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Full-Text Articles in Molecular Biology

Ectopic Expression Of The Arabidopsis Homomeric Acetyl-Coa Carboxylase, Troy Matthew Bunch Jan 2019

Ectopic Expression Of The Arabidopsis Homomeric Acetyl-Coa Carboxylase, Troy Matthew Bunch

Graduate Theses and Dissertations

A number of economic and environmental imperatives have spurred an increased demand for biologically sourced, chemically reduced forms of carbon. Fatty acids, and closely related derivatives, represent an abundant form of such biological carbon. The biosynthesis of these molecules requires acetyl-CoA as a primer substrate, and malonyl-CoA as the extender substrate. Malonyl-CoA is generated by the carboxylation of acetyl-CoA, a reaction catalyzed by acetyl-CoA carboxylase (ACCase), which is thought to be an important determinant of the rate limiting reaction of fatty acid biosynthesis. In Arabidopsis, as in most dicotyledonous plants, two distinct isozymes catalyze this reaction, a cytosolic homomeric form ...


Development And Characterization Of An Immunologically Humanized And Cancer Xenograft Model In Pigs With Severe Combined Immunodeficiency (Scid), Adeline Nicole Boettcher Jan 2019

Development And Characterization Of An Immunologically Humanized And Cancer Xenograft Model In Pigs With Severe Combined Immunodeficiency (Scid), Adeline Nicole Boettcher

Graduate Theses and Dissertations

Swine with severe combined immunodeficiency (SCID) are an emerging large animal model for biomedical research. There have been several SCID pig models described since our first discovery of naturally occurring SCID with mutations in Artemis (DCLRE1C) in 2012. SCID animals are particularly useful in biomedical research due to their lack of T, B, and sometimes NK cells. Absence of the adaptive immune system allows for human cell and tissue xenotransplantation into these SCID animals. The works described within this thesis are categorized under four main goals: (1) further characterization of the immune system of Art-/- SCID pigs, (2) development of ...


Characterization Of A Mucopolysaccharidosis Type I And Galnac Transferase Deficiency Double Knockout Mouse, Karan Gera Jan 2018

Characterization Of A Mucopolysaccharidosis Type I And Galnac Transferase Deficiency Double Knockout Mouse, Karan Gera

Graduate Theses and Dissertations

The mucopolysaccharidoses (MPSs) are a group of lysosomal storage diseases (LSDs) which are characterized by the aberrant primary storage of glycosaminoglycans (GAGs) in lysosomes of multiple organ and tissue systems. The most commonly diagnosed is MPS Type I (MPS I), caused by mutations in the gene which codes for α-L-iduronidase (IDUA). Biochemically, MPS I is characterized by the aberrant primary lysosomal storage of incompletely degraded dermatan and heparan sulfates, along with a secondary accumulation of gangliosides and other compounds in lysosomes. Its clinical manifestation in severe form leads to early death, characterized by progressive central nervous system disease (with behavioral ...


Mutations In Tubb8 And Human Oocyte Meiotic Arrest, Ruizhi Feng, Qing Sang, Yanping Kuang, Xiaoxi Sun, Zheng Yan, Shaozhen Zhang, Juanzi Shi, Guoling Tian, Anna Luchniak, Yusuke Fukuda, Bin Li, Min Yu, Junling Chen, Yao Xu, Luo Guo, Ronggui Qu, Xueqian Wang, Zhaogui Sun, Miao Liu, Huijuan Shi, Hongyan Wang, Yi Feng, Ruijin Shao, Renjie Chai, Qiaoli Li, Qinghe Xing, Rui Zhang, Eva Nogales, Li Jin, Lin He, Mohan L. Gupta Jr., Nicholas J. Cowan, Lei Wang Jan 2016

Mutations In Tubb8 And Human Oocyte Meiotic Arrest, Ruizhi Feng, Qing Sang, Yanping Kuang, Xiaoxi Sun, Zheng Yan, Shaozhen Zhang, Juanzi Shi, Guoling Tian, Anna Luchniak, Yusuke Fukuda, Bin Li, Min Yu, Junling Chen, Yao Xu, Luo Guo, Ronggui Qu, Xueqian Wang, Zhaogui Sun, Miao Liu, Huijuan Shi, Hongyan Wang, Yi Feng, Ruijin Shao, Renjie Chai, Qiaoli Li, Qinghe Xing, Rui Zhang, Eva Nogales, Li Jin, Lin He, Mohan L. Gupta Jr., Nicholas J. Cowan, Lei Wang

Genetics, Development and Cell Biology Publications

BACKGROUND Human reproduction depends on the fusion of a mature oocyte with a sperm cell to form a fertilized egg. The genetic events that lead to the arrest of human oocyte maturation are unknown.

