Life Sciences Commons^™

Considerations For The Use Of Mycorrhizal Fungi Inoculants In Coal Mine Reclamation And Reforestation In Appalachia: A Guide Of Best Practices And Management Recommendations, Cole Hansen Reistrup

Master's Projects and Capstones

Nearly all terrestrial plants form symbiotic relationships with mycorrhizal fungi. These beneficial soil microbes support plant health by increasing plants’ access to nutrients and water, protecting them from pathogens, improving soil properties, and more. A challenge to restoring forests on coal mined land in Appalachia is that mine soils are often degraded in a number of ways, including lacking mycorrhizal fungi, which makes mine soils difficult for trees to grow upon. Substantial improvements to coal mine reforestation success have been made since the development of the Forestry Reclamation Approach in 2005 that provides guidelines for how to create a suitable …

See The Forest For The Leaves: Foliar Fungal Community Composition And Volatile Organic Compound Production, Joshua Copeland

Master's Theses

Endophytes are microbes that live within plants without causing detectable illness. These microorganisms can emit volatile organic compounds (VOCs), airborne signaling molecules that can significantly affect plant growth. Despite the prevalence of endophytes and their VOCs, little is known about these factors important to plant health. With this in mind I isolated fungal endophytes from leaves of Populus trichocarpa and Populus fremontii, tree species with economic and environmental uses, to determine if endophyte communities are reserved between the closely related tree species and over seasons. Surface sterilized leaf fragments were plated onto malt extract agar and the resulting fungi were …

The Contribution Of Foliar Fungi To Agricultural Soil Microbiomes In An Organic Cropping System., Derek R. Newberger

Creative Activity and Research Day - CARD

With finite arable land available to support a growing human population, sustainable agricultural techniques are increasingly needed to provide long-term food security. Traditionally, the focus of using cover crops as a technique in organic agriculture has been on the improved nutrient and physical properties of soil that result from their use. However, recent technological developments in sequencing have enabled a new window into the benefits of plant microbiomes in agriculture that extend beyond these traditionally touted effects. This project will investigate one relatively under-described effect of using cover cropping systems – the transfer of microbes from plants to soil. Specifically, …

Phylogenetic Analysis Of Human Cytomegalovirus Pus27 And Pus28: Ascertaining An Independent Or Linked Evolutionary History, Jessica A. Scarborough

Undergraduate Honors Theses

Human cytomegalovirus (HCMV) is a widespread pathogen that is particularly skilled at evading immune detection and defense mechanisms, largely due to extensive co-evolution with its host’s immune system. One aspect of this co-evolution involves the acquisition of four virally encoded GPCR chemokine receptor homologs, products of the US27, US28, UL33 and UL78 genes. G protein-coupled receptors (GPCR) are the largest family of cell surface proteins, found in organisms from yeast to humans. In this research, phylogenetic analysis was used to investigate the origins of the US27 and US28 genes, which are adjacent in the viral genome. The results indicate that …

Development Of A Non-Invasive Liquid Biopsy For Detection Of Cmvil-10, Alexander Shin, Margarette Mariano, Juliet Spencer

Creative Activity and Research Day - CARD

Human cytomegalovirus (HCMV) is a prevalent type of herpes virus in our population. HCMV infection has no effect on the majority of people, but in some cases HCMV is strongly correlated with various medical outcomes, such as breast cancer. We focus on the UL111A gene product of HCMV, which encodes the secreted protein cmvIL-10. CmvIL-10 is a homolog of human cytokine IL-10 (hIL-10), which has immunosuppressive effects and promotes proliferation and invasion of breast cancer cells in vitro. We are measuring cmvIL-10 in human blood and have found elevated levels of cmvIL-10 in cancer patients. Here, we are investigating the …

Detection Of Hcmv Viral Il-10 (Vil-10) In Healthy Blood Donors, Vivian P. Young

Master's Theses

Human cytomegalovirus (HCMV) is widespread in the general population and can establish lifelong latency with expression of a limited subset of viral genes. The UL111A gene is expressed during both lytic and latent infection, and at least two protein isoforms are produced. During lytic infection, the full length transcript yields cmvIL-10, a potent immunosuppressive viral ortholog of human IL-10 (hIL-10). Alternative splicing of the UL111A transcript yields a truncated protein, LAcmvIL-10, which is expressed during both lytic and latent infection but with a limited range of immunosuppressive functions. The two viral cytokines, collectively termed viral IL-10 (vIL-10), are identical in …

Effect Of Cmvil-10 On Exosome Production By Human Breast Cancer Cells, Susanna N. Basappa

Undergraduate Honors Theses

Human cytomegalovirus (HCMV) is a ubiquitous virus that infects 70-90% of the general population, primarily the immunocompromised, but has been implicated in several forms of cancer, including breast cancer. Breast cancer is the second leading cause of cancer related deaths in women in North America, usually from metastasis. Exosomes are 30-100nm vesicles produced by most cells which carry protein and RNA to cells in their microenvironment. The aim of this study is to investigate the impact of HCMV-infection of a secreted viral cytokine, cmvIL-10, on exosome production by highly metastatic breast cancer cells.

