Biotechnology Commons^™

Evaluation Of Intestinal Microbial Diversity And A New Antibiotic Regimen In Crohn's Disease Patients, Karel Alcedo

Electronic Theses and Dissertations

Crohn's disease (CD) is a chronic granulomatous inflammatory bowel disease involving Mycobacterium avium subspecies paratuberculosis (MAP). Other microorganisms such as adherent-invasive Escherichia coli (AIEC) have also been proposed in CD association. To date, only one study investigated both MAP and AIEC simultaneously using peripheral blood but not in affected intestinal tissues. A standardized and effective antibiotic therapy against MAP and/or AIEC is needed for better treatment. Three antibiotic drugs – Clarithromycin (CLA), Rifabutin (RIF), and Clofazimine (CLO) have been used to treat CD patients suspected with MAP infection. However, the outcome has been controversial. The treatment dosage is high, the …

Genetically-Programmed Suicide Of Adrenergic Cells In The Mouse Leads To Severe Left Ventricular Dysfunction, Impaired Weight Gain, And Symptoms Of Neurological Dysfunction, Aaron Owji

Electronic Theses and Dissertations

Phenylethanolamine-N-methyltransferase (Pnmt) catalyzes the conversion of noradrenaline to adrenaline and is the last enzyme in the catecholamine biosynthetic pathway. Pnmt serves as a marker for adrenergic cells, and lineage-tracing experiments have identified the embryonic heart and hindbrain region as the first sites of Pnmt expression in the mouse. Pnmt expression in the heart occurs before the adrenal glands have formed and prior to sympathetic innervation, suggesting that the heart is the first site of catecholamine production in the mouse. The function of these Pnmt+ cells in heart development remains unclear. In the present study, we test the hypothesis that (i) …

Differential Expression Of Proteins Involved In Vldl Trafficking Causes Reduced Vldl Secretion In Male Ames Dwarf Mice, Faisal Ahmed Moinuddin

Electronic Theses and Dissertations

Cardiovascular diseases (CVDs) have been recorded as the number one cause of death worldwide, accounting for 32% of total deaths annually. More than two-thirds of all CVD cases are associated with atherosclerosis, which is the accumulation of fats and other substances causing plaque formation in the interior walls of major arteries. This leads to narrowing of the lumen and hardening of the arteries, ultimately resulting in angina, heart attack and/or stroke. Studies have shown that the pathogenesis of atherosclerosis and associated CVDs is strongly linked to elevated secretion of liver-specific lipoproteins called very-low-density-lipoprotein (VLDL). VLDLs are crucial lipoproteins responsible for …

Heterosandwich Assay Of Nicotinic Acetylcholine Receptors, Augustine J. Pagan Iv

Theses and Dissertations

Using the technology afforded by Winschel et al., cyclen-1, a high affinity, strong complexation agent for 8-hydroxypyrene-1,3,6 trisulfonate and derivatives, a new assay has been developed for fluorescently labeling proteins of interest (POIs). Ligation of the endogenous ligand for nicotinic acetylcholine receptors (nAChRs), acetylcholine, using click chemistry afforded the triazole derivative of an alkynyl-acylcholine (compound 1) with 8-azidopyrene-1,3,6 trisulfonate (compound 2). Liposomes encapsulated with Rhodamine B were used to strengthen the initial fluorophore response of compound 2, using an anchored form of cyclen-1 complex. Using a palmitoyl tail as the lipophilic moiety for liposomal amplification, the …

Integrated Nanoscale Imaging And Spatial Recognition Of Biomolecules On Surfaces, Congzhou Wang

Theses and Dissertations

Biomolecules on cell surfaces play critical roles in diverse biological and physiological processes. However, conventional bulk scale techniques are unable to clarify the density and distribution of specific biomolecules in situ on single, living cell surfaces at the micro or nanoscale. In this work, a single cell analysis technique based on Atomic Force Microscopy (AFM) is developed to spatially identify biomolecules and characterize nanomechanical properties on single cell surfaces. The unique advantage of these AFM-based techniques lies in the ability to operate in situ (in a non-destructive fashion) and in real time, under physiological conditions or controlled micro-environments.

First, AFM-based …

Synthesis And Characterization Of Core-Shell Zinc Silica Nanoparticles And Zinc Silica Nanogels For Agricultural Applications., Megan Berroth

Electronic Theses and Dissertations

Plant pathogens are a serious problem facing the agricultural industry today. Current methodologies use copper based biocides as the main form of defense. Unfortunately this can lead to damaging environmental effects and increased rates of antimicrobial resistance. In this study, antimicrobial activity of multiple alternative zinc-based nanoformulations were tested against three important plant pathogens: Xanthomonas alfalfae, Pseudomonas syringae, and Clavobacter michiganensis. Xanthomonas sub species cause Citrus canker, a devastating disease that affects millions of citrus trees worldwide while the latter two affect tomato crops. Materials synthesis was completed and the resulting nanoformulations were characterized by Atomic Absorption Spectroscopy, Scanning Electron …

Fmf Assay For Assessing Vaccine Generated Antibodies In A Biomimetic Manner, Vipra Dhir

Electronic Theses and Dissertations

Traditional functional assays such as hemagglutination inhibition (HAI) and micro-neutralization (MN) assays have been routinely used for assessing the vaccine response, since influenza vaccine has been administered in people (1940). Such assays are not always predictive regarding the protection conferred by the influenza vaccine and are not able to monitor neutralization related to stem region of influenza hemagglutinin responsible for virus membrane fusion in the endosomes. In order to study Influenza vaccine response in a more biomimetic manner and overcome the deficiencies of the traditional functional assays, we developed a fluorescent membrane fusion assay (fMF). The assay uses viruses labeled …

Functional Identification Of Nucleus Tractus Solitarius (Nts) Barosensitive Neurons: Effect Of Chronic Intermittent Hypoxia (Cih), Jenya Kolpakova

Electronic Theses and Dissertations

Chronic Intermittent Hypoxia (CIH) is a model used to study obstructive sleep apnea (OSA). Previously, we showed that baroreflex control of heart rate (HR) (baroreflex sensitivity) is reduced in CIH rats. While afferent function and HR in response to vagal efferent stimulation are enhanced, the effect of CIH on the central components, in particular NTS, is still not completely understood. F344 rats (3-4 mo) were exposed either to CIH or room air (RA) for 35-50 days. Following CIH exposure, rats were anaesthetized with Ket/Ace. Using single-unit extracellular recording technique, we recorded NTS barosensitive neurons in response to arterial pressure (AP) …

Mechanism Of Inactivation By High Voltage Atmospheric Cold Plasma Differs Between Escherichia Coli And Staphylococcus Aureus, Lu Han, Sonal Patil, Daniela Boehm, Vladimir Milosavljević, Patrick J. Cullen, Paula Bourke

Articles

Atmospheric cold plasma (ACP) is a promising non-thermal technology effective against a wide range of pathogenic microorganisms. Reactive oxygen species (ROS) play a crucial inactivation role when air or other oxygen containing gases are used. With strong oxidative stress, cells can be damaged by lipid peroxidation, enzyme inactivation and DNA cleavage. Identifying ROS and understanding their role is important to advance ACP applications to a range of complex microbiological issues. In this study, the inactivation efficacy of in-package, high voltage (80 kV_RMS) ACP (HVACP) and the role of intracellular ROS were investigated. Two mechanisms of inactivation were observed …

Biodiesel Production From Castor Oil And Its Application In Diesel Engine, Ismail S, Abu S. A, Rezaur R, Sinin H

ASEAN Journal on Science and Technology for Development

In this study, the optimum biodiesel conversion from crude castor oil to castor biodiesel (CB) through transesterification method was investigated. The base catalyzed transesterification under different reactant proportion such as the molar ratio of alcohol to oil and mass ratio of catalyst to oil was studied for optimum production of castor biodiesel. The optimum condition for base catalyzed transesterification of castor oil was determined to be 1:4.5 of oil to methanol ratio and 0.005:1 of potassium hydroxide to oil ratio. The fuel properties of the produced CB such as the calorific value, flash point and density were analyzed and compared …

Development Of Suspended Particulate Matter Empirical Equation For Tropics Estuary From Landsat Etm+ Data, Razak Z, Zuhairi A, Shahbudin S, Rosnan Y

ASEAN Journal on Science and Technology for Development

Suspended Particulate Matter (SPM) essentially related to the total scattering of particles in the water column. It plays the role as a transport medium for pollutants, total load of organic and inorganic substance in the water phase. In this study, we have developed empirical relationship based on a strong relationship between Landsat near infrared (NIR) band and archived SPM data. The following were the power equations:SPM NE (mg/l) = 11.68x0.666SPM SW (mg/l) = 18.61x0.493where, x (TM4) is radiance of ETM+ NIR band 4 was developed under tropical atmospheric conditions. Using the above equation, the SPM concentration for Northeast and Southwest …

