Microbiology Commons^™

Diagnostic And Therapeutic Potential Of The Over-Expression Of Nucleotide Salvage Pathway Enzymes To The Plasma Membrane Of Solid, Cancerous Tumors, Daniel Sharp, Dr. Kim O'Neill

Journal of Undergraduate Research

Immunotherapy is a developing field in cancer treatment that relies upon the bodies natural defenses to target and eliminate cancerous cells. In order for the immune system to differentiate cancerous tissue from normal tissue there must be some mechanism for targeting only cancerous cells. Without a targeting mechanism, stimulation of the immune system can lead to attack of normal tissue. The use of antibodies to cancer-specific antigens is a viable option to solve this targeting issue. If a cancerous cell specifically expresses a protein on its surface, an antibody to that protein could direct immune cells to the cancerous cell, …

Osteoarthritis And Cartilage, David Holt, Dr. L. C. Bridgewater, Dr. R. E. Seegmiller

Journal of Undergraduate Research

Objective: To test the hypothesis that the spondyloepiphesial dysplasia congenita (sedc) heterozygous (sedc/þ) mouse, a COL2A1 mutant, is a model for the study of osteoarthritis (OA) in the absence of dwarfism and to investigate the presence of HtrA1, Ddr2, and Mmp-13 and their possible involvement in a universal mechanism leading to OA.

Orca Project By Danny Catts And Dr. Johnson, Danny Catts, Dr. Johnson

Journal of Undergraduate Research

After we successfully characterized our virus, we proceeded to investigate the variant’s viral behavior in hopes of finding a difference in comparison to that of the standard. After preliminary steps showed little variation, we took a different direction with the project.

Discovering Protein Interactions Of Hr120gcryab, Kent Jarvis, Dr. Julianne Grose

Journal of Undergraduate Research

The original goal of my project was to find substrates of hR120GCryAB using a yeast two-hybrid model. Over the past few months I was able to achieve this goal. I was able to find over 11 novel substrates that bind with hR120GCryAB. This is new knowledge that had not ever been known before and will hopefully help aid in future treatments of cardiomyopathy.

Novel Nanoinjection Of Yeast Artificial Chromosomes In Mouse Zygotes, Jordan Kitchen, Dr. Sandra Burnett

Journal of Undergraduate Research

Transgenic mice are used to create animal models and are typically generated by the addition of transgenes (4-14 Kb pairs) or artificial chromosomes (150-300 Kb) to the pronucleus of a fertilized mouse egg [1][2]. Because of the size of chromosomes, using them to produce transgenic mice can be very problematic. Current methods for inserting transgenes include delivery through a microinjection device into the pronucleus. Transgenic mice are then created by transplantation of injected eggs into a surrogate female [3]. Larger DNA or yeast artificial chromosomes (YAC) due to the increased amount of genetic material could be used to express an …

Identification Of A Consensus Dna Binding Sequence For Nuclear Gdf5, Michael Baldwin, Dr. Laura Bridgewater

Journal of Undergraduate Research

Nuclear Gdf5 (nGdf5) and nuclear Bmp2 (nBmp2) are newly discovered proteins (1); however, their function has not been determined. They were first found while studying enhancer elements of the Col11a2 gene, and a yeast-one hybrid screen suggested that they could bind to the D/E enhancer of this gene. Further experiments confirmed the presence of these proteins in the nuclei of cells and determined the specific mechanism by which they were translocated to the nucleus (1). This work led to the hypotheses that they function as transcription factors, binding to regulatory regions of target genes to modulate transcription. Two years ago, …

The Unfolded Protein Response Is It The Real Culprit In Osteoarthritis?, Benjamin Fisher, Dr. Laura Bridgewater

Journal of Undergraduate Research

Osteoarthritis (OA) is a disease that affects 26 million of Americans, yet its cause it not understood.1 It is a degenerative disease that affects cartilage at joints, and there are several risk factors that give people a higher probability of developing OA; these risk factors include acute joint injury, obesity, and family history of OA. Scientists are still trying to discover the genetic cause of OA, but no one has been successful at finding the link. In our research, we attempted to use 2 mouse models with genetic mutations to study the proteins involved in OA. Dmm (Disproportionate Micromelia) and …

