Primary cilia are microtubule-based organelles that function in the transduction of signals for cell growth, cell differentiation, and environmental sensing. Cilia are dynamic structures which utilize intraflagellar transport (IFT) to move material along the cilia for their construction and maintenance. Genetic mutations that affect retrograde IFT are known as ciliopathies. IFT is mediated by dyneins, motors that use ATP energy to target cargo. Dynein light chain TCTEX1D2 associates with proteins that are important for ciliogenesis and has been implicated in ciliopathies. Novel candidate dynein light chains in the same family, TCTEX1D1, TCTEX1D3, and TCTEX1D4 are thought to associate with dynein complexes in retrograde IFT, however, their characterization is limited and so it is not known whether they do have roles in ciliogenesis. To identify and characterize the novel light chain proteins, bioinformatics work was performed to understand domains, protein structure, disease associations, protein interactions, protein modifications, gene expression levels, and conservation. The approach included aligning sequences through Clustal Omega to analyze protein modification similarity, congregating known disease associations and protein interactions through bioinformatic databases, and analyzing conservation through HomoloGene and sequence alignment. Results indicate that all proteins are involved in varying diseases associated with cilia , such as Duchenne Muscular Dystrophy for TCTEX1D1, and cancer. They have similar protein interactions and gene expression levels are most significant in reproductive tissues and the brain. Alignment of protein modifications indicated modification in similar regions for TCTEX1D1 and TCTEX1D2. Continuing research for protein molecular characterizations is required through analysis of mammalian stable cell lines.

Somatostatin-producing (SST-producing) neurons in the tuberal nucleus affect feeding in mice. However, when these neurons were ablated (caspase), there was no change in feeding in male mice, while feeding in female mice decreased without any change in body weight. This sex difference may be due to estradiol. To test this hypothesis, we compared feeding in caspase mice and non-caspase mice with or without ovariectomy. We hypothesized that when compared to non-ovariectomized females, feeding would not differ between control and caspase mice in the absence of estradiol. Our inconclusive results suggest that the effects of caspase and estradiol on feeding may be dependent on body weight. Resistin, a gene linked to estradiol and body weight, and its receptor CAP1 was identified. There are high levels of CAP1 in the tuberal nucleus, and mice with high levels of resistin tend to have more visceral adipose tissue (VAT) with larger adipocyte size. Lower levels of estradiol leads to higher levels of resistin, which contribute to higher levels of VAT. We hypothesize that adipocyte size in the VAT is correlated with high levels of resistin which interacts with estradiol and SST-producing neurons to affect feeding in female mice: mice that weigh more will have lower levels of estradiol and higher levels of resistin and will eat less compared to their counterparts. To test this, we are performing a haematoxylin and eosin stain and an RNA analysis of adipocyte tissue. This experiment will answer the etiology of sex differences in the physiology of eating.

In the United States alone, approximately one million people suffer from Parkinson’s disease (a movement disorder). In many cases, these movement disorders cannot be cured, and the goal of treatment is to minimize symptoms and relieve pain. With the goal of improving overall quality of life and giving some level of control back to the individuals, treatments like transcranial magnetic stimulation (TMS) have been one of the few options available for treating these disorders in affected individuals.The goal of this literature review is to explore how successful TMS is for studying in vitro brain slice recordings. A cross-reference study was conducted to analyze some of the most common types of movement disorders. This analysis included an exploration into some of the most prominent and characteristic symptoms for each disorder as well as the treatments that exist (or the lack thereof) as well as the effectiveness of TMS in these different movement disorders. No treatment is completely effective or efficient and it is important to note that these treatments are only temporary fixes that target individual symptoms. Transcranial Magnetic Stimulation (TMS) shows potential in that it may lead to a better understanding of these disorders which in turn can offer better treatments.

Stroke is a leading cause of death and disability, primarily affecting older individuals. While the majority of current research is conducted on young animals for practical reasons, many stroke-affected cellular processes may be changing with age. A functionally-related group of cells essential to brain homeostasis, the neurovascular unit (NVU), includes but is not limited to: endothelial cells, neurons, pericytes, astrocytes, and microglia. Current NVU research on stroke has mostly been conducted on gray matter models, but little is known about the white matter response. White matter strokes (WMS) are typically asymptomatic small vessel occlusions that accumulate over time, which are a leading cause of dementia. Moreover, although sex has a notable impact on functions that maintain brain homeostasis, how these differences factor into WMS have not been well studied. In this study, we sought to more thoroughly understand how age, WMS, and sex interact to affect NVU cell dynamics by examining different cell types using immunostaining. After undergoing stroke and sham surgeries, aged male and female mice were evaluated for components of the NVU. To assess NVU proliferation, we observed the quantity and distance from the infarct border of overlapping signals of endothelial cells (CD31), microglia (Iba1), and pericytes (CD13), and a marker of proliferation (EdU). We also evaluated the size of the fibrotic scar (Col1a1) and axon (NF160) density in the peri-infarct region. Analysis is still ongoing, but this data will provide greater insight into NVU cell changes between sexes post-WMS and potential targets for treatment.

