Biomedical Engineering and Bioengineering Commons^™

In Vitro Simulation Of Microgravity Induced Muscle Loss Successfully Increases Expression Of Key In Vivo Atrophy Markers, Charles P. Harding

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Muscle loss from lack of activity is a serious issue for immobilized patients on Earth and in human spaceflight, where the low gravity environment prevents normal muscle activity. Simulating muscle loss in cultured cells is an important step in understanding how this condition occurs. This work evaluates different means of simulating muscle loss and selects the one that most closely mimics the cellular responses seen in animals and humans.

To simulate the microgravity environment of spaceflight, mouse skeletal muscle cells were grown in a rotary cell culture system (RCCS). Growing the cells within a natural gelled substrate was compared against …

Effect Of Physical Stimuli On Angiogenic Factor Expression In Retinal Pigment Epithelial Cells, Farhad Farjood

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Age-related macular degeneration (AMD) is a major cause of blindness in adults. Abnormal growth of blood vessels in the eye during the course of AMD causes damage to the retina, resulting in irreversible blindness. The goal of this research was to determine whether physical pressure on retinal cells can contribute to the increased blood vessel formation. To replicate the tears in the cell layers, a micropatterning method was used as a means of detaching cells from each other. Two new devices were also developed to mimic slow and fast increases in mechanical pressure on cell layers of the eye. After …

Use Of Surface-Enhanced Raman Scattering (Sers) Probes To Detect Fatty Acid Receptor Activity In A Microfluidic Device, Han Zhang, Wei Zhang, Lifu Xiao, Yan Liu, Timothy A. Gilbertson, Anhong Zhou

Biological Engineering Faculty Publications

In this study, 4-mercaptobenzoic acid (MBA)-Au nanorods conjugated with a GPR120 antibody were developed as a highly sensitive surface-enhanced Raman spectroscopy (SERS) probe, and were applied to detect the interaction of fatty acids (FA) and their cognate receptor, GPR120, on the surface of human embryonic kidney cells (HEK293-GPRR120) cultured in a polydimethylsiloxane (PDMS) microfluidic device. Importantly, the two dominant characteristic SERS peaks of the Raman reporter molecule MBA, 1078 cm⁻¹ and 1581 cm⁻¹, do not overlap with the main Raman peaks from the PDMS substrate when the appropriate spectral scanning range is selected, which effectively avoided the …

Improved Production Of Antifungal Angucycline Sch47554 By Manipulating Three Regulatory Genes In Streptomyces Sp. Scc-2136, Ozkan Fidan, Riming Yan, Du Zhu, Jixun Zhan

Biological Engineering Faculty Publications

Sch47554 and Sch47555 are two angucyclines with antifungal activities against various yeasts and dermatophytes from Streptomyces sp. SCC‐2136. The schgene cluster contains several putative regulatory genes. Both schA4 and schA21were predicted as the TetR family transcriptional regulators, whereas schA16shared significant similarity to the AraC family transcriptional regulators. Although Sch47554 is the major product of Streptomyces sp. SCC‐2136, its titer is only 6.72 mg/L. This work aimed to increase the production of this promising antifungal compound by investigating and manipulating the regulatory genes in the Sch47554 biosynthetic pathway. Disruption of schA4and schA16 led to a significant increase …

Develop A 3d Neurological Disease Model Of Human Cortical Glutamatergic Neurons Using Micropillar-Based Scaffolds, Cheng Chen, Xin Dong, Kai-Heng Fang, Fang Yuan, Yao Hu, Min Xu, Yu Huang, Xixiang Zhang, Danjun Fang, Yan Liu

Biological Engineering Faculty Publications

Establishing an effective three-dimensional (3D) in vitro culture system to better model human neurological diseases is desirable, since the human brain is a 3D structure. Here, we demonstrated the development of a polydimethylsiloxane (PDMS) pillar-based 3D scaffold that mimicked the 3D microenvironment of the brain. We utilized this scaffold for the growth of human cortical glutamatergic neurons that were differentiated from human pluripotent stem cells. In comparison with the 2D culture, we demonstrated that the developed 3D culture promoted the maturation of human cortical glutamatergic neurons by showing significantly more MAP2 and less Ki67 expression. Based on this 3D culture …