Humanized Mouse Model For The Study Of Human Herpesvirus 6 Infections, Stephanie Carlson, Dr. Bradford Berges

Journal of Undergraduate Research

While working on this project, we had several unforeseen setbacks. Human herpesvirus 6 (HHV- 6) proved much more difficult to work with than we had supposed. It was tricky to propagate (make it replicate) and hard to quantify. It does not behave like other viruses we have worked with and had a few tricks up its sleeve. We were expecting a dormant virus but were surprised to see a lytic strain. The cells we used to propagate HHV-6 kept dying, so we spent time looking for protocols and reading up on what others have done to maintain the virus. I …

Determining Nucleosome Positioning In Varying Developmental Stages Of Caenorhabditis Elegans, Kellie Bollenbach, Dr. Steve Johnson

Journal of Undergraduate Research

This project consisted of four main parts. The first was to isolate mononucleosome cores from C. elegans that had been pelleted and flash frozen in liquid nitrogen using micrococcal nuclease. The second part was to use an antibody specific to GFP, which was expressed in the germline of the strain of C. elegans we are using and selectively pull out the nucleosomes bound to the GFP using immunoprecipitation, namely those germline cells. From there, we were to sequence the DNA wrapped around those nucleosomes in order to determine where on the DNA those nucleosomes lie and compare it to the …

Effects Of Urban Stream Burial On Organic Matter Dynamics And Reach Scale Nitrate Retention, Jake J. Beaulieu, Paul M. Mayer, Sujay S. Kaushal, Michael J. Pennino, Clay P. Arango, David A. Balz, Timothy J. Canfield, Colleen M. Elonen, Ken M. Fritz, Brian H. Hill, Hodon Ryu, Jorge W. Santo Domingo

All Faculty Scholarship for the College of the Sciences

Nitrogen (N) retention in streams is an important ecosystem service that may be affected by the widespread burial of streams in stormwater pipes in urban watersheds. We predicted that stream burial suppresses the capacity of streams to retain nitrate (NO₃ ⁻) by eliminating primary production, reducing respiration rates and organic matter availability, and increasing specific discharge. We tested these predictions by measuring whole-stream NO₃ ⁻ removal rates using ¹⁵NO₃ ⁻ isotope tracer releases in paired buried and open reaches in three streams in Cincinnati, Ohio (USA) during four seasons. Nitrate uptake lengths were 29 times …

Selected Durability Studies Of Geopolymer Concrete With Respect To Carbonation, Elevated Temperature, And Microbial Induced Corrosion, Mohammad Sufian Badar

Doctoral Dissertations

This thesis reports a comprehensive study related to the experimental evaluation of carbonation in reinforced geopolymer concrete, the evaluation of geopolymer concretes at elevated temperature, and the resistance of geopolymer concrete to microbial induced corrosion (MIC).

Carbonation: Reinforced concretes, made of geopolymer, prepared from two class F fly ashes and one class C fly ash, were subjected to accelerated carbonation treatment for a period of 450 days. Electrochemical, microstructure and pore structure examinations were performed to evaluate the effect of corrosion caused due to carbonation. GPC specimens prepared from class F fly ash exhibited lower corrosion rates by a factor …

Section Abstracts: Biology With Microbiology And Molecular Biology

Virginia Journal of Science

Abstracts of the Biology with Microbiology and Molecular Biology Section for the 92nd Annual Meeting of the Virginia Academy of Science, May 13-15, 2014, Virginia Commonwealth University, Richmond, Virginia

Biochar And Vermicompost Amendments In Vegetable Cropping Systems: Impacts On Soil Quality, Soil-Borne Pathogens And Crop Productivity, Nathan Lee Shoaf

