Master of Science (MS), Bowling Green State University, 2018, Biological Sciences

To our knowledge there are no current infectious retroviruses found in canines or wild canids. It has been previously thought that the canine reference genome consists only of about 0.15% of sequence of obvious retroviral origin, present as endogenous retroviruses (ERVs) within contemporary canids. In recent analyses of the canine reference genome, a few copies of ERVs were identified with features characteristic of recent integration, for example the presence of some ORFs and near-identical LTRs. Members of this group are referred to as CfERV-Fc1(a) and have been identified to have sequence similarity to the mammalian ERV-Fc/W groups. We have recently discovered and characterized a number of non-reference Fc1 copies in dogs and wild canids, and identified unexpectedly high levels of polymorphism among members of this ERV group. Some of the proviruses we have identified even possess complete or nearly intact open reading frames, identical LTRs, and derived phylogenetic clustering among other CfERV-Fc1(a) members. Based on LTR sequence divergence under an applied dog neutral mutation rate, it is thought these infections occurred within as recently as the last ~0.48 million years. There have been previous, but unsubstantiated, reports of reverse transcriptase activity as well as gamma-type C particles in tumor tissues of canines diagnosed with lymphoma. We hypothesize that expression of members of the CfERV-Fc1(a) lineage is responsible for those observations in canine cancers. We investigated the expression of individual proviruses in canine cancer cell lines, specifically the pol and env gene. There was expression of both genes in three canine-cancer derived cells lines that cluster with CfERV-Fc1(a) members. Clustering of these sequences also suggest that there is either a new sub lineage of CfERV-Fc1(a) or possibly missed polymorphic proviral insertions that are currently assembled as solo LTRs.

Committee: Julia Halo Ph.D. (Advisor); Raymond Larsen Ph.D. (Committee Member); Zhaohui Xu Ph.D. (Committee Member) Subjects: Molecular Biology

PhD, University of Cincinnati, 2014, Engineering and Applied Science: Environmental Science

Cyanotoxins, a family of emerging contaminants, have become a great public and environmental concern due to their extremely high toxicity and widespread presence of cyanotoxin-producing bacteria in aquatic systems. Cyanotoxins and cyanobacteria have been included in the Drinking Water Contaminant Candidate List (CCL 1 and 2) by U.S. Environment Protection Agency (EPA). Cylindrospermopsin (CYN) and microcystin-LR (MC-LR) are two of most commonly found cyanotoxins and have been listed in the newest CCL 3, along with anatoxin-a. TiO2 photocatalysis is one of the effective technologies for cyanotoxins treatment. This dissertation explored the potential of polymorphic TiO2 based photocatalysts for the degradation of cyanotoxins as well as the corresponding reaction mechanisms. The photocatalytic destruction of CYN was investigated using a novel polymorphic TiO2 photocatalyst (PM- TiO2, containing anatase, brookite, and rutile phases) under UV-Vis irradiation. The PM-TiO2 was characterized using several characterization techniques. The impacts of operation parameters, such as photocatalyst loading, pH and presence of natural organic matter, on the treatment efficiency were also examined in this work. The finding of this work can provide fundamental information for the properties of PM-TiO2 and its environmental applications for CYN treatment. The reaction byproducts of CYN during the TiO2 photocatalysis process were further studied using mass spectrometry. Since hydroxyl radical (•OH) is considered to be the main reactive agent of TiO2 photocatalysis, the reaction pathways were proposed based on hydroxyl radical chemistry and detected reaction byproducts. Hydroxylation, sulfate elimination and ring opening reactions on the hydroxymethyl uracil moiety and tricyclic guanidine moiety were proposed to be the main reaction mechanisms of photocatalytic reaction. Besides, detoxification studies based on the cytotoxicity of CYN proved the effective toxicity removal of CYN-con (open full item for complete abstract)

Committee: Dionysios Dionysiou Ph.D. (Committee Chair); Mallikarjuna Nadagouda Ph.D. (Committee Member); Margaret Kupferle Ph.D. P.E. (Committee Member); George Sorial Ph.D. (Committee Member) Subjects: Environmental Science

Master of Science in Biological Sciences, Youngstown State University, 2008, Department of Biological Sciences and Chemistry

Wangiella dermatitidis is a melanized, pathogenic, polymorphic fungus that in vitro grows primarily in a yeast form, but is capable of morphological changes resulting in growth as either hyphae (filaments) or swollen, multicellular bodies. The latter morphology strongly resembles the in vivo form of tissues infected with W. dermatitidis and other closely-related fungi. Each of these three morphologies can be readily induced through varying growth conditions of the wild-type strain or by culture of developmental mutants. Of particular interest is the conversion of the yeast phase to the multicellular growth form, which is the focus of the present study. Whole-cell, protein profiles of the wild-type strain and the temperature-sensitive mutant, Mc3, were developed following incubation at both 25°C and 37°C (body temperature). At both temperatures, the wild type grew as yeast cells, whereas strain Mc3 also grew as a yeast at 25°C, but formed the multicellular growth form at 37°C. Protein profiles showed consistency in landmark proteins found within all four study groups, as well as temperature dependent and strain dependent proteins. Fifty protein spots were excised and sequenced by capillary-liquid chromatography-nanospray tandem mass spectrometry. A Mascot search of established databases revealed putative identities of these proteins. The identified proteins included: heat shock protein 60 mitochondrial precursor, NADP-dependent mannitol dehydrogenase, and copper-zinc superoxide dismutase (Cu-Zn SOD). The results presented in this study indicate the relevance of proteomics as a tool in interpreting the morphological and physiological conditions of phase changes exhibited by W. dermatitidis.