MDA-MB-231 cells were cultured in vitro, …

The Role Of Nitrogen And Phosphorus In The Growth, Toxicity, And Distribution Of The Toxic Cyanobacteria, Microcystis Aeruginosa, James Parrish

Master's Projects and Capstones

Microcystis aeruginosa is among the most common harmful algal-blooming species in the world. Potent microcystins released by M. aeruginosa have been linked to liver failure and death in aquatic mammals, like the endangered California sea otter, and provide a serious public health risk to humans. Once characterized as a freshwater problem, M. aeruginosa is expanding on a global scale, making persistent returns in freshwater, brackish, and coastal marine ecosystems. Though commonly observed dominating aquatic ecosystems in low N:P atomic ratios less than 44:1, the reliability of N:P ratios as a tool for managing and predicting M. aeruginosa blooms is explored …

The Unseen World: Environmental Microbial Sequencing And Identification Methods For Ecologists, Naupaka B. Zimmerman, J. Izard, C. Klatt, J. Zhou, E. Aronson

Biology Faculty Publications

Microorganisms inhabit almost every environment, comprise the majority of diversity on Earth, are important in biogeochemical cycling, and may be vital to ecosystem responses to large-scale climatic change. In recent years, ecologists have begun to use rapidly advancing molecular techniques to address questions about microbial diversity, biogeography, and responses to environmental change. Studies of microbes in the environment generally focus on three broad objectives: determining which organisms are present, what their functional capabilities are, and which are active at any given time. However, comprehending the range of methodologies currently in use can be daunting. To provide an overview of environmental …

Hcmv Promotes Breast Cancer Metastasis: Impacts Of Cmvil-10 In The Tumor Microenvironment, Robin Bishop

Master's Theses

Human cytomegalovirus (HCMV) is a highly species-specific, common human pathogen. While a large majority of people are infected with HCMV worldwide, infection is typically asymptomatic in immune competent individuals. To enhance virus persistence and avoid immune detection by the host, HCMV exploits the strategy of encoding homologs of cellular cytokines. One factor produced by HCMV-infected cells is the viral cytokine cmvIL-10, which acts through the cellular IL-10 receptor to induce downstream cell signaling. In instances where HCMV-infected cells infiltrate the tumor microenvironment, the presence of cmvIL-10 in may initiate events that promote a more invasive tumor phenotype. Cancer cells that …

The Us27 Protein Of Human Cytomegalovirus Enhances Signaling By Human Chemokine Receptor Cxcr4, Kathleen Arnolds, Juliet V. Spencer

Creative Activity and Research Day - CARD

Human cytomegalovirus (HCMV) is a prevalent pathogen that causes little disease except in immune-compromised hosts. HCMV establishes life-long latency by manipulating host immune responses. We have found that the US27 protein profoundly alters the activity of host chemokine receptor CXCR4. Cells expressing US27 and CXCR4 exhibited greater migration and calcium signaling than cells expressing CXCR4 alone. The mechanism for this enhanced signaling appears to be increased expression of CXCR4, and we are continuing to investigate how US27 causes up-regulation of CXCR4. This work is expected to clarify the role of US27 in immune evasion and provide insights into treatment strategies.

Relationship Between Udp-Glucose 4-Epimerase Activity And Oligoglucose Glycoforms In Two Strains Of Neisseria Meningitidis, F K. Lee, B W. Gibson, William Melaugh, A Zaleski, M A. Apicella

Chemistry Faculty Publications

Sodium dodecyl sulfate-polyacrylamide gel analysis of lipooligosaccharide (LOS) from Neisseria meningitidis has demonstrated considerable microheterogeneity in the variable region of LOS due to the presence of novel glycoforms. As a step toward understanding the basis for the expression of these novel glycoforms, we have examined the LOS structures and UDP-glucose 4-epimerase (epimerase) activity levels in two strains (NMB and MA-1) and their respective galE mutants. Strain NMB was found to have low epimerase activity and to contain multiple glycoforms, some of which appear to contain only glucose sugars. The galE mutant had only the oligoglucose glycoforms. Strain MA-1 had higher …