Open Access Theses

Amending soils with biochar and vermicompost has the potential to provide multiple benefits for specialty crops including pathogen suppression and increased crop productivity. Oomycete pathogens such as Phytophthora capsici can devastate crop fields and they are difficult to control presenting significant management challenges for Midwestern growers. Soils amended with a microbially-inoculated softwood biochar increased carrot root growth relative to the control and corn stover biochar in 2011, while the corn stover biochar also increased carrot root growth relative to the control, but only in two varieties. Neither biochar treatment influenced carrot growth in 2012. In greenhouse trials, vermicompost amendments provided …

Effect Of Carvacrol-Loaded Nanoemulsions On A Bioluminescent Strain Of Escherichia Coli O157:H7, Clara Maria Vasquez Mejia

Open Access Theses

Nanoemulsions have been shown to be effective delivery vehicles for poorly water soluble natural antimicrobials. In this study the antimicrobial activity of carvacrol (5-isopropyl-2-methylphenol) nanoemulsions against a bioluminescent Escherichia coli O157:H7 was investigated using light emission as an indicator of cell viability. Different emulsifiers (Ultralec Lecithin and Tween 20), oils (Palm stearin and Coconut oil) and various carvacrol concentrations (0, 1, 2 and 2.5%) were evaluated. Bioluminescence was monitored in situ using a Hamamatsu (photo multiplier tube) sensor module integrated with a Programmable Logic Controller interfaced with a PC for data acquisition. Bioluminescence decreased rapidly with the addition of emulsions …

Biosorption Of Heavy Metals Onto The Surface Of Bacteriophage T4, Zheng Huan Tan

School of Biological Sciences: Dissertations, Theses, and Student Research

Biosorption of heavy metals by bacterial and eukaryotic cell surfaces and the subsequent transport in aqueous environments is well recognized. However, very little is known about the roles viruses play in biosorption. Viruses outnumber prokaryotes and eukaryotes in environmental systems. These organisms represent abundant nanoparticulate organic colloids with reactive surfaces. Here we conducted a series of experiments to assess the biosorption potential of Escherichia coli bacteriophage T4. Adsorption of a heavy metal, Zn²⁺, to the surface of phage T4 was tested in a series of purified phage/metal solutions (0 µM – 1000 µM at 23°C). The Langmuir isotherm …

Comparison Of Two Separation Methods For Biological Particles: Field-Flow Fractionation (Fff) And Sucrose Density Gradients, Cody E. Garrison

OES Theses and Dissertations

Sucrose gradient centrifugation and Field-flow fractionation (FFF) are two different particle separation methods that overcome the problems of similar size microorganisms clumping together during standard filtration methods. FFF separates particles based on size and density via the parabolic velocity profile of laminar flow in a ribbon-like channel. The sucrose method separates particles via centrifugation in a density gradient. Both techniques worked well in separating eukaryotic from prokaryotic microbes, with the preferred method depending on the type and relative abundance of organisms to be separated. Minicells were separated from mother cells in transformed Escherichia coli cultures, heterotrophic flagellates (e.g., Diplonema papillatum …

Reciprocity In Microbiome And Immune System Interactions And Its Implications In Disease And Health., Enayat Nikoopour, Bhagirath Singh

Microbiology & Immunology Publications

Adaptation of the whole microbial normal flora residing in a host to its natural habitat over an evolutionary period has resulted in peaceful coexistence with mutual benefits for both microbiota and host in steady state. This symbiotic relationship between host and microbiota has a significant impact on shaping the immune response in the host to achieve an immune tolerance to microbiota but retaining the ability to respond to invading pathogens. Perturbation of this balance by manipulation of microbial communities in the host can lead to immune dysregulation and susceptibility to diseases. By studying the host in the absence of microbiota …

Deletion Mutant Library For Investigation Of Functional Outputs Of Cyclic Diguanylate Metabolism In Pseudomonas Aeruginosa Pa14, Dae-Gon Ha, Megan E. Richman, George A. O'Toole

Dartmouth Scholarship

We constructed a library of in-frame deletion mutants targeting each gene in Pseudomonas aeruginosa PA14 predicted to participate in cyclic di-GMP (c-di-GMP) metabolism (biosynthesis or degradation) to provide a toolkit to assist investigators studying c-di-GMP-mediated regulation by this microbe. We present phenotypic assessments of each mutant, including biofilm formation, exopolysaccharide (EPS) production, swimming motility, swarming motility, and twitch motility, as a means to initially characterize these mutants and to demonstrate the potential utility of this library.

