Insects have superlative capabilities over contemporary robots: increased mobility, redundancy, coverage area, and can utilize different sensors. Perhaps most importantly, having reduced mass, launch costs can be exponentially lower. The goal of my research project was to create a simulation to compute the pathway/logistics of an insect robot landing on Mars, code a simulation, linearize data and arrays, and learn more about insect robots and space to investigate how to land insect-sized flying robots on Mars. I first reviewed existing information on spacecraft transiting from Earth to Mars, including the need to protect insect-sized robots from space and radiation while in transit. Of great interest would be potentially different de-orbiting and landing scenarios given a lower mass spacecraft, which is still traveling a hundred times faster than a bullet during initial deorbiting. My research speculated that simpler landing strategies like Spirit and Opportunity vs Endurance might be employed for the carrier spacecraft, and once on the surface, a carrier could deploy a small rover to act as a home base supplying power and communications to the flying insect-sized robots, greatly extending the range of the science data collection. My research captured the general characteristics of insect robotics and using a Python program I created, simulated reentry paths and maximum heating rates, which were still high as expected. My next steps would be to test different ballistic coefficients to see if a small payload direct from deorbit landing is possible. The broader implication is the potential for delivering many tiny distributed sensors on Mars to dramatically improve our understanding of the planet and at a lower cost.

Development and testing of sensors and power methods for insect-based robots is a difficult task. Due to the high cost of manufacture with regards to both training time and funds, finding a sustainable and easy-to-produce method to test sensors and power options is essential. Previously, the only option for testing new sensors and power options was using one of the robotic insects, which is risky considering their high costs. Drawing from prior results, we believe a lightweight quadcopter would be faster, easier to produce, more robust, and able to serve as a suitable replacement in sensor testing and development. My goal will be to create the world's lightest and smallest quad-rotor helicopter, “TinyQuad,” with a target mass of 1–2 g. The new helicopter I have designed will enable the testing of new sensors such as cameras and power options such as radio frequency-based charging. This design allows for testing a variety of sensors and electronics configurations very quickly, with the potential to rapidly speed up the prototyping process. I will demonstrate flight capabilities through utilizing wireless charging, and sensor-based feedback control to improve flight stability and duration. I completed calculations and design of this hardware, and anticipate seeing that the collected flight data supports the utilization of a lightweight quadcopter in insect robotics development. This will allow us to rapidly develop and refine sensors for use onboard the RoboFly robotic insect platform. Creating a working quadcopter would result in accelerated prototyping that allows for more unusual sensor and payload designs, and for further research in developing new sensors and power methods for insect robotics, smaller quadcopters, and improved design of micro aerial vehicles.

In this study we developed a nonhuman primate model of simian immunodeficiency virus (SIV)-Zika virus (ZIKV) co-infection to understand how HIV infection impacts ZIKV pathogenesis and test our hypothesis that ZIKV pathogenesis is enhanced in people living with untreated HIV. Previously, we have found delayed viral clearance, as well as delayed and dampened expansion of whole blood monocytes, the ZIKV cellular targets, during SIV infection. Here, we sought to further characterize the innate immune responses of SIV-ZIKV co-infection, by assessing cytokine and chemokine release. Pigtail macaques (n=7) were infected with SIVmac239M and co-infected with ZIKV at 9 weeks post-SIV infection. Co-infected animals were compared to control animals (n=7) infected with ZIKV only. Longitudinal plasma and cerebral spinal fluid (CSF) were collected at timepoints pre- and post-infection and assayed using a multiplex immunoassay to quantify 24 different cytokine and chemokines ex vivo. SIV and ZIKV both induced pro-inflammatory responses, characterized by transient increases in interleukin-17 (IL-17A) and monocyte chemoattractant protein-1 (MCP-1), with no major differences between experimental groups. Plasma MCP-1 concentrations were also found to be consistent with dampened and delayed whole blood monocyte frequencies. Pro-inflammatory interleukin-8 (IL-8), a chemokine needed for recruitment of neutrophils, increased in the plasma during SIV infection but not following ZIKV infection, a result that is in contrast to our previous findings. Transient increases in IL-8 were detected in a few animals in the CSF after ZIKV infection, which may be evidence for neuroinflammation. Overall, no significant differences between SIV+ vs SIV- groups were found for any analytes detected in plasma or CSF during ZIKV infection. Collectively, our results demonstrate that both SIV and ZIKV infections induce a pro-inflammatory response, that is not enhanced by SIV-ZIKV co-infection. This suggests SIV induced immunosuppression does not impair pro-inflammatory cytokine responses during ZIKV infection.

