Biochemical and Biomolecular Engineering Commons^™

Cloning, Fed-Batch Expression And Purification Of A Novel Anti-Candida Peptide And Development Of A Cleavage Resistant Variant Of Green Fluorescence Protein, Rudra Palash Mukherjee

Graduate Theses and Dissertations

This work illustrates that the intelligent design of a bioprocess for the production of peptides of various length is possible by having a prior knowledge of desired product, cell line, fermentation conditions, and choices for downstream recovery. Cloning, expression, and recovery of a novel 12-mer anti-Candida peptide served as an illustrative case for a proposed Escherichia coli platform. The antifungal peptide, expressed as a fusion to GFPUV during high call density fed-batch, was recovered from cell lysate, concentrated using ion exchange chromatography, and finally cleaved with cyanogen bromide. The platform was capable of producing active peptide capable of arresting the …

Antioxidant Potential Of Juglans Nigra, Black Walnut, Husks Extracted Using Supercritical Carbon Dioxide With An Ethanol Modifier, Johnathon Wenzel, Cheryl Storer Samaniego, Lihua Wang, Laron Burrows, Evan Tucker, Nathan Dwarshuis, Michelle Ammerman, Ali Zand

Chemical Engineering Publications

The black walnut, Junglas nigra, is indigenous to eastern North America, and abscission of its fruit occurs around October. The fruit consists of a husk, a hard shell, and kernel. The husk is commonly discarded in processing, though it contains phenolic compounds that exhibit antioxidant and antimicrobial properties. For this study, black walnut husks were extracted using supercritical carbon dioxide with an ethanol modifier. The effects of temperature, ethanol concentration, and drying of walnut husks prior to extraction upon antioxidant potential were evaluated using a factorial design of experiments. The solvent density was held constant at 0.75 g/mL. The optimal …

Sustained Cell Differentiation Of 2d H9 Human Embryonic Stem Cells Into Mesenchymal Stem Cells, Hannah M. Christian

UCARE: Research Products

This experiment consisted of the controlled differentiation of H9 embryonic stem cells to mesenchymal stem cells. Though this experiment was repeated twice and improvement was seen in these repetitions, the cells were only able to be partially differentiated. However, the morphology of the differentiated cells is similar to those of healthy adult mesenchymal stem cells.

The progression of the differentiation can be seen in the microscope slide photos below. Throughout the differentiation, there occurred a decrease in cell survival and reduction of cell growth, but an increase in mesenchymal stem cell morphology. Throughout the last week of the differentiation, very …

Effect Of Packaging Material On Color Properties Of Catsup Tomato Sauce, Naser Kohan-Nia, Babak Pakbin, Razzagh Mahmoudi, Omid Fakhri

Journal of Applied Packaging Research

Catsup and tomato sauce products currently are one of the most popular and commercially available table sauces particularly consumed by young people and families around the world in recent decades. In this study colorimetric properties of catsup sauce samples including L*, a* and b* values in different packaging materials including PP, PE, PET, Glass and Sachet have been investigated during and after 180 days storage time at 22̊ C (environmental temperature). Hunter Lab system has been used to evaluate the color values of sauce samples. According to obtained results of experimental measurements and sensory evaluations, PP and PE packaging materials …

Immobilization Of Cellulase For Large Scale Reactors To Reduce Cellulosic Ethanol Cost, Dezhi Zhang

Doctoral Dissertations

Cellulosic ethanol is an alternative renewable energy source. Cellulase used in the production of cellulosic ethanol is very expensive. The difficulty in separating cellulase from the cellulose solution after the hydrolysis process limits the reusability of the cellulase, which highly precludes the scales of this application because of the high cost of the enzyme. Immobilization of cellulase provides a promising approach to allow the enzyme to be recycled, thus reducing the production cost. This research focused on immobilizing cellulase for reuse to reduce the cellulosic ethanol cost.

Four immobilization techniques were explored for the immobilization of cellulase on four different …

Brain Tumor In A Dish: Glioma/Astrocyte Co-Cultures As A Model For In Vitro Studies, Erin Eickman, Christina Wilson, Srivatsan Kidambi

UCARE: Research Products

This study seeks to engineer an in vitro co-culture model to elucidate the role of glioma-astrocyte interactions on molecular changes in the tumor microenvironment. The use of patterned co-cultures created with polyelectrolyte multilayers and micromolding in capillaries will allow tthe investigation of cell-cell communication. This study will lead to better understanding of the role of healthy cells in cancer progression and potential treatment options.

