Biology Commons^™

Development And Characterization Of Monoclonal Antibodies Directed Against Yaba-Like Disease Virus, Scott Haller

Masters Theses

The hybridoma technique has been used for the production of monoclonal antibodies (mAb) directed against yaba-like disease virus (YLDV). Spleen cells, obtained from female BALB/c mice immunized with YLDV, were fused with P3x63.Ag8.653 myeloma cells and hybrids were selected. Hybridoma clones were screened for production of anti-YLDV antibodies by Indirect Immunofluorescence (IF) microscopy. Positive hybridoma clones were recloned via limiting dilution and characterized. A single clone (H12) has been selected that specifically reacted with YLDV infected cells and not uninfected cells. Immunodiffusion technique determined H12 monoclonal antibody as an IgG1 isoptype immunoglobulin. H12 antibodies recognized a late YLDV protein, and …

Identification And Characterization Of Zinc-Responsive Genes In Pseudomonas Fluorescens, Thomas L. Wilson

Masters Theses

Genes responsive to elevated concentrations of zinc were identified via transposon mutagenesis in the common soil bacterium, Pseudomonas fluorescens ATCC13525. Some of these genes were essential to maintain metal homeostasis in the bacterial cell. DNA sequences of the transposon-tagged genes were determined by DNA sequencing of arbitrary PCR products. DNA sequence analysis indicated that one gene was similar to P-type ATPases responsible for transporting metal ions out of the cell. The corresponding mutant was sensitive to zinc, cadmium, and lead indicating that this gene may be responsible for defending Pseudomonas fluorescens against these metal ions. Four zinc-induced genes shared similarity …

Cadmium Induced Gene Expression And Signal Transduction Pathway, Daisuke Maki

Masters Theses

Cells respond to cadmium (Cd), a common environmental contaminate, by potentially decreasing reduced intracellular glutathione, as well as, increasing the expression of several antioxidant genes. One of these antioxidant genes, glucose-6- phosphate dehydrogenase (G6PDH), is the rate limiting enzyme of the pentose phosphate pathway. This pathway produces NADPH which is used for maintenance of reduced glutathione so, it is hypothesized that Cd should increase G6PDH expression. In this study, we demonstrate that Cd increases the level of G6PDH mRNA in primary rat hepatocytes in culture. In the presence of the antioxidants, N-acetylcysteine (NAC), Trolox, or glutathione monoethyl ester an attenuation …

Role Of Dopamine D4 Receptor In The Genomic And Behavioral Effects Of Psychostimulants: Relevance To Schizophrenia, Bina Garimella

Masters Theses

The role of dopamine D4 receptors in the induction and expression of behavioral sensitization to amphetamine, cocaine and PCP was investigated in this study. Behavioral changes and genomic responses accompanying sensitization were examined using a highly selective D4 receptor antagonist, PNU-101387G. Induction and expression of behavioral sensitization to cocaine was blocked by PNU-101387G. However, the D4 antagonist did not modulate the induction and expression of behavioral sensitization to PCP. Neuroadaptive genomic responses accompanying acute and chronic effects of amphetamine and their modulation by PNU-101387G was examined by looking at alterations in the expression of two immediate-early genes: c-fos and NGFI-A. …

The Role Of Nf-Кb Activation In Breast Carcinoma And Glioblastoma Cell Lines, Kelly T. Murphy

Masters Theses

The nuclear factor-КB (NF-КB)/Rel family of inducible dimeric transcription factors regulates numerous genes associated with the immune, mitogenic, and apoptotic responses. Investigations were made into the role of NF-КB in breast carcinoma (MCF-7, T47D, MDA-MB-231) and glioblastoma (U87MG, U373MG, Ul 18MG) cell lines. NF-КB binding to DNA was quantified using both gel mobility shift assays and transient transfections of a vector containing a luciferase gene under the control of 4 tandem copies of the NF-КB binding sequence. To determine the correlation between NF-КB activation, cellular proliferation, and foci formation, the tumor cells were cultured and analyzed in the presence of …

Isolation And Characterization Of A Genetic Locus That Confers Tolerance To Cadmium And Zinc In Pseudomonas Fluorescens, Szi Fei Feng

Masters Theses

This thesis describes the isolation and characterization of a region containing several genes involved in metal tolerance in the common soil bacterium, Pseudomonas fluorescens ATCC 13525. The DNA region was identified in a cadmium and zinc sensitive mutant, 45-B10, which contains a lacZ-based reporter gene transposon Tn5-B20 inserted in the chromosome. The DNA surrounding the transposon was cloned from this mutant strain and DNA sequencing of this region revealed seven genes that share similarity with genes whose products are involved in the cation-proton antiporter efflux system of other Gram-negative bacteria. Complete DNA sequence was obtained from four of these …

Effects Of Elevated Carbon Dioxide And Nitrogen Fixation On A Legume-Based System: A Test With Soybean And The Soybean Looper, Pseudoplusia Includens, Angela M. Spickard

Masters Theses

Wild-type and non-nodulated soybean were grown at 367 ± 10 ppm [ambient] and 756 ± 10 ppm [elevated] CO₂ to determine whether nitrogen fixation could minimize the effects of elevated CO₂ on plant nutritional and defensive chemistry. Relative to non-nodulated soybean, wild-type soybean had higher concentrations of leaf nitrogen and proteinase inhibitors, as well as lower C:N ratios and lower phenolics. Elevated CO₂ resulted in decreased leaf nitrogen and proteinase inhibitor contents, and increased C:N ratios and phenolics in both soybean strains. Constitutive levels of phenolics and proteinase inhibitors occurred as predicted by the Carbon/Nutrient Balance hypothesis …

Target-And Stage-Dependent Influences On Axonal Outgrowth Of An Identified Motorneuron In Leech Cns Explants, April Crowley

Masters Theses

The neural circuit that controls rhythmic contraction of the hearts in the medicinal leech, Hirudo medicinalis, provides an amenable model to investigate pathfinding and target selection. The bilaterally paired heart tubes (HT) in leech are eventually innervated by the heart excitor motomeuron (HE) following a period of remodeling that requires the presence of the HT. To begin characterizing possible HT-derived cues, co-cultures of CNS with innervated HT have been used. Previous work demonstrated that young, but not adult, HT was able to support profuse general outgrowth, but the HE extended few projections onto HT of any age. Current studies …

Reproduction Partitioning In Polygyne Nests Of The Yellowjacket Vespula Squamosa, Stephanie A. Stewart

Masters Theses

No abstract provided.

Reactive Oxygen Species Generated By Depletion Of Cellular Glutathione Mediate Cadmium Cytotoxicity, Jun Xu

Masters Theses

Studies were put forth to test whether or not reactive oxygen species, which are influenced by the cellular level of reduced glutathione, mediate the toxicity of cadmium. Cadmium has been a focal point in the health care community in the past few years because it has been shown to induce the development of tumors, interfere with cellular signaling, and cause damage to organs, tissues, and biomolecules. Although various studies and theories have been proposed, little direct evidence, especially from studies utilizing primary cells in culture, has accumulated to show the mechanism by which the cadmium induced toxicity occurs.

Our studies …

Characterization Of Nih-3t3 Mouse Fibroblast Cells Transfected With The Somatostatin Receptor Subtype 2 And The Protein Tyrosine Phosphatase Shp-1, Scott Bradford Cross

Masters Theses

The hormone Somatostatin (SMS) and its analogs have been shown to mediate a variety of cellular actions including inhibition of cellular growth and differentiation of endocrine tumors such as insulinomas and glucagonomas. The mechanisms by which SMS exerts its antitumorigenic effects are poorly understood but are thought to include one of the five Somatostatin receptors (SSTR1-5), and the protein tyrosine phosphatase SHP-1. To determine if the antiproliferative effects of SMS were due to the presence of the SSTR2 receptor or the stimulation of SHP-1, we introduced cDNA's of the SMS receptor SSTR2, SHP-1, and a dominant negative mutant of SHP-1 …

Mechanism By Which Insulin And Its Mimetics, Selenate And Vanadate, Regulate Glucose-6-Phosphate Dehydrogenase Gene Expression, Sanjay M. Jivraj

Masters Theses

Glucose-6-phosphate dehydrogenase (G6PDH) is a key enzyme of the pentose phosphate pathway. It controls the carbon flow through this pathway, producing reducing equivalents in the form of NADPH to meet the cellular need for reductive biosynthesis and the maintenance of the cellular redox state. Hepatic expression of G6PDH has been shown to be regulated by hormones, nutrients and some growth factors, however the mechanism by which these factors regulate G6PDH gene expression has not been characterized. Here we investigate the mechanism by which insulin and its mimetics, selenate and vanadate, regulate G6PDH gene expression.

