Chimeric antigen receptor (CAR) T-cell therapy is a revolutionary cancer treatment that utilizes the body’s immune system to recognize and fight malignancies. It describes the process of extracting T cells from a patient and genetically engineering them ex vivo to express CARs that direct T cells to kill cancer cells. Multiple FDA-approved CAR T-cell products have shown promising clinical efficacy in treating cancers such as relapsed/refractory CD19+ B-cell leukemia and lymphoma. However, these products still have limitations in targeting other hematological and solid cancers. Solid tumors are difficult to treat due to their heterogeneity and ability to down-regulate or mutate antigen expression in response to treatment. Also, T-cell exhaustion and treatment associated toxicity result from the inability to precisely control CAR T-cell activity with these current treatments. Biological intermediates such as antibodies may be used to address these barriers, but they are expensive and can elicit an immune response. We aim to develop a universal CAR T-cell system that uses synthetic, high-affinity biomaterials to bind receptors expressed on cancer cells for targeted T-cell killing. For this project, we engineered CD8+ T cells, a cytotoxic subset of T cells, with lentivirus to extracellularly express universal CARs. Using flow cytometry, we confirmed strong expression of CARs on the T-cell surface and demonstrated their ability to interact with a synthetic cancer-targeting intermediate. Lastly, we show that CD8+ CAR T cells pre-armed with the targeting intermediate were capable of selectively lysing cancer cells in vitro that express the receptors of interest. For future studies, I hypothesize that we can expand this universal CAR system by utilizing other synthetic intermediates, such as a heterobifunctional small molecule peptide intermediate, for the treatment of solid tumors.

Despite heavy exposure to Mycobacterium tuberculosis (Mtb), the bacteria that causes Tuberculosis (TB), some individuals show no evidence of infection and by defining these resistance mechanisms, we may identify novel treatment strategies. Among Mtb resistant individuals, our lab identified the Rab5a protein as differentially expressed as compared to controls with Mtb infection. By regulating vesicle trafficking, Rab proteins modulate a variety of cellular pathways including inflammatory signaling, antigen presentation, and autophagy, likely playing a role in Mtb clearance. We hypothesized that loss of Rab5a would alter IFN-êžµ gene expression. Monocyte-like THP-1 cells were electroporated with siRNA targetting Rab5a and yielded 70-90% knockdown at 24 hours versus scrambled siRNA control. Cells were then stimulated with DNA ligands for four hours before RNA analysis. Loss of Rab5a resulted in lower levels of IFN-êžµ gene expression after stimulation with Sheared Calf Thymus DNA (p=0.002, 53.9% reduction), Poly(I:C) (p=0.01, 42.8% reduction), supercoiled plasmid (p=0.03, 45.3% reduction), and cGAMP (p=0.008, 45.7% reduction). We conclude that Rab5a expression is required for Type I IFN production through the DNA-sensing pathway. By characterizing the pathways by which Rab5a modulates the macrophage Mtb response, we may identify host targets to augment protective responses that may serve as adjuncts to current TB treatments and vaccines.

The goal of the SQRLab is to develop technologies for a fully fault tolerant quantum computer capable of processing 10,000 qubits of information. One necessary component is a Paul Trap, consisting of two pairs of blades delivering both direct current (DC) and radio frequency (rf) alternating signals in order to trap and stabilize a string of ions. In order for the modes of the quantum states of these ions to be stable, the stability of power delivery to the device is crucial. Given these design specifications a rf resonator circuit is necessary. The goal of my project is to design, simulate, build, and test a low-loss rf resonator to deliver rf signals to the ion Paul Trap efficiently and stably. Using a toroid shape, I experiment with the material of the wiring and the method of winding the toroid to find a wiring scheme that creates the least loss in delivering power to the experiment. The material used in the wire and dielectric material inside the coil directly affects the efficiency of translating signals. It is desirable that this resonator generates a voltage gain at a specified power level, has impedance matching of the coil with the driving circuit, and contains a method of monitoring this voltage. Thus far, I have measured the voltage gain generated by the coil and the quality factor of the coil (the quality of the resonator frequency as it interacts with the rest of the circuit). I have also designed a capacitive “pickup” to sample the voltage level of the delivered rf power. I will produce noise figures to stabilize the trap power to a set-point voltage level. This research contributes to SQRL’s objective of designing useful scalable quantum systems to simulate nature and perform complex algorithms not permitted by classical computers.

