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Development Of An Electrotransformation Technique For Streptococcus Iniae And Preliminary Characterization Of The Hemolysin Associated With This Bacterium, Kimberly Ann Hahn

Biological Sciences Theses & Dissertations

Streptococcus iniae is a gram-positive organism responsible for causing disease in both freshwater as well as saltwater fish. Every year fisheries lose fish by the tons due to diseases caused by this organism. In 1991, the first reported human case of disease associated with this organism was described. Since this initial case, there have been numerous other reported cases of S. iniae infections. This organism is a catalase negative, facultatively anaerobic organism that produces a capsule and when plated onto blood agar demonstrates β-hemolytic activity.

In this study, the relationship between the hemolysin produced by S. iniae and the lactate …

Map Kinases In Cynomolgus Monkey Sperm Hyperactivation, Emily Nicole Haynes

Biological Sciences Theses & Dissertations

Mammalian sperm exhibit characteristic motility patterns, termed hyperactivated (HA) motility, associated with completion of capacitation. In cynomolgus monkey (Macaca fascicularis) sperm, this HA motility is dependent in vitro upon the addition of exogenous cyclic nucleotide mediators, caffeine and dibutyryl-cyclic adenosine monophosphate (dbcAMP). Previous reports have shown protein tyrosine phosphorylation to be an integral component of this caffeine- and cAMP-stimulated HA motility. This study investigated the involvement of the mitogen-activated protein (MAP) kinase-signaling cascade. Semen specimens were collected in Talp-HEPES medium from proven breeders via electroejaculation. After washing, sperm were incubated in the presence and absence of the MAP …

Fine Root Dynamics In Atlantic White Cedar (Chamaecyparis Thyoides) Wetlands: Patterns Of Ecosystem Development In Natural And Restoration Settings, Harvey Leroy Rodgers

Biological Sciences Theses & Dissertations

This work addresses seasonal and successional factors of root dynamics in natural and restoration Atlantic white cedar (AWC) wetlands. Using minirhizotrons, fine root dynamics in a chronosequence of naturally-regenerating and restoration AWC wetlands were measured to compare ecosystem development trends with restoration conditions. A second study was conducted to compare fine root dynamics in AWC wetlands with contrasting hydroperiods. In addition, a technical study was conducted to determine the feasibility of estimating root biomass with indirect minirhizotron measurements.

Root biomass estimates using minirhizotrons were significantly higher in naturally regenerating AWC wetlands and significantly lower in the restoration site relative to …

Systematics Of The Subterranean Amphipod Genus Stygobromus (Crangonyctidae) In Western North America, With Emphasis On Species Of The Hubbsi Group, Daqing Wang

Biological Sciences Theses & Dissertations

This dissertation presents a study of the systematics of the subterranean amphipod genus Stygobromus of family Crangonyctidae in western North America. A total of 53 species of Stygobromus from western North America (including one species from Wisconsin) are recognized, 28 of which are new to science. All species are described and illustrated using external morphological features. A key to the western North America species of Stygobromus is given, as well as a summary of habitats of the species.

Cladistic analyses are conducted using 32 external morphological characters and two categories of outgroup choices: “all-zero” theoretical ancestor and three non-Stygobromus …

Trichodesmium Spp.: Numerical Studies Of Resource Competition, Carbohydrate Ballasting, And Remote-Sensing Reflectance, Tonya Denise Clayton

OES Theses and Dissertations

In recent years, a new appreciation for the role of diazotrophy in the oceans has emerged. This dissertation reports on three modeling studies designed to investigate ecological processes associated with Trichodesmium spp., the most conspicuous marine diazotroph: (1) characterization of a generalized model Trichodesmium and issues of macronutrient resource competition; (2) carbohydrate ballasting by Trchodesmium and implications for the formation of surface accumulations; and (3) the vertical distribution of Trichodesmium and implications for detection from space.

The first study focuses on issues of nitrogen and phosphorus competition and ecosystem structure. It utilizes a simple ecosystem model that includes dissolved nitrogen …

Positive Regulation Of Pka On Human Gonadotropin-Releasing Hormone (Hgnrh) Gene Expression In Human Placental Jeg-3 Cells, Zhaoyang Wen

Chemistry & Biochemistry Theses & Dissertations

Using the human placental choriocarcinoma JEG-3 cell line as an in vitro human placental model, we studied the mechanisms of the PKA positive regulation of the hGnRH gene expression in the human placenta. Studies in JEG3 cells showed that through the PKA catalytic subunit a, human GnRH upstream promoter activity was stimulated by PKA signaling pathway in a cAMP dependent mechanism. The sequence between —202 (Afl II) and —554 (BamH I) base pair in the human GnRH upstream promoter region appeared to be responsible for the PKA positive regulation of the gene expression. Furthermore, Western blot analysis demonstrated the involvement …

Estrogen Regulation Of The Pregnant Baboon Placental 11Β-Hydroxysteroid Dehydrogenase (11Β-Hsd)-Catalyzed Metabolism Of Cortisol And Cortisone And Its Effect On The Development Of The Fetal Hypothalamic-Pituitary-Adrenal Axis, William Adair Davies

Theses and Dissertations in Biomedical Sciences

Pepe and Albrecht previously demonstrated that the estrogen-regulated change in transuteroplacental metabolism of cortisol (F) and cortisone (E) from preferential reduction (E to F) at midgestation to oxidation (F to E) near term results in a decline of bioactive cortisol crossing the placenta and reaching the fetus culminating in activation of the hypothalamic-pituitary adrenal axis of the baboon and the ontogenesis of rate-limiting steroidogenic enzymes culminating in de novo F secretion. Protein kinase A (PK-A) activity in the baboon fetal adrenal gland cytosolic fraction was increased 2-fold both at term [day 165 of gestation (term = 184 days)] and following …

Interactions Among Murine Cytomegalovirus Us22 Family Gene Products That Influence Viral Pathogenesis, Zaruhi Karabekian

Theses and Dissertations in Biomedical Sciences

Cytomegalovirus (CMV) is a complex, ubiquitous herpesvirus that is characterized by acute, chronic, and latent infections. Monocytes-macrophages are the key target cell type involved in pathogenesis, which is most effectively studied using the murine model of CMV infection. Previously three murine CMV (MCMV) genes (M139, M140, and M141) were identified to regulate viral expression in cultured macrophages and in mice. These genes are members of the US22 gene family with respect to HCMV homology. There is no function assigned to the proteins encoded by these genes. However, deletion of M139, M140, and M141 significantly curtails growth of MCMV in macrophages …

Regulation Of The Hcmv Ul98 Promoter By Cooperation Of The Cyclic Amp Response Element And Gamma Interferon Response Element, Tabmitha Patricia Young Jervey

Theses and Dissertations in Biomedical Sciences

The HCMV UL98 early alkaline exonuclease gene promoter was examined to determine the DNA sequences as well as viral and/or cellular proteins functional in the regulation of this early gene. To assess promoter activation, UL98 promoter sequences were first cloned upstream of the chloramphenicol acetyltransferase (CAT) reporter gene and cotransfected with expression plasmids which express the HCMV major immediate early (IE) proteins 1E72 and 1E86. To more specifically determine the importance of individual cis-acting elements in UL98 promoter activation, the promoter region underwent mutagenesis to delete or alter sequences. The variant promoters were again cloned into a reporter-CAT construct and …

The Effect Of Nitrogen Fertilization On Short-Term Fine Root Dynamics In A Barrier Island Dune Community, John Walter Hutton

Biological Sciences Theses & Dissertations

Fine root dynamics are an important yet poorly understood component of terrestrial ecosystems. In recent years a number of researchers have focused on fine root dynamics; their work has looked at patterns across one or multiple growing seasons on a scale of monthly measures. The purpose of this study was to add to the overall understanding by looking at fine root dynamics on a shorter temporal scale. Minirhizotron observation tubes were used to assess the effect of nitrogen fertilization on short-term fine root dynamics in a nutrient limited, barrier island dune system. Root length elongation and mortality rates followed expected …

A Comparison Of Phytoplankton Communities In Lake Prince And The Western Branch Reservoir, Suffolk, Virginia, Cara Marie Muscio