METHODS We sequenced the exomes of five members of a four-generation family, three of whom had infertility due to oocyte meiosis I arrest. We performed Sanger sequencing of a candidate gene, TUBB8, in DNA samples from these members, additional family members, and members of 23 other affected families. The expression of TUBB8 and all other β-tubulin isotypes was assessed in human oocytes, early embryos, sperm cells ...


Prokineticin-2: A Novel Anti-Apoptotic And Anti-Inflammatory Signaling Protein In Parkinson’S Disease, Matthew Neal Jan 2016

Prokineticin-2: A Novel Anti-Apoptotic And Anti-Inflammatory Signaling Protein In Parkinson’S Disease, Matthew Neal

Graduate Theses and Dissertations

The mechanisms behind the development and progression of Parkinson’s disease (PD) are still not fully understood. Several pathophysiological mechanisms have been implicated in the progression of the disease including oxidative stress, mitochondrial dysfunction, excitotoxicity, protein aggregation, and neuroinflammation. Since current therapeutic options for PD treat only the symptoms without influencing disease progression, recent translational studies have focused on identifying factors that protect against putative pathophysiological mechanisms of PD. We show herein, that the chemokine-like signaling protein, Prokineticin-2 (PK2), is expressed and secreted at higher levels in dopaminergic neurons during the acute phase of inflammatory or neurotoxic stress. Furthermore, we ...


Analysis Of The Induction Of Autophagy During Er Stress And The Vesicle Fusion Machinery At The Trans-Golgi Network In Arabidopsis Thaliana, Xiaochen Yang Jan 2016

Analysis Of The Induction Of Autophagy During Er Stress And The Vesicle Fusion Machinery At The Trans-Golgi Network In Arabidopsis Thaliana, Xiaochen Yang

Graduate Theses and Dissertations

The vacuole in plant cells occupies more than 80% of the cellular volume and is involved in development, detoxification, and degradation of proteins. In this dissertation, I focused on the two vesicle trafficking pathways, autophagy and vacuolar trafficking, which deliver cellular components to the vacuole.

Autophagy is an evolutionarily conserved mechanism in eukaryotic cells for degradation and recycling of cytoplasmic materials and damaged cell components during development or upon encountering stress conditions. Previous studies showed that autophagy is activated by endoplasmic reticulum (ER) stress and delivers ER fragments to the vacuole. ER stress is defined as accumulation of unfolded or ...


Modeling Pediatric Brain And Central Nervous System Cancer In Zebrafish, Staci Lyn Solin Jan 2015

Modeling Pediatric Brain And Central Nervous System Cancer In Zebrafish, Staci Lyn Solin

Graduate Theses and Dissertations

Brain and central nervous system (CNS) cancers are the leading cause of cancer-related death in children (Ostrom QT et al., 2015). Low-grade brain and CNS tumors that require minimal surgical resection due to their location in critical regions are associated with long-term morbidity throughout the life of the child (Armstrong GT et al., 2011). Children diagnosed with high-grade, aggressive brain and CNS tumors generally have a poor outcome and suffer significant deficits in neurological and neuroendocrine function as a result of intensive therapy (Fangusaro J et al., 2012). Effective, targeted therapeutics for the treatment of pediatric brain and CNS cancer ...


Molecular Characterization Of Human, Bovine, And Dolphin Parainfluenza Viruses: Viral Genome Sequencing And Epithelial Cell Signaling Responses Following In Vitro Infection, Kirsten Charlotte Eberle Jan 2015

Molecular Characterization Of Human, Bovine, And Dolphin Parainfluenza Viruses: Viral Genome Sequencing And Epithelial Cell Signaling Responses Following In Vitro Infection, Kirsten Charlotte Eberle

Graduate Theses and Dissertations

This dissertation aims at examining immunological response(s) to PIV-3 infection using in vitro cell culture systems. Based on previous publications, TtPIV-1 was found to be similar to, but distinct from, both human and bovine PIV-3 isolates. These initial findings were based only upon partial sequences of the F and HN viral genes. In Chapter 2 TtPIV-1 is further characterized on cytokine production, viral growth over time, and full genome sequencing as compared to BPIV-3 and HPIV-3. Chapter 3 is aimed at examining the type III IFN response during TtPIV-1, BPIV-3, and HPIV-3 infection. Because paramyxoviruses are known to affect ...