Lignocellulosic biomass, composed of cellulose, hemicellulose, and lignin, has become an attractive renewable carbon source for microbial bioproduction of value-added chemicals. Lignin is relatively difficult to depolymerize, and its enzymatic hydrolysate contains mostly aromatic compounds known to inhibit microbial growth when used as a carbon source. Pseudomonas putida (P. putida), a soil bacteria known for its tolerance to aromatics, has been engineered to catabolize lignin hydrolysate. Engineered microbes have also been encapsulated in hydrogels for on-demand bioproduction and exhibited enhanced tolerance to harsh processing conditions, i.e. freeze-drying and exposure to organic solvents. This research aims to create microbe-laden hydrogels from encapsulating engineered P. putida KT2440 cells in F127-bisurethane methacrylate (F127-BUM) hydrogels for robust on-demand bioproduction when using lignin hydrolysate as a substrate. To mimic growth rate inhibition that often happens in hydrolysate environments, we first examine if P. putida-laden hydrogels remain active when grown in a less-ideal medium, such as M9 minimal media (MM9) known to yield a slower microbial growth rate. We find that hydrogel-encapsulated P. putida containing a plasmid for heterologous expression of a green fluorescent protein (sfGFP) maintained its activity in MM9 continuous culture over two days as measured via fluorescence of the expressed sfGFP. This preliminary result on encapsulated P. putida growth and activity in a less desirable culture environment highlights the potential for utilizing aromatics-rich lignin hydrolysate in bioproduction as a more economical and renewable feedstock alternative.

What type of future manufacturing system would satisfy both the economic and environmental needs of the planet? A circular economy could be the answer. A circular economy operates much like a solitary spaceship that must regenerate water, organic nutrients, and technical materials in a constant process to sustain life. We are privileged to live on a much larger, regenerative spaceship, however, running out of resources is still a risk. Opposed to the current linear economy where end-of-life products such as toasters will be disposed of without extraction of the metals, ceramics, and plastic parts, a circular economy would realize the economic and environmental potential of keeping the parts of toasters in circulation. The purpose of this research is to discern the possible advantages and disadvantages of a widely adopted circular economy through critically evaluating peer-reviewed sources on a multitude of circular industrial sectors and applications. Much of the current research focuses on emerging technologies such as the hydrometallurgy process for lithium-ion batteries. These emerging technologies require heavy investment to become activated before the returns may be realized. It is anticipated that further research will describe natural plastic alternatives such as vegetable cellulose, but there will likely be a gap in research covering the amount of arable land required to grow crops to meet demand. Predictions detail larger corporations likely have more capability for designing circular products and a probable lack of small businesses with circular potential. Additionally, public support would be lacking due to the esoteric nature of the system and a lack of real-world circular business models currently. That is why the motivation behind this literature review is to spotlight the circular economies that are already thriving.

Despite a rise in egalitarian views over the past decade, the trend has recently slowed down and scholars are struggling to understand why. Past research shows an unexpected trend of young millennials, specifically men, reverting to traditional gender norms. Childhood family instability is proposed as one possible explanation, but it has not been examined empirically. The goal of this project is to understand how one form of family instability, divorce, impacts men’s view on love, marriage, and gender norms. The research is driven by two questions. First, how does the experience of divorce affect a father's view on gender? Second, did the father’s post-divorce view on gender affect the son’s attitudes, and, if so, how were these attitudes transmitted from father to son? The data will be collected through individual semi-structured interviews with divorced fathers who had custody of their son and the sons of these fathers who are at least 18. The study expects that children of divorce will reject egalitarian views because they have observed their parents separate through their attempt to uphold egalitarian views. While this project only examines males, future research can expand this study through interviewing single mothers and their female children. This project will help address the puzzling phenomenon of why there is a declining trend toward egalitarian gender views.

