Cell and Developmental Biology Commons^™

Phenotypic And Genotypic Analysis Of Amino Acid Metabolism In Lactobacillus Helveticus Cnrz 32, Jason K. Christiansen

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This study investigated genetic predictions for amino acid biosynthesis and catabolism by Lactobacillus helveticus CNRZ 32, a commercial cheese flavor adjunct that reduces bitterness and intensifies flavor notes. Conversion of amino acids into volatile and nonvolatile flavor compounds by L. helveticus and other lactic acid bacteria in cheese is thought to represent the rate-limiting step in the development of mature cheese flavor and aroma. One of the primary mechanisms for amino acid breakdown by these microbes involves the reversible action of enzymes involved in biosynthetic pathways, so our group investigated the genetics of amino acid biosynthesis in L. helveticus CNRZ …

Effect Of Calcium Channel Antagonists And Sex On Bone Metabolism In Aged Brown Norway Rats, Brent Kidd

Morehead State Theses and Dissertations

A thesis presented to the faculty of the College of Science & Technology at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science by Brent Kidd on April 19, 2007.

Stability Of Eukaryotic Elongation Factor One Alpha Protein In The Cell, Jennifer Watry

Honors Theses

Eukaryotic Elongation Factor One Alpha (eEF1A) is an abundant protein found in every cell of every organism and is involved in functions concerning protein synthesis and degradation, the cytoskeleton, and signaling. How these functions are regulated is not yet understood for this protein. Regulation of the protein may be controlled by transcription of the genes encoding the protein or the stability and modification of the protein. The goal of this project was to determine the stability of eEf1A proteins in Arabidopsis thaliana. Protein stability was determined by treating tissues with cycloheximide, an inhibitor of protein synthesis. Gel electrophoresis and …

Down-Regulation Of Natural Killer Cell Activation In Response To Influenza Virus In Older Adults, Yu Jing

Theses and Dissertations in Biomedical Sciences

Immune senescence contributes to influenza-associated high mortality and morbidity and reduced vaccine efficacy in elderly people. Type I T cell (Thl)-mediated immunity plays a significant role in Immune responses to influenza infection and vaccination. Natural killer (NK) cells secrete significant amount of IFN-7 , a hallmark Thl cytokine, in response to influenza infection. How aging influences human NK cell IFN-7 production in response to influenza virus has not been well documented. In this study we employed human peripheral blood mononuclear cells (PBMC) and performed intracellular cytokine staining and flow cytometry primarily to investigate how aging influences NK cell activation with …

A 368-Base-Pair Cis-Acting Hwp1 Promoter Region, Hcr, Of Candida Albicans Confers Hypha-Specific Gene Regulation And Binds Architectural Transcription Factors Nhp6 And Gcf1p, Samin Kim, Michael J. Wolyniak, Janet F. Staab, Paula Sundstrom

Dartmouth Scholarship

To elucidate the molecular mechanisms controlling the expression of the hypha-specific adhesin gene HWP1 of Candida albicans, its promoter was dissected and analyzed using a green fluorescent protein reporter gene. A 368-bp region, the HWP1 control region (HCR), was critical for activation under hypha-inducing conditions and conferred developmental regulation to a heterologous ENO1 promoter. A more distal region of the promoter served to amplify the level of promoter activation. Using gel mobility shift assays, a 249-bp subregion of HCR, HCRa, was found to bind at least four proteins from crude extracts of yeasts and hyphae with differing binding patterns dependent …

The Role Of Stat And The Jak/Stat Pathway In Mediating The Effects Of Interleukin-6 On Star Expression, Janae Strickland

Theses and Dissertations

Cortisol, a hormone produced by a hormone produced by the adrenal gland, is responsible for many regulatory functions in the body. Cortisol release is mediated by adrenocorticotrophic hormone, or ACTH, through the hypothalamus-pituitary-adrenal or HPA axis. This HPA axis is the major release pathway used during acute stress, during which the levels of ACTH parallel those of cortisol. However, in states of chronic stress, the level of ACTH drops dramatically, while cortisol remains high. This study focuses on the pathway of cortisol release during these chronic stress states, specifically examining the role of IL-6 with respect to STATs and the …