Identification Of New Glutamate Decarboxylases From Streptomyces For Efficient Production Of Γ-Aminobutyric Acid In Engineered Escherichia Coli, Haina Yuan, Hongbo Wang, Ozkan Fidan, Yong Qin, Gongnian Xiao, Jixun Zhan

Biological Engineering Faculty Publications

Background

Gamma (γ)-Aminobutyric acid (GABA) as a bioactive compound is used extensively in functional foods, pharmaceuticals and agro-industry. It can be biosynthesized via decarboxylation of monosodium glutamate (MSG) or L-glutamic acid (L-Glu) by glutamate decarboxylase (GAD; EC4.1.1.15). GADs have been identified from a variety of microbial sources, such as Escherichia coli and lactic acid bacteria. However, no GADs from Streptomyces have been characterized. The present study is aimed to identify new GADs from Streptomyces strains and establish an efficient bioproduction platform for GABA in E. coli using these enzymes.

Results

By sequencing and analyzing the genomes of three Streptomycesstrains, …

Improving Wastewater Treatment Using Algal Biofilms And Bioenergy, Nathan Guymon

Research on Capitol Hill

This project represents a collaboration between Utah State University, Central Valley Water Reclamation Facility (CVWRF), and WesTech Engineering, Inc. The goal of the project is to help water treatment plants meet state nutrient standards and reduce operating costs. This is being accomplished in three major sections:

1. Wastewater treatment – microalgae is grown to remove nitrogen and phosphorus from water.

2. Biogas generation – the microalgae is harvested and processed to produce methane gas used to generate power.

3. Fertilizer production – nutrients are also collected from the wastewater and turned into high-value fertilizer.

By creating systems that remove nutrients …

Muscle Atrophy Marker Expression Differs Between Rotary Cell Culture System And Animal Studies, Charles P. Harding, Elizabeth Vargis

Biological Engineering Faculty Publications

Muscular atrophy, defined as the loss of muscle tissue, is a serious issue for immobilized patients on Earth and for humans during spaceflight, where microgravity prevents normal muscle loading. In vitro modeling is an important step in understanding atrophy mechanisms and testing countermeasures before animal trials. The most ideal environment for modeling must be empirically determined to best mimic known responses in vivo. To simulate microgravity conditions, murine C2C12 myoblasts were cultured in a rotary cell culture system (RCCS). Alginate encapsulation was compared against polystyrene microcarrier beads as a substrate for culturing these adherent muscle cells. Changes after culture …

Identification And Engineering Of Nonribosomal Peptide Biosynthetic Systems, Fuchao Xu

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This research focuses on the understanding and engineering of nonribosomal peptide biosynthetic pathways in Streptomyces coelicolor CH999, Escherichia coli BAP1 and Saccharomyces cerevisiae BJ5464-NpgA. The biosynthetic systems of indigoidine from bacteria and beauvericin/bassianolide from fungi were studied in this research. The production of these valuble products was significantly increased by enhancing their synthetic pathway with metabolic engineering approaches.

Indigoidine is a bacterial natural product with antioxidant and antimicrobial activities. Its bright blue color resembles the industrial dye indigo, thus representing a new natural blue dye that may find uses in industry. Indigo is a dark blue crystalline powder and has …

Production And Biocompatibility Of Spider Silk Proteins In Goat Milk, Richard E. Decker Jr

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Due to its strength, flexibility, and biocompatibility, spider silk is a highly appealing material for applications in the medical field. Unfortunately, natural spider silk is difficult to obtain in large quantities because spiders are territorial and cannibalistic, making them impractical to farm. Synthetic spider silk proteins produced by transgenic hosts such as bacteria and goats have made it possible to obtain the quantities of spider silk needed to study it more fully and to investigate its potential uses. The spider silk proteins produced in our laboratory do not have an optimal purification method to remove all of the non-biocompatible contaminants …