Trapped ions are proving to be a promising realization of quantum computation; computation is performed with an ion — such as 40Ca+ — contained by an electric potential and irradiated with laser beams of multiple wavelengths for ion cooling and qubit state control. Advantages to other implementations include repeatability and efficiency in state preparation, gate operation, and state readout. However, a number of ions are required for useful operation, and larger systems introduce complexity and possibility for error, which necessitates modular and scalable trap geometries fabricated using common methods pioneered by the semiconductor industry. Two-dimensional surface traps can suffer from low trapping depth, low efficiency, and higher ion motion, complicating high-fidelity operation. We explore alternative geometries, such as traps with multiple surfaces, and elucidate acceptable misalignment or non-ideal geometries. Our method exploits a multipole expansion of electric potentials produced through COMSOL; we expect to use the resulting multipole coefficients, trap depth, and trap frequency to inform fabrication tolerances and parameters such as electrode spacing, size, or shape. Through proper fabrication and optimal design informed by this work, traps with high depth, high efficiency, and low ion motion can aid the push for modular trapped-ion quantum computing.

 When homeless youth turn 26 years of age many of the organizations and resources they have been utilizing for survival, such as food and shelter, become unavailable as many organizations no longer consider them youth. The purpose of this study is to identify gaps in the transition period for homeless youth moving towards adulthood to explore ways to make the transition more efficient and accessible. I conducted a systematic literature review among databases and ten studies are included. Restrictions on studies included were publication dates after the year 2000, publications in English only and studies done in the United States. This study includes data from both currently and previously homeless youth from ages 18 to 30. Several areas are identified as gaps that affect the ease of transition for homeless youth including lack of consistent support in case management or mentorship, preparing the youth ahead of time for exit out of youth programs, housing models being utilized at time youth are aging-out at twenty-six years old, and how the youth values themselves. Recommendations for programs such as a housing first model that collaborates with mentoring and community support and youth preparation programs that begin at least a year before a youth turns 26 years old are discussed in this study.

Metal–organic frameworks (MOFs) are a class of crystalline, porous extended solids that are formed through coordination between metal cations and bridging organic ligands. These materials have been a topic of acute interest in the scientific community due to their intrinsic porosity, high surface area, and precise tunability. However, MOFs are typically insulating, which limits the scope of their applications. The recent development of electrically conductive MOFs has opened the door to exciting multifunctional applications in electrocatalysis, advanced electrochemical energy storage, chemical sensing, and much more. However, a molecular-level understanding of charge transport in MOFs remains lacking. My research aims to address this knowledge gap through the investigation of one-dimensional (1D) metal–organic chains. In this presentation, I will introduce the synthesis of a series of highly-tunable 1D metal–organic chains that exhibit delocalized π systems and high electrical conductivity along with studies of how structural parameters such as metal/ligand identity and chain geometry influence their overall electrical and magnetic properties. My preliminary results demonstrate trends in these structure-property relationships that may inform how these materials can be rationally designed with specific magnetic and conductive properties. Ultimately, this work will contribute towards a molecular-level understanding of charge transport and magnetism in metal–organic frameworks, enabling the design of new conductive porous materials that can use electricity to drive chemical processes.