Avpv Neurons Containing Estrogen Receptor-Beta In Adult Male Rats Are Influenced By Soy Isoflavones, Edwin D. Lephart, Lihong H. Bu

Faculty Publications

Background Isoflavones, the most abundant phytoestrogens in soy foods, are structurally similar to 17beta-estradiol. It is known that 17beta-estradiol induces apoptosis in anteroventral periventricular nucleus (AVPV) in rat brain. Also, there is evidence that consumption of soy isoflavones reduces the volume of AVPV in male rats. Therefore, in this study, we examined the influence of dietary soy isoflavones on apoptosis in AVPV of 150 day-old male rats fed either a soy isoflavone-free diet (Phyto-free) or a soy isoflavone-rich diet (Phyto-600). Results: The occurrence of apoptosis in AVPV was examined by TUNEL staining. The incidence of apoptosis was about 10 times …

The Yeast Orthologue Of Grasp65 Forms A Complex With A Coiled-Coil Protein That Contributes To Er To Golgi Traffic, Rudy Behnia, Francis A. Barr, John J. Flanagan, Charles Barlowe, Sean Munro

Dartmouth Scholarship

The mammalian Golgi protein GRASP65 is required in assays that reconstitute cisternal stacking and vesicle tethering. Attached to membranes by an N-terminal myristoyl group, it recruits the coiled-coil protein GM130. The relevance of this system to budding yeasts has been unclear, as they lack an obvious orthologue of GM130, and their only GRASP65 relative (Grh1) lacks a myristoylation site and has even been suggested to act in a mitotic checkpoint. In this study, we show that Grh1 has an N-terminal amphipathic helix that is N-terminally acetylated and mediates association with the cis-Golgi. We find that Grh1 forms a complex with …

Following Temperature Stress, Export Of Heat Shock Mrna Occurs Efficiently In Cells With Mutations In Genes Normally Important For Mrna Export, Christiane Rollenhagen, Christine A. Hodge, Charles N. Cole

Dartmouth Scholarship

Heat shock leads to accumulation of polyadenylated RNA in nuclei of Saccharomyces cerevisiae cells, transcriptional induction of heat shock genes, and efficient export of polyadenylated heat shock mRNAs. These studies were conducted to examine the requirements for export of mRNA following heat shock. We used in situ hybridization to detect SSA4 mRNA (encoding Hsp70) and flow cytometry to measure the amount of Ssa4p-green fluorescent protein (GFP) produced following heat shock. Npl3p and Yra1p are mRNA-binding proteins recruited to nascent mRNAs and are essential for proper mRNA biogenesis and export. Heat shock mRNA was exported efficiently in temperature-sensitive npl3, yra1 …

Intermolecular Interactions Of Homologs Of Germ Plasm Components In Mammalian Germ Cells, Mark S. Fox, Amander T. Clark, Mohammed El Majdoubi, Jean-Louis Vigne, Jun Urano, Chris E. Hostetler, Michael D. Griswold, Richard I. Weiner, Renee A. Reijo Pera

Collected Faculty Scholarship

In some species such as flies, worms, frogs, and fish the key to forming and maintaining early germ cell populations is the assembly of germ plasm, croscopically-distinct egg cytoplasm that is rich in RNAs, RNA-binding proteins and ribosomes. Cells which inherit germ plasm are destined for the germ cell lineage. In contrast, in mammals, germ cells are formed and maintained later in development as a result of inductive signaling from one embryonic cell type to another. Research advances, using complementary approaches, including identification of key signaling factors that act during the initial stages of germ cell development, differentiation of germ …

Glycaemic Index, Glycaemic Load And Risk Of Endometrial Cancer: A Prospective Cohort Study, Stephanie A. Navarro Silvera, Thomas E. Rohan, Meera Jain, Paul D. Terry, Geoffrey R. Howe, Anthony B. Miller

Department of Public Health Scholarship and Creative Works

Objective

High-glycaemic-load diets may increase endometrial cancer risk by increasing circulating insulin levels and, as a consequence, circulating oestrogen levels. Given the paucity of epidemiological data regarding the relationship between the dietary glycaemic index and glycaemic load and endometrial cancer risk, we sought to examine these associations using data from a prospective cohort study.

