Life Sciences Commons^™

Role Of Immunodominant T Cell Responses To Minor Histocompatibility Antigens In A Mouse Model Of Graft-Versus-Host Disease, Marc Adam Berger, B.S. Thomas Jefferson University

Full-Text Theses & Dissertations

Graft-versus-host-disease (GVHD) is a major complication of major histocompatibility complex (MHC) matched bone marrow transplantation. GVHD is the result of an immune-mediated attack, by T lymphocytes derived from donor bone marrow, against recipient tissues expressing minor histocompatibility antigen (miHA) differences. There is a lack of knowledge concerning the nature of in vivo immune responses to miHA during GVHD. C57BL/6By (B6/By) and BALB.B mouse strains both express the H-2B MHC haplotype but differ at more than 40 miHA loci. Both the in vitro and in vivo T cell responses to miHA expressed by BALB.B and CXB recombinant inbred (RI) strains of …

A New High Pressure Sapphire Nuclear Magnetic Resonance Cell, Shi Bai, Craig M. Taylor, Charles L. Mayne, Ronald J. Pugmire, David M. Grant

Faculty Publications

A new version of a single-crystal sapphire high pressure nuclear magnetic resonance (NMR) cell is described that is capable of controlling the sample pressure independent of the temperature. A movable piston inside the cell adjusts and controls the sample pressure from ambient conditions to 200 atm within plus or minus 0.3 atm. The linewidth at half-height for a 13C spectrum of carbon dioxide at 15degrees C and 57.8 atm is found to be 0.5 Hz. The carbon dioxide gas/liquid phase transition is clearly observed by measuring 13C chemical shifts as the sample pressure approaches equilibrium. The time required for this …

Analysis Of The Mechanisms Controlling Phospholipid Biosynthetic Gene Expression In The Yeast Saccharomyces Cerevisiae, Brian Patrick Ashburner

Dissertations

No abstract provided.

Developmental, Tissue, And Sub-Tissue Specific Expression Of The Mammalian Apurinic/Apyrimidinic Endonuclease, Teresa Mari Wilson

Dissertations

No abstract provided.

Modulation Of Queuine Uptake And Incorporation Into Trna By Protein Kinase C And Protein Phosphatase, Rana C. Morris, Bonnie J. Brooks, K. Lenore Hart, Mark S. Elliot

Chemistry & Biochemistry Faculty Publications

It has been suggested that the rate of queuine uptake into cultured human fibroblasts is controlled by phosphorylation levels within the cell. We show that the uptake of queuine is stimulated by activators of protein kinase C (PKC) and inhibitors of protein phosphatase; while inhibitors of PKC, and down-regulation of PKC by chronic exposure to phorbol esters inhibit the uptake of queuine into cultured human fibroblasts. Activators of cAMP- and cGMP-dependent kinases exert no effect on the uptake of queuine into fibroblast cell cultures. These studies suggest that PKC directly supports the activity of the queuine uptake mechanism, and that …

Application Of A Modified Ziehl-Neelsen's Carbol-Fuchsin Stain For Observing Nuclei In Vegetative Hyphae Of The Puffball Species Lycoperdon Pyriforme, Martin J. Huss

Journal of the Arkansas Academy of Science

Nuclei of vegetative hyphae of the puffball species Lycoperdon pyriforme are small and difficult to stain. A simple mitotic staining technique utilizing a modified version of Ziehl-Neelsen's carbol-fuchsin stain, originally developed for examining bacteria and small chromosomes of plants, has proven useful for observing fungal nuclei in this gasteromycete species. Cells of vegetative hyphae were pre-treated for 2 hours with an aqueous solution saturated with naphthalene, fixed in a 3:1 mixture of 100% ethanol and glacial acetic acid, and hydrolyzed for 10 minutes in a 1:1 mixture of 100% ethanol and 12 M hydrochloric acid. Cells were rinsed with distilled …

Membrane Topology Of The Colicin A Pore-Forming Domain Analyzed By Disulfide Bond Engineering, Denis Duche, Jacques Izard, Juan M. Gonzalez-Manas, Michael W. Parker, Marcel Crest, Daniel Baty

Department of Food Science and Technology: Faculty Publications

Four colicin A double-cysteine mutants possessing a disulfide bond in their pore-forming domain were constructed to study the translocation and the pore formation of colicin A. The disulfide bonds connected a-helices 1 and 2, 2 and 10, 3 and 9, or 3 and 10 of the poreforming domain. The disulfide bonds did not prevent the colicin A translocation through the Escherichia coli envelope. However, the mutated colicins were able to exert their in vivo channel activity only after reduction of their disulfide bonds. In vitro studies with brominated phospholipid vesicles and planar lipid bilayers revealed that the disulfide bond that …