In regions with high HIV prevalence, periods of pregnancy and postpartum are associated with higher risk for HIV acquisition. The WHO recommends oral tenofovir (TFV)-based pre-exposure prophylaxis (PrEP) as an effective method of reducing HIV acquisition risk for pregnant people. Although most pregnant patients in Kenya with identified HIV risk factors accept PrEP when offered, >50% discontinue PrEP within the first 30 days.​ Few studies to date examine factors that contribute to PrEP discontinuation in this population and there are no existing intervention studies aimed at improving PrEP adherence during pregnancy/postpartum. We utilized data from the ongoing Mobile Women and Children (mWACh)-PrEP study, a randomized control trial testing a two-way short messaging service (SMS)-based platform to facilitate communication between peripartum patients taking PrEP and remote nurses. In the parent study of 379 HIV-negative, cisgendered Kenyan women taking PrEP, 188 were randomly assigned to the intervention (mWACh-PrEP) group. The purpose of our sub-analysis was to evaluate message contents and identify frequently discussed topics. System messages were tagged into categories based on subject. Of weekly automated messages, 42.1% were responded to by participants, with an average response time of 87 minutes; topics of responses included PrEP concerns (55.1%), antenatal concerns (22.4%), maternal health concerns (17.8%), and infant concerns (17.2%). A total of 366 spontaneous messages were received, with >50% of participants sending at least one spontaneous message; topics of spontaneous messages most frequently included antenatal concerns (22.9%) and PrEP concerns (19.7%). These results indicate that a two-way SMS based intervention provides important support to pregnant and postpartum patients taking PrEP; the mWACh-PrEP system could be an effective solution not only to promote PrEP adherence within this population, but also to increase access to maternal healthcare.

Millions of people go hungry and are insecure about their access to food. Charities and non-profit organizations run numerous food assistance programs to help those in need. Previous research suggests that public services are positively associated with housing prices, demonstrating that intra-jurisdictional differences are capitalized through market processes. However, food assistance services bring negative externalities, such as attracting non-residents to gather in the community, disrupting our previous understanding of housing prices. In this research, we employed a hedonic price modeling approach to estimate the impact of the presence of food assistance centers on housing sales, controlling for property attributes, neighborhood, and city characteristics in Baltimore City, Maryland. Two main measurement methods were used to label the attributes of a property: marking all properties within a neighborhood where the food pantry is present as accessible properties and marking all properties within a one-kilometer radius around the food pantry as accessible properties. We tested two hypotheses in this study. The first hypothesized that the presence of a food assistance center is an important indicator of housing prices. The second hypothesized that the impact of food assistance centers varies by the type of property: condo, townhouse, and single-family residents. Under the one-kilometer radius measurement method, the overall effect of the food pantry on house prices is negative, showing that housing prices are lower for accessible properties than properties without food pantry service. Single-family residents and townhomes are the types of properties that are most affected by the presence of food pantries. These findings have the potential to inform urban economics research devoted to the nature of public services. In particular, they show that the food pantry, as one kind of public service, is important not only in the public health field but also plays a role in affecting housing markets and urban planning.

Many standard oxidants and reductants are non-reusable and toxic, so it is important to pursue cleaner alternatives. In this project, we have synthesized and characterized two metal-bipyridyl supramolecular cages and have studied their application as catalysts for the electrochemical reduction of organic substrates. Supramolecular cages are formed from the self-assembly of organic ligands and metal ions in solution, and they contain internal cavities with unique electronic microenvironments, similar to the interior of enzymes. While these polyhedral structures have been investigated as catalysts for traditional synthetic pathways, their role in electrosynthesis remains underexplored. Electrosynthesis involves the transfer of electrons to and from substrates using an applied potential, rather than chemical redox agents. This method is often hindered by a high kinetic barrier at the electrode-substrate interface, but catalysts can lower this barrier. We hypothesize that redox-active supramolecular cages – cages that can readily interconvert between charge states – can serve as effective electrocatalysts by encapsulating and transferring charge to substrates. To understand the effect of ligand geometry on electrocatalysis, I have synthesized two redox-active ligands with bipyridyl chelating groups. One contains a highly conjugated perylene core, while the other contains a compact core formed from pyromellitic dianhydride. We have metalated these ligands with iron ions to form two tetrahedral supramolecular cages. We then utilized cyclic voltammetry to assess cage-facilitated charge transfer to vicinal dihalide substrates. We observed that the reduction of multiple substrates, including 1,2-dibromo-1,2-diphenylethane, occurred at milder voltages in the presence of the cages, indicating a reduced kinetic barrier. For these substrates, we then performed bulk electrolysis, from which we determined that the percent conversion to the desired product was significantly higher when a cage was present, supporting our hypothesis. Ultimately, we aim to use these cages to enable electrosynthesis of organic feedstocks at lower voltages and with fewer byproducts.

X inactivation is a mechanism of dosage compensation that equalizes gene expression between males (XY) and females (XX). It is mediated by the long non-coding RNA (lncRNA) Xist (X-inactive specific transcript). However, some genes escape X chromosome inactivation(XCI) in females, and thus potentially contribute to sex differences. Escape genes are often identified by allelic expression analysis based on RNA sequences and RNA FISH (fluorescent in situ hybridization) based on imaging. In addition, RNA FISH allows the examination of escape status and the location of the transcript in individual cells. Allelic expression analysis has shown that one X-linked gene Car5b escapes XCI in mouse cells. However, it is not clear whether it escapes XCI in every cell or a portion of the cells. The goal of this project is to use RNA FISH to detect and quantify the escape status of Car5b in wild-type cells and in cells carrying a deletion of an important DNA element required for Car5b escape. In order to achieve the goal, I first prepared the fluorescent probes using the cDNA plasmids of Car5b and Xist. Next, dual RNA FISH was performed for Car5b and Xist in mouse cells to visualize the location of Car5b RNA signals whereas Xist RNA signals serve as the marker of inactivated X chromosome. Analyzing the FISH results is ongoing and we expect to visualize Car5b escape in the wild-type cells but not in the mutated cells. This method can be applied to escape studies in other X-linked genes which could enhance our understanding of X-linked gene regulation and sex difference.

To study genetic factors and the molecular mechanism underlying the development rate of mouse embryos, two mouse subspecies were studied. I have collected mouse embryos from crosses between C57BL6/JxCAST/EIJ (B6xCast), CastxB6 (the reciprocal cross), and intercrosses within either strain to examine the effects of maternal and paternal genomes on the developmental rate. I used mouse embryos every 24 hours at gestational stages E11.5-E14.5. Staging by the Embryonic Mouse Ontogenetic Staging System (eMOSS) is used to estimate the actual developmental stage based on limb bud morphology and compared to the gestational stage based on the estimated time of conception by observation of a vaginal plug. I also used gDNA to determine the sex of each embryo to investigate whether sexes play a role in developmental rate. I observed an interesting pattern that the mouse subspecies and the source of each parental genome affect the developmental rate of embryos. I observed embryos with at least one Cast parent, have faster development than B6xB6 starting at gestational stage E13.5. The developmental rate is independent of the sex of embryos or the litter size. Single-cell transcriptomic analysis is ongoing to determine the genes and mechanisms behind the change in developmental rate of mouse embryos. It is important because the result will contribute to human development in early stages, and help to solve problems such as preterm or underdeveloped infants.

Iron centers which feature metal ligand multiple bonds can be powerful group transfer agents, for example, terminal Fe-oxo intermediates in soluble methane monooxygenase can perform oxo-atom transfer for the selective oxidation of methane to methanol. Abiologically, ligand constructs which enforce desirable electronic and structural configurations have been shown to enhance group transfer to a range of organic substrates. We developed and studied an iron (Fe) molecular complex with two aminophosphine selenide ligands (Se=PPh₂NTol; Ph=Phenyl, Tol=4-Tolyl) that chelate the metal center via the selenium and nitrogen. The iron complex (FeL₂) was synthesized by a reaction between Fe(HMDS)₂ (HMDS = bis(trimethylsilyl)amide) and the aminophosphine selenide. Characterization shows a tetrahedral, high spin, symmetric compound. We hypothesized that FeL₂ can activate and transfer heteroatoms and explored the reactivity of FeL₂ with oxidants, oxo atom donors, and organic azides. Treatment with iodine (I₂) results in oxidation of the iron center (Fe(II) to Fe(III)) and coordination of the iodide counterion results in structural reorganization to a five-coordinate square pyramidal complex. Reactivity with oxo atom donors shows that either the ligand or Fe center are oxidized, and we identified a µ₂-oxo dimer, which is the first Fe-O-Fe dimer to have selenium in its first coordination sphere. We found that FeL₂ forms Fe-nitrenoid intermediates and can perform nitrene transfer to form diazos or do C-H amination, when treated with aromatic and aliphatic azides, respectively. The presented complexes are characterized by single crystal X-ray diffraction (XRD), Evan’s method, nuclear magnetic resonance (NMR), and Ultraviolet-Visible Spectroscopy (UV-Vis). This research builds upon the knowledge of transition metal complexes for heteroatom transformations.