Design, setting, and subjects

We examined the association between dietary glycaemic load and endometrial cancer risk in a cohort of 49 613 Canadian women aged between 40 and 59 years at baseline who completed self-administered food-frequency questionnaires between 1982 and 1985. Linkages to national mortality and …

Mechanism Of Action And Regulation Of Membrane Serine Protease Prostasin In The Prostate And Prostate Cancer, Mengqian Chen

Electronic Theses and Dissertations

The glycosylphosphatidylinositol (GPI)-anchored serine protease prostasin (PRSS8) is expressed at the apical membrane surface of epithelial cells and acts as a suppressor of tumor invasion when re-expressed in highly invasive human prostate and breast cancer cell lines. To better understand the molecular mechanisms underlying the anti-invasion phenotype associated with prostasin re-expression in prostate cancer cells, we expressed wild-type human prostasin or a serine active-site mutant prostasin in the PC-3 human prostate carcinoma cells. Molecular changes were measured at the mRNA and the protein levels. The expression of several invasion-promoting molecules is regulated by prostasin re-expression, mediated by a protein-level down-regulation …

A Pollen Chase Experiment; Examining Varying Levels Of Embryonic Inbreeding Depression, Emily Wilson, Judy Stone

Undergraduate Research Symposium (UGRS)

A pollen chase experiment was performed upon three Costa Rican populations of Witheringia solanacea to examine the breakdown of genetically enforced self incompatibility (SI) and the extent of embryonic inbreeding depression. Self-pollen was applied in the bud, with outcross pollen applied one day later, and outcross pollinations at both intervals as a control. A variety of responses were found among the populations. BOHS readily accepted self pollen and suffered from very low inbreeding depression. Monteverde and Las Cruces both have lower fruit set with self-pollination precedence indicating that bud pollinations can overcome the self-incompatibility response and that embryonic death due …

Ecological Correlates Of Buggy Creek Virus Infection In Oeciacus Vicarius, Southwestern Nebraska, 2004, Amy T. Moore, Eric A. Edwards, Mary Bomberger Brown, Nicholas Komar, Charles R. Brown

School of Natural Resources: Faculty Publications

Buggy Creek virus (family Togaviridae, genus Alphavirus, BCRV) is an alphavirus within the western equine encephalitis virus complex whose primary vector is the swallow bug, Oeciacus vicarius Horvath (Hemiptera: Cimicidae), an ectoparasite of the colonially nesting cliff swallow, Petrochelidon pyrrhonota, that is also a frequent host for the virus.We investigated ecological correlates of BCRV infection in 100-bug pools at 14 different swallow colony sites in southwestern Nebraska from summer 2004, by using plaque assay on Vero cells to identify cytopathic virus and reverse transcription-polymerase chain reaction to identify noncytopathic viral RNA. We found 26.7% of swallow bug pools …

The Expression And Function Of Ppar And Hif-1 In Human Melanoma, Caroline Mills

Theses, Dissertations and Capstones

The first part of my dissertation focuses on the expression and function of PPARs in human melanoma. I found that the A375 cells were significantly growth inhibited in response to _PGJ2 and troglitazone treatment. HEMn-LP showed significant growth inhibition in response to troglitazone. I found that PPARγ and PPARδ mRNA is present in both the SK-Mel 28 and A375 cells. The relative level of PPARα mRNA expression is highest in SK-Mel 28 cells, ~3 fold higher relative to both the normal human melanocytes and A375 cells. PPARγ protein was ~50% higher in both SK-Mel 28 and A375 cells relative to …

Arsenic Toxicity In Plhc-1 Cell Line And The Distribution Of Arsenic In Central Appalachia, Yeong Nam Jeong

Theses, Dissertations and Capstones

Our ecosystem has geobiophysical relationships. Environmental pollution can lead to immediate consequences like organism death or lack of reproduction. It can also lead to long-range consequences through mutations and DNA damage. Fish can be affected very directly by environmental contamination in water but the study of fish is not performed well. We are developing a quantitative assay to assess DNA damage in vitro due to heavy metal contamination using a tissue culture line of Poeciliopsis lucida, the desert pupfish, which is used for other toxicology assays. To detect DNA damage, Single Cell Gel Electrophoresis (Comet Assay) is very useful. …