Committee: Chester R. Cooper PhD (Advisor); Gary R. Walker PhD (Committee Member); John Caguiat PhD (Committee Member) Subjects: Microbiology; Molecular Biology; Public Health

Master of Science (MS), Wright State University, 2010, Microbiology and Immunology

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a disrupter, of B-cell differentiation, induces binding of the aryl hydrocarbon receptor (AhR) nuclear complex to dioxin responsive elements (DRE) within the mouse immunoglobulin heavy chain regulatory region (3'IgHRR), and produces a marked inhibition of 3'IgHRR activation, IgH expression, and antibody secretion in a well-characterized mouse B-cell line (CH12.LX). The mouse 3'IgHRR consists of at least four enhancers (hs3a; hs1,2; hs3b; hs4), and is highly homologous with the three enhancers (hs3; hs1,2; hs4) of the human 3'IgHRR. A polymorphism of the human hs1,2 enhancer (resulting in varying numbers of tandem repeats containing a DRE and κB site) has been correlated with several autoimmune diseases. Although the human and mouse hs1,2 enhancers are share a ~90% identity, luciferase reporter studies in mouse CH12.LX B-cells showed that TCDD inhibited LPS stimulation of the mouse hs1,2 enhancer but co-treatment with TCDD and LPS synergistically activated human hs1,2 enhancer activity. To evaluate transcriptional differences between the human and mouse hs1,2 enhancers, our objectives were to characterize the IM-9 cells as a potential human B-cell model, and to evaluate TCDD-induced transcriptional regulation of the polymorphic human hs1,2 enhancer in a human cell line. We confirmed AhR expression and TCDD-induced CYP1A1 induction in IM-9 cells. Then we transiently transfected IM-9 cells with the human hs1,2 reporters and determined that TCDD activates the human hs1,2 enhancer in IM-9 B-cells, as seen in CH12.LX B-cells. However, the TCDD-induced fold-activation in human IM-9 cells appeared less compared to results in mouse CH12.LX B-cells perhaps due to differences between the mouse and human AhR. Our data suggests that the TCDD-induced inhibition of the mouse hs1,2 enhancer versus the activation of the human hs1,2 enhancer may be related to an inhibitory BSAP site located on the mouse hs1,2 enhancer that is absent from th (open full item for complete abstract)

Committee: Courtney E Sulentic PhD (Advisor); Michael Leffak PhD (Committee Member); Nancy Bigley Phd (Committee Member) Subjects: Biochemistry; Biology; Biomedical Research; Cellular Biology; Environmental Science; Immunology; Microbiology; Molecular Biology; Pharmacology; Toxicology

MS, Kent State University, 2009, College of Arts and Sciences / Department of Biological Sciences

SUMMARY In this study, I examined genetic diversity and levels of population differentiation between eight populations of the invasive plant multiflora rose (Rosa multiflora Thunberg Ex. Murray) located in Portage and Summit Counties, in northeastern Ohio. I used six microsatellite marker loci to determine allelic diversity, percent polymorphic loci, and expected and observed heterozygosity. My results show that multiflora rose populations included have moderate levels of genetic variation. I found a total of twenty five alleles in the six microsatellite loci that were examined. On an average, there were 4.16 alleles per locus, but the average number of alleles per locus observed in each population ranged between 1.66 and 3.33. Effective number of alleles ranged between 1.38 and 2.17, indicating variation in allele frequencies among loci. Average observed heterozygosity (HO = 0.15) was less than expected heterozygosity (HE = 0.43) across all but one population. This trend of heterozygosity deficiency and F statistics results indicate significant level of inbreeding in the studied populations. Most of the loci failed to conform to Hardy-Weinberg expectations in all populations indicating forces of evolution interacting. Analysis of molecular variance revealed significant levels of genetic differentiation between populations. Nei's unbiased estimators of genetic distance range from nearly 0 to 0.59, indicating high genetic differentiation. These findings are then discussed in the light of ecological and historical considerations.

Committee: Oscar Rocha (Advisor); Barbara Andreas (Committee Member); Andrea Case (Committee Member) Subjects: Biology

Master of Science in Chemistry, Cleveland State University, 2007, College of Science

The polymorphs of Tolbutamide and Nifedipine were differentiated by measuring their unique physical properties. Differential Scanning Calorimetry (DSC) and Dielectric Analysis (DEA) verified the variations in these compounds with different crystal structures but the same chemical constituents. The crystalline-amorphous content of the polymorphs were determined based on a new DEA protocol. Solubility (w/w%) and spectroscopic methods clearly distinguished these drugs and compared favorably with the photomicrography of the polymorphs. Powder X-ray Diffraction (PXRD) analysis succintly showed differences in the various types of polymorphs. Scanning Electron Microscopy (SEM) supported the structural distinction and solubility. The FTIR spectra of all the polymorphs were reproducible and illustrated differences between tolbutamide and nifedipine but not within its polymorphs. The analytical techniques developed in this study were also used in analyzing blends of drugs and excipients, e.g. Aspirin and Polyethylene Glycol (PEG). Co-crystallization of drugs with PEG, an excepient, was measured with increases in the PEG heat of fusion beyond its blended concentration. The extra heat associated with the PEG heat of fusion is due to the crystallization of the amorphous drugs in the PEG matrix. These unique co-crystallized blends will yield enhanced drug delivery since PEG is very water soluble.

Committee: Xue-Long Sun (Advisor) Subjects: