Current sociopolitical climates for LGBTQ+ individuals in South Asian countries are hostile, and South Asian communities within America will often reflect similar beliefs. Likewise, there might be an unfavorable atmosphere for LGBTQ+ individuals at faith-based institutions. As such, for South Asian college students enrolled in Christian institutions, their religious context and South Asian cultural practices and values might combine to shape their attitudes towards LGBTQ+ people. To explore this understudied topic, I utilized semi-structured interviews with South Asian students at a Christian institution. Thus far, I have interviewed four participants and intend to complete at least eight interviews to meet the qualitative analysis standard. Preliminary examination reveals our participants’ tendency to avoid conversations about the LGBTQ+ community, and they connected this tendency to South Asian culture. In contrast, they noted that LGBTQ+ topics were actively discussed on their Christian campus. Our study hopes to expand on the research surrounding intersectional identities and their impact on the individual.

Additive manufacturing, also referred to as 3D printing, enables the fabrication of objects of any design based on a computer-aided design model. 3D printed structures comprising biodegradable protein-polymer networks have potential use for biomedical applications. The Nelson lab has developed a resin for vat photopolymerization 3D printing based on the protein bovine serum albumin (BSA). In my work, I investigated the response of these materials under different pH environments in order to simulate physiological conditions and gain an understanding of how these hydrogels respond to these different environments. I chose the protein-polymer network MABSA-PEGDA (Methacrylated Bovine Serum Albumin-Poly(ethylene glycol) diacrylate), a functionalized version of BSA that protects the globular structure of the protein. When altering the pH of given MABSA-PEGDA resins they retain their low viscosity, based on rheological measurements, and thus they retain printability. When printed, MABSA-PEGDA hydrogels have altered swelling and water holding capacities in pH 2 conditions as well as altered compressive moduli depending on the pH used to make the resin. Additionally, we performed CD spectroscopy and found that the alpha helicity of the protein was maintained, meaning secondary structure is not altered. The results suggested that there must be a change in the tertiary structure of the protein which induced changes in the protein-polymer matrix and altered the mechanical properties of the hydrogel. The next set of studies will include protein analysis techniques to understand the structure of BSA within the hydrogel constructs. The cumulative results of these studies will enable the use of these BSA-based materials for applications such as oral drug delivery that requires survival in harsh gastrointestinal environments.

The choice to terminate a pregnancy is rarely an easy one. It is critical that the experience of pregnancy termination is made as comfortable and convenient as possible as women grapple with the mental and physical challenges that arise from their choice to abort. The current standard protocol for at-home medical termination of pregnancy involves the patient-mediated oral misoprostol delivery 24-48 hours after mifepristone. This timed delivery ensures that misoprostol can trigger contractions after the cervix is dilated by mifepristone. This presents a challenge for patients, who have to manage the side effects of mifepristone while also timing the delayed dosage window for misoprostol. Combining these two medications into the single dose modality that instantly releases mifepristone and ensures the delayed release of misoprostol will improve the at-home medical abortion process for patients globally. The nanoparticle encapsulation of misoprostol for oral delivery offers the prospect of delayed release and release modulation through the alteration of variables such as molecular weight and co-polymer polymerization and other formulation parameters. Herein, we aim to use established protocols for nanoparticle encapsulation to fabricate and characterize misoprostol nanoparticles. By varying molecular weight and copolymerization parameters, we aim to tailor misoprostol release and evaluate the efficacy of different encapsulation approaches. We intend to investigate the particle size, entrapment efficiency, shelf-stability, and in vitro release of misoprostol nanoparticles in PBS and simulated gastrointestinal fluid. In doing so, we aim to provide a proof-of-concept of formulating misoprostol into nanoparticles to demonstrate encapsulation and sustained release. This understanding can contribute to the development of a single-dosage modality to meaningfully improve the comfort and ease of at-home medical abortions.

Endophytes are microbes that reside within plants, forming a mutualistic relationship where they improve host physiology in exchange for plant carbohydrates. Inoculating plants with a cultivated consortia of endophytes originally isolated from trees in plant family Salicaceae is proposed as a sustainable strategy for increasing crop yield, plant health, and climate change resilience. However, this plant-microbe relationship may turn parasitic if endophytes become over abundant and their carbohydrate demand exceeds the plant’s capacity. Inoculation method is an important yet underexplored factor in predicting the outcome of this relationship and endophyte colonization success. This study investigates how inoculation methods influence the number of endophytes that colonize different plant tissues. A consortium of Salicaceae endophytes was introduced into hybrid poplar plants grown in sand through different inoculation methods. The methods tested are (1) no inoculation, (2) a pre-planting root soak inoculation, (3) a pre-planting unrooted stem soak inoculation, and (4) a post-planting media soak inoculation. After growth, endophyte density and distribution will be estimated by separately culturing endophytes isolated from leaf, stem, and root tissues and counting their colony forming units per gram. The carbon cost of hosting endophytes will be estimated through in vitro and in planta respiration measurements from the sampled tissue. Epidermal wounding from shoot cutting in the stem-soak inoculation method may increase the amount of entry sites for endophyte colonization. Therefore, it is expected that plants receiving inoculum via stem-soak will show increased endophyte distribution and density and higher respiration rates when compared with other inoculation methods. Comparisons of estimated in planta endophyte densities with plant physiological measurements will establish the relationship between endophyte abundance and host health in poplar. Further research will be needed to create a more generalized model of the effect of individual endophyte distribution and density on the benefits and costs to plant health.  

Prostate cancer is the second most prevalent cancer among men in the United States. Characteristic biochemical markers include abundant Androgen Receptor (AR) and Prostate Specific Antigen (PSA), a downstream marker for AR activity. A rare neuroendocrine prostate cancer (NEPC), however, is characterized by low AR and PSA activity and high Synaptophysin (SYP; NEPC marker) expression. AR pathway inhibitors (ARPIs), the first-line treatment for prostate cancer, have minimal therapeutic effects on NEPC. This suggests that suppressed AR activity inhibits ARPI effect, and restoring AR could induce sensitivity to ARPIs, such as Enzalutamide. This study investigates the impact of overexpressing a tRNA, Arg-TCT-1-1, in NEPC. tRNA-sequencing of NEPC cell lines identified reduced expression of the Arg-TCT-1-1 tRNA isodecoder (Arg-TCT). Stable cell lines were generated for adenocarcinoma (AD) and neuroendocrine (NE) phenotypes, with overexpression of Arg-TCT or mutant TCT (mut-TCT). Reverse Transcriptase Quantitative Polymerase Chain Reaction confirmed Arg-TCT and mut-TCT overexpression. Cell lysate was immunoblotted for AR, PSA, and SYP; cell growth assays were then performed with Enzalutamide in DMSO to assess cell proliferation and sensitivity. Enzalutamide inhibited AR activity in LNCaP (1uM) and C4-2B AD (20 uM). Immunoblot suggests that Arg-TCT-1-1 overexpression rescues AR activity in NE cells, while mut-TCT does not affect AR activity. Cell growth assays reveal Arg-TCT-1-1 upregulation induces increased cell proliferation and enzalutamide sensitivity in NE cells. These novel findings demonstrate that upregulated tRNA promotes ARPI sensitivity in NEPC cells. Further research on how Arg-TCT-1-1 regulates AR activity and its transferability to other cancers is needed. These results suggest a promising therapeutic vulnerability if confirmed in murine models.