Apoptosis Of Dedifferentiated Hepatoma Cells Is Independent Of Nf-Jb Activation In Response To Lps, M. Reidy, Janette Ellis, Erin Schmitz, David Kraus, Gary Bulla

Faculty Research & Creative Activity

Dedifferentiated hepatoma cells, in contrast to most other cell types including hepatoma cells, undergo apoptosis when treated with lipopolysaccharide (LPS) plus the protein synthesis inhibitor cycloheximide (CHx). We recently reported that the dedifferentiated hepatoma cells also exhibit a strong and prolonged NF-jB induction phenotype upon exposure to LPS, suggesting that NF-jB signaling may play a pro-survival role, as reported in several other cell systems. To test the role of NF-jB in preventing LPS-mediated apoptosis, we examined the dedifferentiated cell line M38. Results show that antioxidants strongly inhibited LPS + CHx-mediated cell death in the M38 cells, yet only modestly inhibited …

Lim Kinase 1 Modulates Expression Of Matrix Metalloproteinases And Associates With Gamma-Tubulin: Dual Role In Invasion And Mito, Tenekua Tapia

Electronic Theses and Dissertations

LIM kinase 1 (LIMK1) is a unique dual specificity serine/threonine kinase containing two N-terminal LIM domains in tandem, a PDZ domain and a C-terminal catalytic domain. LIMK1 is involved in modulation of actin cytoskeleton through inactivating phosphorylation of the ADF (actin depolymerization factor) family protein cofilin. Recent studies have shown that LIMK1 is upregulated in breast and prostate cancer cells and tissues, promotes metastasis in animals and induces acquisition of an invasive phenotype when ectopically expressed in benign prostate epithelial (BPH) cells. Furthermore, overexpression of LIMK1 was associated with altered sub cellular localization of the membrane type 1 matrix metalloprotease …

Apoptosis Of Dedifferentiated Hepatoma Cells Is Independent Of Nf-Jb Activation In Response To Lps, M. Ryan Reidy, Janette Ellis, Erin A. Schmitz, David M. Kraus, Gary A. Bulla

Faculty Research & Creative Activity

Dedifferentiated hepatoma cells, in contrast to most other cell types including hepatoma cells, undergo apoptosis when treated with lipopolysaccharide (LPS) plus the protein synthesis inhibitor cycloheximide (CHx). We recently reported that the dedifferentiated hepatoma cells also exhibit a strong and prolonged NF-jB induction phenotype upon exposure to LPS, suggesting that NF-jB signaling may play a pro-survival role, as reported in several other cell systems. To test the role of NF-jB in preventing LPS-mediated apoptosis, we examined the dedifferentiated cell line M38. Results show that antioxidants strongly inhibited LPS + CHx-mediated cell death in the M38 cells, yet only modestly inhibited …

Characterization Of Afn1, A Gene Associated With Cereal Grain Germination, Tenzin Tsewang, Russell Johnson

Undergraduate Research Symposium (UGRS)

The AFN1 gene is transiently expressed in germinating oat grains. As AFN1 is not expressed in dormant oat grains during imbibition, we hypothesize that AFN1 may be involved in stimulating the germination process. Sequence analysis of an AFN1 cDNA clone indicates that the AFN1 polypeptide is similar to a previously identified abscisic acid (ABA) glucosyl transferase. This suggests that AFN1 may be acting to glucosylate ABA, thereby inactivating it. As the hormone ABA is known to inhibit germination, ABA glucosylation/inactivation could lead to germination in grains expressing AFN1. To test this hypothesis, we have constructed an expression plasmid that encodes …

The Effects Of Nematode Infection And Mi-Mediated Resistance In Tomato (Solanum Lycopersicum) On Plant Fitness, Brandon P. Corbett