Biological Sciences Theses & Dissertations

Lake Prince and the Western Branch Reservoir are two eutrophic bodies of water located in Suffolk, Virginia. Lake Prince and its two small tributaries join the Western branch via a constructed spillway. This lake system is a source of water for the surrounding municipalities, and a recreational area for community citizens. In the past, these bodies of water had repeated incidents of low oxygen and nuisance algal blooms, particularly cyanobacteria species. As a result aerators have been installed in the main body of both Lake Prince and the Western Branch Reservoir. In addition, a pipeline has been installed from North …

Investigations Into The Innate Resistance Of The Tick, Dermacentor Variabilis, When Challenged With The Bacterium, Escherichia Coli, Shane Michael Ceraul

Biological Sciences Theses & Dissertations

In addition to the soluble response, many invertebrates control bacterial infections by means of phagocytosis or melanotic encapsulation. In some insects, Escherichia coli growth is reported to be inhibited by aggregation/encapsulation. Soluble and phagocytic responses to bacterial challenge have been reported in ticks, but evidence of an aggregation/encapsulation response was reported only for inanimate (araldite) implants. This study was done to determine how ticks control infection by E. coli. Ticks were challenged by direct inoculation of bacteria into the hemocoel cavity. Using plate counts, no viable E. coli were detected I hour post-inoculation. A direct fluorescence assay (DF A) …

Effect Of Magnesium On The Production Of Scna And Scnr By Streptococcus Pyogenes, Strain Ff22, Laura E. Quinn

Biological Sciences Theses & Dissertations

The lantibiotic SA-FF22, produced by Streptococcus pyogenes strain FF22, was the first described antimicrobial peptide produced by a Streptococcus species. S. pyogenes is a group A Streptococcus responsible for such human illness as strep-throat, rheumatic fever, necrotizing fasciitis, endocarditis, and meningitis. Previous studies of SA-FF22 have shown that antimicrobial activity is lost in the presence of lmM magnesium. We hypothesize that the lack of SA-FF22 activity in the presence of magnesium is due to an absence of transcription of scnA, the gene encoding SA-FF22. The lack of transcriptional activation of scnA may be due to an absence of ScnR, a …

Detection Of Aneuploidy For Chromosomes 7 And 8 Using Fluorescence In Situ Hybridization In Patients With Aplastic Anemia And Sequencing Of The Mitotic Checkpoint Gene Hbub1, Laura Jane Aridgides

Theses and Dissertations in Biomedical Sciences

Aplastic anemia (AA) is characterized by complete bone marrow failure. Progression to myelodysplastic syndromes (MDS) and acute nonlymphocytic leukemia (ANLL) occurs frequently. At the time of transformation, cytogenetic abnormalities are common. Detection of cytogenetic abnormalities prior to leukemic transformation may indicate future disease progression. Karyotype analysis is the current method of choice to evaluate chromosome aberrations. However, fluorescence in situ hybridization (FISH) is more sensitive in detecting these abnormalities.

hBUB1, a mitotic spindle checkpoint gene, was shown to be mutated in two colorectal cancer cell lines with high levels of aneuploidy (Cahill, et al., 1998). Although theoretically possible, conclusive …

Characterization Of Murine Cytomegalovirus M142 And M143 Essential Gene Products, Bridget Linette Dalton

Theses and Dissertations in Biomedical Sciences

Cytomegalovirus (CMV) is a species-specific virus belonging to the Herpesviridae family. This DNA virus causes severe disease or even death in newborns and immunosuppressed patients. Murine CMV (MCMV) provides an opportunity to study the role of various viral products in replication and pathology in the natural host. Successful replication of CMV in the host depends upon the expression of a cascade of viral genes: immediate early (IE), early (E), and late (L). To date only three MCMV IE proteins have been characterized (IE1, IE2 and IE3). Our laboratory recently identified a novel IE gene region within the Hind III I …

Effects Of Human Disturbances On The Behavior Of Dabbling Ducks, Melissa Lynn Pease