Bovine Cell Line Resistant To In Vtro Infection By Bovine Viral Darrheavirus And All Other Known Pestiviruses, Ruben O. Donis, Eduardo F. Flores

School of Veterinary and Biomedical Sciences: Faculty Publications

Bovine cell line resistant to infection by the pestiviruses Bovine Viral Diarrhea Virus (BVDV), Hog Cholera Virus (HCV) and Border Disease Virus (BDV) and all progeny and mutants thereof. A bovine cell line (CRIB) that is resistant to infection by cytopathic and non-cytopathic BVDV and by other Pestiviruses due to a stable, recessive genetic defect which blocks infection at the level of viral entry.

Red Blood Cell Stability In Uremic Rats, Cecilia Mermel Maccallum

Theses and Dissertations

Determining the fragility of the red blood cell (RBC) is important for the diagnosis of and evaluation for treatment of several RBC diseases. In part RBC production is controlled through the hormone erythropoietin secreted by the kidney. In a previous study from this laboratory, it was found that RBC were more stable in uremic male rats compared to controls. In this experiment, uremia was induced in four groups of female rats through a two stage nephrectomy. The nephrectomy involved the removal of two-thirds of the left kidney, followed by the removal of the entire right kidney one week later. The …

Mechanistic Studies On Malate Dehydrogenase From Escherichia Coli, S. Kirk Wright

Masters Theses

Kinetic studies and chemical modification studies were performed on malate dehydrogenase from Escherichia coli (eMDH). Chemical modification studies using diethylpyrocarbonate and iodoacetic acid, along with log V/K profiles indicate that one ionizing group with a pK_a between 7.9 and 8.5 acts as a general acid/base in the catalytic mechanism. Log V profiles suggest that malate binds to the correctly protonated form of the enzyme. These results imply that a histidine residue is required for catalysis.

A primary deuterium kinetic isotope effect of 1.43 (±0.14) was observed on V/K using malate-2-d, while no isotope effect was measured on V_{max …}

Regulation Of Phospholipid Biosynthesis In The Yeast Saccharomyces Cerevisiae By The Ume6 Gene, John Charles Jackson

Dissertations

No abstract provided.

Reproductive Ecology Of Pinckneya Bracteata (Bartram) Raflnesque (Rubiaceae), George H. Rountree

College of Graduate Studies: Theses & Dissertations (1964–2006)

Pinckneya bracteata is a small tree or large shrub that is indigenous to the coastal plain of extreme southern South Carolina, Georgia, and northern Florida. It in occurs in sunny, wet habitats and in small populations that are usually separated by distances of kilometers. This study examined the mating, dispersal, and germination systems of the species. Pinckneya bracteata was found to be protandrous and self incompatible. Ruby-throated Hummingbirds and bumblebees ( Bombus spp. appear to be the primary pollinators. Seed fall velocity is important to the wind dispersal of seeds and decreases as seed size increases. Fall velocity is also …

The Chlorophyll Biosynthetic Enzyme Mg-Protoporphyrin Ix Monomethyl Ester (Oxidative) Cyclase, David Bollivar

Scholarship

No abstract provided.

Characterization Of The Rnasep Sequence In Chlorobium Limicola, Emily J. Hoeger

Presidential Scholars Theses (1990 – 2006)

RNAse P is a ribonucleoprotein involved in cutting out pieces of RNA. Specifically, it is involved in tRNA processing as it makes a single endonucleolytic cleavage in all precursor tRNA's to produce mature 5' ends. The sequence encoding the RNA subunit (rnpB gene) of RNAseP has been previously determined in one strain of Chlorobium limicola. A second strain was obtained from Carolina Biological Supply to test for conservation of sequence characteristics. This second strain was used in a PCR with the primers described in Haas. The amplified rnpB gene sequence appeared to have fewer base· pairs than the Haas …

Tunable Picosecond Infrared Laser System Based On Parametric Amplification In Ktp With A Ti:Sapphire Amplifier, D. E. Gragson, D. S. Alavi, G. L. Richmond

Chemistry and Biochemistry

A picosecond laser system that will generate high-power tunable IR pulses with bandwidths suitable for spectroscopic applications is discussed. The system is based on white-light continuum generation in ethylene glycol and optical parametric amplification in potassium titanyl phosphate. The nonlinear-optical processes are driven by a regeneratively amplified Ti:sapphire laser that produces 1.7-ps pulses at a repetition rate of 1 kHz. Energies as high as 40 and 12 µJ have been achieved over the signal (1.02–1.16-µm) and idler (2.6–3.7-µm) tuning ranges, respectively. The IR beam temporal and spatial characteristics are also presented.

