Life Sciences Commons^™

Factors Influencing The Oxidation Of Lipoproteins And Plasma Lipids, Yanshan Ma

Electronic Theses and Dissertations

The hypothesis that antioxidant vitamins (ascorbate and tocopherols) along with urate protect blood plasma lipids from oxidation was tested. Dietary fat is also an important factor influencing plasma lipid peroxidation. The purpose of this study was to investigate the role of plasma antioxidants and dietary fat on low density lipoprotein (LDL) and plasma lipid oxidation. In the first part of this study, we compared the ability of urate and ascorbate to protect human LDL from in vitro oxidation. LDL oxidation was initiated by 15 mM of a water soluble azo-initiator in the presence or absence of ascorbate or urate. The …

Role Of Heat Shock Protein 70 Kda Cognate In Limiting Thermal Inactivation And Refolding Of Heat-Denatured Nuclear Type I Topoisomerase, Kuo-Kuang Wen

Theses and Dissertations in Biomedical Sciences

Previous studies (Ciavarra et al., 1994) demonstrated that the constitutive 70 kDa heat shock protein (hsc70) protected purified topoisomerase I from thermal injury. In addition, hsc70 was capable of regenerating catalytic activity of heat-denatured topoisomerase I. A whole cell lysate was also active in this reaction assay. The present study demonstrates that heat-denatured topoisomerase I is reactivated by a cytosolic fraction and that this activity is dependent on the presence of cytosolic hsc70. The efficacy of hsc70-mediated refolding of heat-denatured topoisomerase I is greatly enhanced by a cytosolic cofactor(s). In all these refolding reactions, exogenous ATP is not required. Size …

In Situ Regulation Of Cytosolic Phospolipase A₂, Beverly A. Rzigalinski

Theses and Dissertations in Biomedical Sciences

The 85 kDa cytosolic phospholipase A₂ (cPLA₂) is an agonist-responsive effector for intracellular signal transduction through the arachidonate cascade. In vitro studies have demonstrated that this enzyme is regulated by sub-micromolar calcium and is specific for arachidonate as the sn-2 fatty acyl group of phospholipid substrates. However, very little data is available regarding in situ mechanisms which govern the activity of cPLA₂. The primarily objective of these studies was to develop an in situ system for the study of cPLA₂, and investigate mobilization of arachidonate during signal transduction events.

Dimethylsulfoxide differentiation of the …

Determination Of Pancreatic And Salivary Amylase By Enzyme Immunoassay And Their Prevalence In Hyperamylasemic Patients, Sabdra Borgens Ward

Theses and Dissertations in Biomedical Sciences

Currently, amylase determinations are nonspecific for the organ source and are based entirely on the enzymatic properties of amylase to produce a measurable product or byproduct. The determination of pancreatic amylase is important in the diagnosis of acute pancreatitis. Most commercially available tests for amylase employ the measurement of the change in NADH absorbance at 280 nm or of the p-nitrophenol released from a maltotetrose substrate. These are nonspecific measurements of pancreatic amylase and often necessitate other tests to be run such as a serum lipase.

The two predominant isoenzymes of amylase are pancreatic (p-amylase) and salivary (s-amylase); the most …

Structure-Substrate Binding Relationships Of Hiv-1 Reverse Transcriptase, Steve Chien-Wen Huang

Theses and Dissertations in Biomedical Sciences

Human Immunodeficiency Virus, type 1 (HIV-1), is the causative agent of the Acquired Immunodeficiency Syndrome (AIDS). HIV-1 reverse transcriptase (RT), a heterodimer p66/p51, has been the major target for treatment of AIDS. The significance of the p51 subunit and the RNase H domain of p66 in terms of their influence on the RNA-dependent DNA synthesis was investigated. Clones of the wildtype HIV-1 RT subunits, p66 and p51, and a recombinant C-terminal deletion mutant, p64, [Barr, P. J. (1987) Bio/Technoloav 5, 486-489] were employed to study the structure-substrate binding relationships of HIV-1 RT. The activity assays of RNA-dependent DNA synthesis on …

Mechanism Of Mitogenic Action Of Aluminum Ion On Bone Cells : Potential Involvement Of The Insulin-Like Growth Factor Regulatory System, Aera Yoo

