Clinical Detection of Diagnostic Biomarker Panels Using Microfluidic Electrochemical Immunoarrays

Cancer is a worldwide infliction. Cancer does not discriminate. Cancer does not care if you are young or old, rich or poor, disabled or in the prime of your life. It can be caused by genetics, environmental factors and/or lifestyle. The challenge, how do we diagnose and treat such a dynamic disease? Cancer detection is expensive, invasive, inaccurate and lacks sensitivity. New methods that rely on measurements of analytes in solution for detection and quantification are promising alternatives. Panels of protein biomarkers may aid in personalized diagnosis as protein levels in patients are often upregulated or down regulated in relation to a specified disease. For the medical field this provides opportunities to bring cancer detection to clinical practice as it will enable physicians’ access to blood, saliva, or urine bioassays for screening, as well as monitoring progression and response to therapy. The objectives of this thesis are to utilize new technology in microfluidic fabrication, 3D printing and nanomaterial synthesis for ELISA alternatives. The sensors are developed to be sensitive, rapid, inexpensive and multiplexed for point of care diagnostics. These technically facile immunoassays in human patient serum comprise a multivariable approach for statistically improving the probability of diagnosing and differentiating forms of cancer

Determining Which Taxonomic Resolution Provides Greatest Insight to Impact of Mass Morality Events on Early Carrion Colonizers

Mass mortality events (MME) impact local ecosystems especially in systems where insects play a major role in recycling carrion. Insects may be unable to efficiently recycle the carrion. Selecting the appropriate taxonomic resolution will be advantageous especially when assessing sensitivity of arthropod communities to variably sized MMEs. A collaborative study examining an artificial MME using feral swine (Sus scrofa Linnaeus) was conducted at Mississippi State University (MSU). Five different masses of swine carcasses ranging from 24.9 kg to 725.7 kg were placed in a pine forested area, starting in the spring of 2017. Different plots (1 km apart) were created based on density, carrion presence or absence and accessibility by scavengers. Sticky traps were placed among the plots to collect insects and were replaced as needed (between 24 hours and two weeks). Sticky traps were sent to Texas A&M University for identification of insects. This project involves determining the most useful and practical taxonomic level to identify insects using the MME experiment. Based on the different biomass sites, the hypothesis is the diversity and richness of the arthropod community will not vary at each site over time regardless of taxonomic resolution and the best taxonomic resolution is the species level

The Use of Surface Cytokines as Biomarkers in B Cell Mediated Autoimmune Diseases

RA and SLE are B cell-mediated autoimmune diseases dominated by autoantibodies that affect over 1.5 million Americans. Together RA and SLE contribute to over 29 billion in healthcare costs, therefore due to the high financial burden and physical toll of these diseases on the population, there is a critical need to effectively and efficiently diagnose and treat RA and SLE patients. The aim of our studies was to identify biomarkers and drug targets to improve the identification and treatment of RA and SLE patients. As discussed above, APRIL, BAFF and TSLP have been implicated in the pathogenesis of RA and SLE. Current therapeutics used to treat RA and SLE provide sub-optimal response in a third of patients treated. Therefore, identifying subsets of patients based on their surface and soluble biomarkers and identifying which therapeutics that will provide the most benefit for these patient cohorts has the potential to rationalize the treatment of RA and SLE patients. The cytokine APRIL has been implicated as a potential disease mediator in B cell mediated autoimmune diseases. Our studies show that surface APRIL is expressed on circulating myeloid cells and on total CD19+ B cells and correlates with increased disease in RA patients. Our findings suggest that surface APRIL could provide an easily detectable biomarker and be a useful selection criterion for the administration of drugs that antagonize APRIL. Another cytokine, TSLP has been implicated in the pathogenesis of RA. We found that TSLP was elevated in the synovial fluid of RA and in the plasma of SLE patients compared to non-inflammatory OA patients indicating that TSLP may act locally in the joints of RA patients and systemically in SLE patients. These studies provide insight into the role of APRIL and TSLP in B cell mediated autoimmune diseases and provide a rationale to guide treatment strategies based upon the expression of APRIL and TSLP. This work is significant because it has the potential to provide clinicians with easily accessible biomarkers to identify the best treatment plan to target these cytokines and their function in pathways involved in RA and SLE pathogenesis