Development Of An In Vitro Assay To Predict Patient Response To Radiotheraphy, Mary Sheridan

Doctoral

At the present time, the treatment plan for a patient with cancer is usually based on parameters such as tumour site, histological type, and tumour stage and performance status. However, it is well know that the radiosensitivity of human cancers varies widely from one patient to another (Fertil et al., 1981), and that even within these broad categories some tumours will show greater response to radiotherapy than others. If those patients unlikely to be cured by radiotherapy could be identified prior to commencement of treatment, alternative or more aggressive therapies might be selected which may give a better chance of …

Competitive Enzyme Immunoassay Of Catechin In Human Blood Serum, Jay Fedorowicz

Chemistry & Biochemistry Theses & Dissertations

( +)-Catechin is a type of polyphenolic compound, and it is enriched in fruits, plants, and various beverages. Polyphenolic compounds also play an important role in the human diet. Many scientists, especially wine researchers, have proposed that(+)catechin and its polymers could have many positive health effects. Thus, it is essential to learn more about (+)-catechin and other polyphenolics. Presently, high performance liquid chromatography is the only method to detect ( + )-catechin levels in human blood serum, but this method has the high cost of instrumentation and does not provide good sensitivity. The objective of this project is to develop …

Anaerobic Protoporphyrin Biosynthesis Does Not Require Incorporation Of Methyl Groups From Methionine, David Bollivar

Scholarship

No abstract provided.

Biochemical And Molecular Characterization Of The Prostate-Specific Membrane Antigen (Psma), John Karl Troyer

Theses and Dissertations in Biomedical Sciences

Prostate cancer is the most common malignancy and the second leading cause of cancer death in males in the United States. Additionally, the number of deaths attributed to prostate cancer is increasing at a rate of approximately 8% a year. Development of new diagnostic and therapy strategies are needed in order to improve the life expectancy of patients with this disease. One tool which may allow for improvements in prostate cancer diagnosis and therapy is the monoclonal antibody (MAb) 7E11-C5.3 which was first described in 1987. Since then, the antigen recognized by MAb 7E11-C5.3 has been named the prostate specific …

Three-Phase Intersection Points In Monolayers, John P. Hagen, Harden M. Mcconnell

Chemistry and Biochemistry

Some phospholipid/dihydrocholesterol Langmuir monolayers form coexisting liquid phases. Gas domains form at the interface between the phospholipid-rich and dihydrocholesterol-rich liquid phases when these monolayers undergo expansion to low surface pressure. Analysis of the domain shapes thus formed yields the relative line tensions of the gas/phospholipid, gas/dihydrocholesterol, and phospholipid/dihydrocholesterol phase interfaces.

Pattern Of Disease After Murine Hepatitis Virus Strain 3 Infection Correlates With Macrophage Activation And Not Viral Replication, M. Pope, O. Rotstein, E. Cole, S. Sinclair, Rebecca D. Parr, B. Cruz, R. Fingerote, S. Chung, R. Gorczynski, L. Fung, J. Leibowitz, Y. S. Rao, G. Levy

Faculty Publications

Murine hepatitis virus strain (MHV-3) produces a strain-dependent pattern of disease which has been used as a model for fulminant viral hepatitis. This study was undertaken to examine whether there was a correlation between macrophage activation and susceptibility or resistance to MHV-3 infection. Peritoneal macrophages were isolated from resistant A/J and susceptible BALB/cJ mice and, following stimulation with MHV-3 or lipopolysaccharide (LPS), analyzed for transcription of mRNA and production of interleukin-1 (IL-1), tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta), mouse fibrinogen-like protein (musfiblp), tissue factor (TF), leukotriene B4, and prostaglandin E2 (PGE2). Macrophages from BALB/cJ mice produced …

Basic Medical Research As A Means Of Solving National Health Problems, M. Perwaiz Iqbal, M Anwar Waqar

Department of Biological & Biomedical Sciences

No abstract provided.

