensor Data”

Conclusion and Future Direction

While our current testing has started to grant some results, much more testing is required before anything definitive can be said. As this project continues into the next semester or two, much more testing in the water tunnel should be done with more aspect ratios and a wider variety of piezo placement. Researchers should also use other methods like computer simulations to determine exactly why certain piezo placements give higher voltage outputs. This combined with our current piezo experiments may lead to more accurate predictions of power output during varied conditions without the need for experimental testing.

Reference

[1] A.K. Pandey, G. Sharma, R. Bhardwaj, Flow-induced reconfiguration and cross-flow vibrations of an elastic plate and implications to energy harvesting, Journal of Fluids and Structures 122 (October 2023), 103977. https://doi.org/10.1016/j.jfluidstructs.2023.103977.

RESEARCH IN BIOENGINEERING

Recreating a Recombinant R.opacus Bacteria that Can Use Chitin

By Jackie Ramirez

 

Introduction

In 2022, roughly 119 million pounds of American lobster (Homarus americanus) were landed, and this catch was valued at around $515 million. With this gigantic haul of seafood, consumers will eat <50% of the animal, which makes up the lobster meat. The majority of lobster biomass is inedible and is discarded by homes, restaurants and other facilities, and the majority of that waste is lobster shell. The lobster shell contains three main constituents: minerals like calcium carbonate (CaCO3), proteins and chitin/chitosan polysaccharide. Of these shell components, chitin and chitosan have shown value in bio-based processes. Chitin and chitosan are carbohydrate polymers consisting of the amino sugars N-acetyl-D-glucosamine (glcNAc) and/or D-glucosamine (glcN) monomer units. Depending on the degree of acetylation of the polysaccharide, the polymer may be called chitin or chitosan, where the majority of the monomer concentration of chitosan is D-glucosamine. Chitin and chitosan are very attractive biomaterials with a range of household and industrial uses. Regardless, there remains a large percentage of lobster shells that are discarded or underutilized.

Chitin as a biomaterial for biofuel production is a promising and new area of research that will contribute to solving the global climate crisis. Chitinase ChiA, ChiB, and ChiC break down chitin into monomers of N-acetyl glucosamine (NAG). ChiA is an endochitinase that breaks down chitin within a polymer. ChiB and ChiC are exo-chitinases that cleave monomers at the end of a polymer. The monomers produced by these enzymes are used to produce triacylglycerols (TAG). From here, the triacyclglycerols can be trans-esterified into biodiesel. The bioengineering department here at UMass Dartmouth has looked to the surrounding South Coast of Massachusetts as a source of chitin for biofuel production. The shells of crustaceans comprise of 40% chitin by weight. Through research efforts at UMass Dartmouth, chitin has been derived and separated from the protein components of lobster shells (1). This ecofriendly extraction method has given researchers here the ability to utilize crustacean waste from human consumption to isolate chitin and use it for biofuels in conjunction with Rhodococcus Opacus (R. Opacus). R. Opacus, strain PD630, is a gram-positive microbe which will accumulate TAG in the presence of a steady carbon source. It’s high lipid storage ability and rapid turnover rate make it an excellent candidate for biofuel production (2). Chitin is a proposed carbon source for the bacterium. R. Opacus, which is unable to produce the chitinases necessary to break down chitin into its monomer counterparts for biofuel production, therefore this project will make a recombinant strain of R. Opacus to express and secrete chitinase enzymes.

Soon after receiving an OUR grant, my mentor and her collaborators changed the strategy to use the shells. R.opacus is a difficult bacteria to genetically manipulate, so they decided to get another bacteria that is easier to manipulate and has a better chance of taking up plasmids that have the chitinase genes on them. We switched to pseudomonas aeruginosa.

Methods

Genomic DNA isolation using the Promega gDNA isolation kit.

Design primers specific for ChiA from the bacteria S. marcescens and amplify the gene from the genomic DNA.

Initial PCR conditions: Using 5ul gDNA, 1ul of ChiA-For primer and 1ul ChiA-rev primer plus Taq Supermix. The reaction proceeded with standard PCR cycle parameters with annealing at 59 degrees Celsius and 45 cycles.

Second Attempt using a temperature gradient to see what temperature is ideal for primers to anneal to the template.

Figure 1: We tested 57 degrees upto 62 degrees. Each bar indicates the temperature in that well.

Third attempt PCR: We switched to using Q5 high-fidelity Taq DNA polymerase.

Figure 2: These are the PCR Parameters we used with High Fidelity Taq.

Results

Initial attempts to amplify the ChiA, ChiB and ChiC genes from s.marcescens gDNA (Figure 3). The faint bands at the bottom of each lane are primer dimers. We are expecting bands between 1.0kb and 1.5kb. After some research we decided to try using a Taq polymerase that had High Fidelity. The reason was because we are trying to find one gene in a genome of 5,241,455 bp, and we figured that a DNA polymerase that could stay associated with the template better might allow us to get the genes. The High fidelity Q5 DNA polymerase resulted in the expected products between 1kb and 1.5 kb (Figure 4).

                        

Figures 3 (L) and 4 (R): First attempts to amplify ChiA, B and C. 

Having figured out how to get the correct bands I focused on Chitinase B. I was able to amplify ChiB and gel purify only the correct sized band (Figure 5).

Figure 5: Gel purified ChiB genes

The project is being continued by another student. The next steps are to cut the ChiB insert with enzymes and insert it into the vector.