Have you ever watched a true crime show? I’d be surprised if you said no. Stories of violent crime inundate the entertainment offered to individuals whether you’re looking for a TV show or a podcast. People’s worst moments are retold for TV audiences under the guise of investigation. While crime media has always been popular, true crime media, television in particular, is having a moment of unprecedented popularity. Drawing from my own experiences as a white middle-class viewer of true crime I wonder what the dangers of this genre's popularity are. This project investigates this by asking how the consumption of others' trauma through true crime television impacts how individuals who have no contact with the criminal justice system understand violence and crime. It asks whether or not this creates additional distance between individuals who are not system impacted and those who are. And finally, it wonders if true crime's popularity can be used to put its viewers in conversation with abolition. Using Dateline NBC episodes as its archive, this project hopes to navigate the complicated nature of true crime viewership and its harms. The archive of Dateline episodes provides examples of key narratives within true crime as well as facilitates a true crime viewing experience informed by abolitoinist politic. The patterns observed in Dateline are put in conversation with my academic research to both attempt to answer my research questions as well as articulate critiques of true crime television. These intentions are creatively rendered into a zine that synthesizes my research as well as my relationship to the genre and Dateline specifically. I hope that this project inspires individuals to engage critically with their entertainment and understand that entertainment is a means of perpetuating hegemony.

Previous work takes inspiration from 2D metal–organic frameworks to synthesize 0D metal–organic macrocycles which maintain the conductivity of the original material while introducing solution processability. These macrocycles self-assemble into nanotubes through π-π stacking of the aromatic core but, the nanotubes do not have a preferred orientation when imaged by atomic force microscopy (AFM). We hypothesize that by adjusting solvent, drying conditions, and organic ligand functionality we can create a preferred orientation of the macrocycle nanotubes on common substrates which will improve charge carrier mobilities through the aromatic core. This work demonstrates the formation of large domains of nanotube alignment which can lead to greater charge carrier mobility. With unique ambipolar charge carrier transport, metal–organic macrocycles have applications in energy storage, chemical sensing, and electrocatalysis.

In recent years the field of global health has been plagued by accusations of “neocolonialism.” Rather than genuine involvement in health research, physicians and public health researchers in Global South countries are relegated to menial positions in the research pipeline, and local priorities are not reflected in the research that receives the bulk of international funding. The multilateral donors which fund and manage this research are said to devalue the contributions of Southern researchers, who, many studies of global health journals have shown, are not allowed to take on leadership roles in public health research “collaborations” with Western institutions. Moreover, a large critical literature has characterized the field of global health as taking a narrowly conceived, “vertical” view of health in developing countries. This biomedical/technological bias leads donor-funded research to neglect the social determinants of health and illness. The current project contributes to our understanding of the origins and practical manifestations of these institutional biases of health research in Uganda. A historical analysis of colonial medicine in Uganda was conducted and it was found that medical research in Uganda indeed has historically neglected to address the social determinants of health such as poverty, labor conditions, and the presence or absence of social infrastructure (e.g., roads and sanitation systems). In addition, a review of recent COVID-19, Ebola, and HIV/AIDS research confirms the hypothesis of other scholars that donor-driven public health research tends to treat health in isolation from the larger political economy of development, and is biased toward a highly biomedicalized view of health. I conclude by offering an interpretation of these biases and how they vary by individual donor, as well as some suggestions as to what can be done to make Ugandan health science genuinely equitable and responsive to local, rather than Western, needs.