Nucleic-acid based vaccines, including RNA and DNA, provide protective immunity by eliciting antibody (Ab), and cytotoxic T lymphocyte (CTL) responses. FDA approval of mRNA vaccines against SARS-CoV-2 (COVID-19) provide ample evidence that mRNA vaccines are a viable vaccine platform. Once a mRNA vaccine enters a cells cytoplasm, mRNA encoded antigens are produced rapidly inducing an immune response. The production of mRNA encoded antigens will wane over time as mRNA degrades and transfected cells die. Alpha viruses, a RNA virus, have a unique replication process whereby this virus amplifies its RNA genome upon entering a cell. We, as well as others, have developed RNA vaccines that, like alpha viruses, self-amplify once inside of a cell to create more copies of mRNA than entered cell. This self-replicating RNA vaccine is called a replicon RNA vaccine (repRNA). RepRNA induces robust and sustained antigen production and immune responses. Currently, mRNA vaccines are administered as a homologous prime/boost vaccine regimen where the same mRNA vaccine is given as a priming vaccine and boosting vaccine, but antibody titers wane over time allowing for potential infections. This project aims to evaluate If utilizing a heterologous prime/boost regimen with DNA and repRNA vaccines confers more robust and longer lasting antibody and CTL responses than homologous regimens with DNA or repRNA. Mice will be vaccinated with DNA as a priming vaccine and repRNA as a boosting vaccine that encode SARS-CoV-2 spike protein. Anti-SARS-CoV-2 spike IgG antibody titers will be measured at 3-4 different timepoints, and CTL responses will be evaluated using an interferon gamma (IFN-γ) enzyme-linked immune absorbent spot (ELISpot) assay. The insights gained from this project will help to inform future DNA vaccine formulations and regimens as more DNA vaccines and repRNA vaccines enter clinical trials.

Merkel cell carcinoma (MCC) is an aggressive skin cancer with a high (20%) rate of distant metastases, 80 percent of which occur within 2 years of diagnosis. Metastatic MCC (mMCC) to the heart is rare and presents a management challenge. Our systematic literature review revealed only 11 cardiac mMCC case reports. Most (n=6) patients received chemotherapy which is now known to lack durable response in MCC, and 2 received no treatment due to advanced disease and comorbidities. Hence, to better understand cardiac mMCC we queried an MCC data repository of patients diagnosed between 2011-2021. Progression-free survival (PFS) was measured from date of cardiac mMCC to progression or death. Among 582 MCC patients with distant metastases, 9 developed cardiac mMCC. Median age at initial MCC diagnosis (stage I (1), stage III (6), stage IV (2)) was 69 years. Most (n=8) patients  developed mMCC to the right atrium, except for 1 patient (initial stage pIIIA) who had metastasis to the left atrium. Treatment for cardiac mMCC varied: 5 patients received immunotherapy combined with radiotherapy, while the reminder received immunotherapy alone, somatostatin analog, or chemotherapy. Five patients had a complete response in the cardiac lesion after immunotherapy, with or without radiotherapy. Median PFS and overall survival (OS) was 114 and 325 days, respectively. To explore whether presence of cardiac mMCC impacts OS, we matched cardiac mMCC patients to non-cardiac mMCC patients by age, sex, stage, immunosuppression status, and number of prior metastatic episodes. Using Kaplan-Meier statistical analysis, we found no difference in OS for the matched cohort (p=0.96). Taken together, these data indicate the emerging role of immunotherapy and radiotherapy in controlling cardiac mMCC. Furthermore, the location of mMCC to the heart does not appear to confer a worse prognosis compared to non-cardiac sites.

