Caused by organisms in the Apicomplexa phylum, toxoplasmosis and malaria are parasitic diseases with significant global health impacts. An estimated 1 in every 3 individuals worldwide are infected with toxoplasmosis, and over 600,000 die annually from malaria. Apicomplexa are a group of single-celled parasites distinguished by specialized organelles in their apical region. Among these structures are micronemes and rhoptries, secretory organelles essential for host cell invasion. Though the process of microneme and rhoptry synthesis is not well understood, T. gondii Dynamin Related Protein B (TgDrpB) has been identified as a necessary component in their formation. While TgDrpB is implicated in the biogenesis pathway of micronemes and rhoptries, its structure and molecular mechanism are currently unknown. The goals for this project are therefore to investigate TgDrpB structure and mechanism in microneme and rhoptry synthesis. It is hypothesized that TgDrpB initiates the process of organelle biogenesis by budding vesicles from the Golgi, similar to that of human dynamin. Completion of this study will establish the structural basis by which TgDrpB accomplishes membrane fission necessary for the biogenesis of secretory organelles and that work to facilitate TgDrpB activity.

Kawasaki disease (KD) is a rare autoimmune disorder affecting nearly 20 in every 100,000 U.S. children aged 5 years and under. KD causes a harmful immune response towards blood vessels, which can lead to coronary artery aneurysm (CAA). The causes and mechanism of action of KD remain largely a mystery, despite decades of etiological research posing a range of biological, environmental, and genetic causative factors. Due to this lack of information, there are no specific diagnostic tests for the disease, and it is diagnosed only by the presentation of a combination of symptoms. This represents a major deficiency, considering that timely and specific diagnosis is critical for preventing life-threatening complications. However, original research has suggested the existence of several subtypes with distinct clinical features under the broad categorization of KD. These subtypes associate specific variables of KD patients to 4 disease subtypes through hierarchical clustering on principal components (HCPC). Yet, my findings in the process of reproducing this work demonstrated significant shortcomings in the previous methods. Upon extensive testing, the HCPC method proved too sensitive to produce convincing clusters using the available variables. Therefore, future KD research must not rely on HCPC, given its demonstrated instability and risk of misleading subtyping. I thus employ unsupervised k-means clustering and produce 3 putative distinctive, characteristic patient subgroup clusters, which appear markedly less sensitive, providing a more stable framework for following KD subgrouping research. These results suggest that KD may be a syndrome with several distinct variants, requiring targeted care approaches.

The mammalian gut microbiome is composed of trillions of bacteria that play essential roles in host metabolic functions and immune responses. Changes in the composition of the microbiome have been implicated in diseases including colitis, metabolic disorders, and cancer. The intestinal tract in which the microbes reside is a regionalized structure with distinct cell types lining the intestinal epithelium that produce and secrete antimicrobial proteins (AMPs). The bactericidal activity of these secreted AMPs, together with a mucus layer, form a barrier that protects the host epithelium from bacterial invasion, thus maintaining host-microbiome homeostasis. Prior studies have shown that distinct bacterial species inhabit different regions of the intestine, but whether AMPs have a role in driving bacterial localization patterns is not clearly established. In this work, I build on results acquired from spatial transcriptomics analysis that show that AMPs exhibit differential expression along the length of the large intestine. One such AMP is RELMβ, which is expressed in the large intestine and targets Gram-negative bacteria. Here, I find that localized RELMβ expression is necessary for the spatial separation of bacteria from the host epithelium in the proximal large intestine and that bacterial localization patterns are disrupted in the absence of RELMβ using wild-type and knockout mouse models. These findings suggest that microbial localization is driven by differential host AMP expression and provide novel insight into the spatial regulation of host-microbiome interactions, which has implications for our understanding of intestinal inflammation and disease.