Microbiota, Oral Microbiome, And Pancreatic Cancer, Dominique S. Michaud, Jacques Izard

Department of Food Science and Technology: Faculty Publications

Only 30% of patients diagnosed with pancreatic cancer survive one year post-diagnosis. Progress in understanding the causes of pancreatic cancer has been made, including solidifying the associations with obesity and diabetes, and a proportion of cases should be preventable through lifestyle modifications. Unfortunately, identifying reliable biomarkers of early pancreatic cancer has been extremely challenging, and no effective screening modality is currently available for this devastating form of cancer. Recent data suggest the microbiota may play a role in the disease process, but many questions remain. Future studies focusing on the human microbiome, both etiologically and as a marker of disease …

Cd46-Mediated Transduction Of A Species D Adenovirus Vaccine Improves Mucosal Vaccine Efficacy, Zenaido T. Camacho, Michael A. Barry, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

The high levels of preexisting immunity against Adenovirus type 5 (Ad5) have deemed Ad5 unusable for translation as a human vaccine vector. Low seroprevalent alternative viral vectors may be less impacted by preexisting immunity, but they may also have significantly different phenotypes from that of Ad5. In this study we compare species D Ads (26, 28, and 48) to the species C Ad5. In vitro transduction studies show striking differences between the species C and D viruses. Most notably, Ad26 transduced human dendritic cells much more effectively than Ad5. In vivo imaging studies showed strikingly different transgene expression profiles. The …

The Wnt/Planar Cell Polarity Pathway Component Vangl2 Induces Synapse Formation Through Direct Control Of N-Cadherin, Tadahiro Nagaoka, Riuko Ohashi, Ayumu Inutsuka, Seiko Sakai, Nobuyoshi Fujisawa, Minesuke Yokoyama, Yina H. Huang, Michihiro Igarashi, Masashi Kishi

Dartmouth Scholarship

Although regulators of the Wnt/planar cell polarity (PCP) pathway are widely expressed in vertebrate nervous systems, their roles at synapses are unknown. Here, we show that Vangl2 is a postsynaptic factor crucial for synaptogenesis and that it coprecipitates with N-cadherin and PSD-95 from synapse-rich brain extracts. Vangl2 directly binds N-cadherin and enhances its internalization in a Rab5-dependent manner. This physical and functional interaction is suppressed by β-catenin, which binds the same intracellular region of N-cadherin as Vangl2. In hippocampal neurons expressing reduced Vangl2 levels, dendritic spine formation as well as synaptic marker clustering is significantly impaired. Furthermore, Prickle2, another postsynaptic …

Cluster M Mycobacteriophages Bongo, Pegleg, And Rey With Unusually Large Repertoires Of Trna Isotypes, Welkin H. Pope, Kirk R. Anders, Madison Baird, Charles A. Bowman, Michelle M. Boyle, Gregory W. Broussard, Tiffany Chow, Kari L. Clase, Shannon Cooper, Kathleen A. Cornely, Randall J. Dejong, Veronique A. Delesalle, Lisa Deng, David Dunbar, Nicholas P. Edgington, Christina M. Ferreira, Kathleen Weston Hafer, Grant A. Hartzog, J. Robert Hatherill, Lee E. Hughes, Khristina Ipapo, Greg P. Krukonis, Christopher G. Meier, Denise L. Monti, Matthew R. Olm, Shallee T. Page, Craig L. Peebles, Claire A. Rinehart, Michael R. Rubin, Daniel A. Russell, Erin R. Sanders, Morgan Schoer, Christopher D. Shaffer, James Wherley, Edwin Vazquez, Han Yuan, Daiyuan Zhang, Steven G. Cresawn, Deborah Jacobs-Sera, Roger W. Hendrix, Graham F. Hatfull