Toxoplasma gondii is an obligate intracellular parasite in the phylum Apicomplexa that causes serious disease in immunocompromised patients and congenitally infected neonates. T. gondii can infect a wide array of mammalian species and infection is widespread geographically, thus it is considered to be one of the most successful parasites on the planet. Neospora caninum is a closely related pathogen that causes abortion in cattle and neurological disease in dogs, but does not infect humans. T. gondii and N. caninum both serve as model systems for the study of less amenable apicomplexans, including Plasmodium spp., the causative agent of malaria, and Cryptosporidia spp., which causes diarrheal disease in children. We have utilized an organelle isolation and monoclonal antibody approach to develop an array of probes for subcellular localization. We have determined localizations that include the unique organelles of apicomplexans (e.g. inner membrane complex, rhoptries, and apicoplast) and standard eukaryotic organelles (e.g. the mitochondrion). A significant number of these probes cross-react between T. gondii and N. caninum, highlighting the similarity between these organisms. We additionally characterized many of these antibodies by western blot analysis using both reducing and non-reducing conditions to determine the approximate size of the proteins they detect. We then used immunoprecipitation and mass spectrometry to identify the protein targets of a subset of these antibodies. Characterization of these antibodies will establish new molecular tools for the field and identify new putative drug targets in these important pathogens.

Encoders and decoders in communications systems are critical for the accurate and efficient transmission of information over noisy channels. Our research is focused on encoders and decoders for tail-biting convolutional codes used in conjunction with cyclic redundancy check (CRC) codes. We implement encoders and decoders that correct errors in the received message when possible. Often, when an error cannot be corrected, the CRC informs the decoder that the selected codeword is unreliable. In our research, we extend the work of Liang et al., who demonstrated that the use of distance spectrum optimal cyclic redundancy checks (DSO CRCs), along with list decoding, offered significant improvements in signal to noise ratio (SNR) with minimal additional computational cost for low rate convolutional codes of the form 1/n, which have n output bits for every 1 input bit. Our research applies this approach to high rate convolutional codes of the form (n − 1)/n, which have n output bits for every n − 1 input bits. Specifically, we implemented the decoder for a rate-3/4 tail-biting convolutional encoder, and used the dual trellis approach proposed by Yamada et al. for efficient decoding, along with the tree-trellis list decoding algorithm proposed by Roder and Hamzaoui. By implementing this system in C++, we have the ability to simulate its performance at low frame error rates and compare it to both the random coding union bound and the performance of a standard maximum likelihood decoder.

Anxiety-related disorders are the most prevalent forms of mental illnesses today, affecting over 40 million adults in the United States alone. A fundamental component of these disorders is the heightened and unwanted expression of fear. Animal work suggests that fear response is regulated by communication between the basolateral amygdala (BLA), medial prefrontal cortex (mPFC), and ventral hippocampus through fluctuating neuronal oscillations. However, whether this same activity occurs in humans is unknown. Thus, we present a method of eliciting fear response in humans in an image task conducted during simultaneous neural recording. Positive, negative, and neutral emotional images were displayed to four participants while collecting iEEG recordings throughout the task. We demonstrate that there are varying responses to emotional images belonging to differing valence categories. These findings represent one of the first characterizations of fear response with concurrent intracranial recording. Preliminary results lay the foundation for future work in both refining and improving therapies for anxiety-related disorders.