Early in development, cells receive information about their location in an embryo through external cues, which they must interpret to output specific transcriptional responses required for the proper development of the body structures at each position. The same set of signaling cues can give rise to a range of cell fates depending on the specific timing and concentration of the signal, thus the delivery dynamics of an input are essential to inform a cell’s response. Despite their importance in development, these dynamics are difficult to study as small changes can have multi-faceted effects, and signaling pathways often involve complex feedback mechanisms and redundancy in regulation. However, the use of optogenetics to manipulate the amount of signal delivered through precisely timed light inputs offers a novel approach to investigate complicated regulatory networks and directly examine how signaling dynamics impact transcriptional outputs. One system in which these signaling dynamics are incredibly important, yet incompletely understood, is the terminal extracellular signal-regulated kinase (Erk) network in the Drosophila embryo. Early in development, a very low level of Erk activation patterns a narrow stripe of Abdominal-B (Abd-B) expression near the embryo’s posterior, which is required to form posterior spiracle “tail structures” in the adult fly. Abd-B expression requires tailless (tll), a direct Erk target gene and known transcriptional repressor, but it is unclear how this repressor mediates activation of Abd-B expression. In this thesis, I took advantage of the power of optogenetics to dissect the regulatory network that produces a stripe at the correct time in development and position along the embryo’s anterior-posterior axis. I applied specific optogenetic Erk inputs to embryos and then measured the effects on tll and Abd-B expression. I also built new MS2 biosensors for measuring endogenous Abd-B transcription in live embryos to characterize its expression dynamics in relation to tll. I found that Abd-B transcription is promoted by low levels of tll but repressed by higher levels of tll. To make sense of this tll concentration-dependent activation to repression switch, I used optogenetic stimulation to further examine the regulatory relationships between tll, Abd-B, and the gap genes, hypothesizing that a gap gene might be the missing link between a low level of tll expression and activation of Abd-B. I characterized the gap gene giant (gt) as a direct Abd-B repressor and found that expression of tll at even low levels silences gt expression. These findings informed a model for Abd-B regulation in which low levels of tll repress gt to promote Abd-B expression in a narrow stripe. The borders of the stripe are set through repression by gt at the anterior and high levels of tll at the posterior. Overall, this thesis offers a mechanism through which a precisely low level of a signaling input can produce a particular developmental fate.

Animals sense signals from their environment to inform their behavior and decision-making. Many species have evolved to detect alarm cues released from injured or deceased conspecifics, triggering defensive responses that help them to avoid the same threats that led to the demise of other individuals in their population. Understanding the molecular nature of these signaling molecules and the methods of signal transduction provides insight into the mechanisms behind such behaviors. In Caenorhabditis elegans, the Murphy Lab identified a novel signaling molecule released from lysed worms that living conspecifics avoid. We termed this necrotaxis cue “Todstoff” (death substance). This thesis seeks to characterize the molecular identity of Todstoff, as well as its sensory signal transduction. Analysis of the Todstoff signal suggests that it is proteinaceous and smaller than 20 kDa in size, distinct from known ascaroside signals and alarm pheromones. Building upon previous work by the Murphy lab, which established that Todstoff is sensed by the ASH sensory neuron, I further elucidate the intracellular signal processing pathways involved in Todstoff detection. I demonstrate that necrotaxis requires G-protein-coupled receptor signaling, mediated by the ODR-3 G alpha subunit, as well as synaptic transmission via glutamatergic signaling. Finally, I found that C. elegans avoid lysate from other species of Caenorhabditis, but are attracted to protein precipitated from these worms, indicating that necrotaxis in response to related nematode species is likely directed by a signal separate from Todstoff. Together, the results of this thesis illustrate the existence of a novel, post-mortem inter- animal necrotaxis cue that promotes organismal longevity and survival.