Tol2 Gene Trap Integrations In The Zebrafish Amyloid Precursor Protein Genes Appa And Aplp2 Reveal Accumulation Of Secreted App At The Embryonic Veins, Hsin-Kai Liao, Ying Wang, Kristin E. Watt, Qin Wen, Justin Breitback, Chelsy K. Kemmet, Karl J. Clark, Stephen C. Ekker, Jeffrey J. Essner, Maura Mcgrail Feb 2012

Tol2 Gene Trap Integrations In The Zebrafish Amyloid Precursor Protein Genes Appa And Aplp2 Reveal Accumulation Of Secreted App At The Embryonic Veins, Hsin-Kai Liao, Ying Wang, Kristin E. Watt, Qin Wen, Justin Breitback, Chelsy K. Kemmet, Karl J. Clark, Stephen C. Ekker, Jeffrey J. Essner, Maura Mcgrail

Genetics, Development and Cell Biology Publications

Background—The single spanning transmembrane amyloid precursor protein (APP) and its proteolytic product, amyloid-beta (Aβ) peptide, have been intensely studied due to their role in the pathogenesis of Alzheimer’s disease. However, the biological role of the secreted ectodomain of APP, which is also generated by proteolytic cleavage, is less well understood. Here, we report Tol2 red fluorescent protein (RFP) transposon gene trap integrations in the zebrafish amyloid precursor protein a (appa) and amyloid precursor-like protein 2 (aplp2) genes. The transposon integrations are predicted to disrupt the appa and aplp2 genes to primarily produce secreted ectodomains of the corresponding proteins ...


Dissection Of The Mitotic And Nuclear Functions Of Chromator, A Nuclear-Derived Spindle Matrix Component In Drosophila, Yun Ding Jan 2008

Dissection Of The Mitotic And Nuclear Functions Of Chromator, A Nuclear-Derived Spindle Matrix Component In Drosophila, Yun Ding

Retrospective Theses and Dissertations

A spindle matrix has long been proposed to serve as a stationary or elastic molecular matrix substrate for the organization and activities of microtubules and motors, based on the consideration of a mechanical and functional support for the stabilization of microtubule spindle during force generation from mitotic motors. Recently, the identification of four Drosophila proteins, Skeletor, Megator, EAST and Chromator has provided molecular evidence for the existence of this macromolecular matrix structure during mitosis. All of these four proteins have been shown to interact with each other within a protein complex and redistribute from the nucleus at interphase to form ...


Functional Analysis Of The C-Terminal Domain In The Drosophila Jil-1 Histone H3 Kinase, Xiaomin Bao Jan 2008

Functional Analysis Of The C-Terminal Domain In The Drosophila Jil-1 Histone H3 Kinase, Xiaomin Bao

Retrospective Theses and Dissertations

Epigenetic regulation is an important process utilized by biological systems to control gene expression and organize development, with histone modification enzymes among the most important components in this regulatory network. The predominant interphase H3 Serine10 kinase in Drosophila is JIL-1, which plays an important role in regulating chromosome structure and modulating gene expression. The full-length JIL-1 protein is composed of four domains including a N-terminal domain (NTD), two kinase domains (KDI and KDII), and a unique C-terminal domain (CTD). In order to study the biological importance of these individual domains, a domain analysis research approach was taken and different functions ...


