Biotechnology Commons^™

Developent Of A Phospholipid Encapsulation Process For Quantum Dots To Be Used In Biologic Applications, Logan Grimes

Master's Theses

The American Cancer Society predicts that 1,665,540 people will be diagnosed with cancer, and 585,720 people will die from cancer in 2014. One of the most common types of cancer in the United States is skin cancer. Melanoma alone is predicted to account for 10,000 of the cancer related deaths in 2014. As a highly mobile and aggressive form of cancer, melanoma is difficult to fight once it has metastasized through the body. Early detection in such varieties of cancer is critical in improving survival rates in afflicted patients. Present methods of detection rely on visual examination of suspicious regions …

Is Biofuel A Feasible Long-Term Chief Energy Source? A Global Perspective, Rajee Olaganathan, Aston Lee, Debra Tong, Mameegate Zheng Jun Cheston, Zack Ho Xuan Yi

Publications

This scientific paper examines the feasibility of officiating biofuels as the future chief energy source of the world. It weighs the pros and cons of the four different generations of biofuels that have been discovered over time, explaining how the raw materials used in each generation determines each individual level of long-term sustainability. It then further analyses both general advantages and disadvantages, comprising of economic, social, health and environmental impacts; giving the reader an unbiased perspective. Few case studies have also been reviewed to better understand the implications of drawing energy from biofuels, as well as the current state of …

Identifying, Monitoring, Quantifying And Converting Algae To Bio-Fuels In Bio-Reactors, Alice C. Jernigan

Graduate Theses and Dissertations

Growing algae as a source for bio-fuels has become an area of interest due to concerns about global warming and the reliability and ecology of the production of fossil fuels. Dried algae harvested from a pilot water quality improvement technology at the Rockaway Wastewater Treatment Facility in New York were examined as a source of carbohydrates and lipids for the production of bio-fuels in bio-reactors. The length of storage time, storage conditions, sugar and lipid extraction processes, and fuel production were studied. The results show that if the algae is stored dry (0.015 g/g algae even after a year in …

Comparative Efficiency And Driving Range Of Light- And Heavy-Duty Vehicles Powered With Biomass Energy Stored In Liquid Fuels Or Batteries, Mark Laser, Lee R. R. Lynd

Dartmouth Scholarship

This study addresses the question, "When using cellulosic biomass for vehicular transportation, which field-to-wheels pathway is more efficient: that using biofuels or that using bioelectricity?" In considering the question, the level of assumed technological maturity significantly affects the comparison, as does the intended transportation application. Results from the analysis indicate that for light-duty vehicles, over ranges typical in the United States today (e.g., 560-820 miles), field-to-wheels performance is similar, with some scenarios showing biofuel to be more efficient, and others indicating the two pathways to be essentially the same. Over the current range of heavy-duty vehicles, the field-to-wheels efficiency is …

Physiological And Transcriptional Memory In Guard Cells During Repetitive Dehydration Stress, Laetitia Virlouvet, Michael E. Fromm

Nebraska Center for Biotechnology: Faculty and Staff Publications

Arabidopsis plants subjected to a daily dehydration stress and watered recovery cycle display physiological and transcriptional stress memory. Previously stressed plants have stomatal apertures that remain partially closed during a watered recovery period, facilitating reduced transpiration during a subsequent dehydration stress. Guard cells (GCs) display transcriptional memory that is similar to that in leaf tissues for some genes, but display GC-specific transcriptional memory for other genes. The rate-limiting abscisic acid (ABA) biosynthetic genes NINE-CIS-EPOXYCAROTENOID DIOXYGENASE 3 (NCED3) and ALDEHYDE OXIDASE 3 (AAO3) are expressed at much higher levels in GCs, particularly during the watered recovery interval, relative to their low …

The Pseudomonas Syringae Type Iii Effector Hopd1 Suppresses Effector-Triggered Immunity, Localizes To The Endoplasmic Reticulum, And Targets The Arabidopsis Transcription Factor Ntl9, Anna Block, Tania Y. Toruno, Christian G. Elowsky, Chi Zhang, Jens Steinbrenner, Jim Beynon, James R. Alfano

