Engineering Commons^™

Retargeting The Clostridium Botulinum C2 Toxin To The Neuronal Cytosol, Benjamin J. Pavlik, Elizabeth J. Hruska, Kevin E. Van Cott, Paul H. Blum

Department of Chemical and Biomolecular Engineering: Faculty Publications

Many biological toxins are known to attack specific cell types, delivering their enzymatic payloads to the cytosol. This process can be manipulated by molecular engineering of chimeric toxins. Using toxins with naturally unlinked components as a starting point is advantageous because it allows for the development of payloads separately from the binding/translocation components. Here the Clostridium botulinum C2 binding/translocation domain was retargeted to neural cell populations by deleting its non-specific binding domain and replacing it with a C. botulinum neurotoxin binding domain. This fusion protein was used to deliver fluorescently labeled payloads to Neuro-2a cells. Intracellular delivery was quantified by …

Noninvasive Measurement Of Electrical Events Associated With A Single Chlorovirus Infection Of A Microalgal Cell, Seung-Woo Lee, Eun-Hee Lee, Gerhard Thiel, James L. Van Etten, Ravi Saraf

Department of Chemical and Biomolecular Engineering: Faculty Publications

Chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) contains a viral-encoded K⁺ channel imbedded in its internal membrane, which triggers host plasma membrane depolarization during virus infection. This early stage of infection was monitored at high resolution by recording the cell membrane depolarization of a single Chlorella cell during infection by a single PBCV-1 particle. The measurement was achieved by depositing the cells onto a network of one-dimensional necklaces of Au nanoparticles, which spanned two electrodes 70 μm apart. The nanoparticle necklace array has been shown to behave as a single-electron device at room temperature. The resulting electrochemical field-effect transistor …

Pch1 Integrates Circadian And Light- Signaling Pathways To Control Photoperiod-Responsive Growth In Arabidopsis, He Huang, Chan Yul Yoo, Rebecca Bindbeutel, Jessica Goldsworthy, Allison Tielking, Sophie Alvarez, Michael J. Naldrett, Bradley S. Evans, Meng Chen, Dmitri A. Nusinow

Nebraska Center for Biotechnology: Faculty and Staff Publications

Plants react to seasonal change in day length through altering physiology and development. Factors that function to harmonize growth with photoperiod are poorly understood. Here we characterize a new protein that associates with both circadian clock and photoreceptor components, named PHOTOPERIODIC CONTROL OF HYPOCOTYL1 (PCH1). pch1 seedlings have overly elongated hypocotyls specifically under short days while constitutive expression of PCH1 shortens hypocotyls independent of day length. PCH1 peaks at dusk, binds phytochrome B (phyB) in a red light-dependent manner, and co-localizes with phyB into photobodies. PCH1 is necessary and sufficient to promote the biogenesis of large photobodies to maintain an …

Direct Current Stimulation Modulates Ltp And Ltd: Activity-Dependence And Dendritic Effects, Gregory Kronberg

Dissertations and Theses

Transcranial direct current stimulation (tDCS) has been reported to improve various forms of learning in humans. Stimulation is often applied during training, producing lasting enhancements that are specific to the learned task. These learning effects are thought be mediated by altered synaptic plasticity. However, the effects of DCS during the induction of endogenous synaptic plasticity remain largely unexplored. To model endogenous plasticity I induced long-term potentiation (LTP) and depression (LTD) at Schaffer collateral synapses in CA1 of rat hippocampal slices. When induction was paired with concurrent DCS, the resulting plasticity was biased towards potentiation, such that LTP was enhanced and …

Ultrahigh Field Magnetic Resonance Imaging – Technical Development And Translational Applications, Judy Alper

Dissertations and Theses

Magnetic resonance imaging (MRI) may be used to provide detailed images of the human body with excellent soft tissue contrast. Alongside its current widespread clinical applications for diagnosis and treatment, MRI allows researchers to measure structure and function of different tissue types in order to advance our understanding of human biology and enable new medical applications of MRI. In particular, diseases affecting nerves and vessels, such as trigeminal neuralgia, with uncertain etiology can be studied using multiple MRI modalities so that treatment planning can we more effective and patient outcomes can be improved. Ultrahigh field MRI scanners, such as those …

Aconitic Acid, Glycerol And Cinnamic Acid Polyester Characterization For Bone Tissue Engineering, Katie Joanna Hogan

Honors Capstones

No abstract provided.

