Misguidance in Path finding Mechanisms in Twitch-Twice-Roundabout3 Mutant Zebrafish

Hearing loss emanating from damage to the hair cells of the inner ear has significantly impacted the quality of life of over 48 million Americans. Ranging from newborns all the way to the geriatric population, hearing loss is an indiscriminate condition currently affecting every demographic. A common antidote to debilitating conditions that have biological roots, science, may hold a latent answer. Unfortunately, regeneration of the sensory hair cells of the inner ear in mammals does not occur as the damage is permanent. Thus, in our scientific exploration of the biological function and nature of hair cells, we will turn to a model organism that ascertains a definitive regenerative capacity. Zebrafish, a non-mammalian vertebrate, is inherently capable of regenerating sensory hair cells, which exist within the organism’s ear as well as in its sensory lateral line system. Comprehension of the molecular basis of this unique capability has potential to define therapy for hearing loss in human beings.

The lateral line is a mechanosensory system comprised of small sensory patches called neuromasts that are linked together by an interneuromast chain. Each sensory neuromast possesses hair cells extending their stereocillia beyond the epidermis where they may be deflected by water from which they can transduce external stimuli. With respect to the hair cells of human beings, the lateral line hair cells are functionally as well as morphologically similar. Mutations impacting the function and behavior of lateral line hair cells also result in hearing ailments and hearing loss in human beings. With such distinct biological similarity to humans and glaring potential for very sound experimental design, Zebrafish are our model organism of choice.

We intend to focus specifically on the regenerative capacity of the interneuromast chain following the conduction of ablation experiments through Confocal Microscopy. The Roundabout (Robo) family is a major family of cellular receptors that act in conjunction with their Slit ligands, and have been previously implicated in axonal guidance. Previous studies have cited the Twitch Twice gene in encoding Robo3 within Zebrafish, and have been able to correlate this Robo3 receptor to the mechanism of “guidance” in regeneration. The ends of developing interneuromast chains contain a specialized structure called growth cones. Growth cones send out dynamical filamentous projections called filopodia that respond to adhesion and guidance chemically-based sensory stimuli in the external environment. Following characterizing the regenerative process of the Interneuromast chain, we intend to demonstrate that a mutation in the Twitch Twice gene encoding Robo3 results in misguidance during the pathfinding mechanism in regrowth.

Our experiment is centered around a specific transgenic line of Zebrafish that have a genetic construct defined by a Myo6b promoter driving the expression of beta-actin-GFP (green fluorescent protein). Acting as a reporter component, GFP enables us to visualize the entire interneuromast chain of the Zebrafish within the dimensions of Confocal and Flourescent Microscopy. In completing our first major experimental step, we have been consistently screening Zebrafish that were 2 days post-fertilization under our Flourescent microscope. Zebrafish that were positive for our Myo6b genetic construct displayed a complete interneuromast chain that was fluorescing green as a result of our GFP reporter component. We have been isolating these positive Zebrafish on a weekly basis, and have moved to the ablation component of our experiment. Following our weekly screening of Zebrafish that are 2 days post Fertilization, we have been mounting three Zebrafish onto our Confocal Microscope.

Upon mounting this Zebrafish onto to a cover slide via Tricaine solution that anesthetizes them and low melting point agarose, we were able to begin our ablation experiments. Each week, we begin with capturing pre-ablation images of the interneuromast chain between neuromasts L3 and L4 of the Zebrafish. The pre ablation image is very precise in capturing the specific region of the interneuormast chain including the cell bodies that we plan to ablate. After capturing the pre-ablation image, we immediately move to ablating the interneuromast chain through a DAPI Laser, a component of the Confocal microscope. We have been annihilating the interneuromast chain at a power of 405 for exactly 30 seconds, and capture post-ablation images immediately after. This previous week, we moved to conducting a 24 hour time lapse microscopy experiment that was intended to capture the complete regenerative process of the interneuromast chain following ablation. However, after review of the 24 hour time lapse experiment, we noticed that we had only bleached the interneuormast chain, and had not completely ablated it. All of our other settings held up soundly, thus this coming week we plan to carry out the same experimental procedure with enhanced DAPI laser power.