Nebraska Center for Biotechnology: Faculty and Staff Publications

Pseudomonas syringae type III effectors are known to suppress plant immunity to promote bacterial virulence. However, the activities and targets of these effectors are not well understood. We used genetic, molecular, and cell biology methods to characterize the activities, localization, and target of the HopD1 type III effector in Arabidopsis. HopD1 contributes to P. syringae virulence in Arabidopsis and reduces effector-triggered immunity (ETI) responses but not pathogen-associated molecular pattern-triggered immunity (PTI) responses. Plants expressing HopD1 supported increased growth of ETI-inducing P. syringae strains compared with wild-type Arabidopsis. We show that HopD1 interacts with the membrane-tethered Arabidopsis transcription factor NTL9 and …

Nanometer-Scale Membrane Electrode Systems For Active Protein Separation, Enzyme Immobilization And Cellular Electroporation, Zhiqiang Chen

Theses and Dissertations--Chemical and Materials Engineering

Automated and continuous processes are the future trends in downstream protein purification. A functionalized nanometer-scale membrane electrode system, mimicking the function of cell wall transporters, can selectively capture genetically modified proteins and subsequently pump them through the system under programmed voltage pulses. Numerical study of the two-step pulse pumping cycles coupled with experimental His-GFP releasing study reveals the optimal 14s/1s pumping/repel pulse pumping condition at 10 mM bulk imidazole concentration in the permeate side. A separation factor for GFP: BSA of 9.7 was achieved with observed GFP electrophoretic mobility of 3.1×10^-6 cm² s^-1 V^-1 at 10 …

Electrotransfer Of Single-Stranded Or Double-Stranded Dna Induces Complete Regression Of Palpable B16.F10 Mouse Melanomas, Loree Heller, Vesba Todorovic, Maja Cemazar

Bioelectrics Publications

Enhanced tumor delivery of plasmid DNA with electric pulses in vivo has been confirmed in many preclinical models. Intratumor electrotransfer of plasmids encoding therapeutic molecules has reached Phase II clinical trials. In multiple preclinical studies, a reduction in tumor growth, increased survival or complete tumor regression have been observed in control groups in which vector or backbone plasmid DNA electrotransfer was performed. This study explores factors that could produce this antitumor effect. The specific electrotransfer pulse protocol employed significantly potentiated the regression. Tumor regression was observed after delivery of single-stranded or double-stranded DNA with or without CpG motifs in both …

Near-Infrared Surface-Enhanced Fluorescence Using Silver Nanoparticles In Solution, Michael D. Furtaw

Department of Mechanical and Materials Engineering: Dissertations, Theses, and Student Research

Fluorescence spectroscopy is a widely used detection technology in many research and clinical assays. Further improvement to assay sensitivity may enable earlier diagnosis of disease, novel biomarker discovery, and ultimately, improved outcomes of clinical care along with reduction in costs. Near-infrared, surface-enhanced fluorescence (NIR-SEF) is a promising approach to improve assay sensitivity via simultaneous increase in signal with a reduction in background. This dissertation describes research conducted with the overall goal to determine the extent to which fluorescence in solution may be enhanced by altering specific variables involved in the formation of plasmonactive nanostructures of dye-labeled protein and silver nanoparticles …

Detection Of Foodborne Pathogens By Micro-Filtration Using A Continuous Cell Concentrator Device, Klaire E. Jeffries, Eduardo Ximenes, Michael R. Ladisch

The Summer Undergraduate Research Fellowship (SURF) Symposium

Protecting consumers from foodborne illness is an important health concern facing the food industry today. An important deficiency exposed by foodborne illness is the inability to track contaminated food back to the source in a timely manner. Although there are established methods that detect bacterial pathogen contamination, they are limited in distinguishing viable bacteria reliably and quickly. Currently, food pathogen testing requires lengthy culture steps, which many times are delayed even longer due to the lack of in-house testing labs. Typically, two to three days elapses between when the food is sampled and the test results are available. This study …