Intracellular Ros Mediates Gas Plasma-Facilitated Cellular Transfection In 2d And 3d Cultures, Dehui Xu, Biqing Wang, Yujing Xu, Zeyu Chen, Qinjie Cui, Yanjie Yang, Hailan Chen, Michael G. Kong

Bioelectrics Publications

This study reports the potential of cold atmospheric plasma (CAP) as a versatile tool for delivering oligonucleotides into mammalian cells. Compared to lipofection and electroporation methods, plasma transfection showed a better uptake efficiency and less cell death in the transfection of oligonucleotides. We demonstrated that the level of extracellular aqueous reactive oxygen species (ROS) produced by gas plasma is correlated with the uptake efficiency and that this is achieved through an increase of intracellular ROS levels and the resulting increase in cell membrane permeability. This finding was supported by the use of ROS scavengers, which reduced CAP-based uptake efficiency. In …

Biom 3110: Medical Physiology (Syllabus), Bradford D. Pendley

Biomedical Engineering Syllabi Archive

Course Description: Basic principles of human physiology and how these are applied in medicine and biomedical engineering problem and devices; physiological principles using appropriate mathematical and engineering models to construct an understanding of how the body works as an integrated system.

Biom 4110/6110: The Science Of Medicine (Syllabus), Bradford D. Pendley

Biomedical Engineering Syllabi Archive

Course Description: Integration of fundamental principles from physics, chemistry, biology and mathematics, and applications of these principles to solve problems in medicine.

Microbubble Assisted Polyhydroxybutyrate Production In Escherichia Coli, Kadriye Innan, Fulya Ay Sal, Asif Rahman, Ryan J. Putman, Foster A. Agblevor, Charles D. Miller

Biological Engineering Faculty Publications

Background

One of the potential limitations of large scale aerobic Escherichia coli fermentation is the need for increased dissolved oxygen for culture growth and bioproduct generation. As culture density increases the poor solubility of oxygen in water becomes one of the limiting factors for cell growth and product formation. A potential solution is to use a microbubble dispersion (MBD) generating device to reduce the diameter and increase the surface area of sparged bubbles in the fermentor. In this study, a recombinantE. coli strain was used to produce polyhydroxybutyrate (PHB) under conventional and MBD aerobic fermentation conditions.

Results

In conventional …

Rational Design Of Rama-Labeled Nanoparticles For A Dual-Modaility, Light Scattering Immunoassay On A Polystyrene Substrate, Nathan D. Israelsen, Donald Wooley, Cynthia Hanson, Elizabeth Vargis

Biological Engineering Faculty Publications

Background: Surface-enhanced Raman scattering (SERS) is a powerful light scattering technique that can be used for sensitive immunoassay development and cell labeling. A major obstacle to using SERS is the complexity of fabricating SERS probes since they require nanoscale characterization and optical uniformity. The light scattering response of SERS probes may also be modulated by the substrate used for SERS analysis. A typical SERS substrate such as quartz can be expensive. Polystyrene is a cheaper substrate option but can decrease the SERS response due to interfering Raman emission peaks and high background fluorescence. The goal of this research is to …

In-House Fabrication Of Multi-Sized Polystyrene Microcarriers For Neural Stem Cell Expansion, Jessica Forrester

Theses and Dissertations

The clinical demand for neural stem cells drives the need for a reproducible method used to generate a large quantity of well-characterized cells to support regenerative therapies. Microcarriers are currently being used as a scaffold for stem cell culture to aid in this expansion process because they provide a larger surface area while also requiring fewer cell passages compared to monolayer culture. The main objective of this project was to investigate how human neural stem cell attachment, proliferation, and multipotency would respond to three-dimensional (3D) culture using peptide-coated microcarriers of varying diameters. In this study, multi-sized polystyrene microcarriers were fabricated …

The Molecular And Mechanical Mechanisms Of The Age-Associated Increase In The Severity Of Experimental Ventilator Induced Lung Injury, Joseph Ames Herbert

Theses and Dissertations

Abstract

Background

The majority of patients requiring mechanical ventilation are over the age of 65 and advanced age is known to increase the severity of ventilator-induced lung injury (VILI) and mortality. However, the mechanisms which predispose aging ventilator patients to increased mortality rates are not fully understood.