Passive flight has evolved multiple times across different species through the use of gliding membranes called patagia, but there is little known about the development of these structures. Although there is existing research on mammal patagium development that provides insight into its tissue structure and cellular activity patterns, there is no similar data for other gliding organisms. This research aims to uncover the developmental mechanisms underlying patagium formation in reptiles. Specifically, I compare the development of the patagium of a gliding gecko, Hemidactylus platyurus, to the lateral trunk skin of its non-gliding relative, Hemidactylus turcicus. Using histological and fluorescent assays, I characterize the tissue composition, growth patterns, and cell density of developing gecko trunks across embryonic stages. These results reveal that H. platyurus patagium development is marked by the early formation of a dermal condensate of cells early in development, which persists and expands as the patagium grows and is absent in non-gliding trunk tissue. This pattern of increased cell density mirrors the development of the patagium primordium seen in marsupial sugar gliders, suggesting that there exist shared developmental features underlying patagium formation across distant lineages. Although attempts to detect patterns of cell proliferation and apoptosis in H. platyurus tissue were inconclusive, this work highlights potential early developmental constraints in the evolution of gliding membranes and offers new insights into how convergent structures repeatedly arise across vertebrates.

During mitosis, microtubules make up the structural backbone of the mitotic spindle and provide the machinery to segregate chromosomes. The formation of new microtubules (i.e. microtubule nucleation) can happen at centrosomes, near kinetochores of chromosomes, and from pre-existing microtubules (i.e. branching microtubule nucleation) downstream of RanGTP near chromosomes. Branching microtubule nucleation contributes to a majority of microtubules in the mitotic spindle of many cell types and depends on Ran-regulated spindle assembly factors (SAFs) to recruit the universal microtubule template, gTuRC. XRHAMM (Xenopus laevis analog of human RHAMM) is a microtubule associated protein required for early mitotic spindle assembly implied to interact with TPX2 – an essential Ran-regulated SAF – and gTuRC; however, how XHRAMM contributes to mitotic spindle assembly remains to be uncovered. To start addressing this question, my research is focused on if and how XHRAMM is involved in RanGTP-dependent branching microtubule nucleation. Through pull-down assays, I found full-length XRHAMM directly binds to TPX2. To assess how these proteins could be interacting on the microtubule, I performed sequential microtubule binding assays, but my results are inconclusive as to whether TPX2 is recruiting XRHAMM to the microtubule. Also, through pull-down assays, I found the C-terminal domain of XRHAMM directly binds to gTuRC, whereas the N-terminal domain and full-length XRHAMM do not bind to gTuRC, potentially indicating XRHAMM autoinhibition. These results suggest XRHAMM is part of a TPX2-XRHAMM-NEDD1-gTuRC complex and may have a role in branching microtubule nucleation. Understanding XRHAMM’s binding partners and function is important to explain mitotic spindle assembly, and how overexpression of human RHAMM is connected to various forms of cancer.

Hepatitis B Virus (HBV), the major etiologic agent of acute and chronic hepatitis and end-stage liver disease, constitutes a major global health crisis, with over 296 million chronic carriers worldwide. HBV infection can be prevented through prophylactic vaccination, but currently, there is no cure for the disease due to the persistence of the highly stable covalently closed circular DNA (cccDNA). cccDNA serves as the template for the majority of HBV transcripts and its persistence is the root cause for HBV chronicity. By hijacking host DNA repair machinery, cccDNA is formed from lesion-bearing viral relaxed circular DNA (rcDNA), which enters the cell as part of the HBV virion. Our lab has previously identified 5 host factors sufficient for cccDNA biogenesis in vitro. However, in cellulo, it is conceivable that other host factors and redundant pathways may function in this process. In this thesis, we aimed to define all critical host factors essential for rcDNA to cccDNA conversion in cellulo. We first established a CRISPRi platform to downregulate host factors of interest and study its impact on HBV infection. We then validated this platform by silencing DNA Damage Binding Protein 1 (DDB1), a host factor necessary for robust HBV infection, and observed a robust decrease in HBV infection. We utilized this platform to study the impact of silencing 4 out of the 5 factors previously demonstrated to be sufficient for cccDNA biogenesis in vitro as well as several host cellular polymerases. Finally, we developed two novel reporters: one that monitors HBV infected cells by taking advantage of a key viral mechanism, and the other that monitors rcDNA repair by expression of a fluorescent reporter when repaired. Ultimately, the results from this thesis suggest the presence of redundant pathways and factors that function in rcDNA repair in cellulo, and the methodologies established will be utilized in future studies to define a comprehensive set of host factors key for rcDNA repair. These results thus will aid towards a better understanding of key steps in HBV infection and a curative treatment for HBV.