This coming week marks a very exciting time for us, as we strongly believe that with our previous learnings from our attempted 24 hour time lapse experiment, we will be successful this time around. With enhanced DAPI Laser power, we hope to successfully complete the 24 hour time lapse experiment that captures the complete regenerative process of the interneuromast chain. If the experiment is successful, we will maintain the experimental conditions and carry out the same procedure an additional 9 times to validate our findings. Statistical measures will be conducted as well to further strengthen the results of the first major phase of our work. Following the completion of the Part 1, we will move to working with the Twitch Twice/Robo3 mutant Zebrafish. We plan to demonstrate through our 24 Time Lapse Microscopy following ablation that the mutation in Robo3 definitvely results in sensory misguidance during the regenerative process of the interneuromast chain. When we successfully complete this final part of the experiment, we will be able to validate our hypothesis that mutation in ROBO3 affects the sensory based guidance of the growth cone of the interneuromast chain during regeneration.

 

The Impact of Agriculture on Water Quality in Southern Trinidad

I am spending my summer conducting field work in Southern Trinidad to find out the impact of agriculture on water quality. This area and these river systems are very important to me personally, because my family is from Trinidad and they interact with these rivers quite a bit. The rivers here are not only the recipient of a lot of agricultural runoff (which can contain animal waste and fertilizers, among other contaminants), but of great cultural and economic importance to Trinidadians. People practicing the Hindu faith use the river for religious ceremonies, and a big part of the economy here is tourism, which means that the beaches and marine wildlife need to be clean and healthy.

Due to the importance of both agriculture and water quality to the human population living there, southern Trinidad is considered an appropriate model to study human-induced water pollution and its effects on ecosystem and human health. For this reason, I am spending this summer examining how agricultural runoff contributes to microbial and nutrient (nitrogen, phosphorus) pollution.  I will also be sampling and characterizing macroinvertebrate communities in the next few weeks.  These macroinvertebrate communities consist of organisms such as worms, shellfish, and insects (e.g. dragonflies and mosquitoes) that spend their larval phase in water. The composition of these communities is an indicator for overall pollution and ecological health. I will elaborate on this more in my next blog post.

Right now, it is the “rainy season” in Trinidad, which is the time of year when most of Trinidad’s average yearly rainfall occurs. Large amounts of rainfall during the rainy season potentially will flush a hoard of pollution into nearby river systems. For the past few weeks, I have been conducting water sampling, both during rain events and between storms. I have chosen to collect water samples in two different river systems in Southern Trinidad. One is heavily developed by agriculture (the South Oropouche river) and the other has low levels of human land use (the Moruga River). By comparing levels of fecal contamination and nutrients in the two rivers, I hope to make some conclusions about the impact that agriculture is having, and how future agricultural development might influence the ecology of river systems here. Before starting my field work, I walked along each river to find sampling sites.  I found three sites on each river, determining them by accessibility.

Once samples are collected, they need to be tested for fecal contamination and for nutrients (nitrogen, ammonium, and phosphorus). Fecal contamination is found by testing for the presence of bacteria (called coliform bacteria) that are typically found only in the gut of warm-blooded animals. There are many types of coliform bacteria and not all are harmful, but they do indicate that animal waste is making its way into a river. Coliform bacteria produce acid and gas from lactose, and you can take advantage of this to perform a simple test on water to see if they are there. I add 1 mL of the river water sample into a test tube with lactose and a color solution that is sensitive to pH (i.e., acidity) – kind of like litmus paper. I then agitate the solution for about 30 seconds, then leave it for 72 hours at room temperature to let the bacteria do their thing. If the broth does not change color, fecal coliforms were not present in high enough numbers to change the acidity of the water. If it turned yellow, that means that fecal coliforms consumed the lactose, lowering the pH of the water and causing the color solution to change. Interestingly enough, fecal coliforms have been indicated in all the water samples I tested, including the samples from the Moruga River (i.e., the “pristine” river in my study). Although I am not sure of the extent of contamination (only that the samples are contaminated with some level of fecal waste), it was surprising to find that both rivers have this contamination, and that contamination was found even when it wasn’t raining.

My nutrient analysis of river samples will require the use of analytical machines, so unlike the coliform testing, I can’t perform these analyses at the house I am staying at. Nutrient concentrations can change very quickly in water if algae and bacteria are present, so I have to freeze my samples right after collecting them, and keep them stored in the freezer until they can be analyzed. Luckily, I have been in touch with a Professor at the University of Trinidad and Tobago, where I will run a portion of my samples to measure concentrations of nitrate and phosphate, to get some experience with the analysis. When I get back to NYC, I will test all samples for nutrient concentrations (nitrate, phosphate, ammonium).