Loma Linda University Electronic Theses, Dissertations & Projects

Micromolar concentrations of aluminum ion (oxidation state of 3 +) consistently stimulated [3H]thymidine incorporation into cell DNA and increased cellular alkaline phosphatase activity (an osteoblastic differentiation marker) in osteoblast-like cells of chicken and human origin. Although biphasic, dose-dependent stimulations were highly reproducible, the maximal stimulatory dose varied among different experiments. Mitogenic doses of aluminum ion also stimulated collagen synthesis in cultured human osteosarcoma TE85 cells. In addition, the same mitogenic concentrations of aluminum ion enhanced the 1,25 dihydroxy vitamin D3-dependent stimulation of osteocalcin secretion. Together, these findings indicate that aluminum at micromolar concentrations, stimulates the proliferation, differentiation, and activity of …

The Invariant Arginine In Motif 2 Of Escherichia Coli Alanyl-Trna Synthetase : Is Important For Catalysis But Not For Substrate Binding, Ying Lu

Loma Linda University Electronic Theses, Dissertations & Projects

Structural motifs 2 and 3, located in the active site of class II aminoacyl-tRNA synthetases, each contain an invariant arginine thought to participate in interactions with ATP. For Escherichia coli alanyl-tRNA synthetase (AlaRS), sequence comparisons indicate that Arg69 should be aligned with the invariant arginine in motif 2 of other class II synthetases. Site-directed random mutagenesis has been employed to generate a set of proteins containing amino acid substitutions in a portion of motif 2 of AlaRS. In this set, only mutations at position 69 caused the enzyme to lose ability to complement growth of an alaS deletion strain, …

Signal Transduction In Bacterial Chemotaxis, Edward Heath Rowsell

Loma Linda University Electronic Theses, Dissertations & Projects

This study investigated the site of ATP utilization in the signal transduction pathway of bacterial chemotaxis and localized the point of convergence of a methylation-independent system of chemotaxis with the methylation-dependent system. The identity of the signal originating from the phosphotransferase system was investigated by substituting the fructose-inducible HPr-like protein FPr for HPr in transport and chemotaxis. In addition, a novel chemoattractant, glycerol, was identified for Salmonella typhimurium. Histidine-auxotrophic S. typhimurium strains ST23 (hisF) and ST171 (hisF cheB) were depleted for ATP. The times required for the bacteria to adapt to a step increase in serine, …

The Effects Of Unpaired Bases In An Oligo Deoxy (A) * Deoxy (T) Tract On Dna Bending, Vasavi Malineni

Honors Theses

No abstract provided.

Characterization Of The Vasoactivity Of Tachykinins In Isolated Rat Kidney: Functional Studies And In Vitro Receptor Autoradiography, Yuejin Chen

Electronic Theses and Dissertations

Although tachykinins have potent vascular actions, their effect on renal resistance blood vessels is currently unknown. The vasoactive properties of tachykinins and related analogs were assessed in isolated perfused rat kidney. At a basal perfusion pressure (PP) of 75 $\pm$ 6 mm Hg (n = 5), bolus injections of substance P (SP) had no significant vasoactive effect. Following a sustained increase in baseline PP (134 $\pm$ 10 mm Hg) produced by phenylephrine (1 $\mu$M), SP evoked a dose-dependent increase in PP. The largest dose of SP increased PP by 60 $\pm$ 5 mm Hg. The vasoconstrictor response to SP was …

The Role Of Tocopherol, Cholesterol, And Oleic Acid In Lipid Peroxidation, Jherie Jean Demouy Blazier

Honors Theses

No abstract provided.

Translational Regulation Of The C-Jun Proto-Oncogene, Anil Sehgal

Theses and Dissertations in Biomedical Sciences

The v-jun oncogene was originally isolated from the ASV17 virus in 1987. Ever since its isolation, extensive work has been done to understand the role of the v-jun oncogene in cell transformation. The c-Jun protein is a transcription factor which binds to the DNA target TGACTCA. The c-Jun protein binds to DNA in the form of dimers. It can form homodimers with itself and heterodimers with Jun family (JunB and JunD), Fos family (FosB, Fra1 and Fra2), or with CREB family members through the leucine zipper motif. Because the c-jun proto-oncogene plays an important role in cell transformation, extensive work …

Investigation Of Selenium Status In Hypo-, Hyper- And Euthyroid Children, Ping Sun

Chemistry & Biochemistry Theses & Dissertations

Recently, it has been reported that human type I iodothyronine deiodinase, an enzyme important in the conversion of T₄ to T₃ , is a selenoenzyme. Several studies have reported alterations in plasma selenium level in hypo- and hyperthyroid patients.

