Engineering Commons^™

Analyzing Pseudomonas Aeruginosa With Bacteriophage Tags Using Photoacoustic Flow Cytometry, Jennifer C. Schinke

Electronic Theses and Dissertations

The number of daily bacterial infections is climbing and the CDC explains that this is due to the antibiotic-resistant threat in the United States. Finding a faster way of bacterial identification is necessary as it currently takes 1-4 days for a medical lab to culture and identify bacteria. Photoacoustic flow cytometry (PAFC) can be used as an alternative method resulting in swift identification within an hour (Edgar, 2019). Pseudomonas aeruginosa, cell line PA01, will be coated in up to a few hundred red dyed phages making it detectible by the photoacoustic flow cytometry system. Bacteriophages (phages) are viruses that …

Labeling Melanoma Cells With Black Microspheres For Improved Sensitivity In Detection Via Photoacoustic Flow Cytometry, Tori Kocsis

Electronic Theses and Dissertations

Melanoma is an aggressive form of skin cancer known for developing into metastatic disease. Current clinical diagnostics, including medical imaging and tissue biopsy, provide a poor prognosis since the cancer is in the late stages of disease progression. In recent years, photoacoustic flow cytometry has allowed for the detection of circulating melanoma cells within patient blood samples in vitro. Although this method exploits the naturally-produced melanin within the cells, it has only successfully detected highly-pigmented melanoma cell lines. Since various forms of melanoma exist, each with varying melanin concentrations, this research aims to provide a novel method for detecting lightly-pigmented …

Investigating The Effect Of Dissolved Oxygen-Assisted Corneal Cross-Linking (Cxl) On Porcine Corneas, Julianni Dar

Electronic Theses and Dissertations

Corneal cross-linking is a clinical procedure that is known to stop the progression of keratoconus, an eye disease that affects the cornea’s structure, ultimately leading to vision loss in its advanced stages. The typical treatment plan includes riboflavin and UV-A exposure in the hope to increase the mechanical properties of the cornea. There are two types of CXL pathways, with Type-II CXL requiring oxygen. Naturally, the dissolved oxygen is limited in the cornea; therefore, limiting the effect of Type-II CXL. This study proposes to improve the Type-II CXL contribution by integrating dissolved oxygen during the standard CXL treatment used in …

A Microfluidic Assay For Single Cell Bacterial Adhesion Studies Under Shear Stress, Amanda Trusiak

Electronic Theses and Dissertations

The study of bacterial adhesion to host cells is important in understanding bacterial pathogenesis and developing new therapeutic approaches. Here, we studied bacterial adhesion under shear stress using a novel microfluidic method. Specifically, the adhesion of a uropathogenic E. coli strain (FimHOn, ATCC 700928/CFT073) to mannose-modified substrates was studied under flow conditions. The FimHOn E. coli strain expresses FimH which is a mannose-specific adhesin found on the fimbriae that binds to glycoproteins on the epithelium. We developed a microfluidic method that mimics bacterial adhesion to urothelial cells. First, the microfluidic channels were modified by sequentially adsorbing BSA-mannose and BSA onto …

Detecting Viral Particles In Vitro Using Photoacoustic Flow Cytometry, Anie-Pier Samson

Electronic Theses and Dissertations

The Covid-19 pandemic is a powerful example of just how damaging the rapid spread of an unknown virus can be. Viruses have the ability to spread rapidly amongst individuals if not treated and controlled. The first key step towards treatment is the timely and specific diagnosis of the virus causing the infection. An innovative method for the rapid detection of viral particles in solution consists of tagging the viral particles paired with using photoacoustic flow cytometry to irradiate the particles and get a signal. The high affinity of Streptavidin for Biotin can be used to bind Streptavidin-coated microspheres to biotinylated …

Real-Time Monitoring Of Cell Death Progress Using Capacitance Spectroscopy, Suyang Wu

Electronic Theses and Dissertations

Biologics, including the monoclonal antibody (mAb), has experienced rapid development in the last decade. However, the price of biologics is often prohibitively high because of the low process efficiency. Delaying the inevitable cell death improves the productivity of upstream bioprocessing, whose success relies on monitoring the cell death onset that indicates the timing for preventive actions.