 

Doing my field work has been an amazing experience, and has confirmed to me the importance of the small study I am doing. Before I came to Trinidad for the summer, I tried to find other water quality studies of the rivers here, and didn’t find any. It also seemed like most residents on the island, even though they use the rivers quite a bit, are largely unaware of the presence or repercussions of river pollution there. These findings have been reflected in my experiences in the field. While collecting samples from the South Oropouche, some of the local people noticed I was collecting water samples near their homes and asked what I was doing and where I was from. I explained to them what my research was on, and they were shocked to find out what was happening in the river system they use daily. They were glad to find out that someone was conducting water quality tests on the river system they rely on for food and water. Hearing this made me happy because I felt that my research can potentially help better people’s everyday lives.

A Summer of Theatre & Community Building

Please describe the title and purpose of your project as well as the goals and objectives of your research.

Our research is titled The Art of a Community. The purpose of our research is to examine the relationship between non-for-profit  community outreach theatre programs and marginalized inner-city communities, particularly Black and Brown communities. We will explore the ways in which these theatre programs have affected these communities and the lives of their participants.

Highlight what you expect to achieve or learn from this project.

We expect to gain a more complex understanding of the significance of community outreach theatre programs in marginalized communities of the 21st century. Additionally, we intend to understand the role which changing demographics has played in the theatre programs in the NYC area with many longtime residents being pushed out due to gentrification. Additionally, this research will highlight the role of race and class on the creation and support of theatre in relation to community building, especially those which are non-for-profit.

Explain what methods you will use to answer your research questions.

In order to answer the questions we pose in this research (which will be mostly qualitative) we intend to interview staff from Community theatre programs in Brooklyn, Manhattan, Harlem, Queens, Staten Island and the Bronx. In these interviews we will uncover challenges that the programs face, the effects that the outreach programs have had on local communities, examples of successful alums, and projections as to where they see the program going in the near future.

Blog #1 “More Than a Footnote- Kuno van der Horst’s Anne Frank Connection.”

The title of the project Prof. Hearst and I are working on is, “More Than a Footnote- Kuno van der Horst’s Anne Frank Connection.” This project aims to shed light on Kuno van der Horst and his wife Hendrika and what they went through during this time, but also their unknown connection to the helpers and Anne Frank. Through this information, we want to shed light on the different experiences that happened during World War II and simple unknown ways these people helped others during the war. To spread the story of Kuno and what he went through and the things that happened in Holland during the war will hopefully shed more light on the struggles of the everyday people.

Kuno’s family contacted Prof. Hearst and have given us more information on Kuno and Hendrika to share with our research. This is a continuation of our research from last summer and with that, we have past research, but also the ability to contact the family and gain more information. This will help expand the information since there is very little information online. We have this special source to spur us on with our research and any questions we have. This also gives us a more personal look into their lives and the lives of people who lived through that time.

Determining the Gene Expression of Latent BCG in infected Caenorhabditis elegans

The World Health Organization (WHO) recently reported that approximately one-third of the world’s population is thought to be infected with Mycobacterium tuberculosis.  At least half of those infected individuals are infected with drug/multi-drug resistant strains. A major setback in researching tuberculosis is the limited options for in vivo experimenting. This project focuses on evaluating tuberculosis infection in the nematode worm, Caenorhabditis elegans, to try to determine if the worm can be used as a new model to study the disease.

The title of this project is “Determining the Gene Expression of Latent BCG in infected Caenorhabditis elegans.”  The worm C. elegans has an immune system in which is similar to that of humans. Along with attempting to prove that this worm is an ideal model for studying TB, the purpose of this project is to determine the gene expression of latent BCG in vivo. More specifically, we hope to pin point certain genes which are upregulated. From there, we may be able to identify the upregulated genes as the genes that force BCG into dormancy.

