Engineering Commons^™

Enhancing Single Walled Carbon Nanotube Deposition For The Study Of Extracellular Analytes, Joseph A. Stapleton

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

Extracellular signaling is a dynamic process responsible for coordinating large scale biological processes. As such, understanding extracellular signaling is important to our determination of normal function and pathophysiological development. High resolution spatial and temporal information are critical to completely understanding these processes. Unfortunately, current methods of detection are lacking in either spatial or temporal resolution of extracellular products, limiting researchers’ ability to understand complex biological processes. A new group of sensors based on fluorescent single walled carbon nanotubes (SWNT) have shown the potential to provide both high quality spatial and temporal resolution for the sensing of analytes. However, while SWNT …

Design, Development, And Field Testing A Visnir Integrated Multi-Sensing Soil Penetrometer, Nuwan K. Wijewardane

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

The research community in soil science and agriculture lacks a cost-effective and rapid technology for in situ, high resolution vertical soil sensing. Visible and near infra-red (VisNIR) technology has the potential to be used for such sensor development due to its ability to derive multiple soil properties rapidly using a single spectrum. Such efforts must, however, overcome a few challenges: (i) a dry ground soil spectral library that can be used to predict the target soil properties accurately, (ii) a robust design which can acquire high quality VisNIR spectra of soil, (iii) an effective method that can link field intact …

Bioproduction Of Adipic Acid Using Engineered Pseudomonas Putida Kt2440 From Lignin-Derived Aromatics, Howard Willett

Department of Chemical and Biomolecular Engineering: Theses and Student Research

Current industrial synthesis of adipic acid is nonrenewable and depends on a carcinogenic starting material, benzene. Biocatalysis with an engineered microorganism could turn a renewable feedstock into a value-added chemical such as adipic acid. Here we engineered P. putida KT2440 to transform lignin-derived aromatics, coumarate and ferulate, into adipic acid. Lignin is a recalcitrant plant biopolymer burned for thermal energy. Conversion of lignin into a value-added chemical will improve the efficiency of lignocellulose processing plants. The best performing engineered KT2440 strain produces 2.52 mM adipate at a 9.5% (mole/mole) yield. This was achieved by the genetic insertion of non-natural biosynthetic …

Fibrinogen, Factor Xiii And Fibronectin: A Biophysical And Kinetic Characterization Of Their Interactions, Frank Fabian

Department of Chemical and Biomolecular Engineering: Theses and Student Research

The development of recombinant-based liquid fibrin tissue sealants having enhanced hemostatic and wound healing properties will involve understanding as yet not well characterized interactions between fibrinogen, fibrin (Fbn) factor XIII, thrombin and fibronectin. We study these phenomena in the context of comparing plasma derived fibrinogen to recombinant fibrinogen (rFI) produced in the milk of transgenic cows. An abundance of purified γγ and γγ’ FI subspecies enables detailed study of γγ or γγ’ biomonomer and their respective Fbn biopolymer formation as having different substrate behaviors of activated plasma derived factor XIII (pFXIIIa₂b₂). High pressure size exclusion (HPSEC) …

Bio-Desalination Of Brackish And Seawater Using Halophytic Algae, Endalkachew Sahle-Demessie, Ashraf Aly Hassan, Amro El Badawy

Department of Civil and Environmental Engineering: Faculty Publications

Global demand for water is rising. A sustainable and energy efficient approach is needed to desalinate brackish sources for agricultural and municipal water use. Genetic variation among two algae species, Scenedesmus species (S. sp.) and Chlorella vulgaris (C. vulgaris), in their tolerance and uptake of salt (NaCl) was examined for potential bio-desalination of brackish water. Salt-tolerant hyper-accumulators were evaluated in a batch photobioreactors over salinity concentration ranging from 2 g/L to 20 g/L and different nutrient composition for their growth rate and salt-uptake. During algae growth phase, the doubling time varied between 0.63 and 1.81 days for S …

Predictive Modeling Of Fate And Transport Of Three Prevalent Contaminants In Midwest Agroecosystem Surface Waters: Nitrate-N, Atrazine, And Escherichia Coli, Samuel Hansen

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

The majority of streams and rivers in the United States (U.S.) are ecologically impaired, or threatened by anthropogenic stressors. Recent reports have found atrazine in drinking water to be associated with increased birth defects and incidences of Non-Hodgkin’s Lymphoma, with higher levels of significance from exposure to both atrazine and nitrate-N. In contrast, recent illnesses from E. coli contaminating vegetables that originated from irrigation water has increased awareness of identifying sources of E. coli entering irrigation reservoirs.