Bicistronic Design For Precise And Reliable Gene Expression, Nidhi N. Menon, Jenna Rickus

The Summer Undergraduate Research Fellowship (SURF) Symposium

Despite having progressed extensively in the field of synthetic biology in terms of DNA synthesis, analysis and transplanting, we still cannot reliably, quantitatively measure expression of new genetic constructs. We engineered a biobrick compatible expression cassette to control transcription and translation initiation which can be reused in new genetic contexts. Previous research has shown that the Bicistronic design have much lesser variations in expression with varying genes of interest as compared to the regular monocistronic design.(Mutalik, Endy, Guimaraes, Cambray, Lam, Juul, Tran & Paull, 2013) The Bicistronic design(BCD) consists of two Shine-Dalgarno sequences in its translation element which when combined …

Artificial Yeast Polarization Controlled By Chemical Gradient, James K. Nolan, Bernard Tao

The Summer Undergraduate Research Fellowship (SURF) Symposium

Engineering synthetic multicellular systems will lead to new synthetic biology technological platforms, inform developmental biology through recapitulation of natural systems and possibly unveil novel morphologies with practical applications not before reached throughout natural history (Maharbiz, 2012). Creating an exogenous molecular circuit that will polarize unicellular cells into “apical” and “basal” domains relative to a substrate plane would fulfill a missing component towards fully multicellular synthetic cellular communities (Maharbiz, 2012). To this end, a PIP₃ polarization network previously designed by Chau and associates (Chau, Walter, Gerardin, Tang, Lim 2012) was coupled to the specific activation by niacin of a recombinant …

Biofuel From Microalgae – A Review On The Current Status And Future Trends, Rajee Olaganathan, May Zaw Htet, Lim Yan Ling, Sek Hui Yun

Publications

The constant reliance on fossil fuel energy resources is unsustainable, due to both depleting world reserves and increasing green house gas emissions associated with their use and thus there are dynamic research at the global level envisioned at developing alternative renewable and potentially carbon neutral solid, liquid and gaseous biofuels as alternative energy resources. The contemporary knowledge and technology predictions have proved that among the third generation biofuels especially those derived from microalgae are considered the best reasonable alternative energy resource compared to undeniable drawbacks of first and second generation biofuels. Moreover, its efficiency to sequester carbon from the atmosphere …

Algae To Propellants: Biosynthesis Of 1,2-Propanediol From Algal Derived Sugars, Lauren Rachelle Merriman

Graduate Theses and Dissertations

Propylene glycol dinitrate (PGDN) is an important component used in the formulation of energetic materials. A renewable, environmentally-friendly and effective production technique for the production of 1,2-propanediol, an important precursor to PGDN, is desired. While fermentation of 1,2-propanediol has been achieved, it suffers from major limitations such as expensive feed sugars and relatively low yields. It is hypothesized that algae sugars could serve as a feedstock for the microbial synthesis of 1,2-propanediol, thereby reducing production costs and enhancing sustainability.

Algal biomass produced from an Algal Turf Scrubber® has proven to have significant potential as a source of fermentable materials. However, …

Efferent Copy And Corollary Discharge Motor Control Behavior Associated With A Hopping Activity, Wangdo Kim, António P. Veloso, Filipa João, Sean S. Kohles

Mechanical and Materials Engineering Faculty Publications and Presentations

Hoppers respond not only to stimuli from the ground surfaces but also to cues generated by their own behaviors. This leads to desensitization because although the afferent and reafferent signals have distinct causes, they are carried by the same sensory channels. From a behavioral viewpoint, it may be necessary to distinguish between signals from the two causes especially when monitoring changes in the external environment separate from those due to self-movement. We were able to separate afferent sensory stimuli from self-generated, reafferent signals using an action oriented perception system and dynamic programming approach. This effort addressed the question of how …