For Saccharomyces cerevisiae (S. cerevisiae) to survive, they must either synthesize nitrogenous bases or reside in a nitrogenous base-rich environment. If there is a mutated enzyme along the pathway that makes the nitrogenous base uracil, then the organism would be unable to synthesize uracil and die without uracil. When S. cerevisiae is grown in 5-Fluoroortic Acid (5-FOA) media, the URA3 enzyme will catalyze 5-FOA into a toxic intermediate that causes cell death when incorporated into RNA. In URA3 mutations, the intermediate is not produced, and S. cerevisiae can survive. In an experimental setup where we explored mutation patterns in aging S. cerevisiae cells, we used URA3 to select mutants that grow on 5-FOA media when uracil is provided. However, when analyzing the mutant sequences, there were fewer URA3 mutations than expected. We performed Sanger sequencing on the URA3 gene in individual mutants and were surprised to find that some did not have any mutations. We, therefore, sequenced the whole genome of those mutants and found that they had missense mutations in the URA6 gene. We then analyzed the URA6 locus from the original pool. Across the URA6 gene, the mutations appeared randomly spread with the possibility of some mutation hotspots, indicating that they could be loss of function mutations. In contrast with the URA3 mutants, we observed URA6 mutants were able to grow 5-FOA media when uracil was not provided. There may exist an alternative pathway for URA6 S. cerevisiae mutants to synthesize uracil and survive. To further study this observation, we plan on analyzing the function of the URA6 gene in 5-FOA media compared to the URA3 gene and the components of the URA6 pathway. Ultimately, the result of this study could clarify nitrogenous base production pathways in S. cerevisiae and the impacts of 5-FOA on S. cerevisiae.

 Solid tumor cancers are capable of releasing chemicals that are detectable in body fluids. These cancers can cause severe acidosis within the patient, resulting in the change of structure and function of Complement serum proteins, membrane-bound regulators, and receptors that interact with various cells and mediators. A better understanding of the mechanistic interaction between the complement system and tumor-induced acidosis can provide a new direction in cancer immunotherapy research. Here, we investigate the effect of pH on binding between Heparin, Complement Factor H (CFH), and C3b. We found that in controlled in vitro assays conducted through biolayer interferometry, CFH and Heparin bind better to C3b in pH 6.0 but only in lower concentrations of CFH. As for pH 7.4, the binding between CFH, Heparin, and C3b is lower in comparison; however, the binding between only CFH and Heparin is higher at pH 7.4 than at pH 6.0. Our results demonstrate how Heparin, Complement Factor H, and C3b are likely to function in a tumor microenvironment where the pH is generally lower. We anticipate this research to be a starting point for investigating the role of the Complement System in tumor growth and researching the implications of its biological actions with respect to the development of anticancer therapy.

Chimeric antigen receptor (CAR) T cells are used to treat blood cancers; however, neurotoxicity is a common complication that can be life threatening. The neurotoxicity patients may experience includes language and cognitive disorders, seizures, and cerebral edema and hemorrhage. In our previous research on a mouse model of CAR T cell toxicity, we found that leukocytes plugged 11.9% of brain capillaries in CAR T cell treated mice along with an increase of Intracellular Adhesion Molecule (ICAM-1) on brain capillary endothelial cells. We hypothesized the effects of capillary plugging may contribute to neurotoxicity, hence to better understand the mechanism, I am exploring the effects of different cytokines we observe in patients with neurotoxicity on ICAM-1 expression on human brain microvascular endothelial cells (HBMECs). I treated cultured HBMECs in 10, 100, and 1000 picograms/mL of IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-15, CXCL10, GM-CSF, TNF-α, and IFN-γ for 24 hours. Then, I used flow cytometry to measure the amount of ICAM-1 and VCAM-1 expressed by HBMECs. Vehicle controls are included, and isotype control, single stains, and live dead stains are used for flow cytometry. I am comparing the change in the median brightness of ICAM-1 and VCAM-1 expression on HBMECs in response to each cytokine. Interestingly, we found IL-1β significantly increases ICAM-1 expression even with doses slightly above normal blood levels (2.5pg/mL). There were significant increases with TNF-α and IFN-γ only at doses reflecting highly elevated levels in the blood (1000 pg/mL), whereas there was no response to high doses of IL-2, IL-6, IL-8, and IL-10, CXCL10, and GM-CSF. Further studies blocking cytokines that greatly induced ICAM-1 expression in a mouse model and test if that will reduce neurotoxicity without affecting effectiveness of CAR T treatments would help us understand the underlying mechanisms of what causes neurotoxicity.