Chemistry & Biochemistry Faculty Publications

Genomic analysis of a large set of phages infecting the common host Mycobacterium smegmatis mc2155 shows that they span considerable genetic diversity. There are more than 20 distinct types that lack nucleotide similarity with each other, and there is considerable diversity within most of the groups. Three newly isolated temperate mycobacteriophages, Bongo, PegLeg, and Rey, constitute a new group (cluster M), with the closely related phages Bongo and PegLeg forming subcluster M1 and the more distantly related Rey forming subcluster M2. The cluster M mycobacteriophages have siphoviral morphologies with unusually long tails, are homoimmune, and have larger than average genomes …

A Genomewide Overexpression Screen Identifies Genes Involved In The Phosphatidylinositol 3-Kinase Pathway In The Human Protozoan Parasite Entamoeba Histolytica, Amrita B. Koushik, Brenda H. Welter, Michelle L. Rock, Lesly A. Temesvari

Publications

Entamoeba histolytica is a protozoan parasite that causes amoebic dysentery and liver abscess. E. histolytica relies on motility, phagocytosis, host cell adhesion, and proteolysis of extracellular matrix for virulence. In eukaryotic cells, these processes are mediated in part by phosphatidylinositol 3-kinase (PI3K) signaling. Thus, PI3K may be critical for virulence. We utilized a functional genomics approach to identify genes whose products may operate in the PI3K pathway in E. histolytica. We treated a population of trophozoites that were overexpressing genes from a cDNA library with a near-lethal dose of the PI3K inhibitor wortmannin. This screen was based on the rationale …

Deep Rna Sequencing Reveals Hidden Features And Dynamics Of Early Gene Transcription In Paramecium Bursaria Chlorella Virus 1, Guillaume Blanc, Michael Mozar, Irina V. Agarkova, James R. Gurnon, Giane Yanai-Balser, Janet M. Rowe, Yuannan Xia, Jean-Jack Riethoven, David D. Dunigan, James L. Van Etten

Nebraska Center for Virology: Faculty Publications

Paramecium bursaria chlorella virus 1 (PBCV-1) is the prototype of the genus Chlorovirus (family Phycodnaviridae) that infects the unicellular, eukaryotic green alga Chlorella variabilis NC64A. The 331-kb PBCV-1 genome contains 416 major open reading frames. A mRNA-seq approach was used to analyze PBCV-1 transcriptomes at 6 progressive times during the first hour of infection. The alignment of 17 million reads to the PBCV-1 genome allowed the construction of single-base transcriptome maps. Significant transcription was detected for a subset of 50 viral genes as soon as 7 min after infection. By 20 min post infection (p.i.), transcripts were detected for …

Chip-Seq And Rna-Seq Reveal An Amrz-Mediated Mechanism For Cyclic Di-Gmp Synthesis And Biofilm Development By Pseudomonas Aeruginosa, Christopher J. Jones, David Newsom, Benjamin Kelly, Yasuhiko Irie, Laura K. Jennings, Binjie Xu, Dominique H. Limoli, Joe J. Harrison, Matthew R. Parsek, Peter White, Daniel J. Wozniak

Biology Faculty Publications

The transcription factor AmrZ regulates genes important for P. aeruginosa virulence, including type IV pili, extracellular polysaccharides, and the flagellum; however, the global effect of AmrZ on gene expression remains unknown, and therefore, AmrZ may directly regulate many additional genes that are crucial for infection. Compared to the wild type strain, a ΔamrZ mutant exhibits a rugose colony phenotype, which is commonly observed in variants that accumulate the intracellular second messenger cyclic diguanylate (c-di-GMP). Cyclic di-GMP is produced by diguanylate cyclases (DGC) and degraded by phosphodiesterases (PDE). We hypothesized that AmrZ limits the intracellular accumulation of c-di-GMP through transcriptional repression …