The Use Of Co-Culturing In Solid Substrate Cultivation And Possible Solutions To Scientific Challenges, Wanying Yao, Sue E. Nokes

Biosystems and Agricultural Engineering Faculty Publications

This perspective systematically summarizes the use of solid substrate co‐cultures in agriculture, food, plant, and industrial biotechnology applications. The summarization is organized by organism, i.e. fungus, bacteria, yeast and then co‐cultivation of either two or three organisms. Generally, in solid substrate co‐culture, the organisms synergistically penetrate and degrade the solid substrate, thereby increasing product yield and productivity over a monoculture. Efforts to increase co‐culture performance include optimizing process parameters (pH, temperature, moisture, and oxygen demand) and defining the acceptable types of substrate. Scientific challenges exist in understanding the interactions between microbial stains, such as viability, suite of products, and bio‐transformations. …

A Morphological Analysis Of A Hybrid Swarm Of Native Ulmus Rubra And Introduced U. Pumila (Ulmaceae) In Southeastern Nebraska, Christian Elowsky, Ingrid E. Jordon-Thaden, Robert B. Kaul

Nebraska Center for Biotechnology: Faculty and Staff Publications

The parental species and hybrid swarm of native Ulmus rubra Muhl. and the introduced, naturalized and weedy U. pumila L. were investigated in a 65-km transect in Lancaster, Saunders, and Butler counties in Nebraska. Thirty-two trees of U. rubra, 32 of U. pumila, and 50 of the hybrid swarm were sampled for leaves and buds and subsampled for flowers and fruits. Leaves were measured for petiole length, blade length, width, primary and secondary teeth per cm, number of secondary teeth per primary tooth, and texture. Buds were scored for color and distribution of trichomes. Flowers were sampled for stamen counts …

Influence Of Media Composition On The Growth Rate Of Chlorella Vulgaris And Scenedesmus Acutus Utilized For Co2 Mitigation, Czarena L. Crofcheck, Xinyi E, Aubrey Shea, Michael D. Montross, Mark Crocker, Rodney Andrews

Biosystems and Agricultural Engineering Faculty Publications

Atmospheric carbon dioxide levels have increased since the industrial revolution due to increasing combustion of fossil fuels. One possible CO₂ mitigation strategy is the use of microalgae for mitigation of CO₂. This paper focuses on the influence of media composition on the growth rate of two microalgae strains, Chlorella vulgaris and Scenedesmus actus. A KNO₃ based medium was found to work better for Chlorella, while a urea based equivalent worked better for Scenedesmus. The urea based media investigated here resulted in growth similar to that found with previously reported KNO₃ based media. …

Biomass Gasification: An Alternative Solution To Animal Waste Management, Hanjing Wu

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

The overall goal of this research was to evaluate gasification of animal waste as an alternative manure management strategy, from the standpoints of syngas production and biochar application.

To meet the overall objective, the thermogravimetric characteristics of dairy manure, as a thermochemical conversion feedstock, were studied firstly. Then, gasification technology was applied to dairy manure and feedlot manure using a fluidized-bed laboratory-scale gasifier. In addition, biochar derived from the feedlot manure was examined for its effects on nutrient leaching as a soil amendment. Finally, a life cycle assessment was conducted to evaluate greenhouse gas emissions of two feedlot manure management …

Expression, Production, And Purification Of Novel Therapeutic Proteins, Mckinzie Shea Fruchtl

Graduate Theses and Dissertations

Interest in the production of recombinant proteins consisting of collagen binding domain (CBD) fused to a bioactive material has increased due to the targeting/attachment capabilities of CBD. For example, CBD fusions can be applied to the reversing of bone density loss and the repair of the eardrum, specifically, by choosing an appropriate fusion partner (parathyroid hormone or epidermal growth factor). The production of CBD fusions was examined using batch and fed-batch culturing of Escherichia coli to express the fusion proteins, and affinity chromatography to isolate the final product.