Insulin exerts its tissue specific affect …

Stage-Dependent Expression Of Target-Derived Influences Stimulates Axon Outgrowth In Hirudo Medicinalis, Tina M. Harik

Masters Theses

Our lab has been investigating the development of neuronal phenotype and circuitry in Hirudo medicinalis. Early in embryogenesis the heart excitor (HE) motor neuron selectively extends multiple arbors into the body wall prior to heart tube (HT) formation. HE contact with HT primordium at later stages of development results in remodeling such that arbors are elaborated on the HT, while those in the body wall are retracted. HT-derived cues are necessary for HE remodeling (Jellies, 1995. Am. Zool. 35:529; Jellies and Kopp, 1995. Invert. Neurosci. 1:145). HT and body wall explants were co-cultured to investigate the possible influences of …

Characterization Of Swinepox Virus Secretory Polypeptides, Takeshi Shimamura

Masters Theses

Swinepox virus (SPV) is the only known member of the genus Suispoxvirus. SPV causes a mild disease in swine characterized by slight fever, and weight loss, associated with lesions restricted to the skin. There have been few studies on SPV. Other poxviruses, such as vaccinia virus, have been well characterized and the biological activities of their secretory polypeptides have been analyzed. This study describes a major SPV secretory polypeptide with an apparent molecular mass of 35kDa. Poxvirus encoded polypeptides secreted during infection are known to exhibit immunomodulatory activities. The 35kDa SPV secretory polypeptide was tested.for its ability to inhibit …

Demographics And Condition Of Coyotes In Illinois, Daniel M. Lloyd

Masters Theses

No abstract provided.

Effect Of Insulin And Vanadate On Sre-Mediated C-Fos Expression And Jnk Activation, Francisco J. Berguido

Masters Theses

Insulin mimetics are compounds that mimic the action of insulin in the body and are currently being studied as possible therapeutic agents against diabetes. Vanadium has been identified as an element with insulin-like properties with regard to metabolic processes such as lowering blood glucose levels. Few studies have implicated vanadium in mitogenic actions such as cell growth and differentiation. We studied the action of vanadate on the expression of the insulin-induced SRE, a cis-element of the well-characterized mitogenic gene c-fos. We measured this expression by means of a plasmid containing the reporter gene luciferase linked to the sequences from c-fos …

Activation Of Signal Proteins By Insulin And Selenate In Primary Rat Hepatocytes, Ginny Lynn Garlock

Masters Theses

Insulin has a wide range of metabolic and mitogenic effects in cells. The mechanism of insulin action in controlling these events is not well understood, but involves, at least in part, a complex phosphorylation cascade. Selenate has been shown to mimic insulin on several metabolic processes, including the regulation of glucose-6-phosphate dehydrogenase (G6PDH) gene expression. Its mechanism of action, however, is not known. To investigate the mechanisms of insulin and selenate action on the regulation of G6PDH, signal proteins from primary rat hepatocytes were first studied. Insulin and sodium selenate similarly increased tyrosyl phosphorylation of the β-subunit of the insulin …

Isolation And Characterization Of Cadmium Hypertolerant Mutants Of Arabidopsis Thaliana, Santiago Xavier Navarro

Masters Theses

Cadmium is an environmental pollutant which accumulates by deposition from anthropogenic activities. Plants readily take-up cadmium from soil and is concentrated in shoot tissues. Plant and animal systems are affected detrimentally by the exposure to cadmium and are known to produce metal binding peptides as part of detoxification mechanisms. In an attempt to find alternative methods of soil cleanup, efforts are being undertaken to isolate heavy metal accumulating plants. Alternatively, to reduce the accumulation of cadmium in food, plants that exclude heavy metals are also being sought. The objective of this project was to isolate mutants which can be used …