Identification And Heterologous Reconstitution Of A 5-Alk(En)Ylresorcinol Synthase From Endophytic Fungus Shiraia Sp. Slf14, Huiwen Yan, Lei Sun, Jinge Huang, Yixing Qiu, Fuchao Xu, Riming Yan, Du Zhu, Wei Wang, Jixun Zhan

Biological Engineering Faculty Publications

A new type III polyketide synthase gene (Ssars) was discovered from the genome of Shiraia sp. Slf14, an endophytic fungal strain from Huperzia serrata. The intron-free gene was cloned from the cDNA and ligated to two expression vectors pET28a and YEpADH2p-URA3 for expression in Escherichia coli BL21(DE3) and Saccharomyces cerevisiae BJ5464, respectively. SsARS was efficiently expressed in E. coli BL21(DE3), leading to the synthesis of a series of polyketide products. Six major products were isolated from the engineered E. coli and characterized as 1,3-dihydroxyphenyl-5-undecane, 1,3-dihydroxyphenyl-5-cis-6'-tridecene,1,3-dihydroxyphenyl-5-tridecane, 1,3-dihydroxyphenyl-5-cis-8'-pentadecene, 1,3-dihydroxyphenyl-5-pentadecane and 1,3-dihydroxyphenyl-5-cis-10'-heptadecene, respectively, …

Novel Devices For Studying Acute And Chronic Mechanical Stress In Retinal Pigment Epithelial Cells, Farhad Farjood, Elizabeth Vargis

Biological Engineering Faculty Publications

Choroidal neovascularization (CNV) is a major cause of blindness in patients with age-related macular degeneration (AMD). Overexpression of vascular endothelial growth factor (VEGF), a potent angiogenic protein, by retinal pigment epithelial (RPE) cells is a key stimulator of CNV. Mechanical stress occurs during different stages of AMD and is a possible inducer of VEGF expression in RPE cells. However, robust and realistic approaches to studying acute and chronic mechanical stress under various AMD stages do not exist.The majority of previous work has studied cyclic stretching of RPE cells grown on flexible substrates, but an ideal model must be able to …

Hydrodeoxygenation Of Aqueous Phase Catalytic Pyrolysis Oil To Liquid Hydrocarbons Using Multi-Functional Nickel Catalyst, Hossein Jahromi, Foster A. Agblevor

Biological Engineering Faculty Publications

Herein we investigated the hydrodeoxygenation (HDO) of aqueous phase pinyon-juniper catalytic pyrolysis oil (APPJCPO) using a new multifunctional red mud-supported nickel (Ni/RM) catalyst. The organic liquid yield after HDO of APPJCPO using 30 wt. % Ni/RM at reaction temperature of 350 °C was 47.8 wt. % with oxygen content of 1.14 wt. %. The organic liquid fraction consisted of aliphatics, aromatics, and alkylated aromatic hydrocarbons as well as small amounts of oxygenates. The RM support catalyzed ketonization of carboxylic acids. The Ni metal catalyzed partial reduction of oxygenates that underwent carbonyl alkylation with aldehydes and ketones on the RM. Catalyst …

An Efficient Process For Co-Production Of Γ-Aminobutyric Acid And Probiotic Bacillus Subtilis Cells, Hongbo Wang, Jinge Huang, Lei Sun, Fuchao Xu, Wei Zhang, Jixun Zhan

Biological Engineering Faculty Publications

This study was to establish an integrated process for the co-production of γ-aminobutyric acid (GABA) and live probiotics. Six probiotic bacteria were screened and Bacillus subtilis ATCC 6051 showed the highest GABA-producing capacity. The optimal temperature and initial pH value for GABA production in B. subtilis were found to be 30 °C and 8.0, respectively. A variety of carbon and nitrogen sources were tested, and potato starch and peptone were the preferred carbon and nitrogen sources for GABA production, respectively. The concentrations of carbon source, nitrogen source and substrate (sodium L-glutamate) were then optimized using the response surface methodology. The …