Ionic Driven Embedment Of Hyaluronic Acid Coated Liposomes In Polyelectrolyte Multilayer Films For Local Therapeutic Delivery, Stephen Lawrence Hayward, David Francis, Matthew J. Sis, Srivatsan Kidambi

UCARE: Research Products

The ability to control the spatial distribution and temporal release of a therapeutic remains a central challenge for biomedical research. Here, we report the development and optimization of a novel substrate mediated therapeutic delivery system comprising of hyaluronic acid covalently functionalized liposomes (HALNPs) embedded into polyelectrolyte multilayer (PEM) platform via ionic stabilization. The PEM platform was constructed from sequential deposition of Poly-LLysine (PLL) and Poly(Sodium styrene sulfonate) (SPS) “(PLL/SPS)4.5” followed by adsorption of anionic HALNPs. An adsorption affinity assay and saturation curve illustrated the preferential HALNP deposition density for precise therapeutic loading. (PLL/SPS)2.5 capping layer on top of the deposited …

The Role Of Poly(Acrylic Acid) In Conventional Glass Polyalkenoate Cements, Adel M.F. Alhalawani, Declan J. Curran, Daniel Boyd, Mark R. Towler

Chemical and Biochemical Engineering Faculty Research & Creative Works

Glass polyalkenoate cements (GPCs) have been used in dentistry for over 40 years. These novel bioactive materials are the result of a reaction between a finely ground glass (base) and a polymer (acid), usually poly (acrylic acid) (PAA), in the presence of water. This article reviews the types of PAA used as reagents (including how they vary by molar mass, molecular weight, concentration, polydispersity and content) and the way that they control the properties of the conventional GPCs (CGPCs) formulated from them. The article also considers the effect of PAA on the clinical performance of CGPCs, including biocompatibility, rheological and …

Influence Of Strong Preformed Particle Gels On Low Permeable Formations In Mature Reservoirs, Mahmoud O. Elsharafi, Baojun Bai

Geosciences and Geological and Petroleum Engineering Faculty Research & Creative Works

In mature reservoirs, the success of preformed particle gel (PPG) treatment rests primarily on the ability of the PPG to reduce and/or plug the high permeable formations but not damage the low permeable formations. Static test models (filtration test model and pressure test model) were used to determine the effect of PPG on low permeable formations. This work used a strong preformed particle gel, Daqing (DQ) gel made by a Chinese company. The particle gel sizes were ranged from 30 to 120 mesh for this work. PPGs are sized in a millimeter or micrometer, which can absorb over a hundred …

Characterization Of Silica-Based And Borate-Based, Titanium-Containing Bioactive Glasses For Coating Metallic Implants, Omar Rodriguez, Declan J. Curran, Marcello Papini, Lana M. Placek, Anthony W. Wren, Emil H. Schemitsch, Paul Zalzal, Mark R. Towler

Chemical and Biochemical Engineering Faculty Research & Creative Works

Bioactive glasses have found applications in diverse fields, including orthopedics and dentistry, where they have been utilized for the fixation of bone and teeth and as scaffolds for drug delivery. The present work outlines the characterization of two novel titanium-containing glass series, one silica-based and one borate-based. For the silica-based series, titanium is added at the expense of silicon dioxide whereas for the borate-based series, it is added at the expense of boron oxide as confirmed by Energy Dispersive Spectroscopy. Amorphous structures are obtained for silica-based glass at 15 mol% TiO₂ and for borate-based glasses at 0 mol% and …

State Estimation Of Glucose And Insulin Dynamics, Morgan Nicholas Miller

ETD Archive

Process simulation and state estimation have very important applications in chemical engineering as well as the biomedical field. Diabetes is a rapidly growing disease in the United States with 29 million people already diagnosed. The estimation of glucose and insulin concentration in patients is necessary in order to effectively treat diabetes. The Bergman Minimal Model is a popular process model that is used to simulate glucose and insulin dynamics. A simulation of this model was created based on estimated parameters for the model from historical data. This thesis investigated the estimation of glucose concentration, insulin concentration, and effect of active …

Characterization Of Biochars Produced From Peanut Hulls And Pine Wood With Different Pyrolysis Conditions, James W. Lee, Bob Hawkins, Michelle K. Kidder, Barbara R. Evans, A. C. Buchanan, Danny Day

Chemistry & Biochemistry Faculty Publications

Background

Application of modern biomass pyrolysis methods for production of biofuels and biochar is potentially a significant approach to enable global carbon capture and sequestration. To realize this potential, it is essential to develop methods that produce biochar with the characteristics needed for effective soil amendment.

Methods

Biochar materials were produced from peanut hulls and pine wood with different pyrolysis conditions, then characterized by cation exchange (CEC) capacity assays, nitrogen adsorption–desorption isotherm measurements, micro/nanostructural imaging, infrared spectra and elemental analyses.

Results

Under a standard assay condition of pH 8.5, the CEC values of the peanut hull-derived biochar materials, ranging from …

Exploration Of Compound Viability As An Organ-Selective Anti-Metastatic Therapeutic Treatment For Triple Negative Breast Cancer, Ashley E. Noblin

Honors Theses

Phenformin and metformin are compounds that are currently used to treat Type II Diabetes and have been shown to suppress breast cancer metastasis. These compounds were tested, after extensive research and screening of current approved oncology drugs, in various experiments with the cell lines MDA-MB-231, LM, and BoM, with MDA-MB-231 being the parental cell line, LM being the lung metastatic line, and BoM being the bone metastatic line. The cells were treated with compound addition at various concentrations and subject to a two-day concentration response viability study in order to determine any inhibitory effects, followed by a five-day assay in …