The Role Of Free Radicals In The Plant's Response To Pathogen Attack In Tobacco, Phillip L. Peters

Masters Theses

Previous studies have implicated the involvement of free radicals, specifically reactive oxygen species (ROS), as a key component in plant response to pathogen attack. In this study we examined the effect free radical scavenging compounds have on lesion area, salicylic acid levels, catalase, and superoxide dismutase in response to inoculation with Tobacco Mosaic Virus (TMV), a viral pathogen. Hydroponically grown Nicotiana tabacum (Xanthi nc) were pretreated with one of two antioxidants, either purpurogallin (PPG) or N-acetyl cysteine (NAC). Antioxidants were supplied to the plant via the root system and the leaves were subsequently challenged by inoculation with theTMV. Leaf material …

Life History Characteristics Of Daphnia Lumholtzi And Its Susceptibility To An Invertebrate Predator, Eden L. Rawski

Masters Theses

No abstract provided.

Expression And Regulation Properties Of Pseudomonas L-Arabinose Utilization Plasmids, Mary E. Barecki

Masters Theses

The L-arabinose utilization pathway in Pseudomonas fluorescens is a unique metabolic pathway. In Pseudomonas, L-arabinose is converted to the end product α-ketogluterate and does not involve phosphorylated intermediates. This differs from the well studied L-arabinose pathway found in Escherichia coli which contains phosphorylated intermediates.

The enzymes of the L-arabinose pathway have been shown to be induced in the presence of L-arabinose in P. saccharophila. The current study determined regulatory properties of previously cloned chromosomal fragments of P. fluorescens which confer an L-arabinose positive phenotype to the surrogate host, Pseudomonas putida. Cells of P. putida containing plasmids pPZ221, …

The Effects Of Benzo(A)Pyrene And Dimethylbenzanthracene On Cultured Porcine Aortic Endothelial Function, Michelle Lynn Kagey

Masters Theses

Polycyclic aromatic hydrocarbons are environmental contaminants which have been shown to initiate and promote atherosclerosis and carcinogenesis in various animal species. These findings have been based primarily on the increase in incidence and size of smooth muscle cell tumors in these animals were exposed to benzo(a)pyrene (BaP) or dimethylbenzanthracene (DMBA). The endothelium has also been identified as an important mediator in vascular disease and atherogenesis, although the endothelial response to these PAHs has not been investigated. Thus, the focus of this study was on characterization of the endothelial response to BaP and DMBA. Concentration/response relationships for these chemicals were examined, …

Expression Of Α4Β1 Integrin On Tissue Eosinophils In The August Rat, John C. Phipps

Masters Theses

The integrin α4β1 is expressed on a wide variety of peripheral leukocytes, and functions in recruitment of these cells to sites of inflammaiton. We infected august (AUG) rats with the helminth parasite Nippostrongylus brasiliensis and later used leukocytes from the peritoneum of the sensitized animals and antibodies against α4β1 integrin to localize that adhesion molecule on the cell surfaces after antigen challenge. We found that after antigen challenge, eosinophils, monocytes, and small lymphocytes, but not mast cells, expressed α4β1 integrin if the animals had been previously infected, but cells from uninfected animals did not show expression of this molecule. Our …

The Flora And Ecology Of Seidman Park, Kent County, Michigan, Pamela J. Laureto

Masters Theses

A floristic study of Seidman Park, Kent County, Michigan was performed. The study identified 521 vascular plants belonging to 96 families and 284 genera. The plant communities of Seidman Park were delimited by identification of dominant species and observation of the physical environment. It was believed that communities existed within the upland forest that could not be identified by observation alone. The use of Non-Metric Multidimensional Scaling (NMS) ordination techniques identified three communities within the upland forest. These were a xeric community, a dry-mesic community and a nutrient-rich community. Three sub-communities of the dry-mesic community were also identified.