Using polarized Zeeman-effect atomic absorption spectroscopy, we measured selenium, zinc, copper and manganese concentrations in the plasma and/or red blood cells in children with or without thyroid disease being seen at the outpatient clinic at Children's Hospital of the King's Daughters. Children with thyroid disease were subdivided into untreated, treated and nonresponsive groups. Data were analyzed using Student's t-test …

The Endocytosis Of Iodinated Protein Substrates In Cultured Human Skin Fibroblasts, Daniel P. Wilcockson

Master's Theses

No abstract provided.

Transformation Related Defects In Vacuolar Acidification And Mannose 6-Phosphate Receptor Function: Effects On Lysosomal Targeting, Valerie Pracht

Dissertations

No abstract provided.

The Role Of Alternative Splicing In The Regulation Of Cathepsin B Expression In Human Tumors, Qiuming Gong

Dissertations

No abstract provided.

The Effect Of Acute Ethanol Exposure On The Male Rodent Reproductive Axis, Margaret Mary Halloran

Dissertations

No abstract provided.

Ribosomal Protein S3: A Multifunctional Eukaryotic Dna Repair Enzyme Active On Ultraviolet And Oxidative Dna Damage, David M. Wilson

Dissertations

No abstract provided.

Properties Of A Novel Early Lymphoid Activation Gene (Elag) Isolated From A Hodgkin's-Disease Cell Line, John Scott Bennett

Dissertations

No abstract provided.

Spiroplasma (Entomoplasmatales) Associated With Tabanidae (Diptera) And Lampyridae (Coleoptera), Jimmy Wedincamp Jr.

Legacy ETDs

Abstract 1:

To determine the exit points of Spiroplasma from tabanids, 45 naturally infected Tabanus gladiator and 6 T. sulcifrons were restrained and fed 10% sucrose. The flies were allowed to feed for 24 hours and the resulting oral and anal specks were cultured in MID broth. Spiroplasmas were isolated from 21 of 51 oral specks but none of 23 anal specks deposited on plastic. In contrast, when anal specks were deposited in a sucrose solution, 9 of 28 anal specks in sucrose yielded spiroplasma cultures. Air-dried spiroplasma cultures in 5% sucrose survived for up to 30 days in the …

Characterization And Genetic Analysis Of A Pigmentation Mutant Of Neurospora Crassa, Craig West Banks

Legacy ETDs

The mutant overaccumulator of carotenoids, ovc. was obtained from the Fungal Genetics Stock Center in Kansas City, Kansas. This mutant had been isolated following UV irradiation of the colonial mutant col-4 and mapped to Linkage Group IV (R. Harding, 1984 Neurospora Newsletter 34: 23-25). Examination of ovc in our laboratory revealed that in addition to having increased pigmentation, the mutant was osmotic sensitive, failing to grow on 6% NaCl. Morphology of ovc closely resembles that of cut, another osmotic sensitive mutant in LG IV. A cross of ovc to the osmotic sensitive mutant cut failed to produce …

The Role Of Sexual Selection In Structuring West Indian Passerine Communities, C. Lynne Starling

Legacy ETDs

Patterns of plumage dimorphism were examined among species of passerines native to 19 West Indian islands. Based on sexual selection theory, the predictions were made that dimorphic species: (1) occur on fewer and larger islands than monomorphic species due to reduced colonization ability and reduced survivorship imposed by sexual selection pressure, and (2) should be added to island communities disproportionately as area increases, an idea which was tested using linear models.

No significant difference was observed between number of islands or minimum island area occupied by dimorphics versus monomorphics. The traditional ANCOVA model best fits the data and suggests that …

Susceptibility Of Eumeces Species Skinks To A Southeastern Isolate Of The Lyme Disease Spirochete (Borrelia Burgdorferi), Gregory A. Cummins

Legacy ETDs

The SI-4 strain of Borrelia burgdorferi obtained from field collected Ixodes scapularis on Sapelo Island, Georgia, was infective to seven of eight cotton mice (Peromyscus gossypinus) by needle inoculation. I. scapularis larvae were then fed on them, and nymphs molting from these larvae were then fed on laboratory mice, Mus musculus, in an attempt to transmit the spirochete. One of three mice was successfully infected; confirmation of the presence of B. burgdorferi was made using BSK II culture and the polymerase chain reaction (PGR). A subsequent attempt was made to determine if SI-4 was infective for Eumeces …

Regulatory Analysis Of Vimentin Expression In Metastatic Versus Nonmetastatic Breast Cancer Cell Lines, Danielle N. Bird

Theses and Dissertations

The intermediate filament gene family, composed of six classes, shows both tissue and development-specific expression. Vimentin is a unique member of the intermediate filament protein family: although it is expressed in cells of mesenchymal origin, vimentin may also be expressed with other intermediate filament proteins during early stages of development. In some cases, as differentiation continues, vimentin is normally down-regulated whereas other intermediate filament proteins, like desmin, glial fibrillary acidic protein, or neurofilaments are turned on in muscle, glial cells, or neurons, respectively. In some metastatic cancers, including breast and prostate cancers, vimentin is aberrantly expressed, despite the embryonic origin …

Expression, Purification And Characterization Of Hepatitis C Virus Core Protein From E.Coli Using A Chemically Synthesized Gene (1), And Cloning, Expression, Purification And Characterization Of The Major Core Protein (P26) From Equine Infectious Anaemia Virus, Ashley James Birkett

Theses and Dissertations

The hepatitis c virus (HCV) core gene has been chemically synthesized and used to direct the expression of core protein in Escherichia coli (E.coli). When cloned downstream of the inducible T7 promotor the core gene directed the expression of a soluble protein with a molecular weight of 22 kDa. Under native conditions the protein behaved in a manner consistent with the formation of a multimeric structure, which may represent assembled core particles. Further, when examined by electron microscopy a heterogeneous mixture of nucleocapsid-like particles were visible. Core protein was specifically recognized by antibodies present in HCV infected serum, suggesting that …

Cloning, Expression, Purification And Characterization Of Domain B Of Wheat Germ Agglutinin, Ann C. Rice

Theses and Dissertations

Wheat germ agglutinin (WGA) belongs to a family of dimeric chitin binding lectins specific for N-acetyl-D-glucosamine (GlcNAc) and N-acetyl-D-neuraminic acid (NeuNAc). The polypeptide chain consists of a tandem repeat of four conserved 4.3 kDa domains (A, B, C, D) stabilized by four disulfide bonds. Saccharide binding occurs in the dimer interface where domains of different subunits are in contact (A with D, B with C). An aromatic amino acid rich pocket on one domain (sufficient for saccharide binding) and a polar region on the contacting domain constitute a complete binding site. Saccharide binding affinities may differ among the four unique …

Uptake Routes Of Benzo[A]Pyrene In An Estuarine Killifish: Cytochrome P4501a Immunohistochemistry Using Whole Fish, Maria Karin Cochran

Dissertations, Theses, and Masters Projects

No abstract provided.

Global Phylogeography Of Yellowfin Tuna, Thunnus Albacares, And Mackerels Of The Genus Scomber, Daniel R. Scoles

Dissertations, Theses, and Masters Projects

Intraspecific genetic relationships within yellowfin tuna, Thunnus albacares, and three mackerels of the genus Scomber were studied by restriction fragment length polymorphism (RFLP) analysis of mitochondrial DNA (mtDNA). The comparison of these scombrids, with different larval distributions, adult distributions, and vagilities, served to investigate the hypothesis that population structure in marine fishes results from geographic and physical oceanographic processes that limit dispersal of early life history stages. Samples of 20 yellowfin tuna were examined from each of five Pacific locations and one Atlantic location. MtDNA analysis with 12 informative restriction endonucleases demonstrated considerable genetic variation, as evidenced by overall nucleon …

Mitochondrial Dna Variation In Natural And Cultured Populations Of The Bay Scallop, Argopecten Irradians (Lamarck) And The Calico Scallop, Argopecten Gibbus (Dall), Sandra Grace Blake

Dissertations, Theses, and Masters Projects

No abstract provided.

Novel Suicide Elements For Gene Disruption In Escherichia Coli, Brian Christopher Nicholson

Dissertations, Theses, and Masters Projects

No abstract provided.