Pulmonary edema is a hallmark of VILI and the severity of edema increases with age. Ventilation with conservative fluid management decreases mortality rates in acute respiratory distress (ARDS) patients, but has not been investigated in VILI. We hypothesized that age-associated increases in pulmonary edema promote age-related increases in ventilator-associated mortality. Endoplasmic reticulum (ER) …

Effects Of Antidepressants On Human Mesenchymal Stem Cell Differentiation On Clinically Relevant Titanium Surfaces, Nancy B. Ayad

Theses and Dissertations

Selective Serotonin Reuptake Inhibitors (SSRIs) are the most frequently prescribed class of drugs worldwide and are implemented in the treatment of depression and other psychiatric disorders. SSRIs relieve depressive symptoms by modulating levels of the neurotransmitter serotonin in the brain. SSRIs block the function of the serotonin transporter, thereby increasing concentrations of extracellular serotonin. However, serotonin levels in the neurons of the brain only account for 5% while the remaining 95% is present outside the brain. Serotonin receptors and transporter are located on bone resident cells (mesenchymal stem cells (MSCs)), osteoblasts and osteoclasts, and serotonergic activity is believed to affect …

Bioactive Poly(Beta-Amino Ester) Biomaterials For Treatment Of Infection And Oxidative Stress, Andrew L. Lakes

Theses and Dissertations--Chemical and Materials Engineering

Polymers have deep roots as drug delivery tools, and are widely used in clinical to private settings. Currently, however, numerous traditional therapies exist which may be improved through use of polymeric biomaterials. Through our work with infectious and oxidative stress disease prevention and treatment, we aimed to develop application driven, enhanced therapies utilizing new classes of polymers synthesized in-house. Applying biodegradable poly(β-amino ester) (PBAE) polymers, covalent-addition of bioactive substrates to these PBAEs avoided certain pitfalls of free-loaded and non-degradable drug delivery systems. Further, through variation of polymer ingredients and conditions, we were able to tune degradation rates, release profiles, cellular …

Synthesis And Characterization Of Antioxidant Conjugated Poly(Βeta-Amino Ester) Micro/Nanogels For The Suppression Of Oxidative Stress, Prachi Gupta

Theses and Dissertations--Chemical and Materials Engineering

Oxidative stress is a pathophysiological condition defined by an increased production of reactive oxygen species (ROS), which can result in the growth arrest of cells followed by cell disintegration or necrosis. A number of small molecule antioxidants (e.g. curcumin, quercetin and resveratrol) are capable of directly scavenging ROS, thereby short-circuiting the self-propagating oxidative stress state. However, poor solubility and rapid 1^st pass metabolism results in overall low bioavailability and acts as a barrier for its use as a drug to suppress oxidative stress efficiently.

To overcome this limitation, these small molecule antioxidants were covalently conjugated into poly(β-amino ester) (PβAE) …

Nanosecond Pulsed Platelet-Rich Plasma (Nsprp) Improves Mechanical And Electrial Cardiac Function Following Myocardial Reperfusion Injury, Barbara Y. Hargrave, Frency Varghese, Nektarios Barabutis, John Catravas, Christian Zemlin

School of Medical Diagnostics & Translational Sciences Publications

Ischemia and reperfusion (I/R) of the heart is associated with biochemical and ionic changes that result in cardiac contractile and electrical dysfunction. In rabbits, platelet-rich plasma activated using nanosecond pulsed electric fields (nsPRP) has been shown to improve left ventricular pumping. Here, we demonstrate that nsPRP causes a similar improvement in mouse left ventricular function. We also show that nsPRP injection recovers electrical activity even before reperfusion begins. To uncover the mechanism of nsPRP action, we studied whether the enhanced left ventricular function in nsPRP rabbit and mouse hearts was associated with increased expression of heat-shock proteins and altered mitochondrial …

Modeling Early Stage Bone Regeneration With Biomimetic Electrospun Fibrinogen Nanofibers And Adipose-Derived Mesenchymal Stem Cells, Michael P. Francis, Yas M. Moghaddam-White, Patrick C. Sachs, Matthew J. Beckman, Stephen M. Chen, Gary L. Bowlin, Lynne W. Elmore, Shawn E. Holt

School of Medical Diagnostics & Translational Sciences Publications

The key events of the earliest stages of bone regeneration have been described in vivo although not yet modeled in an in vitro environment, where mechanistic cell-matrix-growth factor interactions can be more effectively studied. Here, we explore an early-stage bone regeneration model where the ability of electrospun fibrinogen (Fg) nanofibers to regulate osteoblastogenesis between distinct mesenchymal stem cells populations is assessed. Electrospun scaffolds of Fg, polydioxanone (PDO), and a Fg:PDO blend were seeded with adipose-derived mesenchymal stem cells (ASCs) and grown for 7-21 days in osteogenic differentiation media or control growth media. Scaffolds were analyzed weekly for histologic and molecular …