RESEARCH IN BIOENGINEERING

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SKOV3 Ovarian Cancer Cells Research

By Ilya Korovaev

 

Abstract

Ovarian cancer stays undetected in 70% of cases until stages II, III, and 5-year survival rate is 36% for stage III, but this rate can have significant improvement if ovarian cancer could be detected at an early stage. It has been proven that cancerous cells actively produce exosomes, specifically SKOV3 ovarian cancer cells produce around 20000 exosomes per day and secrete them into the blood stream or lymph. If there will be a detection technique that could find those exosomes inside the body fluids, then ovarian cancer could be detected in the early stages. My research is focused on SKOV3 ovarian cancer cells culturing with the following exosome extraction and studying.

Introduction

Exosomes are nanoscaled extracellular vesicles secreted by cells with a size from 30 to 150nm. An exosome has phospholipids double-layer with specific protein markers on its surface and can contain DNA, RNA or proteins. Cancer cells use those exosomes to prepare other regions of the body for metastases acceptation. The process is the following: created in cancer cell exosome contains protein and DNA fragments to enter healthy cells and start the process of healthy cells mutation. Next step is to colonize prepared area with the metastases. The number of secreted exosomes by a single cancer cell is around 20000 per day, and they are getting secreted into lymph or blood flow to get to their destinations. SKOV3 ovarian cancer cells secrete exosomes with tetraspanins exosomal markers: CD9, CD63, CD81. CD9, CD8 which can be used to detect them among other exosomes and start the treatment as soon as the exosomes are detected. For easier exosomes detection they will be excreted from the cells and studied.

Methods

Cell Thawing:

Cells were taken from -80°C freezer and placed on the ice. After thawing process is completed, the cells were put into prewarmed cell PBS media at 37°C and centrifuged at 120RPM for 8 min. Pour the media out using a pipet and put 1 mL new media and resuspend the cells in the media using pipet. Transfer cells with media into culturing flask and add another 4 mL. Put culturing flask containing the cells and media into incubator at 37°C.

Media Change:

Remove the media from the flask using pipet and put 5 mL of fresh media into the flask. Put the flask with cells into the incubator at 37°C. Media change has been done every two days for the first culture and every three days for the second culture.

Cell Splitting:

Remove the media from the culturing flask using pipet. Put 1 mL of trypsin and rinse the culturing flask with it, put another 1 mL of trypsin and put the flask into the incubator for 10  minutes. Put media in the ratio 2:1 2- the media, 1 – trypsin. Centrifuge the solution for 6 min at 130RPM and pour the media out. Resuspend the cells in 1mL of media. Count the cells: put 10µL of the resuspended cells and 10 μL Trypan blue and pipet the solution onto counting plates on each side. Use cell counting machine and calculate the number of cells. If it is less than 1 million cell cells were put into culturing flask and follow media change process. If the number of cells exceeds 1 million cells cell can be transversed into 6-well plate 120 µL of cells will be transferred into 4 plates. Into first two wells 2ml of FBS media with exosomes will be added and 2mL of exosome-free FBS media. Next step is to monitor the growth of the cells to determine if exosomes-rich media speeds up the growth of the cells.

Results

The first culture of cells survived very well, and after 5 media changes it reached the 1million cells mark. After transferring those cells into 6-well plate on the second day cells got contaminated and died. The second culture of the cells I decided to change the media replacement. I changed the media every third day, and, after two media changes, the cells showed good growth but after the fourth media change, they all died. For the third and final time, I decided to keep media change on every second day because it showed the best results. After changing the media 4 times I decided to split them. Before splitting I prepared 4 different medias: normal (without synthetic FBS), synthetic (without normal FBS), 50% (with 0.5ml synthetic and 0.5 normal FBS), 25% (with 0.75mL synthetic and 0.25 mL normal FBS. After cell splitting and putting cells into 4 different plates and adding 2 mL of each media into four plates, cells died before next media change.

Conclusion

After running the cell culture for three times, all three times cells died. The first time it happened because of contamination; and the two other times, I assume that they did not have enough nutrients to survive.

 

References

  1. Siegel, R. L.; Miller, K. D.; Jemal, A., Cancer Statistics, 2019. CA: A Cancer Journal for Clinicians 2019, 69 (1), 7-34.
  2. Zhang, X.; Yuan, X.; Shi, H.; Wu, L.; Qian, H.; Xu, W., “Exosomes in Cancer: Small Particle, Big Player.”
    Journal of Hematology & Oncology 2015, 8 (1), 83.
  3. A. N. Böing, E. van der Pol, A. E. Grootemaat, F. A. W. Coumans, A. Sturk, and R. Nieuwland, “Single-step Isolation of Extracellular Vesicles by Size-exclusion Chromatography,” J Extracell Vesicles, vol. 3, no. 1, 2014, doi: 10.3402/jev.v3.23430.
  4. Nowak, M.; Janas, Ł.; Stachowiak, G.; Stetkiewicz, T.; Wilczyński, J. R., Current Clinical Application of Serum Biomarkers to Detect Ovarian Cancer. Przeglad menopauzalny = Menopause review 2015, 14 (4), 254-259
  5. Mashouri, Ladan, et al. “Exosomes: Composition, Biogenesis, and Mechanisms in Cancer Metastasis and Drug Resistance.” Molecular Cancer, vol. 18, no. 1, 2 Apr. 2019, https://doi.org/10.1186/s12943-019-0991-5.
  6. Tai, Yu‐Ling, et al. “Exosomes in Cancer Development and Clinical Applications.” Cancer Science, vol. 109, no. 8, 1 Aug. 2018, pp. 2364–2374, www.ncbi.nlm.nih.gov/pmc/articles/PMC1182327. https://doi.org/10.1111/cas.13697. Accessed 27 May 2020.
  7. Raposo G, Stoorvogel W. “Extracellular Vesicles: Exosomes, Microvesicles, and Friends.” J Cell Biol. Feb 18, 2013; 200 (4): 373-83. doi: 10.1083/jcb.201211138. PMID: 23420871; PMCID: PMC3575529.
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