This study proposes to develop a real-time monitoring model that quantifies the dying cell percentage in lab-scale bioreactors using capacitance spectroscopy. The capacitance spectroscopy contains cell death-related information due to various physical properties changes during the cell death process, e.g., cytoplasmic conductivity change. The …

Development Of A Microfluidic Viscoelastic Hemostatic Assay For Real-Timeviscosity Measurements Of Blood, Shay Kent

Electronic Theses and Dissertations

Blood coagulation disorders are malfunctions in the body’s ability to control blood clotting. It can result in either insufficient clotting causing an increased risk of bleeding or excessive clotting obstructing blood flow. The rapid and accurate diagnosis of coagulopathies is an important, unmet need in the clinical setting. Rapidly identifying the source of bleeding, either acquired or inherited, is critical to reduce the risk of major blood loss and deliver personalized hemostatic therapies. Viscoelastic hemostatic assays, or VHAs, deliver an effective solution to the diagnosis of coagulopathies by evaluating global hemostatic function using whole blood rather than plasma. VHAs are …

Machine Learning Applied To Colloidal Properties Of Perfluorocarbon Nanoemulsions For Imaging In Ards/Ali, Marco Hosfeld

Electronic Theses and Dissertations

Acute Respiratory distress Syndrome (ARDS) and Acute Lung Injury (ALI) are inflammatory lung pathologies consisting of non-hydrostatic pulmonary edema leading to hypoxia and impaired gas exchange in the lungs. ARDS/ALI is both difficult to study and treat as it is not in itself a specific pathology but rather a syndrome consisting of many pathologies that vary case by case. It is, however, consistently characterized by an explosive acute inflammatory response in the lung parenchyma leading to hypoxia. Although time has seen to an increase in the understanding of ARDS/ALI, the mortality rate remains in the range of 30-50%. For these …

Diagnosis Of Melanoma Disease State From Patient Blood Samples Using Photoacoustic Flow Cytometry, Jacob Salvatore

Electronic Theses and Dissertations

Approximately 1 in 50 Americans develop melanoma in their lifetime. Early detection of melanoma is pivotal to the survival of the patient, with a 99% survival rate for 5 or more years after an early diagnosis. Metastasis, or the spread of cancer, increases the chances of cancerous growth in other parts of the body. In this study, we propose the use of photoacoustic flow cytometry as a diagnostic of patient disease state, by closely monitoring the amount of circulating melanoma cells (CMCs) in a patient’s blood before and after chemotherapy treatments. Using this patient data, we were able to make …

Detecting A Heterogenous Sample Of Pigmented Melanoma Cell Lines Using Photoacoustic Flow Cytometry, Margaret Cappellano

Electronic Theses and Dissertations

Metastatic melanoma is the deadliest form of skin cancer, which is in part, attributed to its rapid aggression and lack of response to typical treatment methods. There are far too often cases where a lymph node biopsy does not detect the severity of the cancer, which in turn causes a lack of diagnosis until a mass can be visually detected on a scan, such as a PET, CT, or MRI. Once visible on a scan, the cancer is too progressive for successful treatment. To avoid this, we investigated how a blood sample can be used to negate a missed diagnosis, …

Characterizing The Dormancy And Repair Of A Circumneutral Passive Remediation System Receiving Iron And Sulfate-Rich Amd, Garrett Struble

Electronic Theses and Dissertations

The Wingfield Pines remediation system is a ~20-acre circumneutral, gravity-fed system constructed in 2009 to passively remediate Fe from an ~2000 gallon/minute effluent. In 2017, a fracture to the system’s underground cavern caused the AMD effluent to bypass the system, flowing directly into Chartiers Creek and leaving the system decommissioned between 2017-2019. Soil slurry and water samples were collected in the weeks prior to the system’s reinstalment in September 2019 through January 2020 in order to determine the effect of the repair on remediation efficiency and microbial community composition. This study acted as the first to describe the temporary decommissioning …

Developing A Multimedia Interface For Electrical Biosignal Interpretation, Christopher Cox

Undergraduate Research and Scholarship Symposium

In the last ten years, a great deal of interest has been generated around the practice of using plants’ electrical signals to create sound or music. In order to render biosignals capable of producing sound electronically, some kind of interface is necessary that converts a plant’s natural electrical signals into data that a computer can understand. Existing commercial interfaces cost at minimum $3001,2 . Worse yet, two popular off-the-shelf interfaces conform their output signals to stereotypical human notions of what a plant might “sound like”. In this project we fabricated a simple but high-performance interface for less than $5 in …