Somatostatin Receptors Signal Through Efa6a-Arf6 To Activate Phospholipase D In Clonal Β-Cells, Justin A. Grodnitzky, Nasser Syed, Michael J. Kimber, Tim A. Day, Julie G. Donaldson, Walter H. Hsu May 2007

Somatostatin Receptors Signal Through Efa6a-Arf6 To Activate Phospholipase D In Clonal Β-Cells, Justin A. Grodnitzky, Nasser Syed, Michael J. Kimber, Tim A. Day, Julie G. Donaldson, Walter H. Hsu

Biomedical Sciences Publications

Somatostatin (SS) is a peptide hormone that inhibits insulin secretion in β-cells by activating its Gi/o-coupled receptors. Our previous work indicated that a βγ-dimer of Gi/o coupled to SS receptors can activate phospholipase D1 (PLD1) (Cheng, H., Grodnitzky, J. A., Yibchok-anun, S., Ding, J., and Hsu, W. H. (2005) Mol. Pharmacol. 67, 2162–2172). The aim of the present study was to elucidate the mechanisms underlying SS-induced PLD activation. We demonstrated the presence of ADP-ribosylation factor Arf1 and Arf6 in clonal β-cells, HIT-T15. We also determined that the activation of PLD1 was mediated through Arf6 ...


Characterization Of Three Spindle Matrix Proteins, Megator, East And Asator, In Drosophila, Hongying Qi Jan 2007

Characterization Of Three Spindle Matrix Proteins, Megator, East And Asator, In Drosophila, Hongying Qi

Retrospective Theses and Dissertations

Two previously identified proteins Skeletor and Chromator provide the molecular evidence for the existence of a spindle matrix complex in Drosophila. The spindle matrix has been proposed to be a macromolecular complex, which plays a role in organization and stabilization of the mitotic spindle as well as in providing structural support for counterbalancing force production.;In this dissertation, I present the identification and characterization of three more putative spindle matrix candidate proteins, Megator, EAST and Asator. Megator, an ortholog of the mammalian TPR protein, contains a large coiled-coil domain at its NH2-terminus and an acidic, non-structural COOH-terminus. Immunohistochemistry studies show ...


Map Based Candidate Gene Cloning And Functional Analysis Of Genes Involved In Vlcfas Synthesis, Li Fan Jan 2007

Map Based Candidate Gene Cloning And Functional Analysis Of Genes Involved In Vlcfas Synthesis, Li Fan

Retrospective Theses and Dissertations

VLCFAs are biosynthesized by an endoplasmic reticulum associated enzyme system, referred to as fatty acid elongase. Fatty acid elongase generates VLCFAs by elongating 18C fatty acids. The enzymatic machinery of fatty acid elongation is believed to be composed of 3-ketoacyl-CoA synthase, 3-ketoacyl-CoA reductase, 3-hydroxyacyl-CoA dehydratase, and enoyl-CoA reductase. However, the insoluble nature of the fatty acid elongase system has hampered efforts to purify the system and to identify the genes that encode the component proteins. In the current study a candidate gene cloning approach was used to demonstrate that the previously defined gl26 gene of maize encodes a predicted enoyl-CoA ...


Analysis Of A Nuclear Complex Of Proteins That Form A Spindle Matrix During Mitosis In Drosophila , Uttama Rath Jan 2005

Analysis Of A Nuclear Complex Of Proteins That Form A Spindle Matrix During Mitosis In Drosophila , Uttama Rath

Retrospective Theses and Dissertations

A spindle matrix is a macromolecular structure hypothesized to provide a stationary support or strut for the anchorage of motor proteins during force generation, microtubule sliding, and chromosome segregation in mitosis. Previous work in Drosophila has shown Skeletor, as the first putative molecular candidate of spindle matrix structure since it possesses many features consistent with the proposed spindle matrix structure.;We have identified two more putative spindle matrix candidate proteins, Chromator and Megator, using yeast two-hybrid interaction assay, cross-immunoprecipitation and immunocytochemistry. Chromator is a novel and essential chromodomain containing protein that interacts directly with Skeletor and extensively colocalizes with it ...


Identification Of A Role For Janus Kinase2 (Jak2) In Skeletal Muscle Mitogenesis, Myogenesis, And Hypertrophy , Symantha Ann Anderson Jan 2005

Identification Of A Role For Janus Kinase2 (Jak2) In Skeletal Muscle Mitogenesis, Myogenesis, And Hypertrophy , Symantha Ann Anderson

Retrospective Theses and Dissertations

Skeletal muscle is a dynamic tissue with tight developmental control and the ability to significantly remodel in the postnatal animal. Understanding the molecular mechanisms underlying skeletal muscle development and postnatal growth is critical to potential therapeutic manipulation. Janus kinase2 (JAK2) is a tyrosine kinase critical for growth of numerous cell types and is causative in many cancers, including leukemia. In this study we sought to understand the role of the JAK/STAT signaling cascade in skeletal muscle. In particular, this research focused on the role of JAK2 in skeletal muscle mitogenesis, myogenesis, and hypertrophy. Little work has been done regarding ...