Retinoic Acid Regulation Of Thyroid Hormone Action In Bone Cells, Anjali Babbar

Loma Linda University Electronic Theses, Dissertations & Projects

Retinoic acid and thyroid hormone are known to play key roles in the regulation of endochondral ossification. However, the issue of whether these two hormones interact with each other to regulate bone functions remains to be established. We investigated how thyroid hormone and retinoic acid interact to regulate cells involved in endochondral bone formation. We demonstrate that thyroid hormone treatment stimulates differentiation of ATDC5 chondrocytes and promotes formation of mineralized nodules while retinoic acid treatment at high dose inhibits chondrocyte differentiation and formation of mineralized nodules. Furthermore, thyroid hormone induced mineralized nodule formation is inhibited by co-treatment with retinoic acid …

Vaccines Within Vaccines: The Use Of Adenovirus Types 4 And 7 As Influenza Vaccine Vectors, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

Adenovirus Types 4 and 7 (Ad4 and Ad7) are associated with acute respiratory distress (ARD). In order to prevent widespread Ad-associated ARD (Ad-ARD) the United States military immunizes new recruits using a safe and effective lyophilized wildtype Ad4 and Ad7 delivered orally in an enteric-coated capsule. We cloned Ad4 and Ad7 and modified them to express either a GFP-Luciferase (GFPLuc) fusion gene or a centralized influenza H1 hemagglutinin (HA1-con). BALB/c mice were injected with GFPLuc expressing viruses intramuscularly (i.m.) and intranasally (i.n.). Ad4 induced significantly higher luciferase expression levels as compared with Ad7 by both routes. Ad7 transduction was restored …

Cluster M Mycobacteriophages Bongo, Pegleg, And Rey With Unusually Large Repertoires Of Trna Isotypes, Welkin H. Pope, Kirk R. Anders, Madison Baird, Charles A. Bowman

University Faculty Publications and Creative Works

Genomic analysis of a large set of phages infecting the common host Mycobacterium smegmatis mc2155 shows that they span considerable genetic diversity. There are more than 20 distinct types that lack nucleotide similarity with each other, and there is considerable diversity within most of the groups. Three newly isolated temperate mycobacteriophages, Bongo, PegLeg, and Rey, constitute a new group (cluster M), with the closely related phages Bongo and PegLeg forming subcluster M1 and the more distantly related Rey forming subcluster M2. The clusterMmycobacteriophages have siphoviral morphologies with unusually long tails, are homoimmune, and have larger than average genomes (80.2 to …

The Toxavapa Toxin-Antitoxin Locus Contributes To The Survival Of Nontypeable Haemophilus Influenzae During Infection, Dabin Ren, Alexis A. Kordis, Daniel E. Sonenshine, Dayle A. Daines

Biological Sciences Faculty Publications

Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen that is a common cause of acute and recurrent mucosal infections. One uncharacterized NTHi toxin-antitoxin (TA) module, NTHI1912-1913, is a host inhibition of growth (higBA) homologue. We hypothesized that this locus, which we designated toxAvapA, contributed to NTHi survival during infection. We deleted toxAvapA and determined that growth of the mutant in defined media was not different from the parent strain. We tested the mutant for persistence during long-term in vitro co-culture with primary human respiratory tissues, which revealed that the DeltatoxAvapA mutant was attenuated for survival. We then …

Cluster M Mycobacteriophages Bongo, Pegleg, And Rey With Unusually Large Repertoires Of Trna Isotypes, Welkin H. Pope, Kirk R. Anders, Madison Baird, Charles A. Bowman

University Faculty Publications and Creative Works

Genomic analysis of a large set of phages infecting the common host Mycobacterium smegmatis mc2155 shows that they span considerable genetic diversity. There are more than 20 distinct types that lack nucleotide similarity with each other, and there is considerable diversity within most of the groups. Three newly isolated temperate mycobacteriophages, Bongo, PegLeg, and Rey, constitute a new group (cluster M), with the closely related phages Bongo and PegLeg forming subcluster M1 and the more distantly related Rey forming subcluster M2. The clusterMmycobacteriophages have siphoviral morphologies with unusually long tails, are homoimmune, and have larger than average genomes (80.2 to …