Understanding The Effect Of Adaptive Mutations On The Three-Dimensional Structure Of Rna, Justin Cook

Undergraduate Research and Scholarship Symposium

Single-nucleotide polymorphisms (SNPs) are variations in the genome where one base pair can differ between individuals.¹ SNPs occur throughout the genome and can correlate to a disease-state if they occur in a functional region of DNA.¹According to the central dogma of molecular biology, any variation in the DNA sequence will have a direct effect on the RNA sequence and will potentially alter the identity or conformation of a protein product. A single RNA molecule, due to intramolecular base pairing, can acquire a plethora of 3-D conformations that are described by its structural ensemble. One SNP, rs12477830, which …

Project Alien, Rebecca Mccallin, Madelyn Hoying, Alex Evans, Matthew Nestler, Karli Rae Sutton, Garett Craig, Lucia Secaia Del Cid, Alexander Guy, Rachel Fernandez, Amanda Trusiak, Paige Aley, Ingabire Gakwerere, Nina Dorfner, Maria Mosbacher, Mary Flavin, Selvin Hernandez, Audrey Steen, Benjamin Kazimer

Undergraduate Research and Scholarship Symposium

Project ALIEN is a comprehensive plan to send humans to Mars to look for life on the Martian surface while exploring the viability and adaptability of terrestrial microbes in Martian atmospheric conditions. ALIEN will use a ballistic capture trajectory to get to Mars and stay in aerostationary orbit for a 30-day surface mission, during which two surface crewmembers will perform a variety of experiments to achieve the mission’s goals of Martian microbial discovery within brines of the Gale Crater and terrestrial microbe adaptability and viability to Martian conditions. Experimentation is based on the presumption that Martian microbes are metabolically similar …

Bioprinting Alginate Structures Using The Fresh Method, Amanda Pellegrino

Undergraduate Research and Scholarship Symposium

Three-dimensional bioprinting is a tissue engineering and regenerative medicine technique that utilizes biomaterials to print clinically relevant scaffolds. These scaffolds are used for tissue repair and to treat disease, such as end-organ failure. The main problem with 3D bioprinting is its inability to print soft biomaterials, which collapse due to gravitational forces. In addition, the materials that can be used do not mimic native tissue’s mechanical properties and texture. To combat this, Dr Feinberg’s laboratory at Carnegie Mellon University developed the freeform reversible embedding of suspended hydrogels (FRESH) method. The FRESH method allows for soft biomaterials to be supported in …

Multiphysics Computational Model Of Fluid Flow And Mass Transport In Aneurysm, Tanja Cupac

Electronic Theses and Dissertations

The abdominal aortic aneurysm is progressive, asymptomatic, and can eventually lead to rupture which is a catastrophic event leading to massive internal bleeding and possibly death. AAA cases have been characterized by the development of an intraluminal thrombus (ILT). The ILT correlates with the progression of hypoxia in the arterial wall. The extent that ILT presence reduces oxygen flux to the wall has not been quantified and there is rather a poor understanding of key parameters that can affect thrombus-mediated oxygen transport in AAA. The purpose of this study is to address this gap and to assess the effects of …

Bacteria Analysis By Using A Supervised Machine Learning Algorithm Based On Droplet Microfluidics, Yulder Daniel Angarita

Electronic Theses and Dissertations

Sepsis is a major medical problem and massive resources have been invested in developing and evaluating alternative treatments. Statistics indicate that sepsis causes between one third and one half of all hospital deaths in the United States. Sepsis has a high impact on health care in the US, with direct sepsis costs in 2009 exceeding $15.4 billion. A research study found that a 1-hour delay in appropriate antimicrobial care resulted in a 7% - 10% rise in mortality. Several professional societies seek to reduce sepsis mortality by targeting the timely use of diagnostic tests and antimicrobial therapy. The diagnostic instruments …

A Microfluidic Platform For High-Throughput Screening Of Aquaporin Performance, Adriana Del Pino Herrera, Jordan Hoydick, Rachel Rauh, Elyssa El-Hajj, Madison Burchfield