Structural And Molecular Analysis Of Leech Neuronal Proteins, Calsensin And Filamin , Deepa V. Venkitaramani Jan 2004

Structural And Molecular Analysis Of Leech Neuronal Proteins, Calsensin And Filamin , Deepa V. Venkitaramani

Retrospective Theses and Dissertations

This thesis describes the structural and molecular analysis of two leech neuronal proteins, Calsensin and Filamin. Calsensin is an EF-hand calcium-binding protein expressed by a subset of peripheral sensory neurons which fasciculate into a single tract in the leech central nervous system. Calsensin is a 9 kD protein with two EF-hand calcium-binding motifs. Calsensin can multimerize via disulfide bridge formation involving the two cysteine residues in vitro. Furthermore, Calsensin shows differential gel migration depending on the presence or absence of calcium suggesting a conformational change upon calcium binding. Using multidimensional NMR spectroscopy we have determined the solution structure and backbone ...


The Essential Jil-1 Kinase Regulates Histone H3 Phosphorylation And Maintains Chromatin Structure In Drosophila , Weiguo Zhang Jan 2004

The Essential Jil-1 Kinase Regulates Histone H3 Phosphorylation And Maintains Chromatin Structure In Drosophila , Weiguo Zhang

Retrospective Theses and Dissertations

JIL-1 encodes a chromatin-associated tandem serine/threonine kinase in Drosophila melanogaster. JIL-1 predominantly associates with interbands on polytene chromosomes. It appears at higher levels on the X chromosome than on autosomes in males, and distributes about equally on all chromosomes in females. In addition, in males JIL-1 overlaps with the MSL (for male specific lethal) complex that specifically associates with the male X chromosome on which expression of most genes are upregulated.;We isolated JIL-1 mutants by inducing imprecise excision of an EP element inserted in JIL-1. We found that JIL-1 is required for viability of both females and males ...


Characterization Of Two Novel Spindle Matrix Proteins In Drosophila, Chromator And Megator , Dong Wang Jan 2003

Characterization Of Two Novel Spindle Matrix Proteins In Drosophila, Chromator And Megator , Dong Wang

Retrospective Theses and Dissertations

A spindle matrix has been proposed to help organize and stabilize microtubules during mitosis. This concept has gained new interest due to the identification of Skeletor as a potential component of the spindle matrix. Skeletor is associated with chromosomes during interphase but reorganizes into a spindle-like structure during early prophase before the nuclear envelope breaks down and microtubules enter the nucleus. The Skeletor spindle extends from pole to pole during metaphase coaligning with the microtubule spindle and persists even after nocodazole treatment dissembles the microtubules. These features make Skeletor an ideal candidate as a spindle matrix protein.;In this study ...


Molecular Interactions Of Talin And Actin: A Study On The Specific Domains Of Talin Involved In The Interactions , Ho-Sup Lee Jan 2002

Molecular Interactions Of Talin And Actin: A Study On The Specific Domains Of Talin Involved In The Interactions , Ho-Sup Lee

Retrospective Theses and Dissertations

Talin is an essential cytoskeletal protein present in cell-matrix type adherens junctions where it helps in attachment of the actin cytoskeleton to the plasma membrane. The nature of the interaction between talin and actin was examined in this dissertation by investigating three actin binding sites (ABS1--3), with ABS1 located in the 47 kDa N-terminal head and ABS2 and ABS3 present in the 190 kDa C-terminal tail of the talin molecule. Cosedimentation of expressed GST-fusion deletion constructs from the talin head with F-actin showed that actin binding activity of the talin head is contained within residues 271--320. Talin constructs containing the ...