Paraoxonases (PONs) are a family of three closely related genes found on the long arm of chromosome 7. The genes encode PON1, PON2, and PON3, which are primarily involved in metabolizing oxidized lipids and modulating oxidative stress. However, each of the PON enzymes are involved in important secondary reactions. PON2 is an intracellular enzyme localized in the mitochondria that plays a vital role in modulating oxidative stress and inactivating microbial quorum sensing factors. Individuals with PON2 deficiencies are sensitive to oxidative stress. It may be possible to restore PON2 function by creating an injectable protein for individuals with a PON2 deficiency. Our goal is to actively express recombinant PON2 in an E. coli expression system and inject PON2 into PON2 knockout mice to determine if PON2 function can be restored. To express PON2 in E. coli we designed a synthetic DNA sequence by removing the transmembrane sequence of PON2 and replacing it with the signaling sequence from PON1, which facilitates the purification of the chimeric protein. We transformed the synthetic PON1/PON2 plasmid in E. coli and are currently performing gel electrophoresis and activity assays to analyze the expression of PON2. If we see protein expression as expected, then we will purify the recombinant PON2 for injection using a histidine tag that we added to the end of the protein coding sequence in the construct. The histidine tag allows for single-step purification via affinity chromatography, which we will then inject into PON2 knockout mice to observe their response to oxidative stress. Findings from this experiment will allow for further understanding of PON2 function and its restoration via an injectable protein, as well establishing that E. coli expression systems can be used as a more cost-effective method for pursuing further PON2 related research.

 Twelve million US citizens are currently suffering from visual impairment, many of them with late-stage blindness, which is accompanied by a drop in visual acuity, and the inability to recognize faces (CDC, 2022). Currently, few electronic sight restoration devices (SRDs) exist as treatment options for late-stage impairment that are precise in targeting retinal cells. Current SRDs cannot selectively stimulate on and off-center retinal cells – instead, all cells are stimulated regardless of their biologically natural firing pattern. Our research aims to examine whether neural plasticity can aid in overcoming this distortion in sighted individuals, using dichoptic presentation of stimuli that roughly mimics this simultaneous stimulation of on- and off- cells. Specifically, we convolved Fourier filters (F, and it’s contrast reverse complement F′) with images that are contrasted reversed complements of one another (I and I′) to induce a similar coding distortion caused by SRDs. We hypothesized that, through perceptual learning, sighted participants who received training with these distorted stimuli would be able to adapt to these distorted on-and-off cell responses. To test our hypotheses, participants were assigned to one of two groups. While both groups performed an object discrimination task with the distorted visual input, the experimental group did 25 hours of video game (VG) training, while the control group (CG) completed 5 hours of the object discrimination task. Participants in the VG group played a game that is an adaptation of Fruit Ninja, and tested in the discrimination task every 5 hours of video game play. Preliminary results indicate that individuals assigned to the VG group displayed superior performance in the object discrimination task, compared to the control group. Our results indicate that individuals with SRDs may have the potential to learn to decode unnatural visual cell population responses, which could improve their visual perceptions and enhance quality of life.

As the fields of quantum cryptography and quantum computing grow there is a greater need for training electrical engineers who are interested in these subjects. At the University of Washington, there has been a push in the Electrical & Computer Engineering department to increase the accessibility of these fields. The goal of my work is to generate procedures that can be followed by undergraduate-level engineering students to teach them foundational quantum mechanical processes. As a part of this effort, a new hardware lab has been created, the Quantum Technologies Teaching and Testbed (QT3) Lab. The QT3 lab includes a quED kit that produces polarization-entangled photon pairs and requires minimal laboratory experience to operate. By using the motor and hand controls included in the kit students will be able to follow straightforward step-by-step processes to see the violation of Bell’s inequality. Bell’s Theorem is the idea that parts of quantum theory require non-local effects meaning the correlation between two measurements cannot be accounted for with a signal traveling at the speed of light. One way this has been demonstrated is through the CHSH inequality where two entangled photons are sent through polarizers set to several different angles and then through two detectors. The counts of the number of “coincidences” or simultaneous detections are recorded and then they can be used to calculate a statistic that describes the correlation between the photons. If they are entangled they will show a value that violates the correlation possible within local realism. In my research, I created a setup to replicate the CHSH experiment and verify the result. I then designed an educational lab around this process to allow students to learn from it. This will give students an introduction to many of the fundamental concepts required to understand how quantum computing functions.