Effects of mass death on community structure and ecosystem function

Death and decomposition are natural processes that are generally well-understood. However, large events of death, such as mass mortality events (MMEs) are increasing in frequency and their impacts on the ecosystem are largely unknown. These events may have both bottom-up effects from increased nutrient input as well as top-down effects from loss of an ecological functional group by the affected population. Different functional MMEs may result in different top-down effects, creating cascading effects. In Chapter 1, I test the hypothesis that scavenger and herbivore simulated MMEs generate novel bottom-up and top-down effects. Results indicate that MMEs have a significant effect on communities, including on soil chemistry, plant tissue, soil microbes, and soil arthropods. Carrion effects on the community were both a result of biomass (MMEs vs. single carcasses) as well as functional group exclusion (herbivores, scavengers). Further, MMEs may also generate long-lasting community effects due to the size and nature of the disturbance. In Chapter 3, I evaluated the potential long-term of effects of MMEs by sampling an experimental MME that was conducted four years earlier. I found that MMEs generated long-term asymmetrical effects on ecosystems, with some noticeable changes in increased soil nutrients as well as an unexpected effect of biomass on aboveground arthropod communities, with very little effect on belowground soil arthropods. However, studies of long-term decomposition from mass carcasses may expand beyond studying MMEs. Composting of carrion is a continuous disturbance event, with numerous carcasses being deposited in the same location over a longer period of time. In Chapter 2, I analyzed potential effects on the surrounding community at a unique instance of concentrated carcass disposal (5 years old). Significant differences were revealed between samples taken near the compost pit (0 m, 5 m) compared to further distances (10 m, 25 m, 50 m) with calcium being increased away from the pit, different soil microbial communities at the pit than farther distances and increased aboveground arthropod abundance at the pit. These experiments provide us with a greater, holistic understanding of previously understudied events of mass death on community structure and ecosystem function

Pavlov Patient Database

This project is implementing and deploying a functional patient database with customizable treatment plans and progress notes for Choices!, a counseling service center. To build this application and support the various requirements, several frameworks were considered for this project including Django, Flask, and Ruby on Rails. The Ruby on Rails suite was selected for reasons of existing familiarity for the team. Existing libraries within Rails allow for a choice of several database engines; MySQL, Postgresql, MongoDB, and NoSQL were considered. MySQL was adopted as a result of the extensive documentation and for continuity with the previous solution. Front-end requirements were met with a combination of HTML and Javascript, linked to a Bootstrap 5 framework in order to streamline user accessibility. A key quality desired by the customer was ease-of-use, to encourage higher efficiency and easy adoption by the stakeholders. In the process of development, secure data handling also emerged as a further and highly-desired attribute

Conference fees and accessibility: when doors are barred

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Interferometric Ex Vivo Evaluation of the Spatial Changes to Corneal Biomechanics Introduced by Topographic CXL: A Pilot Study

PURPOSE: To determine the efficacy of interferometry for examining the spatial changes to the corneal biomechanical response to simulated intraocular pressure (IOP) fluctuations that occur after corneal cross-linking (CXL) applied in different topographic locations. METHODS: Displacement speckle pattern interferometry (DSPI) was used to measure the total anterior surface displacement of human and porcine corneas in response to pressure variations up to 1 mm Hg from a baseline pressure of 16.5 mm Hg, both before and after CXL treatment, which was applied in isolated topographic locations (10-minute riboflavin soak [VibeX-Xtra; Avedro, Inc], 8-minute ultraviolet-A exposure at 15 mW/cm2). Alterations to biomechanics were evaluated by directly comparing the responses before and after treatment for each cornea. RESULTS: Before CXL, the corneal response to loading indicated spatial variability in mechanical properties. CXL treatments had a variable effect on the corneal response to loading dependent on the location of treatment, with reductions in regional displacement of up to 80% in response to a given pressure increase. CONCLUSIONS: Selectively cross-linking in different topographic locations introduces position-specific changes to mechanical properties that could potentially be used to alter the refractive power of the cornea. Changes to the biomechanics of the cornea after CXL are complex and appear to vary significantly depending on treatment location and initial biomechanics. Hence, further investigations are required on a larger number of corneas to allow the development of customized treatment protocols. In this study, laser interferometry was demonstrated to be an effective and valuable tool to achieve this