Skeletal muscle progenitor cells (SMPCs) and skeletal muscle stem cells called satellite cells (SCs) differentiate to form skeletal muscle. SMPCs are mainly present during prenatal development and SCs are mainly present in adults. Duchenne Muscular Dystrophy (DMD) is a fatal disease that occurs due to lack of a protein called dystrophin which leads to reduced efficiency of SCs. This leads to reduced ability to regenerate muscle. There is currently no cure for DMD. Reprogramming cells from a patient's biopsy into human pluripotent stem cells (hPSCs) and differentiating them into healthy SCs for transplantation back into the patient is a promising stem cell therapy for DMD. Protocols have been developed to differentiate hPSCs into SMPCs. However, these hPSC-derived SMPCs (hPSC-SMPCs) best resemble immature embryonic-to-fetal SMPCs (week 7-12) that are not as regenerative as SCs, and it is not known how to mature hPSC-SMPCs into SCs for use in therapies. One potential avenue of promoting maturation of hPSC-SMPCs is to manipulate the epigenome of hPSC-SMPCs into that resembling SCs. However, epigenetic differences between SMPCs and SCs are not well understood. Using single cell ATAC sequencing, I am identifying differences in chromatin accessibility between hPSC-SMPCs and SMPCs and SCs derived from tissues. This will provide insight into how to modulate hPSC-SMPCs to become more regenerative SCs.

Modern day mobile applications not only require a low latency, but also incur a high energy cost. However, mobile devices are limited in their battery life and computational capabilities. Mobile Edge Computing (MEC) reduces latency and energy consumption by offloading some or all of the computing tasks to MEC servers. The Mobile Edge Compute Applications (MECA) lab establishes a 5G development environment that allows for experiments and testing with the 5G network and MEC. This particular project is an autonomous car controller which offloads control decisions to a central server, using a camera as input. Using gRPC, the camera sends images to the central server, which processes them and returns a control signal to guide the car. The goal is to compare the latency of this setup with traditional autonomous car controllers and achieve similar or better performance. At the moment, since it is difficult to gain access to physical materials, a simulation that imitates the complexity of video processing was used to gather data on the effectiveness of this strategy. As expected, sending the processing to a server is more time consuming with simple tasks, due to the initial time cost associated with the server-client communication. With more complex tasks, the server processing time is faster than processing at the client, since the server has better computational capabilities. The next step is applying these concepts to a physical setup with proper video processing and a car which takes the control signal.

The COVID-19 pandemic has highlighted the growing need for doctors to assess and treat their patients virtually rather than in person. In order to do this, it is important to be able to take a person’s vitals virtually. Currently, there are algorithms that are able to find heart rate based on a video of a person. However, their accuracy varies based on skin tone, with a much higher accuracy for lighter skin tones than darker ones. These inaccuracies need to be corrected before this technology is allowed to be used in the medical field. Our lab attempts to minimize this bias by incorporating diverse training sets to train deep learning algorithms DeepPhys and PhysNet in order to determine the optimal proportion of lighter to darker skin tones in our training set needed to achieve an accurate ppg reading for all skin tones. Participants were asked to connect to a pulse oximeter and placed in front of a white background, while five one-minute videos were taken in sync with the pulse oximeter ppg readings. These videos were then used to train and test DeepPhys and PhysNet with different proportions of lighter skin tones to darker skin tones. Although the experiment is not yet complete, initial results show that having more darker skin participants in the training set decreases the accuracy of the networks in the testing set, but we believe that this is due to not having enough darker skin toned participants needed to truly test our hypothesis.

A minority of patients with melanoma respond to immunotherapies, potentially due to immune cell exclusion at metastatic melanoma sites such as the liver. Analyses of autopsy-derived melanoma samples illustrate dense fibrotic capsules in and around liver metastases (mets) and lack of immune infiltration. Interest in three-dimensional (3D) tumor modeling has developed rapidly in recent years, motivated by desires to study cellular interactions more accurately in the tumor microenvironment. Here, we modeled 2D and 3D in vitro systems of melanoma in the liver tumor microenvironment to investigate the roles of vitamin D pathway molecules in this context; in our previous RNA Sequencing analysis, we discovered upregulation of vitamin D pathway molecules in melanoma liver mets. We studied the migration and invasion capabilities of melanoma cells under the companionship of hepatocytes and liver stromal cells (separately) in 2D. In addition to culturing 3D melanoma alone, we co-cultured melanoma cells with hepatocytes and liver stromal cells in 3D. This study established a more physiologically relevant in vitro model through which to study melanoma liver mets. Our 2D investigations demonstrated that melanoma cells migrate and invade towards liver stromal cells but not liver cells. Our work thus far has set the stage to illuminate the important role(s) vitamin D pathway molecules play in maintaining the metastatic liver tumor microenvironment, which may subsequently inform future therapeutic directions.