Determination of N-Linked Glycosylation Changes in Hepatocellular Carcinoma and the Associated Glycoproteins for Enhanced Biomarker Discovery and Therapeutic Targets

With hepatocellular carcinoma (HCC) remaining as the fifth most common cancer in the world, causing more than 700,000 deaths annually, the need for reliable, early stage diagnoses and preventive treatments is crucial. While serum glycoproteins are hepatic in origin, making them excellent targets for HCC biomarkers, they can originate from both cancerous and non-cancerous regions and direct analysis of cancerous tissue itself is lacking. To counteract this, I hypothesized that direct tissue analysis combined with proteomic analysis could be utilized to identify more potential targets specific to HCC for early detection. This was done with a primary focus on glycosylation—as most clinically approved biomarkers are glycoproteins—and examined direct tissue glycomics in conjunction with glycoproteomic techniques through two specific aims: 1) Determining patterns of N-linked glycan changes in HCC tissue using MALDI imaging mass spectrometry to compare to previously published serum changes and 2) identifying glycopeptides containing changes in observed patterns of N- linked glycans in HCC samples using a targeted glycoproteomic approach. In Aim 1, HCC tissue was examined using MALDI imaging mass spectrometry to v verify changes in glycosylation via direct tissue analysis. Here, it was found that increased branching and fucosylation were directly associated with the cancerous tissue when compared to normal or cirrhotic. To further identify changes in glycosylation, two methods (one novel and one adapted for imaging) were implemented on tissue to further classify N-linked glycan isoforms through linkage analysis, specifically for sialic acids and core fucose. Again, it was shown that core fucose is most directly related to HCC tissue, thus confirming serum findings in the literature. For Aim 2, the novel method of determining core fucosylation was used in conjunction with glycoproteomic techniques to further elucidate the core fucosylated glycoproteins of interest. With the tag left behind following the enzymatic cleavage, targeted glycoproteomics was used to determine glycoproteins of interest while eliminating some biases inherent in the method, such as low ionization efficiencies for more complex N-glycans. This work outlines the first in-depth analysis of HCC tissue specifically regarding N- glycan changes, a novel application to determine N-glycan isoforms, and the application of these methods for glycoproteomic enhancement. With these findings, new trends in glycosylation related to the disease state could be further uncovered, as well as provide new biomarker candidates or therapeutic targets for future studies

Barriers of Volunteerism for Coaches of Collegiate Division I Swimming Programs

Barriers to volunteerism are not researched often or concretely in specific volunteer populations. The aim of this project was to find agreement among a group of potential executive level volunteers in coaching to identify the most impactful and prevalent barriers preventing them from volunteer service. A Delphi study was utilized to identify and find consensus for these barriers as identified by the 30 collegiate swimming coaches who participated. A verification interview was administered to add further clarity to the results and allow participants to speculate as to methods to overcome the barriers listed. Key findings included the themes of dedicated time, length of time for participation, and timing of the event during the season as being the most impactful and prevalent results. The findings serve as a key in creating mitigation strategies for organizations to overcome these barriers in the future

Characterization of Benzene Laser-Induced Nonthermal Equilibrium via Nitric Oxide Laser Induced Fluorescence Temperature Measurements

A benzene Laser Induced Non-thermal Equilibrium (LINE) technique was characterized for utilization in N₂ supersonic and hypersonic gas flow fields. Characterization of a LINE technique, toward tuning of the rate, magnitude, and geometry of a Collisional Energy Transfer (CET) perturbation to or from the bath is desirable because CET rate has been shown to be coupled with macroscopic flow field properties like turbulent fluctuations in velocity. Therefore, the creation of a tunable LINE technique will allow for testing of the coupling of Non-Thermal Equilibrium (NTE) and turbulence in canonical flow fields in order to create models that can help design more efficient supersonic and hypersonic vehicles. To characterize benzene LINE in this dissertation the relaxation of highly vibrationally excited benzene, generated by pulsed UV laser excitation, was studied using the transient rotational-translational temperature rise of the N₂ bath, which was measured by proxy using two-line Laser Induced Fluorescence (LIF) of seeded NO. The resulting experimentally measured time-dependent N₂ temperature rises were modeled with MultiWell based simulations of CET from benzene Vibration to N₂ Rotation-Translation (V-RT). In flow fields above room temperature we find that the average energy transferred in benzene deactivating collisions depends linearly on the internal energy of the excited benzene molecules and depends approximately linearly on the N₂ bath temperature between 300 K and 600 K. The results are consistent with experimental studies and classical trajectory calculations of CET in similar systems. In low temperature flow fields (140 K-300 K) the CET rate was found to have an inverse temperature dependence which may indicate the turning on of a new CET pathway at low temperatures. Since very little work on the relaxation of highly vibrationally excited molecules at temperatures below ~250 K has been done in the past, more experiments and simulations are needed to determine the mechanism of the increase in CET rate at low temperature. A benzene LINE technique has now been characterized over a diverse range of temperatures. It may now be utilized to generate perturbations in supersonic and hypersonic flow fields