Mechanisms Of Somatostatin-Induced Paradoxical Increase In Insulin Secretion In The Presence Of Arginine Vasopressin In Clonal [Beta]-Cell Hit-T15 , Henrique Cheng Jan 2002

Mechanisms Of Somatostatin-Induced Paradoxical Increase In Insulin Secretion In The Presence Of Arginine Vasopressin In Clonal [Beta]-Cell Hit-T15 , Henrique Cheng

Retrospective Theses and Dissertations

We investigated the mechanism underlying somatostatin-induced increase in [Ca2+]i leading to insulin release in the presence of arginine vasopressin (AVP) in clonal beta-cell HIT-T15. Somatostatin increased [Ca2+]i and insulin release in a biphasic pattern, characterized by a sharp and transient increase followed by a rapid decline to the sub-basal level. Pretreatment with pertussis toxin, which inactivates Gi/Go, abolished the effects of somatostatin. U-73122, an inhibitor of phospholipase C, antagonized somatostatin-induced increase in [Ca2+] i. In Ca2+-free environment, somatostatin still increased [Ca2+]i, whereas depletion of intracellular Ca 2+ stores with thapsigargin, a microsomal Ca2+ ATPase inhibitor ...


Coordination Of Cell Morphology And Nuclear Division By The Bud Neck Control Ring In Saccharomyces Cerevisiae , Courtney Lynn Thomas Jan 2001

Coordination Of Cell Morphology And Nuclear Division By The Bud Neck Control Ring In Saccharomyces Cerevisiae , Courtney Lynn Thomas

Retrospective Theses and Dissertations

Coordination of daughter cell growth and nuclear division are readily observed but not well understood in the yeast Saccharomyces cerevisiae . A screen was performed to identify mutants which lost this coordination such that they exhibited an elongated cell morphology (improper cell growth), but continued to divide. The results of this screen and suppressor analysis suggests the bud neck ring is a control structure which determines cell morphology as well as nuclear division as an upstream regulator of Cdc28p-cyclin complexes.;The effects of mutations in specific components of the bud neck ring on the assembly of other components into the structure ...


Drosophila Jil-1 Kinase Mediates Histone H3 Ser10 Phosphorylation, Maintains Higher Order Chromatin Structure, And Is Implicated In Dosage Compensation , Yanming Wang Jan 2001

Drosophila Jil-1 Kinase Mediates Histone H3 Ser10 Phosphorylation, Maintains Higher Order Chromatin Structure, And Is Implicated In Dosage Compensation , Yanming Wang

Retrospective Theses and Dissertations

The Drosophila JIL-1 tandem Ser/Thr kinase is associated with chromosomes/chromatin throughout the cell cycle in early embryos and localized to hundreds of sites on the open interband regions on third instar larvae polytene chromosomes. Interestingly, the level of JIL-1 is upregulated on the male X chromosome, which is hypertranscribed for dosage compensation. The distribution of JIL-1 overlaps with that of MSL (male specific lethal) proteins and JIL-1 is associated with the MSL complex.;To further study the function of JIL-1 in vivo, a series of JIL-1 mutants from hypomorphs to null were generated. Analyzing the phenotypes of JIL-1 ...


Height Referencing Biochemical Cassette, Marc D. Porter, Vivian W. Jones, Curtis L. Mosher, Eric Henderson Nov 2000

Height Referencing Biochemical Cassette, Marc D. Porter, Vivian W. Jones, Curtis L. Mosher, Eric Henderson

Iowa State University Patents

A height referencing biochemical cassette comprising of a surface suitable to act as a coupling agent, a height referencing indicator, and molecules bonded to the surface. This cassette is a capable of use to test for bonding to these molecules through measuring the height difference between the indicator and the surface. This invention provides an efficient means to quickly and easily test for bonding of molecules to other molecules.


Tissue Specific Expression And Regulation Of The Mitogen Regulated Protein Genes , John Tollen Fassett Jan 2000

Tissue Specific Expression And Regulation Of The Mitogen Regulated Protein Genes , John Tollen Fassett

Retrospective Theses and Dissertations

Mitogen Regulated Protein (MRP; also called Proliferin; or MRP/PLF) is a member of the Prolactin/Growth Hormone gene family. There is evidence that MRP/PLF is a uterine growth factor and an angiogenesis factor. Between four and six highly homologous MRP/PLF genes (mrp/plf ) exist in the mouse, but the significance of multiple genes has not been determined. MRP/Plf is produced at high levels during mid-gestation by the placenta, where expression is restricted to the trophoblastic giant cells. To better understand the regulation of the mrp/ plf genes in vivo, we have developed an assay to analyze ...