Inquiry: The University of Arkansas Undergraduate Research Journal

The Mi gene in tomato (Solanum lycopersicum) is a single, dominant resistance (R) gene that confers resistance against several species of insects and root-knot nematodes. Tis study examined the impact of root-knot nematode infestation and the plant growth and reproduction of near-isogenic tomato cultivars with and without Mi. The objectives of this experiment were to examine the potential fitness costs and benefits of the R gene-mediated herbivore resistance, and to explore the role of nematodes as a selection pressure favoring plants that carry Mi. Mi-mediated resistance dramatically reduced nematode reproduction on tomato. In the presence of nematodes, plants that carried …

Erythropoietin Improves Skeletal Muscle Microcirculation And Tissue Bioenergetics In A Mouse Sepsis Model, Raymod Kao, Anargyros Xenocostas, Tao Rui, Weixiong Huang, James Rose, Claudio M. Martin

Anatomy and Cell Biology Publications

Introduction The relationship between oxygen delivery and consumption in sepsis is impaired, suggesting a microcirculatory perfusion defect. Recombinant human erythropoietin ( rHuEPO) regulates erythropoiesis and also exerts complex actions promoting the maintenance of homeostasis of the organism under stress. The objective of this study was to test the hypothesis that rHuEPO could improve skeletal muscle capillary perfusion and tissue oxygenation in sepsis. Methods Septic mice in three experiments received rHu-EPO 400 U/kg subcutaneously 18 hours after cecal ligation and perforation ( CLP). The first experiment measured the acute effects of rHuEPO on hemodynamics, blood counts, and arterial lactate level. The …

New Microanatomical Characters For Scorpion Taxonomy (Arachnida: Scorpiones), Michael Brewer

Theses, Dissertations and Capstones

Morphological characters used in scorpion systematics are limited to a few reliable structures. The most recent high-level, comprehensive revision of scorpion phylogeny, 38 of the 105 morphological characters were based on trichobothria – socketed mechanosensory seta (Soleglad & Fet, 2003). Other microstructures such as granules, setae, spines, etc. have been found in scorpions but their use for taxonomy was limited. In an effort to better understand the complex relationships among these ancient but similar looking animals, we attempted to first locate new microstructures and then assess their reliability and variability thus determining the levels at which they are diagnostic. The …

Poxviral B1 Kinase Overcomes Barrier To Autointegration Factor, A Host Defense Against Virus Replication, Matthew S. Wiebe, Paula Traktman

Nebraska Center for Virology: Faculty Publications

Barrier to autointegration factor (BAF) is a DNA-binding protein found in the nucleus and cytoplasm of eukaryotic cells that functions to establish nuclear architecture during mito-sis. Herein, we demonstrate a cytoplasmic role for BAF in host defense during poxviral infections. Vaccinia is the prototypic poxvirus, a family of DNA viruses that replicate ex-clusively in the cytoplasm of infected cells. Mutations in the vaccinia B1 kinase (B1) com-promise viral DNA replication, but the mechanism by which B1 achieves this has remained elusive. We now show that BAF acts as a potent inhibitor of poxvirus replication unless its DNA-binding activity is blocked …

Ultrafine Particle Initiation Of Lipid Peroxidation In Mitochondria, Josephine Kimberly Wells

Undergraduate Honors Theses

Studies have shown that car exhaust particles increase cellular oxidative stress (Li, 2002), and people who live in highly polluted areas have shown a higher incidence of cardiovascular disease (Beck, 2005). Recently a theory has been developed that ultrafine particles (< 0.1 µm in diameter) contained in the atmospheric pollution are responsible for initiating the adverse effects of damage and disease (Li, 2003).

Ultrafine particles (UFP), which make up 72% of the particle concentration in the air (Beck, 2005), typically originate from sources involving combustion, such as cars, forest fires, volcanoes and industry (USDHHS, 1995). The small size and abundance of UFPs provide ample surface area with an increased absorption for damaging substances such as iron or reactive organic compounds (Beck, 2005; …

Modulation Of Retroviral Restriction And Proteasome Inhibitor-Resistant Turnover By Changes In The Trim5Α B-Box 2 Domain, Felipe Diaz-Griffero, Alak Kar, Michel Perron, Shi-Hua Xiang, Hassan Javanbakht, Xing Li, Joseph Sodroski