Method For The Destruction Of Endotoxin In Synthetic Spider Silk Proteins, Richard E. Decker, Thomas I. Harris, Dylan R. Memmott, Christopher J. Peterson, Randolph V. Lewis, Justin A. Jones

Biological Engineering Faculty Publications

Although synthetic spider silk has impressive potential as a biomaterial, endotoxin contamination of the spider silk proteins is a concern, regardless of the production method. The purpose of this research was to establish a standardized method to either remove or destroy the endotoxins present in synthetic spider silk proteins, such that the endotoxin level was consistently equal to or less than 0.25 EU/mL, the FDA limit for similar implant materials. Although dry heat is generally the preferred method for endotoxin destruction, heating the silk proteins to the necessary temperatures led to compromised mechanical properties in the resultant materials. In light …

Development And Optimization Of A Produced Water, Biofilm Based Microalgae Cultivation System For Biocrude Conversion With Hydrothermal Liquefaction, Benjamin L. Peterson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Extraction of oil and gas in Utah’s Uintah Basin results in large quantities of wastewater, or produced water, with nutrients and residual organic chemical that represent a significant resource for producing energy-related and value-added products. Produced water was obtained as a biomass producing nutrient source from industries operating in Utah’s Uintah Basin. Within the Uintah Basin (defined as Uintah and Duchesne Counties within Utah) approximately 93 million barrels of water were produced in 2013 while only 11% of the water was disposed of through evaporation, with the national average at 2%. The rest is reinjected into the subsurface.

The goal …

Piezoresponse, Mechanical, And Electrical Characteristics Of Synthetic Spider Silk Nanofibers, Nader Shehata, Ishac Kandas, Ibrahim Hassounah, Patrik Sobolčiak, Igor Krupa, Miroslav Mrlik, Anton Popelka, Jesse Steadman, Randolph V. Lewis

Biology Faculty Publications

This work presents electrospun nanofibers from synthetic spider silk protein, and their application as both a mechanical vibration and humidity sensor. Spider silk solution was synthesized from minor ampullate silk protein (MaSp) and then electrospun into nanofibers with a mean diameter of less than 100 nm. Then, mechanical vibrations were detected through piezoelectric characteristics analysis using a piezo force microscope and a dynamic mechanical analyzer with a voltage probe. The piezoelectric coefficient (d₃₃) was determined to be 3.62 pC/N. During humidity sensing, both mechanical and electric resistance properties of spider silk nanofibers were evaluated at varying high-level …

Biomass And Phycocyanin From Oil And Natural Gas Extraction Produced Water Utilizing A Cyanobacteria Dominated Rotating Algal Biofilm Reactor (Rabr), Jonathan L. Wood

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The production of cyanobacterial biofilm biomass and phycocyanin from Rotating Algal Biofilm Reactors utilizing undiluted produced water from oil and natural gas extraction as a culture medium was investigated in this study. Produced water is the largest waste stream generated by the oil and natural gas industries and represents a large volume of non-potable water that may be available for algae culture with minimal impact on freshwater resources. Combining the use of produced wastewater as culture medium with the production of high value algal pigments, such as phycocyanin, may increase the economic viability of algae culture and wastewater purification. High …

Novel Methods To Produce Large Recombinant Spider Silk Proteins Via Polymerization, Nathan L. Hebert

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Spider silk has long been a subject of scientific research due to its remarkable mechanical properties. Until recently, there has been no way to effectively obtain spider silk except by harvesting it from individual spiders. With advances in technology, the genes that code for the individual spider silk proteins have been isolated and genetically engineered into other hosts to produce recombinant spider silk proteins (rSSp) of varying sizes, Larger rSSp have correspondingly greater mechanical properties in any resulting materials. Using current production methods, larger rSSp cannot be produced in commercially viable quantities while simultaneously being economically viable. The current production …

Development And Characterization Of Aqueous-Based Recombinant Spider Silk Protein Biomaterials With Investigations Into Potential Applications, Thomas I. Harris