Endophytes are mutualistic microbes that promote plant growth and stress tolerance by improving host nutrient uptake and producing phytohormones. Plants grown under drought or nutrient limitations have benefited from inoculation with endophytes isolated from wild poplar. Previously, tissue culture-grown poplar has received a pre-planting inoculation to successfully promote growth under abiotic stress. However, a lag in plant growth and initial negative physiological and morphological response has also been observed in plants inoculated early in their development. These results suggest that a favorable plant-endophyte interaction may depend on the method of inoculation. To further understand the best method to maximize plant growth promotion with endophyte inoculation, I have explored how the timing of inoculum delivery impacts plant physiological processes. In a greenhouse experiment, hybrid poplar plants were split into three treatment groups: an uninoculated control group, a pre-planting group inoculated with an endophyte consortium via a root soak, and a post-planting group inoculated via a soil soak. Measuring changes in root system architecture, plant height, and biomass will determine how the plants allocate their resources in response to inoculation treatments. Leaf chlorophyll fluorescence will be measured to assess the stress put on plants from the association with endophytes, while measurements of leaf photosynthetic and root respiration rates will be used as a proxy for changes in the host’s carbon budget. I anticipate that allowing plants to acclimate to greenhouse conditions before inoculation will allow them to have greater above- and belowground productivity due to the photosynthetic cost of establishing symbiosis. Host plant physiology will be correlated with additional measurements of endophyte density and distribution throughout the plant to relate inoculation timing to estimated intracellular endophyte concentrations. This information will provide a greater understanding of the initial costs and benefits of endophyte-plant symbiosis and optimal timing of inoculation.

Approximate computing provides benefits with respect to logic area, latency, and/or power consumption without significantly affecting the outputs of some applications. Various approximate computing techniques have been applied for floating-point (FP) dividers, as division is resource-expensive compared to other FP operations. Since the accuracy requirement may vary depending on the application, this project uses an accuracy-configurable FP divider design. My design first computes the approximate reciprocal of a divisor in a hardware-friendly way. To compensate for errors, I calculated multiple error biases based on error analyses of possible input value ranges. A specific error correction is then applied using a lookup table to select the correct value from the table as determined by the divisor input. The calculated reciprocal is then multiplied by a dividend using an iterative logarithmic FP multiplier, which features accuracy-configurability. This allows applications to determine the desired level of accuracy, versus latency and power consumption, by selecting the number of iterations the multiplier will use. My FP divider design greatly improves the accuracy of previous approximate logarithmic FP divider designs by only adding a small number of hardware resources to our existing FP multiplier. Compared to the state-of-the-art design, the proposed design reduces the amount of required lookup table logic blocks by 53%, and the number of flip-flops by 90%, in a hardware implementation. As the prevalence of division and multiplication-intensive applications, such as artificial neural networks, continues to increase, improving the efficiency of these operations is becoming more critical. This research demonstrates that approximate computing can be a viable approach for many applications and provides a benchmark for future researchers working on methods to streamline division and multiplication operations.

Aging is associated with shifts in the composition of brain extracellular matrix chondroitin sulfate glycosaminoglycans (CS-GAGs). CS-GAGs are comprised of repeating glucosamine and N-acetylgalactosamine units that are either non-sulfated (0S-CS), mono-sulfated (4S-CS, 6S-CS), or di-sulfated (2S6S-CS, 4S6S-CS, 2S4S-CS/Dermatan) and participate in the regulation of brain plasticity. The mono-sulfated 6S-CS isomer is predicted to play a key role in the induction of circuit plasticity during neurodevelopment. Therefore, we asked whether this isomer also shows consistent age-related changes between wild-type mice and humans in the regions of the hippocampus and cortex. Our preliminary data generated from cohorts of mice ranging in age from 7 days to 2 years (50%M/50% F) reveal that 6S-CS abundance is highest at 7 days of age and declines with increasing age (9-22 mice/group). We analyzed the relative abundance of the 6S-CS isomer in n=57 hippocampal and cortical human tissue samples (age: newborn - 95 years, sex: 50%M/50% F). Initially, the human samples exhibited the highest abundance of 6S-CS isomer following birth (<1 month age) that then declined at >1M to 29 years of age, phenocopying the results from mice. However, in contrast to mice in which 6S-CS abundance decreased progressively with aging, we found that in humans, 6S-CS abundance began to increase starting at 30 to 99 years of age (R² = 0.84, p-0.0001). The biphasic model of changes in 6S-CS abundance in humans throughout normal aging was previously unknown. Collectively, these findings demonstrate that age-associated changes in brain extracellular matrix 6S-CS isomer abundance in human tissue do not reflect the age-related decline of 6S-CS isomers that occur in mice. Therefore, additional research is needed to establish the utility and robustness of using rodent models to study aging and other age-related extracellular matrix diseases in humans.