The development of the immune system begins in utero and maternal exposures to pathogens during pregnancy and lactation influence long-term immunity in offspring, impacting their life-long susceptibility to infection and inflammation. The hygiene hypothesis suggests that modern hygiene practices may hinder proper immune training, raising the risk of immune disorders. Building on this, my thesis investigates how maternal exposure to helminths enhances antiviral immunity in offspring. Specifically, I examined the effects of maternal helminth infection on offspring responses to influenza by monitoring clinical symptoms, immune responses, and inflammatory markers following infection. My findings show that a maternal helminth infection confers protection against influenza in offspring by reducing viral load and lung inflammation. I also found that treatment with the helminth-derived metabolite indole-3-propionate (IPA) recapitulates these protective effects, although at a smaller magnitude. This confirms that the maternal gut microbiota acts as an immune educator during early development. These results build on prior work in our lab showing similar protective effects against respiratory syncytial virus (RSV), indicating that maternal immune education of offspring is broadly important across respiratory viruses. My findings highlight the importance of how maternal microbial exposures influence offspring immune development and underscore the potential of microbiota-derived metabolites to promote tissue immunity. This research provides new insights into maternal-offspring immune education and its implications for maternal and child health in an increasingly hygienic world.

In contrast to invertebrate models of embryonic development, where symmetry breaking is spatially and temporally well-defined, early mammalian embryos lack maternally inherited or fertilization-induced asymmetries that would instruct subsequent cell fate decisions. Instead, cell fates are thought to gradually emerge during mammalian embryo development. A key question in the field is whether cell fates emerge in a stochastic fashion, or whether systematically emerging heterogeneities among cells can influence fate decisions. Previous work has shown that SOX2 expression, a key transcription factor of the inner cell mass lineage, is initiated in a heterogenous manner among inner cell mass cells. Moreover, this heterogeneity was linked to the initiation of subsequent differentiation events. This study investigates whether early morphological and temporal features of cells —blastomere volume, surface exposure, and division timing—correlate with the earliest induction of SOX2. Using live imaging of endogenously tagged SOX2 in preimplantation mouse embryos, lineage trajectories were reconstructed from the 8-cell to 32-cell stages. Morphological parameters including blastomere volume, relative exposed surface area, and division timing were quantified and compared between SOX2-positive and SOX2-negative inner cell mass cells. No consistent associations were found between early SOX2 induction and either volume or relative surface exposure at the 16-cell stage. However, it was qualitatively observed that SOX2-positive ICM cells frequently divided later than their SOX2-negative counterparts, suggesting that delayed division may facilitate transcriptional priming.

Kaposi’s sarcoma-associated herpesvirus (KSHV) is an oncogenic gammaherpesvirus and remains one of the leading causes of cancer among immunocompromised individuals world-wide. Like all herpesviruses, KSHV maintains a biphasic life cycle, consisting of a life-long latency sporadically broken by active lytic replication. Like all viruses, KSHV infection induces the remodeling of multiple organellar compartments to either maintain viral latency or promote lytic replication.However, how KSHV drives these remodeling events and how they promote virus-induced tumor progression remain poorly understood. Recently, our group has uncovered that organelle remodeling during herpesvirus infection is directly linked to virus-induced alterations in membrane contact sites (MCS), protein-to-protein or protein-to-membrane connections that form dynamic organelle-organelle communication networks. Here, we use super resolution confocal microscopy to characterize dynamic KSHV-induced remodeling events of mitochondria, peroxisomes, and endosome populations throughout KSHV infection. Then, using siRNA-based knockdown approaches, we demonstrate that MCS proteins that form contacts between the ER and each of these compartments have a significant impact on KSHV lytic gene expression, virion production, and cell survival during lytic replication. Lastly, we find that knockdown of these MCS proteins can not only partially reverse KSHV induced alterations of mitochondria and peroxisomes but also significantly impact the abundance of multiple oncogenic and antiviral cell signaling receptors. Collectively, these findings demonstrate that membrane contacts play a critical role both in driving organelle remodeling events during both phases of KSHV infection that may promote both oncogenic cell signaling and successful lytic replication. Findings from this study have significantly improved our understanding of how KSHV infection alters subcellular compartments to promote diverse replication states and how these changes may drive viral oncogenesis.