Characterization Of Intracellular Survival Of Mycobacterium Avium Subspecies Paratuberculosis In J774 Murine Macrophages , Jesse Michael Hostetter Jan 2000

Characterization Of Intracellular Survival Of Mycobacterium Avium Subspecies Paratuberculosis In J774 Murine Macrophages , Jesse Michael Hostetter

Retrospective Theses and Dissertations

Mycobacterium avium subspecies paratuberculosis (M. a. ptb) is the causative agent of Johne's disease, a progressive diarrheal disease of wild and domestic ruminants. Like other pathogenic species of mycobacteria, M. a. ptb is an intracellular pathogen of monocytes and monocyte-derived macrophages. In this dissertation we tested the hypothesis that M. a. ptb survives within macrophages by inhibition of acidification and maturation of the phagosomal compartment in which it resides;Chapter 1 consists of a general introduction and a literature review of recent reports concerning normal phagosomal maturation and maturation of the mycobacterial phagosome. Chapter 2 characterizes acidification of the ...


Two Dimensional Models Of Tumor Angiogenesis , Serdal Pamuk Jan 2000

Two Dimensional Models Of Tumor Angiogenesis , Serdal Pamuk

Retrospective Theses and Dissertations

Angiogenesis, the formation of new capillaries from pre-existing vessels, is essential for tumor progression. It is critical for the growth of primary cancers. In this thesis we present a new approach to angiogenesis, based on the theory of reinforced random walks, coupled with a Michaelis-Menten type mechanism. This views the endothelial cell receptors as the catalyst for transforming angiogenic factor into proteolytic enzyme in order to model the first stage. In our model we use a curvature-induced proliferation term for the endothelial cell equation. Our numerical results indicate that the proliferation of endothelial cells is high at the tip. Also ...


Characterization Of Skeletor: A Novel Nuclear Protein Implicated In Mitotic Spindle Assembly , Diana Lee Walker Jan 2000

Characterization Of Skeletor: A Novel Nuclear Protein Implicated In Mitotic Spindle Assembly , Diana Lee Walker

Retrospective Theses and Dissertations

The mAb 2A labels Drosophila embryos in a novel, dynamic staining pattern that identifies a nuclear meshwork during interphase, which is remodeled to form a spindle structure at metaphase. This intriguing pattern made the antigens recognized by mAb 2A interesting targets for cloning and analysis, Skeletor was found to be encoded by a complex locus. Two alternatively spliced transcripts from the Skeletor locus were identified, Orf1b/Orf2 and Orf1a. Orf1b/Orf2 has an unusual structure, containing two overlapping open reading frames with coding potentials of 87 kD and 81 kD respectively. Analysis of Orf2 amino acid content reveals overall high ...


Mechanisms Of Negative Regulation Of Protein Prenylation: Investigation Of Murine Guanylate-Binding Protein 1 , John Thomas Stickney Jan 2000

Mechanisms Of Negative Regulation Of Protein Prenylation: Investigation Of Murine Guanylate-Binding Protein 1 , John Thomas Stickney

Retrospective Theses and Dissertations

Covalent attachment of isoprenoid lipids to proteins is a post-translational modification that occurs on approximately 2% of all cellular proteins. Proteins that contain a CaaX motif as their final four amino acids are presumed to be targets for the addition of C15 or C20 isoprenoids by farnesyl transferase (FTase) or geranylgeranyl transferase I (GGTase I) respectively. This modification is generally believed to occur stoichiometrically, and irreversibly, in vivo. Thus, if prenylation is to be regulated, this control must occur prior to lipid addition. At the moment, there is little evidence that the enzymatic activity of prenyl transferases can be modulated ...


Fatty Acid Modification Of Cells In Culture By Overexpression Of Exogenous Desaturases , Travis Jay Knight Jan 1999

Fatty Acid Modification Of Cells In Culture By Overexpression Of Exogenous Desaturases , Travis Jay Knight

Retrospective Theses and Dissertations

COS-1 cells were transfected transiently with expression vectors encoding stearoyl-acyl-carrier-protein desaturase (SACPD) from castor bean, stearoyl-CoA desaturase (SCD) from mice, and a control vector containing SACPD cDNA in reverse orientation. The hypothesis was that both exogenous desaturases would be active in COS-1 cells, thereby increasing the proportion of monounsaturated fatty acids produced by the cells. Messenger RNA and anti-myc immunoreactive proteins were detected at 12 and 24 hours post-transfection respectively and increased through 48 hours post-transfection. When acetate was used as the fatty acid precursor in cultures for a 24-hour period starting 24 hours post-transfection, the ratio of monounsaturated to ...