An Evaluation Of The Relationship Between Sigma-1 And Sigma-2 Receptors And The Endocannabinoid System, Jenn Miller

Honors Theses

Within the past two decades, sigma receptors have become a popular area for research. Although much has been learned about their structure, subtypes, and functions; there is still much to be learned. Consisting of two receptor subtypes (sigma-1 and sigma-2), it has been discovered that sigma receptor ligands potentiate the analgesic effects of both opiates and cannabinoids, though the exact mechanism and sigma subtype on which this occurs is still unknown. The purpose of this study is to determine if potentiation of opiates and cannabinoids occurs through sigma-1 or sigma-2 receptor signaling. Tetrad assays were performed for: 1. Morphine, an …

The Effect Of Drying On The Release Of Small Hydrophilic Molecular Drugs From Polyelectrolyte Complex Beads, Dounsavanh Letdara

Williams Honors College, Honors Research Projects

The effects of bead composition and dryness on the release rate of small molecules, like a drug or dye, from polyelectrolyte complex (PEC) beads were studied. Release studies were performed on dried and not dried beads prepared using different PEC to drug ratios. The beads were dried for two hours, which allowed them to reach a constant weight. The beads were loaded with a dye, so their release could be detected through the visible light regime. A plate reader was used to determine the absorbance of a sample, and then the amount of dye released was determined. The data presented …

Microbiologically Influenced Corrosion: Rhamnolipid Effects, Andrea M. Wildroudt

Williams Honors College, Honors Research Projects

The purpose of this experiment was to identify if a rhamnolipid could inhibit the growth of a biofilm on carbon steel coupons and result in less corrosion activity. This type of research impacts various industries (with one of the largest being the oil and gas industry). Multiple systems with varying concentrations of the rhamnolipid were created to determine, if any, the optimum concentration to inhibit biofilm growth. The systems were created and exposed to optimum physio-chemical environments to promote SRB induced corrosion. Coupons were immersed for 28 days. Seven systems were created with varying rhamnolipid concentrations; to ensure accurate results, …

Calcium Alginate Microbead Production Via An Air Assisted Shearing Process, Ryan Loftus

Williams Honors College, Honors Research Projects

The purpose of this project was to model the size of calcium alginate microbeads produced by extruding a solution of 1.5 wt.% alginate from a syringe and needle with air shear generated from a pneumatic line flowing in an annular tube around the extrusion needle. The air shear would create microdroplets that would fall into a solution of 2 wt.% calcium chloride in water, crosslinking the droplets into beads. These microbeads can be used in drug dispersion applications or in encapsulation of cells throughout the body due to their biocompatibility. The goal was to create uniform beads that ranged from …

Protein Sequence-Structure-Function Relationship: Testing Ke-50 Modification On Recombinant Green Fluorescent Protein (Acgfp), Anna E. Burkey

Williams Honors College, Honors Research Projects

Protein modification shows promise for improving the effectiveness of protein therapy, including the use of Interferon-beta-1b for the treatment of multiple sclerosis. The addition of a chain of 50 lysine and glutamic acid residues (also called a KE50 modification) was tested on green fluorescent protein (AcGFP) to determine its effect on the bioactivity of the protein. Standard transformation protocol was used to grow the normal and modified proteins in E.Coli, and the bioactivity of the protein was measured using quantum yield and extinction coefficients to determine the reduction in brightness from the native form to the modified form. The …

Construction Of Polyurethane Fabric Nanocomposites For Use In Resistance Temperature Detectors-Effect Of Polyurethane Concentration, Multi-Walled Carbon Nanotubes, And Oxidant, Jordan M. Shaffer

Williams Honors College, Honors Research Projects

Current resistance temperature detectors (RTDs) are rigid and create pressure points when used in contact with the human body. The current RTDs can also not be used in confined areas like the inside of a prosthetic. The research done for this paper explores the possibility of creating a soft fabric like RTD that can accurately measure temperature. This RTD could be used in medical devices, be placed in prosthetic legs, and be worn for a long period of time creating no discomfort for the user. Polyurethane scaffolds were created through an electrospinning process and multiwalled carbon nanotubes were adhered to …

Antibacterial Properties Of Poly (Octanediol Citrate)/Gallium-Containing Bioglass Composite Scaffolds, Ehsan Zeimaran, Sara Pourshahrestani, Ivan Djordjevic, Belinda Pingguan-Murphy, Nahrizul Adib Kadri, Anthony W. Wren, Mark R. Towler

Chemical and Biochemical Engineering Faculty Research & Creative Works

Bioactive glasses may function as antimicrobial delivery systems through the incorporation and subsequent release of therapeutic ions. The aim of this study was to evaluate the antimicrobial properties of a series of composite scaffolds composed of poly (octanediol citrate) with increased loads of a bioactive glass that releases zinc (Zn²⁺) and gallium (Ga³⁺) ions in a controlled manner. The antibacterial activity of these scaffolds was investigated against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria. The ability of the scaffolds to release ions and the subsequent ingress of these ions into hard tissue was evaluated …