Climate change and a rapidly growing world population threaten global agriculture and food stability. As temperatures change and natural disasters like droughts and floods become more common, it is critical to develop technologies that can make crops more robust, less wasteful, and more resistant to changes in climate. One way to achieve this is to target Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the most abundant protein on the planet and the enzyme responsible for forming complex biological molecules out of atmospheric carbon. Rubisco binds to atmospheric CO2 during the Calvin-Benson-Bassham Cycle, a critical step in photosynthesis. Despite the high conservation of Rubisco across thousands of species, it lacks binding specificity and can also bind atmospheric O2 instead of CO2 in a process called photorespiration. This process is harmful to plants and limits the efficiency of overall photosynthesis. To deal with this problem, several species have evolved carbon concentrating mechanisms that create a high CO2/O2 ratio around Rubisco, promoting carbon fixation and preventing photorespiration. One such carbon concentrating mechanism is built around the algal pyrenoid. The pyrenoid is responsible for highly efficient photosynthesis in most types of green algae and constitutes over a third of global carbon fixation. By engineering the pyrenoid into plant chloroplasts, more efficient crops could be produced to combat the challenges presented to the agriculture industry by climate change. Here, we present a highly effective and rapid system for the transient expression and analysis of algal pyrenoid genes in Nicotiana benthamiana and investigate the expression of three essential pyrenoid components in plant chloroplasts.

With the increasing prevalence of metabolic disease globally, fasting has been identified as a potential tool to effectively improve metabolic health. Fasting drives pleiotropic effects across all organ systems, including altered metabolome profiles and gut microbiome composition. Both of these changes on their own can significantly impact metabolic health. Changes in metabolomics and microbiome composition are complex, as the host metabolome shapes microbiome composition via nutrient inputs, and the microbiome can affect the host metabolome via bacterial outputs. However, the interaction between these two areas in fasting has not been studied. Thus, this study sought to investigate potential causal roles of the gut microbiome in the systemic metabolic response to fasting. To carry out this task, we subjected mice with an intact or antibiotic-depleted microbiome to 48 hours of water-only fasting, followed by metabolomic profiling of multiple tissues and serum. We identified fasting-induced increases of fatty acids and acylcarnitines in intestinal content, with microbiome depletion-mediated blunting or exacerbation of the fasting effect. We also found significant changes in microbiome composition and gene expression related to fatty acid and acylcarnitine metabolism, suggesting possible mechanisms for the microbiome-mediated fasting effect. Finally, we found evidence suggesting that our observed murine patterns of fatty acid and acylcarnitine metabolism are relevant for human metabolism. Overall, these results suggest that the gut microbiome plays an important causal role in regulating fatty acid and acylcarnitine metabolism during fasting. This work serves to improve our understanding of how the diet and the microbiome of an individual interact to regulate the metabolic activity of the host.