Biomechanical Evaluation of Decellularized and Crosslinked Corneal Implants Manufactured From Porcine Corneas as a Treatment Option for Advanced Keratoconus

Currently corneal transplantation is the main treatment for late-stage keratoconus; however, transplantation procedures are accompanied by significant risk of post-surgical complications; this in addition to supply limitations imposed by a worldwide shortage of human donor corneas, has driven the development of alternative therapies. One such therapy is the use of corneal implants derived from porcine corneas (Xenia®, Gebauer Medizintechnik GmbH, Neuhausen, DE). In contrast to human donor tissue, these implants can be produced on demand and due to the processes used pose no risks for host-immune rejection. Their use has already been demonstrated clinically in patients for preventing the progression of topographic changes in keratoconus whilst improving visual acuity. The implants are derived from natural tissue and not standardised synthetic material, whilst this likely reduces the risk of issues with bio-incompatibility, there is inevitably variability in their intrinsic mechanical properties which requires investigation. Here, speckle interferometry is employed to examine the biomechanical properties, in response to physiologically representative forces, of native porcine corneal tissue prior to processing and after a proprietary 4-stage process involving decellularization, washing, compression and crosslinking. The control lenticules had an average Young’s modulus (E) of 11.11 MPa (range 8.39–13.41 MPa), following processing average E of the lenticules increased by 127% over that of the unprocessed tissue to 25.23 MPa (range 18.32–32.9 MPa). The variability in E of the lenticules increased significantly after processing suggesting variability in the propensity of the native tissue to processing. In summary, it is possible to produce thin (<90 µm) lenticules from porcine corneas with enhanced stiffness that are effective for treating late-stage keratoconus. Due to the observed variability in the responses of lenticules to processing, interferometry could be a useful technique for ensuring quality control in commercial production via biomechanical screening

A Pilot Qualitative Case Study of Agricultural and Natural Resources Scientists’ Twitter Usage for Engaging Public Audiences

Scientists are frequently asked to broadly share their expertise and research with a variety of audiences, beyond typical academic circles in their home disciplines. That could include developing community engagement programs, school outreach, leveraging online social networks, and other activities. The purpose of this study was to examine U.S. agricultural and natural resources (ANR) scientists’ typical science communication channels, their experiences utilizing Twitter for sharing their knowledge, research, and engaging in online public science discussion. Diffusion of Innovations theory and the model of science in-reach versus outreach guided this study. Researchers used a qualitative case study design. Data collection included ANR scientist interviews (n = 8) and application of Internet-based research methods for observing scientists’ Twitter activities. Four themes emerged from the data: 1) academic journals and conferences as scientists’ typical communication channels, yet Extension efforts help to broaden audiences, 2) scientists expected research to be peer-reviewed before public dissemination to combat misinformation and spreading of ‘junk science’, 3) scientists balanced professionalism, personalization, promotion, and Twitter hashtags for engagement, and 4) scientist-identified barriers to using Twitter included lack of time and avoiding heated discussions. Recommendations include revisiting scientists’ job descriptions and expectations for online science engagement. Also, there should be continual development and implementation of science communication training for scientists targeting best Twitter practices, growing followers for outreach beyond academic colleagues and groups, using visuals for online engagement, intentional scheduling for social media, and how to effectively navigate heated online discussions