Evaluating the provision of school performance information for school choice

We develop and implement a framework for determining the optimal performance metrics to help parents choose a school. This approach combines the three major critiques of the usefulness of performance tables into a natural metric. We implement this for 500,000 students in England for a range of performance measures. Using performance tables is strongly better than choosing at random: a child who attends the highest ex ante performing school within their choice set will ex post do better than the average outcome in their choice set twice as often as they will do worse

Think-tanking the challences in three regions: the Great Basin

The most unique characteristics of the Great Basin are the ecological fragility of the resources, scarcity of water, predominance of federal land, high degree of urbanization, independence of rural people. Factors most likely to impede EM are conflicting goals and missions of agencies, conflicting social values among stakeholders, slow recovery rates of biophysical systems, and difficulty of predicting responses to natural disturbances and management actions. Characteristics most likely to facilitate EM are new political climates promoting consensus, extensive federal lands, social diversity, and improving management technology. A critical need for successful EM is communication and promoting public understanding of the process

The European Pine Shoot Moth (Rhyacionia buoliana Schiff.): With Special Reference to Its Occurrence in the Eli Whitney Forest

The European pine shoot moth has been recognized as a pest .in Europe for over a century. It was first discovered in the United States in 1914, since when it has become a serious enemy of red pine. The insect is becoming increasingly abundant in this country and is known to be present in Massachusetts, Rhode Island, Connecticut, New York, New Jersey, Pennsylvania, West Virginia, Illinois, Ohio, Michigan, possibly Florida, and in the provinces of Ontario and British .Columbia in Canada. Some fifteen species of pines susceptible to injury in varying degree have been reported as hosts .of this insect, including many of the more important timber trees of both Europe and North Anlerica. A brief morphological description of the more important external characters of all stages of the insect has been give

Accounting trends & techniques: not-for-profit organizations: financial statement reporting and disclosure practices

https://egrove.olemiss.edu/aicpa_att/1059/thumbnail.jp

Making and shaping endochondral and intramembranous bones

Skeletal elements have a diverse range of shapes and sizes specialized to their various roles including protecting internal organs, locomotion, feeding, hearing, and vocalization. The precise positioning, size, and shape of skeletal elements is therefore critical for their function. During embryonic development, bone forms by endochondral or intramembranous ossification and can arise from the paraxial and lateral plate mesoderm or neural crest. This review describes inductive mechanisms to position and pattern bones within the developing embryo, compares and contrasts the intrinsic vs extrinsic mechanisms of endochondral and intramembranous skeletal development, and details known cellular processes that precisely determine skeletal shape and size. Key cellular mechanisms are employed at distinct stages of ossification, many of which occur in response to mechanical cues (eg, joint formation) or preempting future load‐bearing requirements. Rapid shape changes occur during cellular condensation and template establishment. Specialized cellular behaviors, such as chondrocyte hypertrophy in endochondral bone and secondary cartilage on intramembranous bones, also dramatically change template shape. Once ossification is complete, bone shape undergoes functional adaptation through (re)modeling. We also highlight how alterations in these cellular processes contribute to evolutionary change and how differences in the embryonic origin of bones can influence postnatal bone repair

Characterization of Benzene Laser-Induced Nonthermal Equilibrium via Nitric Oxide Laser Induced Fluorescence Temperature Measurements

A benzene Laser Induced Non-thermal Equilibrium (LINE) technique was characterized for utilization in N₂ supersonic and hypersonic gas flow fields. Characterization of a LINE technique, toward tuning of the rate, magnitude, and geometry of a Collisional Energy Transfer (CET) perturbation to or from the bath is desirable because CET rate has been shown to be coupled with macroscopic flow field properties like turbulent fluctuations in velocity. Therefore, the creation of a tunable LINE technique will allow for testing of the coupling of Non-Thermal Equilibrium (NTE) and turbulence in canonical flow fields in order to create models that can help design more efficient supersonic and hypersonic vehicles. To characterize benzene LINE in this dissertation the relaxation of highly vibrationally excited benzene, generated by pulsed UV laser excitation, was studied using the transient rotational-translational temperature rise of the N₂ bath, which was measured by proxy using two-line Laser Induced Fluorescence (LIF) of seeded NO. The resulting experimentally measured time-dependent N₂ temperature rises were modeled with MultiWell based simulations of CET from benzene Vibration to N₂ Rotation-Translation (V-RT). In flow fields above room temperature we find that the average energy transferred in benzene deactivating collisions depends linearly on the internal energy of the excited benzene molecules and depends approximately linearly on the N₂ bath temperature between 300 K and 600 K. The results are consistent with experimental studies and classical trajectory calculations of CET in similar systems. In low temperature flow fields (140 K-300 K) the CET rate was found to have an inverse temperature dependence which may indicate the turning on of a new CET pathway at low temperatures. Since very little work on the relaxation of highly vibrationally excited molecules at temperatures below ~250 K has been done in the past, more experiments and simulations are needed to determine the mechanism of the increase in CET rate at low temperature. A benzene LINE technique has now been characterized over a diverse range of temperatures. It may now be utilized to generate perturbations in supersonic and hypersonic flow fields