Coronary artery disease (CAD) is the most common type of heart disease, with about 20.5 million adults having CAD in the U.S, as of 2023. This disease is caused by plaque buildup beneath the endothelium of the arteries that supply blood to the heart, called coronary arteries. Currently, durable polymer drug eluting stents (DP-DES) are most typically used in treatment of this disease, due to lower rates of complications compared to older generation stents. However, fully biodegradable stents have recently been developed and studied within preclinical studies and promising clinical trials. In order to evaluate the association between biodegradable polymer stent use and clinical outcomes, as compared to DP-DES use, a meta-analysis of clinical trials and data available in respect to the in-stent restenosis and target lesion failure (TLF) rates of biodegradable stents, and of DP-DESs, was conducted. Critically, this study has found that the efficacy of fully biodegradable stents is not significantly different from the more commonly used DP-DESs, when analyzed in the general patient population, and in the diabetic patient population. In addition, several novel biodegradable polymers were identified, which demonstrate promise in being used to create new and more effective biodegradable stents. Overall, the findings of this study have highlighted the potential of fully biodegradable stents, encouraging future research in the development and clinical testing of new biodegradable stents.

Biomolecular condensates, membraneless compartments found ubiquitously within eukaryotic cells, regulate cellular organization and biochemistry. Forming via liquid-liquid phase separation (LLPS), these complexes typically behave like droplets, fusing and flowing the way a liquid might. However, issues within the cell, including neurodegenerative diseases like ALS, induce a liquid-to-solid transition within condensates known as aging. Aging is detrimental to condensates as they rely on their liquid properties to adequately function, yet the molecular mechanism that drives these droplets to enter a solid-like, arrested state remains poorly understood. We seek to propose a valid mechanism for condensate aging that is both generalizable and agrees with observed biological behavior. Using coarse-grained molecular dynamics simulations through the program LAMMPS, we show that condensates become dynamically arrested through the formation of laddered constructs within the system that effectively freeze polymer movement. Building on a theoretical framework of conformational entropy, we find that aging is the result of molecular-level choices and dynamics. Crosslinkers between polymers converge to adjacent binding sites in order to minimize the entropic cost of forming bonds. Following the formation of these bonds, we see that more rigid polymers experience fewer fluctuations and can extend the lifetime of these linker ladders. The accumulation of linkers and maintenance of ladders through the simulation ultimately leads to the slowing of dynamics and structural hardening associated with the macroscopic event of condensate aging. By illuminating how simple physical mechanisms motivate complex cell-level outcomes, we hope to lay the groundwork for future research in biomolecular condensates and neurodegenerative diseases.

Influenza A Virus (IAV) infection poses an ongoing threat to human health. The IAV genome is contained in its negative-sense RNA, which replicates using its RNA-dependent RNA polymerase. However, during IAV genome replication, shorter aberrant RNA such as mini viral RNA (mvRNA) can be produced. Previous study has found that mvRNAs PA-60 and PA-66 were detected in mitochondrial fractions of infected cells. However, the method of transportation of mvRNA to mitochondria is not yet understood. In order to detect mvRNA, an LbuCas13abased detection system was optimized for mvRNA detection, testing for conditions including Mg2+ source, Mg2+ concentration, pH-stabilizing and crowding agent buffers, and LbuCas13a:crRNA ratios. It was found that only LbuCas13a:crRNA ratio yielded a significant difference in mvRNA detection. This research also sought to determine the method of localization of mvRNA to mitochondria using components of the innate immune signaling pathway. Retinoic acid-inducible gene 1 or mitochondrial antiviral signaling protein were knocked out in cells expressing the synthetic mvRNA NP61. mvRNA detection found no significant difference in mitochondrial detection in the absence of retinoic acid-inducible gene 1 but potentially suggests an increase in mitochondrial detection in the absence of mitochondrial antiviral signaling protein. These results provide insights into optimal conditions of mvRNA detection using LbuCas13a, and a potential relationship between the innate immune signaling pathway’s involvement in mvRNA mitochondrial localization.