Oritavancin and dalbavancin are mechanistically similar to vancomycin and daptomycin and are among the longest-acting antimicrobials in clinical use with half-lives up to 14-days. This long duration of action increases antimicrobial exposure but prolongs the mutant selection window: the concentration range that selects for resistant mutants. Previous work shows that these drugs readily select for resistance and cross-resistance to vancomycin and daptomycin in Staphylococcus aureus. We hypothesized a similar cross-resistance selection potential against colonizing opportunistic pathogens including enterococci. While less virulent than S. aureus, enterococci are leading causes of multi-drug resistant infections among elderly and immunocompromised patients. To test this hypothesis, we serially passaged two strains of Enterococcus faecalis and three strains of Enterococcus faecium for 21-days in escalating concentrations of oritavancin or dalbavancin to facilitate the evolution of resistance. Minimum inhibitory concentrations (MICs) for oritavancin, dalbavancin, vancomycin, and daptomycin were performed by broth microdilution on isolates recovered from screening plates. Whole genome sequencing (WGS) was performed to characterize genotypic mechanisms of resistance. Resistance phenotype stability was assessed by passaging isolates on drug-free media and repeated MIC testing. Among dalbavancin-passaged enterococci, only daptomycin cross-resistance emerged. Serial passage in oritavancin selected for cross-resistance to dalbavancin in all strains, and cross-resistance to vancomycin among vancomycin-susceptible enterococci. Cross-resistance to daptomycin was not observed. WGS revealed fewer mutations than expected. Despite elevated MICs, no mutations were detected in E. faecalis. Only one strain of E. faecium acquired mutations in the vanZ and rodA gene. The absence of mutations in our evolved strains requires further investigation, including repeating WGS and screening for gene copy number variants that may have been missed by our initial analysis. Our data suggests that clinicians should exercise caution when using oritavancin to treat enterococcal infections due to the potential for cross-resistance to first-line antibiotics, such as daptomycin and vancomycin.

Teacher burnout is a historical issue plaguing schools, but instead of school leadership focusing on decreasing teacher burdens, many teachers experience increased rates of burnout due to their focus being shifted to the added responsibilities to implement evidence-based practices (EBPs). Existing literature has shown that principals’ leadership type (i.e., transactional vs. transformational) and the implementation climate present (i.e., perceptions that EBP use is rewarded, expected, and supported) can impact teacher burnout. My research analyzes teacher burnout rates as they relate to leadership type and implementation climate when implementing Tier 1 (i.e., universal social, emotional, and behavioral programs and practices) services. As part of a larger ongoing study of a leadership-focused implementation strategy (i.e., strategies designed to improve adoption, fidelity, and impact of EBPs), principals, teachers, and school staff from 10 elementary schools within the same district completed a survey battery at the beginning of the school year. Data collected by the research team included things such as leadership type, implementation leadership, teacher burnout, and implementation climate. Preliminary analyses are ongoing to identify trends amongst school-building implementation leadership, leadership types, implementation climate, and teacher burnout when implementing Tier 1 EBPs, which can reveal how key implementation aspects relate to one another.