Methods to accurately predict atrazine and E. coli occurrence and potential sources in waterways continue to limit the identifying appropriate and effective prevention …

Advanced Selection Methodologies For Dnazymes In Sensing And Healthcare Applications, Sandeep Kumar, Shikha Jain, Neeraj Dilbaghi, Amrik Singh Ahluwalia, Ashraf Aly Hassan, Ki-Hyun Kim

Department of Civil and Environmental Engineering: Faculty Publications

DNAzymes have been widely explored owing to their excellent catalytic activity in a broad range of applications, notably in sensing and biomedical devices. These newly discovered applications have built high hopes for designing novel catalytic DNAzymes. However, the selection of efficient DNAzymes is a challenging process but one that is of crucial importance. Initially, systemic evolution of ligands by exponential enrichment (SELEX) was a labor-intensive and time-consuming process, but recent advances have accelerated the automated generation of DNAzyme molecules. This review summarizes recent advances in SELEX that improve the affinity and specificity of DNAzymes. The thriving generation of new DNAzymes …

Metabolomics Of Sorghum Roots During Nitrogen Stress Reveals Compromised Metabolic Capacity For Salicylic Acid Biosynthesis, Amy M. Sheflin, Dawn Chiniquy, Chaohui Yuan, Emily Goren, Indrajit Kumar, Max Braud, Thomas Brutnell, Andrea L. Eveland, Susannah Tringe, Peng Liu, Stephen Kresovich, Ellen Marsh, Daniel P. Schachtman, Jessica E. Prenni

Nebraska Center for Biotechnology: Faculty and Staff Publications

Sorghum (Sorghum bicolor [L.] Moench) is the fifth most productive cereal crop worldwide with some hybrids having high biomass yield traits making it promising for sustainable, economical biofuel production. To maximize biofuel feedstock yields, a more complete understanding of metabolic responses to low nitrogen (N) will be useful for incorporation in crop improvement efforts. In this study, 10 diverse sorghum entries (including inbreds and hybrids) were field-grown under low and full N conditions and roots were sampled at two time points for metabolomics and 16S amplicon sequencing. Roots of plants grown under low N showed altered metabolic profiles at …

The Small Rna Teg41 Regulates Expression Of The Alpha Phenol-Soluble Modulins And Is Required For Virulence In Staphylococcus Aureus, Rachel L. Zapf, Richard E. Wiemels, Rebecca A. Keogh, Donald L. Holzschu, Kayla M. Howell, Emily Trzeciak, Andrew R. Caillet, Kellie A. King, Samantha A. Selhorst, Michael J. Naldrett, Jeffrey L. Bose, Ronan K. Carroll

Nebraska Center for Biotechnology: Faculty and Staff Publications

Small RNAs (sRNAs) remain an understudied class of regulatory molecules in bacteria in general and in Gram-positive bacteria in particular. In the major human pathogen Staphylococcus aureus, hundreds of sRNAs have been identified; however, only a few have been characterized in detail. In this study, we investigate the role of the sRNA Teg41 in S. aureus virulence. We demonstrate that Teg41, an sRNA divergently transcribed from the locus that encodes the cytolytic alpha phenolsoluble modulin (αPSM) peptides, plays a critical role in αPSM production. Overproduction of Teg41 leads to an increase in αPSM levels and a corresponding increase in …

Terpene Synthase Genes Originated From Bacteria Through Horizontal Gene Transfer Contribute To Terpenoid Diversity In Fungi, Qidong Jia, Xinlu Chen, Tobias G. Kollner, Jan Rinkel, Jianyu Fu, Jessy Labbe, Wangdan Xiong, Jeroen S. Dickschat, Jonathan Gershenzon, Feng Chen