Drivers And Barriers For Going Green: Perceptions From The Business Practitioners In Malaysia, Loke S-P, Khalizani K, Rohati S, Sayaka A

ASEAN Journal on Science and Technology for Development

The changes of global environmental conditions have placed great challenges to governments and societies. While it is not easy for the companies to go green, we need a renewed concern for our environment in order to revive the nation’s economic growth, social cohesion and ecological balances. This article identifies the drivers and barriers for the business industry to adopt green practices. A total of 571 business companies from the Perak State participated in this study. Four variables: (1) Regulations (β=0.159, p<0.05); (2) Social responsibility (β=0.201, p<0.05); (3) Pro-environmental organizational culture (β=0.389, p<0.01); and (4) Organizational supports (β=0.369, p<0.01) werefound to significantly affect the company’s green initiatives. The results indicated that the main internal barriers were: it lacked of financial resources (66.2%) and skilled staff (63.9%); whereas the main external barriers were: the penalty imposed were not severe enough for making any extra efforts (64.8%) and the penalty was light for violation of environmental regulations (63.2%). This research had implications for the academics, practitioners and policy makers. It provided greater insights into the green practices in Malaysian firms. The research findings also urged the local governments to greatly enhance regulatory scrutiny on the production and manufacturing industries.

Design, Assessment, And In Vivo Evaluation Of A Computational Model Illustrating The Role Of Cav1 In Cd4+ T-Lymphocytes, Brittany D. Conroy, Tyler A. Herek, Timonthy D. Shew, Matthew Latner, Joshua J. Larson, Laura Allen, Paul H. Davis, Tomáš Helikar, Christine E. Cutucache

Department of Biochemistry: Faculty Publications

Caveolin-1 (CAV1) is a vital scaffold protein heterogeneously expressed in both healthy and malignant tissue. We focus on the role of CAV1 when overexpressed in T-cell leukemia. Previously, we have shown that CAV1 is involved in cell-to-cell communication, cellular proliferation, and immune synapse formation; however, the molecular mechanisms have not been elucidated.We hypothesize that the role of CAV1 in immune synapse formation contributes to immune regulation during leukemic progression, thereby warranting studies of the role of CAV1 in CD4⁺ T-cells in relation to antigen-presenting cells. To address this need, we developed a computational model of a CD4⁺ immune …

Long-Acting Antituberculous Therapeutic Nanoparticles Target Macrophage Endosomes, Benson J. Edagwa, Dongwei Guo, Pavan Puligujja, Han Chen, Joellyn Mcmillan, Xinming Liu, Howard Gendelman, Prabagaran Narayanasamy

Nebraska Center for Biotechnology: Faculty and Staff Publications

Eradication of Mycobacterium tuberculosis (MTB) infection requires daily administration of combinations of rifampin (RIF), isoniazid [isonicotinylhydrazine (INH)], pyrazinamide, and ethambutol, among other drug therapies. To facilitate and optimize MTB therapeutic selections, a mononuclear phagocyte (MP; monocyte, macrophage, and dendritic cell)-targeted drug delivery strategy was developed. Long-acting nanoformulations of RIF and an INH derivative, pentenyl- INH (INHP), were prepared, and their physicochemical properties were evaluated. This included the evaluation of MP particle uptake and retention, cell viability, and antimicrobial efficacy. Drug levels reached 6 μg/106 cells in human monocyte-derived macrophages (MDMs) for nanoparticle treatments compared with 0.1 μg/106 cells for native …

First Proof Of Concept Of Sustainable Metabolite Production From High Solids Fermentation Of Lignocellulosic Biomass Using A Bacterial Co-Culture And Cycling Flush System, Wanying Yao, Sue E. Nokes

Biosystems and Agricultural Engineering Faculty Publications

To improve the lignocellulose conversion for ABE in high solids fermentation, this study explored the feasibility of cycling the process through the cellulolytic or/and solventogenic phases via intermittent flushing of the fermentation media. Five different flushing strategies (varying medium ingredients, inoculum supplement and cycling through phases) were investigated. Flushing regularly throughout the cellulolytic phase is necessary because re-incubation at 65 °C significantly improved glucose availability by at least 6-fold. The solvents accumulation was increased by 4-fold using corn stover (3-fold using miscanthus) over that produced by flushing only through the solventogenic phase. In addition, cycling process was simplified by re-incubating …