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Spider silks are incredible natural materials that possess desirable combinations of strength, elasticity, weight, and robustness. Other properties such as biocompatibility and biodegradability further increase the worth of these materials. The possibility of farming spiders is impractical due to spiders’ natural behaviors. Modern biotechnologies have allowed for recombinant spider silk proteins (rSSps) to be produced without the use of spiders. However, the features responsible for spider silks impressive properties can cause difficulties with producing silk materials. A recently developed water-based and biomimetic solvation method has provided a solution to such difficulties and has also led to novel silk biomaterials. Most …

Whole Cell Cross-Linking To Discover Host-Microbe Protein Cognate Receptor/Ligand Pairs, Bart C. Weimer, Poyin Chen, Prerak T. Desai, Dong Chen, Jigna Shah

Biological Engineering Faculty Publications

Bacterial surface ligands mediate interactions with the host cell during association that determines the specific outcome for the host–microbe association. The association begins with receptors on the host cell binding ligands on the microbial cell to form a partnership that initiates responses in both cells. Methods to determine the specific cognate partnerships are lacking. Determining these molecular interactions between the host and microbial surfaces are difficult, yet crucial in defining biologically important events that are triggered during association of the microbiome, and critical in defining the initiating signal from the host membrane that results in pathology or commensal association. In …

Hydrotreating Of Guaiacol: A Comparative Study Of Red Mud-Supported Nickel And Commercial Ni/Sio2-Al2o3 Catalysts, Hossein Jahromi, Foster Agblevor

Biological Engineering Faculty Publications

Upgrading of bio-oil through catalytic hydrotreating was investigated with guaiacol as a model compound. A nickel supported on red mud (Ni/RM) hydrotreating catalyst was developed and compared to the standard Ni/SiO₂-Al₂O₃ catalysts under similar experimental conditions. The Ni/RM catalyst was characterized by inductively coupled plasma atomic emission spectrometry (ICP-AES), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM), BET specific surface area, and temperature programmed reduction (TPR). The effects of reaction temperature (300, 350, 400 °C) and initial hydrogen pressure (4.83 MPa (700 psi), 5.52 MPa (800 psi), and 6.21 MPa (900 psi)) on products distribution …

New Insights Into The Glycosylation Steps In The Biosynthesis Of Sch47554 And Sch47555, Ozkan Fidan, Riming Yan, Gabrielle Gladstone, Tong Zhou, Du Zhu, Jixun Zhan

Biological Engineering Faculty Publications

Sch47554 and Sch47555 are antifungal compounds from Streptomyces sp. SCC‐2136. The availability of the biosynthetic gene cluster made it possible to track genes that encode biosynthetic enzymes responsible for the structural features of these two angucyclines. Sugar moieties play important roles in the biological activities of many natural products. An investigation into glycosyltransferases (GTs) might potentially help to diversify pharmaceutically significant drugs through combinatorial biosynthesis. Sequence analysis indicates that SchS7 is a putative C‐GT, whereas SchS9 and SchS10 are proposed to be O‐GTs. In this study, the roles of these three GTs in the biosynthesis of Sch47554 and Sch47555 are …

Designing A Synthetic Spider Silk-Based Coating For Urinary Catheters To Reduce The Risk Of Cautis, Alexander Cook

Undergraduate Honors Capstone Projects

Urinary and Intravenous (IV) catheters are two of the most commonly used medical devices for administering vital drugs and obtaining diagnostic samples from patients. Unfortunately, hospital acquired infections (HAIs) occur at alarming rates due to catheter usage. The aim of this project was to design a urinary catheter that would decrease the occurrence of these infections with a catheter coating that utilizes the antimicrobial properties of synthetic spider silk in combination with the antiseptic chlorhexidine. This synthetic spider silk catheter will introduce a novel approach to catheterization in the medical industry.

Designing A Synthetic Spider Silk Based Coating For Urinary Catheters To Reduce The Risk Of Cautis, Tess Marie Armbrust

Undergraduate Honors Capstone Projects

Catheter associated urinary tract infections (CAUTis) are a leading cause of morbidity and mortality in patients with life-threatening conditions. Foley catheters are the most common catheter used when a patient requires hospitalization but can become burdened with microbes which lead to CAUTis, the most common source of hospital acquired infection (HAI). The attached bacteria can form biofilms on the catheter which are difficult to treat using oral antibiotics and can lead to chronic infections. Synthetic spider silk as a catheter coating shows great promise because of demonstrated strength, biocompatibility, and antimicrobial properties. In addition, the antiseptic chlorhexidine can enhance the …

Investigations Of Polyhydroxyalkanoate Secretion And Production Using Sustainable Carbon Sources, Chad L. Nielsen

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Polyhydroxyalkanoates (PHAs) are a type of biologically-produced plastic known for their biocompatibility and biodegradability. They have the potential to replace petroleum-based plastics as an environmentally-friendly alternative. This is beneficial because the release of plastics into environments such as the ocean and the buildup of plastics in landfills are major concerns facing society today. Currently, however, PHAs are significantly more expensive than their petroleum-based counterparts. This is largely due to the cost of carbon sources and of extracting the bioplastics from bacteria. The goal of these studies was to examine replacing traditional carbon sources used in PHA production like sugar and …

Silane Modulation Of Protein Conformation And Self-Assembly, Abul Bashar Mohammad Giasuddin

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This research focused on development of nanoparticle- based therapeutics against amyloid fibrils. Amyloid fibrils are associated with various diseases such as Parkinson’s, Huntington’s, mad cow disease, Alzheimer’s, and cataracts. Amyloid fibrils develop when proteins change their shape from a native form to a pathogenic “misfolded” form. The misfolded proteins have the ability to recruit more native proteins into the pathogenic forms, which self-assemble into amyloid fibrils that are hallmarks of the various protein-misfolding diseases listed above. Amyloid fibrils are highly resistant to degradation, which may contribute to the symptoms of amyloid diseases. Synthetic drugs, natural compounds, and antibodies are widely …

The Use Of Microfluidics And Dielectrophoresis For Separation, Concentration, And Identification Of Bacteria, Cynthia Hanson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Typical bacterial analysis involves culturing and visualizing colonies on an array of agar plates. The growth patterns and colors among the array are used to identify the bacteria. For fast growing bacteria such as Escherichia coli, analysis will take one to two days. However, slow growing bacteria such as mycobacteria can take weeks to identify. In addition, there are some species of bacteria that are viable but nonculturable. This lengthy analysis time is unacceptable for life-threatening infections and emergency situations. It is clear that to decrease the analysis of the bacteria, the culturing and growth steps must be avoided. …

Microfluidic Chip For Non-Invasive Analysis Of Tumor Cells Interaction With Anti-Cancer Drug Doxorubicin By Afm And Raman Spectroscopy, Han Zhang, Lifu Xiao, Qifei Li, Xiaojun Qi, Anhong Zhou

Biological Engineering Faculty Publications

Raman spectroscopy has been playing an increasingly significant role for cell classification. Here, we introduce a novel microfluidic chip for non-invasive Raman cell natural fingerprint collection. Traditional Raman spectroscopy measurement of the cells grown in a Polydimethylsiloxane (PDMS) based microfluidic device suffers from the background noise from the substrate materials of PDMS when intended to apply as an in vitro cell assay. To overcome this disadvantage, the current device is designed with a middle layer of PDMS layer sandwiched by two MgF2slides which minimize the PDMS background signal in Raman measurement. Three cancer cell lines, including a human lung cancer …

3d Tissue Engineering, An Emerging Technique For Pharmaceutical Research, Gregory Jensen, Christian Morrill, Yu Huang

Biological Engineering Faculty Publications

Tissue engineering and the tissue engineering model have shown promise in improving methods of drug delivery, drug action, and drug discovery in pharmaceutical research for the attenuation of the central nervous system inflammatory response. Such inflammation contributes to the lack of regenerative ability of neural cells, as well as the temporary and permanent loss of function associated with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and traumatic brain injury. This review is focused specifically on the recent advances in the tissue engineering model made by altering scaffold biophysical and biochemical properties for use in the treatment of neurodegenerative diseases. …