Undergraduate Research and Scholarship Symposium

Aquaporins are a family of small integral membrane proteins that transport water across cell membranes in response to osmotic gradients. They facilitate fluid secretion and absorption across epithelial surfaces in kidney tubules, exocrine glands, and gastrointestinal tract. Here, we describe a novel microfluidic method to evaluate and screen for aquaporin-based transmembrane permeability in mammalian cells. A microfluidic device was designed and fabricated for the encapsulation of single mammalian and yeast cells in micron-sized droplets. For this purpose, Chinese Hamster Ovarian (CHO) cells were used. CHO cells express AQP1 (aquaporin-1) homologous to human kidney aquaporins. The cells were cultivated and exposed …

Pilot Study Exploring The Effect Of Targeted Cox-2 Inhibition In Macrophages Responding To Neuronal Injury; Promoting Enhanced Axonal Regeneration, Alyssa Brauckmann

Electronic Theses and Dissertations

Celecoxib nanoemulsion (CXB-NE) has been developed as a macrophage targeted analgesics by Dr. Janjic and her team at Duquesne University, (Janjic et al, 2018; Liu et al, 2020; Saleem et al, 2019b; Vasudeva et al, 2014). The CXB-NE nanoemulsion carrying a Nonsteroidal Anti-inflammatory (NSAID) inhibitor of COX-2 activity result in a reduction in PGE2 expression in macrophages. Using CXB-NE in rats that have peripheral nerve injury constricting the sciatic nerve relieves hypersensitivity, a pain-like behavior. The treatment also decreases inflammation associated with this chronic constriction injury (Janjic et al, 2018; Saleem et al, 2019b; Stevens et al, 2019). In this …

Identification Of Human Adrenal Carcinoma Cells Using Photoacoustic Flow Cytometry, Alexis Stahl

Electronic Theses and Dissertations

With consistently high mortality rates in patients with adrenal cancer, lack of early detection and diagnosis are an ever increasing problem in fighting the deadly disease. Currently, very few patients are diagnosed with adrenal cancer within the early stages due to the absence of symptoms until large tumors are formed, which is in most cases is fatal. Because of this, early detection equipment is crucial to optimizing the survival rate in patients. We studied the absorbance wavelength of human adrenal carcinoma cells, as well as the minimum cell concentration needed in samples using photoacoustic flow cytometry (PAFC). PAFC generates ultrasonic …

Characterization Of Photoacoustic Flow Cytometry Signals, Craig Benzinger

Electronic Theses and Dissertations

Photoacoustic flow cytometry has been utilized to clinically determine the presence of melanoma circulating tumor cells (CTCs). Further investigation was conducted into the morphology of detection signals and how they could be manipulated to allow for further classification. Novel features were extracted from waveforms that appear to have strong classification ability. Neural networks were also used to determine classification potential and the creation of feature mapping for future unsupervised classification. Detections were expanded from single waves to a time dependent multiwave event. Waveforms were also determined to be of non-parametric distribution, allowing for classification by neural network but not allowing …

Multiphysics Computational Model Of Fluid Flow And Mass Transfer In Aneurysm, Tanja Cupac

Graduate Student Research Symposium

The abdominal aortic aneurysm (AAA) mostly affects men between the ages 65-85 and is very common with more than 3 million cases in the United States per year. AAA is defined as the weakening of the arterial wall producing a blood-filled bulge in the body’s largest blood vessel. AAA is progressive, asymptomatic, and can eventually lead to rupture which is a catastrophic event leading to massive internal bleeding and possibly death. Of the males within this age range, 1-3% will die from this disease. About 75% of AAA cases will be characterized by the development of an intraluminal thrombus (ILT). …

Fluoride Removal From Water Using A 3d Printed Calcium Carbonate Filter, Sophia Bakar, David Kahler, Benjamin S. Goldschmidt

Undergraduate Research and Scholarship Symposium

Groundwater containing high concentrations of fluoride is the most common source of drinking water in rural areas in parts of east Africa, India, and China. The elevated levels of fluoride cause skeletal and dental fluorosis, which is the weakening and decay of bone structures due to the leeching of calcium from the body as calcium and fluoride bond by the process of adsorption. Over 150 million people are suffering from some form of fluorosis due to the consumption of groundwater. Calcium carbonate has been demonstrated to influence fluoride removal in several forms. To make fluoride removal a cost-effective and user-friendly …