Identification Of The Adp-L-Glycero-D-Manno-Heptose-6-Epimerase (Rfad) And Heptosyltransferase Ii (Rfaf) Biosynthesis Genes From Nontypeable Haemophilus Influenzae 2019, W A. Nichols, B W. Gibson, William Melaugh, N G. Lee, M Sunshine, M A. Apicella

Chemistry Faculty Publications

Haemophilus influenzae is an important human pathogen. The lipooligosaccharide (LOS) of H. influenzae has been implicated as a virulence determinant. To better understand the assembly of LOS in nontypeable H. influenzae (NtHi), we have cloned and characterized the rfaD and rfaF genes of NtHi 2019, which encode the ADP-L-glycero-D-manno-heptose-6-epimerase and heptosyltransferase II enzymes, respectively. This cloning was accomplished by the complementation of Salmonella typhimurium lipopolysaccharide (LPS) biosynthesis gene mutants. These deep rough mutants are novobiocin susceptible until complemented with the appropriate gene. In this manner, we are able to use novobiocin resistance to select for specific NtHi LOS inner core …

Characterization Of A Transposon Tn916-Generated Mutant Of Haemophilus Ducreyi 35000 Defective In Lipooligosaccharide Biosynthesis, B W. Gibson, A A. Campagnari, William Melaugh, M A. Apicella, S Grass, Jing Wang, Katherine L. Palmer, R S. Munson

Chemistry Faculty Publications

To define the role of the surface lipooligosaccharide (LOS) of Haemophilus ducreyi in the pathogenesis of chancroid, Tn916 mutants of H. ducreyi 35000 defective in expression of the murine monoclonal antibody (MAb) 3F11 epitope on H. ducreyi LOS were identified by immunologic screening. One mutant, designated 1381, has an LOS which lacks the MAb 3F11 epitope and migrates with an increased mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gene disrupted by the Tn916 element in strain 1381 was identified by cloning the sequences flanking the Tn916 element. The sequences were then used to probe a lambda DASHII genomic library. …

The Lipooligosaccharides Of Haemophilus Ducreyi Are Highly Sialylated, William Melaugh, A A. Campagnari, B W. Gibson

Chemistry Faculty Publications

The major lipooligosaccharides of the sexually transmitted pathogen Haemophilus ducreyi 35000 have been previously found to terminate in N-acetyllactosamine and sialyl-N-acetyllactosamine, Neu5Ac alpha 2-->3Gal beta 1-->4GlcNAc (W. Melaugh, N. J. Phillips, A. A. Campagnari, M. V. Tullius, and B. W. Gibson, Biochemistry 33: 13070-13078, 1994). In this study, mass spectrometry and composition analyses have shown that the lipooligosaccharides from three other H. ducreyi strains also contain N-acetyllactosamine and are highly sialylated (approximately 30 to 50%), although one African strain was found to contain neither of these structural features.

Use Of Pyocin To Select A Haemophilus Ducreyi Variant Defective In Lipooligosaccharide Biosynthesis, A A. Campagnari, R Karalus, M A. Apicella, William Melaugh, A J. Lesse, B W. Gibson

Chemistry Faculty Publications

Haemophilus ducreyi, a cause of genital ulcer disease in developing countries, appears to facilitate the heterosexual transmission of the human immunodeficiency virus in Africa. Despite an increase in studies of this gram-negative human pathogen, little is known about the pathogenesis of chancroid. Our studies have shown that the lipooligosaccharides (LOS) of H. ducreyi may play an important role in ulcer formation. Monoclonal antibody and mass spectrometric analyses identified a terminal trisaccharide present on H. ducreyi LOS that is immunochemically similar to human paragloboside. This epitope is present on the LOS of Neisseria gonorrhoeae, and it may be the site of …

Investigation Of The Structural Heterogeneity Of Lipooligosaccharides From Pathogenic Haemophilus And Neisseria Species And Of R-Type Lipopolysaccharides From Salmonella Typhimurium By Electrospray Mass Spectrometry, B W. Gibson, William Melaugh, Nancy J. Phillips, M A. Apicella, A A. Campagnari, J M. Griffiss

Chemistry Faculty Publications

Heterogeneity in the lipooligosaccharides (LOS) of pathogenic Haemophilus and Neisseria species is evident from the multiplicity of components observed with electrophoretic analyses. Knowledge of the precise structures that make up these diverse LOS molecules is clearly the key to reaching an understanding of pathogenic processes such as phase variation and molecular mimicry. Except for a few cases, little is known about the specific structural features of LOS that underlie phase variation and molecular mimicry, partly because of the inherent difficulties in the structural elucidation of these complex glycolipids. In the lipopolysaccharides (LPS) from Salmonella typhimurium and Escherichia coli, rough, or …