Effects Of Insulin And Insulin-Mimetics On Mrna Levels Of C-Fos And C-Jun In Primary Rat Hepatocytes In Culture, Guangmin Li

Masters Theses

Insulin is capable of regulating cellular and metabolic processes as well as gene expression. Selenate and vanadate are two compounds that have been found to mimic the action of insulin in several tissue and cell types. cFos and cJun are the products of immediate early responsive genes c-fos and c-jun. They are members of the family of the AP-1 (Activator Protein-1) transcription factors. AP-1 transcription factors bind as cJun.cJun homodimer or cJun.cFos heterodimer to the TRE found in the promoter regions of several genes. AP-1 is characterized by its ability to alter gene expression in response to various peptide …

The Role Of Nitric Oxide In The Modulation Of The Leukocyte-Endothelial Cell Interactions During Ischemia And Reperfusion, Fernando López-Neblina

Masters Theses

Nitric oxide (NO) is involved in several metabolic pathways and physiological phenomena. The role of NO in inflammation after ischemia and reperfusion (I/R) is controversial. The early phases of inflammation include the leukocyte-endothelial (L/EC) cell interactions which have been divided in rolling, activation, adhesion and migration.

The first experiment was to demonstrate the role of NO in a model of renal I/R. We demonstrated that exogenous NO improves survival and renal function tests after renal I/R, while endogenous NO does not appear to be an important contributor to renal I/R. We also demonstrated that NO modulate the infiltration of neutrophils …

Electromyographic Comparison Of Internal And External Obliques Using A Modified Version Of Kendall's Strength Test Positions For Upper And Lower Abdominals, Rebecca A. Currier, Margie L. Johnson

Masters Theses

The purpose of this study was to examine the activity of the internal obliques (upper abdominals) versus the external obliques (lower abdominals) during a modified version of Kendall's upper and lower abdominal strength tests using surface electromyography on adults. The study was not correlated to Kendall's theory due to the necessity to have the internal oblique electrode placed on the anterior abdomen. Twenty-four women and sixteen men participated in the study. All subjects were taught two positions ("easy" and "hard") for both abdominal tests and performed eight trials. A normalized ratio was generated by dividing one "hard" trial by the …

Comparison Of The Effects Of Aerobic Dance To Water Aerobic Training On Maximal Oxygen Consumption, Terri L. Bedford, Tamara A. Dusterwinkle, Darcy J. Hoppman

Masters Theses

Water aerobics is one mode of exercise that is gaining popularity. Water aerobics allows adults who participate in an exercise program to improve their cardiorespiratory fitness. The purpose of this study was to determine if water aerobic training is as effective as land dance aerobic training in improving cardiorespiratory fitness in sedentary healthy adults. Eighteen subjects participated in this study; nine in dance aerobics and nine in water aerobics. Before and after eight weeks of training, subjects performed a graded maximal exercise test on a Schwinn Air-Dyne while maximal oxygen uptake was measured via a Beckman Metabolic Cart. ANCOVA was …

Regulation Of Leukotriene C4 Biosynthesis In Calcium Ionophore A23187-Challeneged August Rat Peritoneal Leukocytes, James M. Huebner

Masters Theses

Leukotriene (LT) C₄ is a potent myotropic product of arachidonic acid 5-lipoxygenation that is thought to play a role in the pathophysiology of immediate hypersensitivity reactions and inflammation. We designed the present study to evaluate the capacity of August (AUG) rat mixed peritoneal leukocytes to generate LTC₄ when challenged with different concentrations of the Ca²⁺ ionophore A23187 and to test the hypothesis that LTC₄ biosynthesis is regulated via a protein kinase C (PKC)-dependent mechanism. The AUG rat, which has a spontaneously high number of peritoneal eosinophils that can be harvested by a simple lavage procedure, is …

Identification Of An Insulin-Responsive Element In The Rat Glucose-6-Phosphate Dehydrogenase Gene, Sheri Lynn Holmen

Masters Theses

The expression of several genes has been shown to be either positively or negatively regulated by insulin at the transcriptional level, however no single insulin responsive DNA element has yet been identified. The transcription factors responsible for stimulation or inhibition of gene expression by insulin also remain to be identified. In recent years, expression of the gene for glucose-6-phosphate dehydrogenase (G6PDH) in primary rat hepatocytes has been found to be influenced by hormones such as insulin, dexamethosone, and glucagon. The molecular mechanism(s) however, underlying hormonal regulation of G6PDH gene transcription remains to be elucidated. The focus of this study was …