Nebraska Center for Biotechnology: Faculty and Staff Publications

Fungi are successful eukaryotes of wide distribution. They are known as rich producers of secondary metabolites, especially terpenoids, which are important for fungi-environment interactions. Horizontal gene transfer (HGT) is an important mechanism contributing to genetic innovation of fungi. However, it remains unclear whether HGT has played a role in creating the enormous chemical diversity of fungal terpenoids. Here we report that fungi have acquired terpene synthase genes (TPSs), which encode pivotal enzymes for terpenoid biosynthesis, from bacteria through HGT. Phylogenetic analysis placed the majority of fungal and bacterial TPS genes from diverse taxa into two clades, indicating ancient …

The Impact Of Extreme Virtual Elevation Above Grade On Construction Workers' Physiological Responses, Physical Responses, And Task Performance, Mahmoud Habibnezhad

Department of Construction Engineering and Management: Dissertations, Theses, and Student Research

On average, in every two work hours, one person dies from work-related injuries at construction sites. Most incidents are due to falling from elevated surfaces. Slips, trips, and loss of balance are the main causes. Studies suggest that instigating visual mismatch and physiological changes are among the most important reasons behind falling from narrow elevated surfaces. By using advanced virtual reality models, this dissertation aims to highlight some of the possible effects of a destabilizing environment (i.e., elevation above grade) on workers’ physiological responses and task performance. More specifically, this dissertation strives to find potential effects of elevation above grade …

Manipulation Of The Precursor Supply For High-Level Production Of Longifolene By Metabolically Engineered Escherichia Coli, Yujin Cao, Rubing Zhang, Wei Liu, Guang Zhao, Wei Niu, Jiantao Guo, Mo Xian, Huizhou Liu

Department of Chemical and Biomolecular Engineering: Faculty Publications

Longifolene is a naturally occurring tricyclic sesquiterpene widely used in many different fields. Up to now, this valuable terpene was mainly manufactured from the high-boiling fraction of certain pine resins. Microbial production can be a promising alternative to the extraction from natural plant sources. Here, we present the metabolic engineering strategy to assemble biosynthetic pathway for longifolene production in Escherichia coli. E. coli was rendered to produce longifolene by heterologously expressing a codon optimized longifolene synthase from Picea abies. Augmentation of the metabolic flux to farnesyl pyrophosphate (FPP) by different FPP synthases conferred a 1.8-fold increase in longifolene …

Comparative Kinetic Modeling Of Growth And Molecular Hydrogen Overproduction By Engineered Strains Of Thermotoga Maritima, Raghuveer Singh, Rahul Tevatia, Derrick White, Yaşar Demirel, Paul H. Blum

Department of Chemical and Biomolecular Engineering: Faculty Publications

Thermotoga maritima is an anaerobic hyperthermophilic bacterium known for its high amounts of hydrogen (H₂) production. In the current study, the kinetic modeling was applied on the engineered strains of T. maritima that surpassed the natural H₂ production limit. The study generated a kinetic model explaining H₂ overproduction and predicted a continuous fermentation system. A Leudking-Piret equation-based model predicted that H₂ production by Tma200 (0.217 mol-H₂ g^–1-biomass) and Tma100 (0.147 mol-H₂ g^–1-biomass) were higher than wild type (0.096 mol-H₂ g^–1 -biomass) with reduced rates of maltose utilization. …

Delineating The Role Of Fanca In Glucose-Stimulated Insulin Secretion In Β Cells Through Its Protein Interactome, Dragana Lagundzin, Wen-Feng Hu, Henry C. H. Law, Kimiko L. Krieger, Fangfang Qiao, Emalie J. Clement, Andjela T. Drincic, Olgica Nedic, Michael J. Naldrett, Sophie Alvarez, Nicholas T. Woods

Nebraska Center for Biotechnology: Faculty and Staff Publications

Hyperinsulinemia affects 72% of Fanconi anemia (FA) patients and an additional 25% experience lowered glucose tolerance or frank diabetes. The underlying molecular mechanisms contributing to the dysfunction of FA pancreas β cells is unknown. Therefore, we sought to evaluate the functional role of FANCA, the most commonly mutated gene in FA, in glucosestimulated insulin secretion (GSIS). This study reveals that FANCA or FANCB knockdown impairs GSIS in human pancreas β cell line EndoC-βH3. To identify potential pathways by which FANCA might regulate GSIS, we employed a proteomics approach to identify FANCA protein interactions in EndoC-βH3 differentially regulated in response to …