Intercellular Cooperation And Competition In Brain Cancers: Lessons From Drosophila And Human Studies, Indrayani Waghmare, Austin Roebke, Mutsuko Minata, Madhuri Kango-Singh, Ichiro Nakano

Biology Faculty Publications

Glioblastoma (GBM) is a primary brain cancer with an extremely poor prognosis. GBM tumors contain heterogeneous cellular components, including a small subpopulation of tumor cells termed glioma stem cells (GSCs). GSCs are characterized as chemotherapy- and radiotherapy-resistant cells with prominent tumorigenic ability. Studies in Drosophila cancer models demonstrated that interclonal cooperation and signaling from apoptotic clones provokes aggressive growth of neighboring tumorigenic clones, via compensatory proliferation or apoptosis induced proliferation. Mechanistically, these aggressive tumors depend on activation of Jun-N-terminal kinase (upstream of c-JUN), and Drosophila Wnt (Wg) in the apoptotic clones. Consistent with these nonmammalian studies, data from several mammalian …

The Exometabolome Of Clostridium Thermocellum Reveals Overflow Metabolism At High Cellulose Loading, Evert K. Holwerda, Philip G. Thorne, Daniel G. Olson, Daniel Amador-Noguez, Nancy L. Engle, Timothy J. Tschaplinski, Johannes P. Van Dijken, Lee R. Lynd

Dartmouth Scholarship

BackgroundClostridium thermocellum is a model thermophilic organism for the production of biofuels from lignocellulosic substrates. The majority of publications studying the physiology of this organism use substrate concentrations of ≤10 g/L. However, industrially relevant concentrations of substrate start at 100 g/L carbohydrate, which corresponds to approximately 150 g/L solids. To gain insight into the physiology of fermentation of high substrate concentrations, we studied the growth on, and utilization of high concentrations of crystalline cellulose varying from 50 to 100 g/L by C. thermocellum. .

Overexpression Of Patatin-Related Phospholipase Aiiiβ Altered The Content And Composition Of Sphingolipids In Arabidopsis, Maoyin Li, Jennifer E. Markham, Xuemin Wang

Department of Biochemistry: Faculty Publications

In plants, fatty acids are primarily synthesized in plastids and then transported to the endoplasmic reticulum (ER) for synthesis of most of the complex membrane lipids, including glycerolipids and sphingolipids. The first step of sphingolipid synthesis, which uses a fatty acid and a serine as substrates, is critical for sphingolipid homeostasis; its disruption leads to an altered plant growth. Phospholipase As have been implicated in the trafficking of fatty acids from plastids to the ER. Previously, we found that overexpression of a patatin-related phospholipase, pPLAIIIβ, resulted in a smaller plant size and altered anisotropic cell expansion. Here, we determined the …

Enhanced Expression Of Codon Optimized Mycobacterium Avium Subsp. Paratuberculosis Antigens In Lactobacillus Salivarius, Christopher D. Johnston, John P. Bannatine, Rodney Govender, Lorraine Endersen, Daniel Pletzer, Helge Weingart, Aidan Coffey, Jim O'Mahony, Roy D. Sleator

Department of Biological Sciences Publications

It is well documented that open reading frames containing high GC content show poor expression in A+T rich hosts. Specifically, G+C-rich codon usage is a limiting factor in heterologous expression of Mycobacterium avium subsp. paratuberculosis (MAP) proteins using Lactobacillus salivarius. However, re-engineering opening reading frames through synonymous substitutions can offset codon bias and greatly enhance MAP protein production in this host. In this report, we demonstrate that codon-usage manipulation of MAP2121c can enhance the heterologous expression of the major membrane protein (MMP), analogous to the form in which it is produced natively by MAP bacilli. When heterologously over-expressed, antigenic determinants …

Sensitivity Analysis Of Biological Boolean Networks Using Information Fusion Based On Nonadditive Set Functions, Naomi Kochi, Tomáš Helikar, Laura Allen, Jim A. Rogers, Zhenyuan Wang, Mihaela T. Matache

Department of Biochemistry: Faculty Publications

Background: An algebraic method for information fusion based on nonadditive set functions is used to assess the joint contribution of Boolean network attributes to the sensitivity of the network to individual node mutations. The node attributes or characteristics under consideration are: in-degree, out-degree, minimum and average path lengths, bias, average sensitivity of Boolean functions, and canalizing degrees. The impact of node mutations is assessed using as target measure the average Hamming distance between a non-mutated/wild-type network and a mutated network.

Results: We find that for a biochemical signal transduction network consisting of several main signaling pathways whose nodes …