Accessible Bioprinting: Adaptation Of A Low-Cost 3d-Printer For Precise Cell Placement And Stem Cell Differentiation, John A. Reid, Peter A. Mollica, Garett D. Johnson, Roy C. Ogle, Robert D. Bruno, Patrick C. Sachs

School of Medical Diagnostics & Translational Sciences Publications

The precision and repeatability offered by computer-aided design and computer-numerically controlled techniques in biofabrication processes is quickly becoming an industry standard. However, many hurdles still exist before these techniques can be used in research laboratories for cellular and molecular biology applications. Extrusion-based bioprinting systems have been characterized by high development costs, injector clogging, difficulty achieving small cell number deposits, decreased cell viability, and altered cell function post-printing. To circumvent the high-price barrier to entry of conventional bioprinters, we designed and 3D printed components for the adaptation of an inexpensive 'off-the-shelf' commercially available 3D printer. We also demonstrate via goal based …

Assessing The Spatial Relationship Between Fixation And Foveal Specializations, Melissa A. Wilk, Adam M. Dubis, Robert F. Cooper, Phyllis Summerfelt, Alfredo Dubra, Joseph Carroll

Biomedical Engineering Faculty Research and Publications

Increased cone photoreceptor density, an avascular zone (FAZ), and the displacement of inner retinal neurons to form a pit are distinct features of the human fovea. As the fovea provides the majority of our vision, appreciating how these anatomical specializations are related is important for understanding foveal development, normal visual function, and retinal disease. Here we evaluated the relationship between these specializations and their location relative to the preferred retinal locus of fixation (PRL). We measured foveal pit volume, FAZ area, peak cone density, and location of the PRL in 22 subjects with normal vision using optical coherence tomography and …

Motor Output Variability Impairs Driving Ability In Older Adults, Neha Lodha, Hwasil Moon, Changki Kim, Tanya Onushko, Evangelos A. Christou

Biomedical Engineering Faculty Research and Publications

Background: The functional declines with aging relate to deficits in motor control and strength. In this study, we determine whether older adults exhibit impaired driving as a consequence of declines in motor control or strength.

Methods: Young and older adults performed the following tasks: (i) maximum voluntary contractions of ankle dorsiflexion and plantarflexion; (ii) sinusoidal tracking with isolated ankle dorsiflexion; and (iii) a reactive driving task that required responding to unexpected brake lights of the car ahead. We quantified motor control with ankle force variability, gas position variability, and brake force variability. We quantified reactive driving performance with a combination …

Lessons Learned From A 10-Year Collaboration Between Engineering And Industrial Design Students In Capstone Design Projects, Jay R. Goldberg, Pascal Malassigné

Biomedical Engineering Faculty Research and Publications

Engineers and industrial designers have different approaches to problem solving. Both place heavy emphasis on identification of customer needs, manufacturing methods, and prototyping. Industrial designers focus on aesthetics, ergonomics, ease of use, and the user’s experience. They tend to be more visual and more concerned with the interaction between users and products. Engineers focus on functionality, performance requirements, analytical modeling, and design verification and validation. They tend to be more analytical and more concerned with the design of internal components and product performance. Engineers and industrial designers often work together on project teams in industry. Collaboration between the two groups …

Comparing Offline Decoding Performance In Physiologically Defined Neuronal Classes, Matthew D. Best, Kazutaka Takahashi, Aaron J. Suminski, Christian Ethier, Lee E. Miller, Nicholas G. Hatsopoulos

Biomedical Engineering Faculty Research and Publications

Objective: Recently, several studies have documented the presence of a bimodal distribution of spike waveform widths in primary motor cortex. Although narrow and wide spiking neurons, corresponding to the two modes of the distribution, exhibit different response properties, it remains unknown if these differences give rise to differential decoding performance between these two classes of cells. Approach: We used a Gaussian mixture model to classify neurons into narrow and wide physiological classes. Using similar-size, random samples of neurons from these two physiological classes, we trained offline decoding models to predict a variety of movement features. We compared offline …

Severity Of Spinal Cord Injury Influences Diffusion Tensor Imaging Of The Brain, Michael B. Jirjis, Aditya Vedantam, Matthew D. Budde, Benjamin Kalinosky, Shekar N. Kurpad

Biomedical Engineering Faculty Research and Publications

Background: The purpose of this study was to determine whether DTI changes in the brain induced by a thoracic spinal cord injury are sensitive to varying severity of spinal contusion in rats.