Quantum sensing platforms based on ensembles of nitrogen-vacancy (NV) centers in diamond enable high-sensitivity magnetic field detection at room temperature, making them uniquely suited for studying biological systems. Our work leverages the advantages of this diamond-NV sensing platform to directly measure the bend stiffness of individual DNA molecules using quantum-enabled magnetic imaging. In our experiment, a single DNA molecule is tethered to the surface of a diamond-NV sensor at one end and adhered to a ferromagnetic nanoparticle at the other end. A magnetic tweezer is used to apply a torque on the magnetic particle which bends the DNA strand in an effort to align the nanoparticle’s magnetic moment with the applied magnetic field. Quantum control and microscopy of the NV centers subsequently allows us to image the particle’s magnetic dipole field and the total applied field. Any deviation of the particle’s magnetic moment vector from the applied field vector can then be attributed to an opposing torque exerted by the DNA on the magnetic particle. Thus, measuring the difference between these vectors allows for the measurement of DNA bend stiffness. With this project, I detail the use of optically detected magnetic resonance and magnetic field image analysis to accurately measure the components of the aforementioned vectors. Additionally, I explore the limits of the diamond-NV platform in sensing the applied magnetic field in a highly symmetric configuration. Prior measurements have inferred the bend stiffness of DNA from statistical measurements such as cyclization. The novel direct measurement of DNA bend stiffness enabled by this project will help the scientific community understand sequence-dependent DNA flexibility, which plays a key role in biological processes such as eukaryotic nucleosomal compaction, viral genome packaging, and transcription regulation.
 

As part of the shared Quantum Technology Teaching and Testbed (QT3) Laboratory, several microscopes are being developed to characterize quantum emitters in various material systems. We want to be able to control the amount of laser power used to excite these quantum emitters. One such microscope being developed is a wide-field microscope that combines optical and microwave excitation to coherently control the spin states of quantum emitters and read them out optically. It is important to be able to control the laser power because the amount of power used will affect the spin initialization and readout in these measurements. To implement laser power control, we use a feedback loop between a variable laser power attenuator and a power meter. The power meter uses a photodiode setup in a reverse-bias configuration. Upon absorbing incident light, the photodiode outputs a current that is proportional to the incident laser power. A load resistor converts that current to a voltage, which is read into a computer by an Arduino. This readout will provide feedback to the variable power attenuator to reach the desired power. The power meter is expected to measure laser power to within 9 μW and is designed to have the dynamic range needed to allow us to coherently control ensembles of quantum bits.

As part of the shared Quantum Technology Teaching and Testbed (QT3) Laboratory, a confocal microscope is being developed to study individual quantum emitters in various solid-state systems, such as nitrogen-vacancy centers in diamond and silicon-vacancy centers in silicon carbide. Laser power is a key control parameter in confocal measurements as the signal to noise ratio of photons collected from the quantum emitters depends on the intensity of the excitation laser. I am developing a laser power control system using a variable power attenuator and a power meter. The power attenuator is composed of a neutral density (ND) filter wheel attached to a stepper motor, which rotates the filter wheel until the desired power output is reached. The power meter uses a photodiode and a reverse-bias circuit to measure the incident light power as a voltage which is read by a computer through an Arduino. The power meter and power attenuator are combined in a feedback configuration where the stepper motor continuously turns the ND filter wheel until the power meter measures the desired power. With incident power 50 mW, I expect to vary the laser power between 5 μW and 45.5 mW by reducing the power by 5.9% every step of the stepper motor. This low-cost laser power control system will allow us to image single quantum emitters in a variety of materials and eventually perform quantum entanglement protocols with these emitters.