Nebraska Center for Virology: Faculty Publications

An intact B-box 2 domain is essential for the antiretroviral activity of TRIM5α. We modeled the structure of the B-box 2 domain of TRIM5α based on the existing three-dimensional structure of the B-box 2 domain of human TRIM29. Using this model, we altered the residues predicted to be exposed on the surface of this globular structure. Most of the alanine substitutions in these residues exerted little effect on the antiretroviral activity of human TRIM5αhu or rhesus monkey TRIM5α_rh. However, alteration of arginine 119 of TRIM5α_hu or the corresponding arginine 121 of TRIM5α_rh diminished the abilities of …

Distribution And Conservation Genetics Of The Cow Knob Salamander, Plethodon Punctatus Highton (Caudata: Plethodontidae), Matthew R. Graham

Theses, Dissertations and Capstones

Being lungless, plethodontid salamanders respire through their skin and are especially sensitive to environmental disturbances. Habitat fragmentation, low abundance, extreme habitat requirements, and a narrow distribution of less than 70 miles in length, makes one such salamander, Plethodon punctatus, a species of concern (S1) in West Virginia. To better understand this sensitive species, day and night survey hikes were conducted through ideal habitat and coordinate data as well as tail tips (10 to 20 mm in length) were collected. DNA was extracted from the tail tips and polymerase chain reaction (PCR) was used to amplify mitochondrial 16S rRNA gene fragments. …

Self-Consistent Analyses For Potential Conduction Block In Nerves By An Ultrashort High-Intensity Electric Pulse, R. P. Joshi, A. Mishra, Q. Hu, K. H. Schoenbach, A. Pakhomov

Bioelectrics Publications

Simulation studies are presented that probe the possibility of using high-field (>100kV ∕ cm), short-duration (∼50ns) electrical pulses for nonthermal and reversible cessation of biological electrical signaling pathways. This would have obvious applications in neurophysiology, clinical research, neuromuscular stimulation therapies, and even nonlethal bioweapons development. The concept is based on the creation of a sufficiently high density of pores on the nerve membrane by an electric pulse. This modulates membrane conductance and presents an effective "electrical short" to an incident voltage wave traveling across a nerve. Net blocking of action potential propagation can then result. A continuum approach based …

Synthesis And Antifungal Properties Of Alpha-Methoxy And Alpha-Hydroxyl Substituted 4-Thiatetradecanoic Acids, Nestor Carballeira, Rosann O'Neill, Keykavous Parang

Pharmacy Faculty Articles and Research

4-Thiatetradecanoic acid exhibited weak antifungal activities against Candida albicans (ATCC 60193), Cryptococcus neoformans (ATCC 6603 1), and Aspergillus niger (ATCC 16404) (MIC = 4.8-12.7 mM). It has been demonstrated that alpha-methoxylation efficiently blocks P-oxidation and significantly improve the antifungal activities of fatty acids. We examined whether antifungal activity of 4-thiatetradecanoic acid can be improved by a-substitution. The unprecedented (+/-)-2-tiydroxy-4-thiatetradecanoic acid was synthesized in four steps (20% overall yield), while the (+/-)-2-methoxy-4-thiatetradecanoic acid was synthesized in five steps (14% overall yield) starting from 1-decanethiol. The key step in the synthesis was the hydrolysis of a trimethylsilyloxynitrile. In general, the novel (+/-)-2-methoxy-4-thiatetradecanoic …

The Zebrafish As A Model Of Ventricular And Arterial Pole Malformations, Adrian Colin Grimes

MUSC Theses and Dissertations

The zebrafish is becoming an almost ubiquitously accepted model for the study of comparative developmental biology, its utility coming to rival that of the fruitfly, frog, chick and mouse. Unfortunately, however, it is widely held belief that the zebrafish cardiac outflow tract makes it unusual as a model of comparative cardiogenesis and this is based, in part, on the report that bulbus arteriosus undergoes a developmental myocardium-to-smooth muscle phenotypic transition. This would imply that the zebrafish may be poorly suited to the study of human cardiac outflow abnormalities. While the use of chamber-specific molecular markers has allowed extensive characterization of …