Methods: A control, mild, moderate, or severe contusion injury was administered over the eighth thoracic vertebral level in 32 Sprague-Dawley rats. At 11 weeks postinjury, ex vivo DTI of the brain was performed on a 9.4T Bruker scanner using a pulsed gradient spin-echo sequence.

Results: Mean water diffusion in the internal capsule regions of the brain and pyramid locations of the brainstem were correlated with motor function (r …

Automated Selection Of The Optimal Cardiac Phase For Single-Beat Coronary Ct Angiography Reconstruction, Daniel Stassi, S. Dutta, H. Ma, A. Soderman, D. Pazzani, E. Gros, D. Okerlund, Taly Gilat-Schmidt

Biomedical Engineering Faculty Research and Publications

Purpose: Reconstructing a low-motion cardiac phase is expected to improve coronary artery visualization in coronary computed tomography angiography (CCTA) exams. This study developed an automated algorithm for selecting the optimal cardiac phase for CCTA reconstruction. The algorithm uses prospectively gated, single-beat, multiphase data made possible by wide cone-beam imaging. The proposed algorithm differs from previous approaches because the optimal phase is identified based on vessel image quality (IQ) directly, compared to previous approaches that included motion estimation and interphase processing. Because there is no processing of interphase information, the algorithm can be applied to any sampling of image phases, …

Assessing Photoreceptor Structure Associated With Ellipsoid Zone Disruptions Visualized With Optical Coherence Tomography, Drew Scoles, John A. Flatter, Robert F. Cooper, Christopher S. Langlo, Scott Robison, Maureen Neitz, David V. Weinberg, Mark E. Pennesi, Dennis P. Han, Alfredo Dubra, Joseph Carroll

Biomedical Engineering Faculty Research and Publications

Purpose: To compare images of photoreceptor layer disruptions obtained with optical coherence tomography (OCT) and adaptive optics scanning light ophthalmoscopy (AOSLO) in a variety of pathologic states.

Methods: Five subjects with photoreceptor ellipsoid zone disruption as per OCT and clinical diagnoses of closed-globe blunt ocular trauma (n = 2), macular telangiectasia type 2 (n = 1), blue-cone monochromacy (n = 1), or cone-rod dystrophy (n = 1) were included. Images were acquired within and around photoreceptor lesions using spectral domain OCT, confocal AOSLO, and split-detector AOSLO.

Results: There were substantial differences in the extent and appearance of the photoreceptor mosaic …

Biomechanics Of Osteogenesis Imperfecta: Current Concepts And Emerging Horizons, Carolyne Albert, Jessica M. Fritz, Gerald F. Harris

Biomedical Engineering Faculty Research and Publications

Osteogenesis inperfecta (OI) is associated with bone fragility. Long bone fractures are a common occurrence in individuals with OI. Although there have been significant advances in understanding the genetic defects associated with OI, the mechanisms behind bone fragility in this patient population are not yet well understood. This fragility is believed to stem in part from characteristic bone mass deficiencies. Research further suggests that the material properties of the bone are also compromised in individuals with this disorder. There is currently on quantitative method available to assess bone fracture risk in individuals with OI. This chapter examines several critical elements …

Fluoroscopic System For Assessment Of In Vivo Hindfoot Kinematics During Gait: Control Data And Applications In Osteogenesis Imperfecta, Ben Mchenry, Jason Long, Peter Smith, Haluk Altiok, Gerald F. Harris

Biomedical Engineering Faculty Research and Publications

No abstract provided.

Motion Analysis Strategy Appropriate For 3d Kinematic Assessment Of Children And Adults With Osteogenesis Imperfecta, Jeffrey D. Kertis, Jessica M. Fritz, Sergey Tarima, Gerald F. Harris

Biomedical Engineering Faculty Research and Publications

Human motion analysis provides a quantitative means of assessing whole body and segmental motion of subjects with musculoskeletal pathologies. This chapter describes a low cost motion analysis appropriate for complete three-dimensional (3D) assessment of upper and lower extremity kinematics. The system has been designed to support lower cost outreach efforts that require accuracy and resolution on the order of classical fixed lot systems such as Vicon. The focus of this work addresses the assessment needs typically seen in adults and children with osteogenesis imperfect (OI) experiencing ambulatory and upper extremity challenges.

Nih Visible Human Project Humerus Numerical And Physical Models: Characterization For Osteogenesis Imperfecta Fracture Study, Prateek Grover, Gerald F. Harris

Biomedical Engineering Faculty Research and Publications

No abstract provided.