Interdisciplinary – Page 3 – UROP Spring Symposium 2021

Interdisciplinary

Identifying neuronal components necessary for cool temperature sensing

Temperature sensing is necessary for homeostatic regulation, probing the environment for pleasant or aversive cues, and must be reliable across a broad range of temperatures. Under normal conditions, cool temperatures are not painful. However, patients suffering from chronic pain perceive normally innocuous cool temperatures as an unbearably painful sensation known as cold allodynia. How innocuous cool temperatures are signaled in the spinal cord has not been well studied. This study aims to uncover the basic neuronal components necessary for cool temperature transmission in the dorsal horn of the spinal cord. To accomplish this, an intersectional genetic strategy was implemented to ablate several different cell-type markers in the dorsal horn, then tested for temperature sensing impairments. Interestingly, Calbindin 1 (Calb1) neurons, which are mostly local interneurons, were necessary for the detection of cool but not cold stimuli. Calb1 is a heterogeneous population. Using In Situ Hybridization and immunohistochemistry, the Calb1 population was characterized based on markers for excitatory/inhibitory neurons, laminae layer, and somatosensory neurons for pain and itch. By understanding these different subpopulations, the role Calbindin 1 neurons play in transmitting cool and cold sensations can be further investigated. This study provides insight into the neural basis of cool temperature transmission in the spinal cord, which may lead to treatments for patients suffering from cold allodynia.

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Characterization of TRPM8-expressing sensory neurons

The cation channel TRPM8 has been implicated in cold detection, yet no identification of distinct subpopulations of TRPM8-expressing sensory neurons responsible for detecting different levels of cold stimuli (noxious vs. innocuous) has been made. The current study takes advantage of Designer Receptors Exclusively Activated by Designer Drugs (DREADD), light-sensitive channelrhodopsin-2 ion channels (ChR2), thermal behavioral assays, immunostaining, and RNAscope technology to investigate the role of different subpopulations of TRPM8 sensory neurons in dorsal root ganglia of mice. First, we want to study the function of TRPM8 sensory neurons in vivo by expressing an inhibitory Gi DREADD selectively in TRPM8 sensory neurons in the dorsal root ganglia (DRG) of mice. Upon injection of the DREADD agonist clozapine-N-oxide (CNO), the DREADD-expressing TRPM8 sensory neurons could be selectively inhibited to infer neuronal function following cold-plate behavioral assays. Sensory neurons in the DRG can be broadly divided into three groups based on cell body diameter: small, medium and large. From our RNAscope results, “small” TRPM8 neurons expressed significantly higher levels of the trpm8 gene than “medium to large” TRPM8 neurons, potentially providing a basis for differentiation between the two subpopulations. Thermal-related behavioral experiments are still ongoing. We hypothesize that the two subpopulations of TRPM8 sensory neurons serve distinct functions in sensing cold stimuli of different intensities.

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Exploring the CRISPR-Cas9 System through Human EMX1 Gene Deletions

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a technology used to edit genomes at very high precision. It enables precise editing of genomic loci with a RNA-guided Cas9 nuclease that can cleave target DNA that is complementary to a guide RNA (gRNA). Then, the CRISPR-Cas9 system can be used to achieve various goals such as treating inherited diseases (e.g., cystic fibrosis). The objective is to test the specificity and function of the CRISPR-Cas9 system in deleting a targeted sequence in mammalian cells. Mammalian 293AD cells were first transfected with the Cas9-expressing plasmid and human EMX1 ()-specific 3.1+4.1 gRNA sequences. 72 hours after transfection, the DNA was extracted and purified from the cells. The hEMX1 modified region was amplified using a set of primers in the polymerase chain reaction (PCR). The PCR products were run in a 2% agarose gel for gel electrophoresis where the sizes of the DNA fragments were compared to a marker. Results from gel electrophoresis showed that the Cas9 system was successfully able to target the hEMX1 gene when 3.1+4.1 gRNA was present. However, pNTAP posi-tag, a plasmid without the hEMX1 sequence, also showed unexpected cuts which lead to DNA fragments of various sizes. The results show that the CRISPR-Cas9 system was propitious in targeting the hEMX1 DNA sequence. The unexpected cuts made in pNTAP posi-tag will require further experimentation, with one possibility being to extract, purify, and sequence the unanticipated DNA fragments in order to explain the results.