Biological Sciences Theses & Dissertations

Disturbance of wintering and migrating birds by human activities can cause birds to expend energy in avoidance of humans and reduces the time available for resting and feeding at a time in the annual cycle when fat deposition and energy conservation are important. Also, human disturbances can effectively cause habitat loss by displacing birds from feeding or resting habitat. Managers of natural resources are increasingly faced with decisions about the types and amounts of public use that should be allowed without lowering the value of the resource for wildlife. In order for managers to make decisions about how to minimize …

Seasonal Phytoplankton Assemblages Of Five Ephemeral Ponds In York County, Virginia, Michelle R. Kokolis

Biological Sciences Theses & Dissertations

Five ephemeral ponds were identified in and around the Grafton Ponds Natural Area Preserve, approximately 56 km north of the City of Norfolk, Virginia. These ponds, which are filled seasonally by precipitation and groundwater discharge, range from 0.1 ha to 2 ha in size and from 4 cm to 3 meters in depth. Ponds for the study were chosen based on variations in their size, depth, basin cover material, canopy openness, and historical inundation period. The purpose of this study is to relate differences in the phytoplankton assemblages to the physical and chemical characteristics of the ponds. All five ponds …

In Support Of A Rationally Managed Fishery: Age And Growth In Patagonian Toothfish (Dissostichus Eleginoides), Julian R. Ashford

Biological Sciences Theses & Dissertations

Patagonian toothfish (Dissostichus eleginoides) occur on the continental shelves and shelf breaks of southern South America and the Southern Ocean. Stock structure, critical to good fisheries management, can be inferred from growth differences between areas, but available growth data are compromised by inconsistencies in age estimation methods, sampling and sample sizes, and techniques used to derive estimates. I asked the scientific question: how is growth in Patagonian toothfish structured spatially within the Southern Ocean? I developed a multi-stage randomized design to sample fish caught by commercial longline, and an age estimation methodology. Because toothfish are difficult to age, …

The Implications Of The Target-Area Hypothesis On The Population Dynamics Of The Spotted Spiny Lobster, Panulirus Guttatus, Denice N. Robertson

Biological Sciences Theses & Dissertations

The target-area hypothesis, based on the theory of island biogeography, predicts that larger islands are more effective at intercepting passive immigrants. Most marine invertebrates have meroplanktonic larvae and open population dynamics, so immigration to populations in isolated benthic habitats is primarily by pelagic larval recruits. Thus, recruitment to isolated habitat “islands” may be more continuous and predictable on large islands than on small ones. Consequently, populations on large islands should not only be larger than those on small islands, but should also have more evenly distributed size structures. These differences in size structure among populations in isolated habitats of differing …

Characterization Of The Biological Activities Of Recombinant Fusion Protein Green Fluorescent Protein/Human Zona Pellucida Protein 3 (Gfp/Hzp3), Zhiyong Lin

Theses and Dissertations in Biomedical Sciences

Despite numerous reports indicating the successful production of bioactive recombinant ZP3, no report has shown the rhZP3 having direct binding activity with human sperm. Recombinant ZP3 generated from our previous study displayed binding activity with human sperm through indirect evidence from hemizona assay (HZA).

This present study focused on the production of recombinant ZP3 with direct binding activity with human sperm. Through the application of a pEGFP expression vector, fusion protein GFP/ZP3 was successfully generated and expressed. The expression of GFP/ZP3 was evidenced by RT-PCR and western blot. The fusion protein was partially purified by Ni-NTA affinity column from cell …

Characterization Of The Human Cytomegalovirus Ul75 Late Gene Promoter, Bernard J. P. Mcwatters

Theses and Dissertations in Biomedical Sciences

Gene expression during productive infection by the human cytomegalovirus (HCMV) occurs in an ordered and sequential manner, beginning with immediate early (IE), then early (E) and finally late (L) gene expression. Significant work has addressed the regulation of IE and E gene expression while relatively little work has addressed the control of late gene expression. In order to further address HCMV late gene expression, the promoter of the HCMV UL75 (glycoprotein H, gH) late gene was characterized. The data obtained in this study were combined with observations made in two other studies that have addressed HCMV late gene expression to …