Different medium formulations, feeding strategies, and induction methods were tested in order …

The Geometry And Sensitivity Of Ion-Beam Sculpted Nanopores For Single Molecule Dna Analysis, Ryan Connor Rollings

Graduate Theses and Dissertations

In this dissertation, the relationship between the geometry of ion-beam sculpted solid-state nanopores and their ability to analyze single DNA molecules using resistive pulse sensing is investigated. To accomplish this, the three dimensional shape of the nanopore is determined using energy filtered and tomographic transmission electron microscopy. It is shown that this information enables the prediction of the ionic current passing through a voltage biased nanopore and improves the prediction of the magnitude of current drop signals when the nanopore interacts with single DNA molecules. The dimensional stability of nanopores in solution is monitored using this information and is improved …

Colloidal Nano-Apatite Particles With Active Luminescent And Magentic Properties For Biotechnology Applications, Rajendra Kasinath, Kumar Ganesan

Environmental Engineering

Colloidal Nano-apatite Particles with Active Luminescent and Magentic Properties for Biotechnology Applications. The synthesis of functional nano-materials is a burgeoning field that has produced remarkable and consistent breakthroughs over the last two decades. Individual particles have become smaller and shown potential for well defined functionality. However, there are still unresolved problems, a primary one being the loss of functionality and novelty due to uncontrolled aggregation driven by surface energy considerations. As such the first design criteria to harness the true potential of nanoparticles is to prevent unwanted agglomeration by: (1) improving, and, if possible, (2) controlling aggregation behavior. This requires …

Response To "Sodium Current Inhibition By Nanosecond Pulsed Electric Field (Nspef) - Fact Or Artifact?" By Verkerk Et Al, Andrei G. Pakhomov

Bioelectrics Publications

It was nice to learn that our studies of nanosecond pulsed electric field (nsPEF) effects on membrane currents [Nesin et al., 2012; Nesin and Pakhomov, 2012] gained the attention of scientists outside the immediate field of bioelectromagnetics.

The Effects Of Hydrostatic Pressure On Early Endothelial Tubulogenic Processes, Ryan M. Underwood

Theses and Dissertations--Biomedical Engineering

The effects of mechanical forces on endothelial cell function and behavior are well documented, but have not been fully characterized. Specifically, fluid pressure has been shown to elicit physical and chemical responses known to be involved in the initiation and progression of endothelial cell-mediated vascularization. Central to the process of vascularization is the formation of tube-like structures. This process—tubulogenesis—is essential to both the physiological and pathological growth of tissues. Given the known effects of pressure on endothelial cells and its ubiquitous presence in the vasculature, we investigated pressure as a magnitude-dependent parameter for the regulation of endothelial tubulogenic activity. To …

Real-Time In Vivo Imaging Of Size-Dependent Transport And Toxicity Of Gold Nanoparticles In Zebrafish Embryos Using Single Nanoparticle Plasmonic Spectroscopy, Lauren M. Browning, Tao Huang, Xiao-Hong Nancy Xu

Chemistry & Biochemistry Faculty Publications

Noble metal nanoparticles (NPs) show distinctive plasmonic optical properties and superior photostability, enabling them to serve as photostable multicoloured optical molecular probes and sensors for real-time in vivo imaging. To effectively study biological functions in vivo, it is essential that the NP probes are biocompatible and can be delivered into living organisms non-invasively. In this study, we have synthesized, purified and characterized stable (non-aggregated) gold (Au) NPs (86.2 +/- 10.8 nm). We have developed dark-field single NP plasmonic microscopy and spectroscopy to study their transport into early developing zebrafish embryos (cleavage stage) and their effects on embryonic development …

Engineering Novel Terpene Production Platforms In The Yeast Saccharomyces Cerevisiae, Xun Zhuang

Theses and Dissertations--Plant and Soil Sciences

The chemical diversity and biological activities of terpene and terpenoids have served in the development of new flavors, fragrances, medicines and pesticides. While terpenes are made predominantly by plants and microbes in small amounts and as components of complex mixtures, chemical synthesis of terpenes remains technically challenging, costly and inefficient. In this dissertation, methods to create new yeast lines possessing a dispensable mevalonate biosynthetic pathway wherein carbon flux can be diverted to build any chemical class of terpene product are described. The ability of this line to generate diterpenes was next investigated. Using a 5.5 L fed bath fermentation system, …

Bile-Acid-Mediated Decrease In Endoplasmic Reticulum Stress: A Potential Contributor To The Metabolic Benefits Of Ileal Interposition Surgery In Ucd-T2dm Rats, Bethany P. Cummings, Ahmed Bettaieb, James L. Graham, Jaehyoung Kim, Fangrui Ma, Noreene Shibata, Kimber L. Stanhope, Cecilia Giulivi, Frederik Hansen, Jacob Jelsing, Niels Vrang, Mark Kowala, Michael L. Chouinard, Fawaz G. Haj, Peter J. Havel

Nebraska Center for Biotechnology: Faculty and Staff Publications

Post-operative increases in circulating bile acids have been suggested to contribute to the metabolic benefits of bariatric surgery; however, their mechanistic contributions remain undefined. We have previously reported that ileal interposition (IT) surgery delays the onset of type 2 diabetes in UCD-T2DM rats and increases circulating bile acids, independently of effects on energy intake or body weight. Therefore, we investigated potential mechanisms by which post-operative increases in circulating bile acids improve glucose homeostasis after IT surgery. IT, sham or no surgery was performed on 2-month-old weight-matched male UCD-T2DM rats. Animals underwent an oral fat tolerance test (OFTT) and serial oral …

Optimizing Electrode Design For Microbial Fuel Cells Used For Wastewater Treatment, Lindsay Nichols, John A. Hogan

STAR Program Research Presentations

Microbial fuel cells (MFC) utilize bacteria to generate an electrical current that can be used in the decomposition of sludge and human urine. In a MFC there is an anode (for oxidation of organic compounds), cathode (reduction of oxygen or carbon dioxide), and a proton exchange membrane (PEM, allows protons to migrate); reduction-oxidation reactions between the anode and cathode produce a measurable current. Bacteria that are found in sludge can be used to produce electrons in a voltaic cell, but optimizing conditions for harnessing the energy is crucial to making a MFC efficient. Research has shown that the ratios of …

Inactivation Of Bacterial Opportunistic Skin Pathogens By Nonthermal Dc-Operated Afterglow Atmospheric Plasma, L. C. Heller, C. M. Edelblute, A. M. Mattson, X. Hao, J. F. Kolb

Bioelectrics Publications

AIMS: Multidrug-resistant opportunistic pathogens are clinically significant and require the development of new antimicrobial methods. In this study, Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus aureus cells were exposed to atmospheric plasma on agar plates and in vitro on porcine skin for the purpose of testing bacterial inactivation.

METHODS AND RESULTS: Microbial inactivation at varying exposure durations was tested using a nonthermal plasma jet generated with a DC voltage from ambient air. The observed reduction in colony forming units was quantified as log₁₀ reductions.

CONCLUSIONS: Direct plasma exposure significantly inactivated seeded bacterial cells by approx. 6 log_{10 …}

Ethanol And Anaerobic Conditions Reversibly Inhibit Commercial Cellulase Activity In Thermophilic Simultaneous Saccharification And Fermentation (Tssf), Kara K. Podkaminer, William R. Kenealy, Christopher D. Herring, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

A previously developed mathematical model of low solids thermophilic simultaneous saccharification and fermentation (tSSF) with Avicel was unable to predict performance at high solids using a commercial cellulase preparation (Spezyme CP) and the high ethanol yield Thermoanaerobacterium saccharolyticum strain ALK2. The observed hydrolysis proceeded more slowly than predicted at solids concentrations greater than 50 g/L Avicel. Factors responsible for this inaccuracy were investigated in this study.