Through this project I expect to better understand the gene expression in latent BCG in comparison to the gene expression in active BCG.  Through this understanding, I hope to be able to isolate specific genes and correlate them to the latency phase. This sort of research does not have many research articles to base experiments off of, however, my research mentors and I were able to create a food source that both the BCG and C. elegans are able to grow on. From there we will allow C. elegans to feed on BCG for a week. In order to ensure that the bacteria infected the worm, we will be utilizing fluorescent BCG. This will give us a view of where the bacteria are in the worm. We will then lyse open the worms and extract the BCG. We will finally run an RNA sequence test on the bacteria to determine the genes in which are upregulated.

The beginning of “A Comprehensive Guide to Volunteerism in Westchester County”

The title of my project is “A Comprehensive Guide to Volunteerism in Westchester County.” The purpose of this project is to provide a guide to “Volunteer! New York” to find ways to improve New York State’s standing within the Corporation for National Service rankings. The goal of this research is to find approximately how many volunteers there are in Westchester County, how many hours they are volunteering, provide recommendations of what volunteer managers in Westchester County need to improve their services, and use the data collected to provide recommendations on ways Westchester County can improve and promote volunteerism. The methods that will be used to answer our research questions include a survey and qualitative interviews with select figures in the county.

I hope to learn how exactly we can improve volunteerism in Westchester County. Being a member of the Pace community, one that is so focused on volunteering and giving back, it can be easy to stay in this bubble and think that the rest of the world around me feels the same way. However, taking a step back, we can quickly see that that is not the case and that not everyone feels as passionate about volunteering as Pace students do. I don’t believe that there are enough people looking into volunteerism in New York from the angle that we are, so I hope that this project has a large impact — preferably one that will move New York State from 49th out of 51 in the ranks of volunteerism.

Persuade to Date: A meta-analyses of advertising appeals of online dating applications from American, European, and Asian countries.

The title of the research is: Persuade to Date: A meta-analyses of advertising appeals of online dating applications from American, European, and Asian countries. The study will examine the advertising appeals and messages used by the online dating services. We will discover persuasiveness of the core messages used by the most popular dating websites and applications within the European, American (Northern and Southern), and Asian countries. What is more, we will do a cross-national and cross-continental analysis which will explore any potential differences—between the types of the advertising appeals—which may exist. If the differences will be discovered, we will consider cultural, economic, and social factors.

To find the most popular dating websites and applications for each country from given continents, I used the Google search where I was typing the following: “online dating application” or “online dating website” and the name of the country I was researching. I assumed that the outcomes (websites and applications) that showed up, were one of the most popular for a certain nation. What is more, I was also using the ratings of the most successful services if they popped out in the Google search results. It was even more helpful in determining which online dating services are popular in a certain society. Subsequently, I collected their advertising appeals and I put all of them in a table. The table was created for each country separately where there were the following columns: name of a dating service, its core message, and the references. Furthermore, I determined the most essential and significant parts of these marketing messages based on different criteria: emotional, wealth, religion, social influence (other do, so do I) etc. On the next step, I had to create an Excel file which will be used as a material for coding. The following information was provided in a given order to each of the excel columns: continent, country, advertising message, added features, the original language of the provided advertising message, determination whether the dating service automatically translated the website into English, determination whether the service is global or local, determination whether the service is an application or the website.

This research is one step towards knowing how to understand the world. It is done by finding the variables whose dependence we examine and by trying to explain why there are correlated, we discover new things, become more open, and develop ourselves. This is also what I expect from this study. However, this experience teaches me how to collect and manage data, have an objective approach, and do coding. Moreover, I have already taken a Statistics class, so I can apply the theory—I got to know from this class—into practice.

Challenges of students with ASD in Higher Education and Beyond

Doing research on the Challenges and Success of students with Autism in Post-Secondary Education and beyond, I begin with a brief discussion about the characteristics of individuals on the Autism Spectrum, which cover the difficulties they have in social communication with others:

  • Low to High Cognitive Potential
  • Rigidity and Repetitive Movements
  • Intense interests on one or two areas
  • Tonal Prosody of Speech
  • Difficulty Interacting and Poor Eye Contact
  • General Inflexibility
  • Theory of Mind
  • Sensitivity to sound, crowds, touch or taste

Individuals with ASD will develop at their own pace by, breaking the barriers they face with their communication skills. Parents who have children with ASD are striving to help their kids become independent as they grow. Diving into the purpose of my research, I have discovered the challenges they face during their undergraduate career:

  • Transition from High School to college
  • Moving into a larger environment that is less structured
  • Classes not being scheduled daily
  • Follow a syllabus
  • Self-Advocacy
  • Executive Functioning Difficulties
  • Time Management
  • Organization
  • Communication with Professors
  • Living away from home
  • Self-Responsibility
  • Difficulty in accessing accommodations

Overall, the main objective is to apply possible strategies that will enable ASD students to reach their full potential as they go to college, get hired into the work place, and become productive members of society.