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Single Molecule Dynamics of RNAs in Stress Granules

Stress granules (SGs) are condensates of RNAs and proteins formed in the cytoplasm of the cell when exposed to acute stress. Though recent studies have concluded that SGs appear to play an important role in tumor progression, gene expression, and neurodegeneration, the exact function of SGs is not completely understood. Furthermore, the dynamics of the mRNAs near the surface of the SGs is an area of active research and may provide important clues about the granules’ function, for example, how they can govern the exchange of materials in the cell. To study the dynamics of the mRNAs near the granules’ boundaries, we developed an algorithm which delineates the boundaries in highly inclined and laminated optical sheet (HILO) microscopy images. The simple but fast and robust algorithm we developed to be used with ImageJ is capable of finding the SGs’ boundaries with diffraction limited accuracy and works even on highly inhomogeneous cell images and for granules of arbitrary shape, size, and internal structure, which is an advancement compared to existing methods. After identifying the mRNA molecules in the same field of view using single molecule microscopy, we will use our approach to compare the mRNAs’ locations and SGs’ boundaries over time to study the dynamics of mRNAs on the SG surface. We are now applying this algorithm to study the dynamics of the mRNAs in the cell in order to better understand the interaction of mRNAs with SGs.

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The role of social media in electoral politics and violence in sub-Saharan Africa

Election violence remains a regular occurrence in many countries around the world. This project aims to identify and better understand the online atmosphere of politically affiliated groups on social media during violent elections in sub-Saharan Africa. We consider specifically the case of Côte d’Ivoire, where multiparty elections have been routinely tainted by violence since the 1990s. In the most recent presidential elections, at least 87 people were killed in election-related violence. Data for this project are collected from CrowdTangle, a Facebook API, that tracks social media posts in public Facebook groups. The dataset (N=2,073,664) includes all posts in 169 public Facebook groups supporting the five main political candidates in Côte d’Ivoire from 1/1/2015 to 12/1/2020. This time period covers two presidential elections (2015 and 2020), one legislative election (2016), and a constitutional referendum (2016). UROP students were tasked with coding photo and video content from the 6,248 posts that received at least 500 likes and the 2,963 posts that had at least 100 shares. The students also collected the names of the posters in order to infer ethnicity. We anticipate finding variation in support for democracy or violence conditional on political party affiliation. Trends in content may help uncover key tensions behind election-related violence in Côte d’Ivoire. Understanding the political dynamic of Ivoirians on social media will better help us understand the proliferation of violent rhetoric around elections, who is likely to use it, and perhaps inform policy interventions on reducing election-related violence.

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Social media, youth, and election violence in Francophone Africa

Election violence remains a regular occurrence in many countries around the world. This project aims to identify and better understand the online atmosphere of politically affiliated groups on social media during violent elections in sub-Saharan Africa. We consider specifically the case of Côte d’Ivoire, where multiparty elections have been routinely tainted by violence since the 1990s. In the most recent presidential elections, at least 87 people were killed in election-related violence. Data for this project are collected from CrowdTangle, a Facebook API, that tracks social media posts in public Facebook groups. The dataset (N=2,073,664) includes all posts in 169 public Facebook groups supporting the five main political candidates in Côte d’Ivoire from 1/1/2015 to 12/1/2020. This time period covers two presidential elections (2015 and 2020), one legislative election (2016), and a constitutional referendum (2016). UROP students were tasked with coding photo and video content from the 6,248 posts that received at least 500 likes and the 2,963 posts that had at least 100 shares. The students also collected the names of the posters in order to infer ethnicity. We anticipate finding variation in support for democracy or violence conditional on political party affiliation. Trends in content may help uncover key tensions behind election-related violence in Côte d’Ivoire. Understanding the political dynamic of Ivoirians on social media will better help us understand the proliferation of violent rhetoric around elections, who is likely to use it, and perhaps inform policy interventions on reducing election-related violence.

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ORBIT: Online Resource for Building Intercultural Teams

The ORBIT project is aimed at launching a platform that encourages collaboration between people pursuing projects or researching similar topics of interest. The platform overcomes international borders and language barriers by encouraging intercultural connections. My contribution to this project has been investigating how to extend the existing research to a new audience of middle school students and their teachers. To this extent, we are developing a version of the platform based on the existing beta platform. Because the current platform was created with faculty in mind, we are conducting new research to reconfigure some of the elements to support the middle school student audience. Working in partnership with middle school teachers and administrators, we seek to provide a medium which effectively creates an engaged virtual or hybrid project-based learning environment where students can collaborate and build interpersonal skills. The literature shows that these activities support social-emotional learning, which is particularly critical for students in this age group. We have found that students enrolled solely in remote learning feel disengaged from their peers and their classes (compared to in-person instruction) causing them to face negative repercussions in their academics and personal lives. We have developed an IRB proposal to further investigate these students’ needs and how to best accommodate them in a collaborative virtual environment. As a next step, once the proposal is approved, we look forward to gaining insights from interviewing the students directly and involving them in a series of design-based research activities.

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Boards of Directors – A Behavioral Analysis

This project is related to the psychology of Boards of Directors. We are attempting to find out why certain companies allow or promote unethical behavior from their employees, and/or engage in it themselves. The purpose is generally to seek knowledge on the subject, but also to begin to lay down a framework for how companies can establish a healthy work environment. Our methodology is literature review, the findings from which have been put into a paper. The driving force behind unethical behavior is broadly capitalistic – our research has shown that many companies are willing to cut corners for the sake of saving money. These results are extremely relevant, especially when considering how to regulate this behavior moving forward. Calling more attention to the ways in which companies take advantage will hopefully help put better protections in place for employees and discourage further unethical behavior in the future.

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Cross Cultural Negotiation

As globalization and foreign policy have dominated international relations throughout the 21st century, the need for understanding how to strengthen and extend these bonds has only grown stronger as well. Sectors such as law, business, government, and trade rely on these inter and intracontinental ties in order to build their networks and provide necessary services that benefit both producers and consumers. Without them, productivity and revenue would fall, and very little chance for expansion or cultural exchange could take place. Studying topics such as etiquette, societal expectations, lifestyles, introversion, hierarchy, and individualism all contribute to how each city, state, and country expect to conduct themselves in cross-cultural settings. Not only do they inform diverse parties in how to approach a given situation, but they also provide a chance to bridge the similarities and differences that characterize each culture. Within the context of negotiations, it is essential that these attributes are well understood in order to produce the best possible outcomes for each contingent. In a world where opportunity is limitless, the information from these findings helps to establish a secure foundation from which a progressive dialogue can take place. By examining the inner-workings of cross-cultural negotiations, society stands to gain a more comprehensive interpretation and appreciation of how life differs in each region of the world, and how those changes influence exchanges.

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Nonviolent Resistance in Evereg and Fenesse

Current research on the Armenian Genocide of 1915 (??? ?????) is thorough and comprehensive, but when it addresses Armenian resistance to these crimes, it tends to focus on armed resistance rather than nonviolent resistance. Additionally, research on individual Ottoman Armenian towns and villages is generally cultural in nature, and while this is extremely important, it also tends to overlook the village’s nonviolent resistance to genocide, when present. This results in the belief that provincial Armenians were either passive to the atrocities they faced, or that they could only protect themselves through reactionary violence. This study aims to show that the Armenians the twin towns of Evereg and Fenesse (modern Develi) were able to nonviolently resist their complete erasure from this world through the foundation of the Evereg Mesrobian and Fenesse Roupinian educational societies by using a combination of historical analysis and interviews of the descendants of the survivors of Evereg and Fenesse. An analysis of these texts could show that the curriculum inspired a revolutionary mentality, or that the subjects studied at these educational institutions allowed for the preservation of the cultures of Evereg and Fenesse as its inhabitants (the few who were lucky enough to survive massacre, that is) made their way to Syria, Lebanon, Iraq, and eventually the United States. Hopefully, this research will show that the existence of these educational societies aided in the survival of Evereg and Fenesse Armenians themselves. These pre-genocide and post-genocide accounts will show that when armed resistance to ethnic cleansing is not an option, nonviolent resistance through the creation and maintenance of educational institutions and societies can ensure not just the survival of human beings, but also the continuation of their culture.

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