Quantifying Iron Overload Using Mri, Active Contours, And Convolutional Neural Networks, Andrea Sajewski, Stacey Levine

Undergraduate Research and Scholarship Symposium

Iron overload, a complication of repeated blood transfusions, can cause tissue damage and organ failure. The body has no regulatory mechanism to excrete excess iron, so iron overload must be closely monitored to guide therapy and measure treatment response. The concentration of iron in the liver is a reliable marker for total body iron content and is now measured noninvasively with magnetic resonance imaging (MRI). MRI produces a diagnostic image by measuring the signals emitted from the body in the presence of a constant magnetic field and radiofrequency pulses. At each pixel, the signal decay constant, T2*, can be calculated, …

Re-Evaluating Performance Measurement: New Mathematical Methods To Address Common Performance Measurement Challenges, Jordan David Benis

Electronic Theses and Dissertations

Performance Measurement is an essential discipline for any business. Robust and reliable performance metrics for people, processes, and technologies enable a business to identify and address deficiencies to improve performance and profitability. The complexity of modern operating environments presents real challenges to developing equitable and accurate performance metrics. This thesis explores and develops two new methods to address common challenges encountered in businesses across the world. The first method addresses the challenge of estimating the relative complexity of various tasks by utilizing the Pearson Correlation Coefficient to identify potentially over weighted and under weighted tasks. The second method addresses the …

Implementation Strategy Report, G Jager, M Musariri

Policy

In essence the Implementation Strategy provides a plan for implementing and maintaining an optimal South African Water Resources Monitoring Network. As such the Strategy includes a number of key aspects considered for each of the surface water, groundwater and water quality monitoring programs, as summarized below:

Finalizing the optimal monitoring network design, including the location, the variables/constituents being monitored, the frequency of observations, as well as the implementation priority for each monitoring site.

Identifying opportunities for the integration of processes involved in the implementation process, such as the coordinated development or upgrading of monitoring sites based on physical location and …

Scientific Review Report, J D. Rossouw, M Musariri

Policy

This report provides the Scientific Review results and recommendations regarding the existing surface and groundwater monitoring sites for all nine Water Management Areas following the Regional WMA Network Design Workshops held in Nelspruit, Cape Town, Durban,King Williams Town, Bela-Bela and Bloemfontein from March to June 2016. During these workshops theoretical monitoring network considerations for each WMA were presented to various stakeholders, and the considerations were used to review the existing networks and obtain changes and improvements to the networks as recommendations. The main objectives of the workshops were to review the existing monitoring networks against the prioritized National Monitoring Objectives …

Network Inventory Map Book 9: Berg-Olifants, South Africa

Policy

The Department of Water and Sanitation (DWS) is the custodian of ten national monitoring programs. The overall aim of this project is to undertake an evaluation of each monitoring network, in its present condition, and to redesign and realign the network based on scientific analysis and the strategic and management objectives of DWS and of the country as a whole. The water resources monitoring network will be optimised to ensure sustainable, relevant and up-to-date data of an acceptable quality. This Network Inventory Task focused on the production of maps to illustrate the spatial distribution of the existing monitoring stations for …

Network Inventory Map Book 4: Pongola-Mtamvuna, South Africa

Policy

The Department of Water and Sanitation (DWS) is the custodian of ten national monitoring programs. The overall aim of this project is to undertake an evaluation of each monitoring network, in its present condition, and to redesign and realign the network based on scientific analysis and the strategic and management objectives of DWS and of the country as a whole. The water resources monitoring network will be optimised to ensure sustainable, relevant and up-to-date data of an acceptable quality. This Network Inventory Task focused on the production of maps to illustrate the spatial distribution of the existing monitoring stations for …

Network Inventory Map Book 8: Breede-Gouritz, South Africa

Policy

The Department of Water and Sanitation (DWS) is the custodian of ten national monitoring programs. The overall aim of this project is to undertake an evaluation of each monitoring network, in its present condition, and to redesign and realign the network based on scientific analysis and the strategic and management objectives of DWS and of the country as a whole. The water resources monitoring network will be optimised to ensure sustainable, relevant and up-to-date data of an acceptable quality. This Network Inventory Task focused on the production of maps to illustrate the spatial distribution of the existing monitoring stations for …