The pyrenoid is a key organelle in the algal carbon-concentrating mechanism (CCM) which is responsible for one-third of global carbon dioxide fixation; it optimizes photosynthesis by channeling concentrated CO2 through membrane tubules into a matrix containing the CO2-fixing enzyme, Rubisco, and the linker protein EPYC1. Engineering a pyrenoid into crop plants lacking native CCMs would increase yields and heat resilience while minimizing water usage, providing a reliable means of food production in the face of climate change. Previous work identified MITH1 as a protein required for proper pyrenoid tubule formation in the model alga Chlamydomonas reinhardtii (Chlamydomonas), but the mechanisms underlying MITH1 function remain uncharacterized. This thesis aims to determine which domains of MITH1 are responsible for its role in tubule biogenesis. In vitro liposome flotation assays demonstrate that the MITH1-VIPP1-like domain is sufficient to bind and shape liposomes, suggesting that it is critical for membrane interactions. By transforming MITH1 constructs lacking different domains into wildtype and mith1 mutant Chlamydomonas, I discovered that MITH1’s long coiled-coil (CC) domain is necessary for binding Rubisco, and that halving the CC domain creates disorganized tubules that retain pyrenoid function. These findings suggest that MITH1 functions as a multivalent adaptor protein, with its N-terminal domain mediating membrane interaction and its coiled-coil region facilitating Rubisco matrix integration, providing critical insights for bioengineering efforts to introduce pyrenoid-based CCMs into C3 crop plants and increasing our understanding of this important organelle.

Studies of genome organization in Drosophila melanogaster have revealed that physical interactions between DNA loci influence gene expression. Recently, ultra-long interactions between loci megabases apart have been discovered, defined as meta-loops. Notably, genes brought into proximity by these meta-loops are enriched for those involved in axon guidance and synapse formation, suggesting a role in shaping Drosophila behavior. Building on these insights, studying genome organization in mosquitoes may help uncover genetic mechanisms underlying behaviors relevant to vector-borne disease transmission. This project aimed to characterize the three-dimensional genome organization in Anopheles coluzzii and Culex quinquefasciatus mosquitoes through Micro-C chromosome conformation capture technology. This technique enabled the creation of high-resolution maps that revealed both local and broad-scale chromatin architecture. Analysis of genes near Anopheles meta-loops revealed enrichment of gene ontology terms related to neural development, consistent with findings in Drosophila. Comparisons of chromatin structure across sexes showed minimal differences, suggesting that sex-specific gene expression may rely on other regulatory mechanisms. Comparisons between the Anopheles brain and gonads revealed notable variation in chromatin structure, implicating both local and long-range interactions in tissue-specific gene expression. Together, these findings highlight the possible regulatory role of genome organization in mosquitoes, which may be leveraged to address the global burden of vector-borne disease.

Chronic kidney disease (CKD) is an imminent public health crisis posing a disproportionate threat to Hispanic populations. While previous studies have acknowledged environmental and lifestyle risks contributing to CKD, there remains a critical gap in literature regarding the role of oxidative stress in genetic susceptibility in this population. This study investigated the interplay of oxidative stress and genome-wide susceptibility to CKD through a genome-wide association study (GWAS) by seeking to identify single nucleotide polymorphisms (SNPs) within the glutathione peroxidase (GPX) gene family thought to be involved in renal protection from reactive oxygen species (ROS). No significant SNP associations were found within the GPX gene family; however, candidate SNPs were identified. Given the inextricable link between disease and systemic oppression, the 2024 Dialysis Facility Report was used to evaluate healthcare disparities in ESRD treatment between Medicaid expansion and non-expansion states while quantifying Hispanic representation. Findings reveal significant disparities in CKD prevalence outcomes for Hispanics which could be attributed to historical oppression in the field of nephrology and barriers to receiving care leading to late diagnosis. This research supports a pivot in nephrology that accepts preventative strategies and structural change. Advancing health equity in renal care strongly supports inclusive genomics, expanding Medicaid, instituting screening programs and simultaneously dismantling institutional inequities to guarantee rigorous healthcare in all at-risk populations.