As far as expectations, I want to find out what supports are necessary in Post Secondary Education for students with ASD to overcome the difficulties they have nation wide. In addition, I aim to create Post-Secondary Education is a safe learning environment all over the world for students with learning differences. Moving forward into the process, I will be learning how different programs operate in helping students succeed in Post-Secondary Education.

The next step of the project is to do to a comparative analysis of the national averages of students with ASD in colleges and universities as opposed to the students with ASD or learning differences at Pace University who receive comprehensive support. My biggest hope is that the statistics I present will encourage educators and administrators to think about how they can assist students with learning differences to succeed during their college career and provide some necessary tools that will help them overcome their weaknesses.

Empowerment in Action: An Analysis on Effective Employee Empowerment in a Corporate Environment

 

As the title suggests, I will be conducting research on employee empowerment and strong leadership styles. This analysis will further provide insight into how empowerment techniques in a corporate space will enable employees to increase productivity, which in turn, alters the overall atmosphere of the company’s culture. Throughout this research, I hope to identify the methods and strategies used by corporate figures to enhance the employee experience. By analyzing these leadership styles, I aspire to understand which approach is ideal for a corporate culture and how it can be implemented in diverse offices. This will provide an in-depth insight into how typical leadership and managerial styles influences intersectional employees.

My goal is to be able to identify how most folks would like to be led, while also analyzing the contrast of how folks have been led in the past. I hope that this correlation will provide information on which leadership style has the most positive impact on employees. I also hope to garner knowledge on whether or not that leadership increases productivity and staff socialization. A primary objective is to take the analysis on leadership and continue to present it at conferences in the next year. Leadership and management have always been important to me. Working as a Resident Assistant, Orientation Leader and a plethora of other on-campus jobs have taught me the true essence of leadership and how its implementation can truly alter how staff members work with one another and how much they are able to achieve as a unit.

In terms of my methodology, I plan to do a lot of literary research. I have recently checked out literary works from the New York Public Library to understand leadership more in-depth. I am currently reading Creating Magic and Creative Confidence, by Lee Cockerell and David Kelley, respectively. Upon doing this research, I hope to gain primary insight by administering surveys to a plethora of individuals regarding leadership styles. I also aspire to set up interviews with corporate officials in order to gain a further understanding of leadership. Being an active member in the student affairs community, I am confident that I will be able to connect with a plethora of individuals who have insight on managing leaders and creating an environment for leaders to develop their full potential.

Free Fatty Acid Profiles of Milk Products

Through the use of several techniques, Dr. Mojica and I will analyze the compositional components of breast milk. In this research project, Free Fatty Acid Profiles of Milk Products, I will first be conducting research on a variety of databases to compile current publications on breast milk samples. This will allow me to perform a comparative analysis of already existing files, which will allow me to form stronger hypothesis when I formally conduct my own hands-on research. By gathering data that already exists, I will be able to analyze how breast milk samples differ by year (when the data was collected), sample size and group, location (which country the milk samples were collected), fat content, sample preparation, and instrument. This collection of data will allow me to shape how I conduct my own breast milk analysis, and what methods appear to be the most efficient in reaching thorough, accurate profiles.

After collecting multiple, varying publications, I will then conduct my own experiment on breast milk. I will use breast milk samples and machinery in the chemistry department to analyze free fatty acid profiles. Using the publications I collected as a framework and guide for my own experiment, I will analyze the same factors that I observed in the publications, then compare my findings against the other compiled publications. I will analyze how the results differ, and why, whether it be due to difference in location, experimental error, or other factors.

From this project, I hope to grasp a stronger understanding of how research is conducted, proper methods to use in different circumstances, and expand my knowledge on how comparative studies function. In terms of my own research, I aim to better understand breast milk composition in relation to outside factors.

All in all, I will use comparative analysis, self-conducted experimentation, and gathering of similar research publications to conduct my research with Dr. Mojica on free fatty acid profiles of breast milk.