Ellie Cook |

Research in Bioengineering

October 9, 2024 Ellie Cook Leave a comment

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

College of Engineering

Research in Sociology

October 9, 2024 Ellie Cook Leave a comment

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.

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

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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

College of Arts and Sciences

Research in Nursing

October 9, 2024 Ellie Cook Leave a comment

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

College of Nursing and Health Sciences

Research in Bioengineering

October 9, 2024 Ellie Cook Leave a comment

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.

College of Engineering

Research in Biology

October 9, 2024 Ellie Cook Leave a comment

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: