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Understanding the expression and role of pros-1 in the male gonad of C. elegans
Get PDFThe gene pros-1 is a transcription factor that is highly expressed within neuronal sheath cells, glial cells, and excretory canal cells. pros-1 plays a role in cell determination of those cell types in the nematode C. elegans, which promotes organismal development. But the degree to which pros-1 presence is important is still not fully understood, because there are many genes involved in development that when mutated or damaged can result in unexpected phenotypes or even total loss of function to a certain developmental mechanism. What makes pros-1 valuable to research is that it is a functional homologue to a gene found in humans called prox-1, a transcription factor that is vital in the proper neurogenesis of the central nervous system in the human body and altered levels of the PROX-1 protein have been linked to several forms of cancer. Recent data has shown that pros-1 is highly expressed in the somatic gonads of male C. elegans.
We want to understand whether the pros-1 genes plays a role in the development of the somatic gonad, however this is complicated by the fact that pros-1 expression in the excretory canal cell and nervous system is required for the survival of the animal. Our project has developed a system of gonad-specific protein silencing to investigate the essential factor of target genes on the development of the somatic gonad in C. elegans. This is achieved by crossing genetically altered nematode strains to generate progeny of C. elegans that contain a protein silencing tool that allows for the degradation of a green fluorescent protein (GFP) tag that is attached onto pros-1.
Degradation is performed by a tissue specific targeting system called a degron system that utilizes the proteosome. Degrons utilize small nanobodies to target a fluorescent protein fused to the target protein and then bring the fluorescently tagged protein to the proteosome, silencing the expression of our target gene pros-1. The degron system is only expressed in the gonad to target the pros-1-GFP fusion protein. This system will allow for the removal of the essential gene pros-1 in the male gonads, and to determine the loss of function phenotype.
GFP is a versatile biological marker because its protein structure fluoresces a bright green light when excited by a certain light emission, and is commonly used to visualize protein localization, and detecting transgenic expression in vivo. We will be able to confirm the effectiveness of this protocol by the amount of GFP fluorescence in degraded vs nondegraded strains of C. elegans as well as by determining if the male gonad is misinformed during development when pros-1 is removed. Once the C. elegans have the pros-1::GFP fusion and the ckb-1::degron fusion, PROS-1 will fluoresce in all locations it is expressed in except the somatic gonad because that is the only location the degron construct is also being expressed and thus binding to the GFP tag and subjugating the protein to degradation. This overall expression system provides an efficient and testable system for tissue specific protein silencing in within C. elegans which will work to understand the role of pros-1 in the function of the male somatic gonad
National Residency Matching Program: Looking at the Data through Linear Regressions
Get PDFThe National Residency Matching Program (NRMP) oversees the process of medical school graduates being matched to a residency program. The NRMP determines both the hospital and residency program for medical students. Prior to matching, both hospital programs and students rank each other. The NRMP uses these lists to determine the matches. Four distinct models using data from hospitals and applicants were used to determine what characteristics lead to a chance of being matched. Each model went through multiple rounds of testing to determine the importance of the different independent variables. In each data set, the dependent variable is either the percentage of positions filled, or the percentage of applicants matched. The first model looks at data from 1987 to 2022. The second data set looks at the data without the switch involved. Finally, the third and fourth data sets break down applicants by their preferred specialty or the type of medical school they attended. After analyzing the four models, it was determined that volunteer experience and USMLE Step 2 score are the characteristics most significant in determining an applicant’s chance of matching to a residency program
Black Maternal and Infant Mortality Rates and Food Apartheid in Louisville
Get PDFThe purpose of this thesis is to determine whether there is a correlation between the Black maternal and infant mortality rates and food apartheid or food deserts. This thesis will highlight that further research is needed in understanding how food insecurity may influence maternal and infant mortality. This thesis collected data from several midsize cities to assess accessibility to healthy food options and maternal and infant mortality rates. The cities were selected based on similarities in demographic and geographic makeup and having experienced a similar pattern of redlining. St. Louis, Memphis, Knoxville, and Pittsburgh were cities that were chosen for comparison to Louisville. Since Louisville is the main city of focus for this thesis, Louisville will be the benchmark used to compare the other chosen cities. Factors such as food insecurity, food deserts, grocery store areas, and maternal and infant mortality rates were collected. The data from each city was extensively analyzed and interpreted. The data shows there may be a correlation between redlining, food insecurity, and high rates of Black maternal mortality or Black infant mortality rates. The information from the different cities shows that neighborhoods that were colored red during the redlining policies were more likely to have high rates of food insecurity. These neighborhoods are more likely to have high rates of maternal and infant mortality for people of color, specifically Black women and children. Further research is needed to explore food apartheid as another factor that may exacerbate Black maternal and infant mortality rates
An Analysis of Burnout and Wellness of Healthcare Workers Based on Patient Population and Healthcare Worker Location
Get PDFHealthcare workers, even with all of the positive work that they do, often suffer from increased burnout and decreased wellness/quality of life. This problem is not new and has been steadily worsening for many years, however, there are factors which impact burnout and wellness that have not been analyzed or have produced mixed results in prior burnout and wellness literature. There has been little to no research done on the impact of patient population demographics on burnout and wellness in healthcare workers, and the research analyzing the impacts of demographic healthcare worker location has consistently produced mixed results often based on specialty and timing. This study used a 39 question survey to analyze personal, work-related, and patient-related burnout along with overall psychological wellness in healthcare workers across 15 different demographic categories, namely demographic patient population and demographic healthcare worker location. 79 participants from 18 different healthcare worker occupations completed the survey. Subsequent data analysis found that healthcare workers on average experience moderate personal and work-related burnout, and that the average healthcare worker has a low enough quality of life to warrant screening for major depressive disorder. Additionally, there was a significantly higher personal burnout compared to patient-related burnout and work-related burnout compared to patient-related burnout (p \u3c 0.001). Overall, the impacts of patient population demographics and healthcare worker location demographics were negligible for burnout and wellness except for patients in rural areas being associated with increased patient-related burnout (p = 0.045). Shift length (p = 0.19, p = 0.003, and p = 0.005), type of facility (p = 0.017, p = 0.049, and p = 0.028), and intent to leave profession (p = 0.003, p = 0.010, and p = 0.093) were correlated with all three dimensions of burnout (each p value listed is for each type of burnout in this order: personal, work-related, and patient-related). Intent to leave profession (p = 0.012) and intent to leave position (p = 0.013) were both significantly correlated with psychological wellness. This research indicates that there are several factors associated with increased burnout and decreased wellness in healthcare workers, and indicates a need to offer additional resources and support for our healthcare workers to prevent further decline in the mental health and wellbeing of healthcare workers
Gonads without glp-1: Silencing glp-1 in the Male Somatic Gonad in Caenorhabditis elegans
Get PDFIn C. elegans, the gene glp-1 encodes for a Notch receptor called GLP-1, one of two found in C. elegans’ genome. The gene has been previously implicated in the development of the hermaphroditic germline as well as playing a role in the mitosis/meiosis decision. Genetic screening has further identified it as potentially playing a role in the development of the male somatic gonad, making it an ideal candidate for a reverse genetic. We did this by silencing glp-1 and observing if any alterations to the gonad’s phenotype occur.
Normally this could be done by performing a gene knockout. However, due to the nature of Notch receptors’ overall role in the regulation of tissue development, it is an essential gene, and silencing it would result in worms dying as embryos, long before a gonad capable of being studied could form. In order to conduct a knockdown study, CRISPR-Cas was used to tag GLP-1 at its C-terminus with green fluorescent protein (GFP). In a separate strain of worms, degron, a method used for targeting proteins tagged with GFP for ubiquination and lysis, was placed under a promoter specific to the gonad. When the two strains are crossed, they will result in a worm that has functional GLP-1 in all tissues except for the gonad.
In order to resolve the sex ratio issue and focus on the development of the male gonad, which has been neglected in the literature, a mutant strain of him-8 worms that produced offspring with a ratio of three males for every ten viable was cultivated. The bulk of our work was spent crossing the degron strain with the him-8 strain, resulting in a new strain of worms that express degron in the gonad and are disproportionately likely to be male. By coupling this strain with the use of CRISPR to tag any gene of interest with GFP, a gonad-specific knockdown study can be performed not only for glp-1 but for any gene expressed in the gonad, essential or not.
We crossed the new strain with degron and the mutant him-8 with the glp-1::GFP worms, generating both a produce a worm homozygous for all three genes, which should thus contain adult male worms without GLP-1 in their gonads and a strain with him-8 and glp-1::GFP but not degron, allowing us easy access to a source of a male control group. These we will examine for alterations against the N2 male gonad. In doing so, we will hopefully further understand Notch receptors’ role both in the development of the male gonad as well as in general tissue development
How Does Social Media Consumption Relate to a College Student\u27s Mental Health and Body Image
Get PDFMental Health and Body Image issues have become an epidemic, especially among teenagers and young adults. Obsession with social media is also growing among younger audiences. With the increase of users using social media, research has shown that there is a growth in body image issues which has been directly influenced by social media. Despite the research already performed, there hasn’t been a lot of traction surrounding this topic. College students are thrown into constant exposure to the ‘ideal’ college life through social media sharing apps. Between classes, extracurriculars, and meetings, college students are on their phones a lot. Most college students now grew up in an age where being on all social media apps is the norm. Because of this, we see students who are already struggling emotionally and mentally with class load, financials, and other factors being influenced by what people around them or people that show up on their ‘for-you-page” are posting. Although social media is not necessarily a new topic, there hasn’t been copious amounts of research done on how it impacts specifically college students (both male and female) and how this can influence their mental health and body image. I think we can learn a lot from performing this study and continue to draw awareness to how important researching this is. We can learn the impact of quality-of-life social media on younger generations, especially those already going through life stages
The Pressure to \u27Do More with Less\u27: Exploring Donor Misconceptions about Nonprofit Financial Health and Performance
Get PDFAs the nonprofit sector has grown in size and importance over the years, so has competition for donations and attention to the financial behaviors of organizations. Donors and information intermediaries have long expected nonprofits to remain financially lean - there is an expectation for organizations to direct all revenues directly to program areas, and any other spending is viewed as a misappropriation of funds. Nonprofit organizations attempt to satisfy this widely held donor expectation at the expense of organizational infrastructure and growth potential. I reviewed the Economic Research Institute’s Form 990 data and NCCS CORE data for 2013/2014 and 2018/2019 to analyze how donors respond to nonprofit financial measures through their donative behaviors. I analyze the two periods to determine if donative behaviors, and thus donor expectations, have shifted with increased research and discourse on the negative implications of NPO financial leanness. This study looks at the root of the discussion about misguided donor expectations, the need for greater impact disclosure, and the need for more discussion with donors about the importance of organizational growth and capacity building to further program impact. Donor aversion to administrative spending and emphasis on financial leanness is evident in the findings. This reaffirms the need for more discussion about the importance of overhead spending and capacity building in the nonprofit sector and for more transparent and uniform means of reporting organizational impact