Nutrient Limitation In A Southwestern Desert Reservoir: Eutrophication Of Las Vegas Bay, Lake Mead, Nevada, Davine M. Lieberman

Publications (WR)

Algal bioassay tests were conducted with Selenastrum capricornutum and natural algae on inner Las Vegas Bay, Lake Mead, Nevada, from December 1992 through September 1993, to identify any nutrient limitation in an area of the reservoir that has experienced problems associated with severe nutrient enrichment. Three areas were sampled based on a gradient of water quality conditions that existed in Las Vegas Bay (LVB). Disodium ethylenedinitrilotetraacetate (EDTA) significantly stimulated algal growth compared to non-EDTA treatment. Algal bioassays indicated that phosphorus (P) was the primary limiting nutrient at all stations for most of the test dates. Chl a response with EDTA …

The Cytoplasmic Tail Domain Of The Vacuolar Protein Sorting Receptor Vps10p And A Subset Of Vps Gene Products Regulate Receptor Stability, Function, And Localization, Joan Lin-Cereghino, Eric G. Marcusson, Scott D. Emr

College of the Pacific Faculty Articles

VPS10 of Saccharomyces cerevisiae encodes a type I transmembrane receptor protein required for the sorting of the soluble vacuolar hydrolase carboxypeptidase Y (CPY). To characterize the essential structural features and intercompartmental transport itinerary of the CPY receptor, we have constructed mutant forms of Vps10p that alter the carboxyterminal cytoplasmic tail of the protein. In addition, we have analyzed the effect these mutations as well as mutations in several VPS genes have on the function, stability, and localization of Vps10p. Although wild-type Vps10p is very stable over a 3-h chase period, overproduction of Vps10p results in PEP4-dependent degradation of the receptor. …

Ph And Calcium Dependence Of Hemolysis Due To Rickettsia Prowazekii: Comparison With Phospholipase Activity, David M. Ojcius, M. Thibon, C. Mounier, Alice Dautry-Varsat

All Dugoni School of Dentistry Faculty Articles

Rickettsia prowazekii invades nucleated cells through phagocytosis and subsequently proliferates in the cytoplasm of the host cell. Hemolysis and a phospholipase A2 (PLA2) activity at neutral pHs have previously been reported; even though the phagosomal environment is most likely acidic. We here show that R. prowazekii and R. typhi also lyse erythrocytes at mildly acidic pHs, compatible with an early phagosomal compartment. For R. prowazekii, hemolysis at an acidic pH but not a neutral pH is enhanced by Ca2+, raising the possibility that more than one membranolytic factor may be produced by the rickettsiae. The rickettsiae alone display PLA2 activity, …

Environments Of The Four Tryptophans In The Extracellular Domain Of Human Tissue Factor: Comparison Of Results From Absorption And Fluorescence Difference Spectra Of Tryptophan Replacement Mutants With The Crystal Structure Of The Wild-Type Protein, C. A. Hasselbacher, E. Rusinova, R. Rusinova, R. A. Kohanski, W. Lam, A. Guha, J. Du, T. C. Lin, I. Polikarpov, C. W. G. Boys, Y. Nemerson, W. H. Konigsberg, J. B. A. Ross

Chemistry and Biochemistry Faculty Publications

The local environments of the four tryptophan residues of the extracellular domain of human tissue factor (sTF) were assessed from difference absorption and fluorescence spectra. The difference spectra were derived by subtracting spectra from single Trp-to-Phe or Trp-to-Tyr replacement mutants from the corresponding spectrum of the wild-type protein. Each of the mutants was capable of enhancing the proteolytic activity of factor Vila showing that the mutations did not introduce major structural changes, although the mutants were more susceptible to denaturation by guanidinium chloride. The difference spectra indicate that the Trp residues are buried to different extents within the protein matrix. …

Investigation Of The Substrate Recognition Characteristics And Kinetics Of Mammalian Mitochondrial Dna Topoisomerase I, Zeki Topcu

Theses and Dissertations in Biomedical Sciences

Topoisomerases are DNA-modifying enzymes found in prokaryotes, eukaryotes, viruses and organelles such as chloroplast and mitochondria. Information about these enzymes in eukaryotic systems is mostly limited to nuclear enzymes, although our laboratory has been characterizing the biochemical and biophysical properties of the mammalian mitochondrial topoisomerases. We have determined the polarity of the attachment of mitochondrial topoisomerase I to its substrate DNA. To study the substrate preference and kinetic parameters of mitochondrial topoisomerase I, selected regions of mammalian mitochondrial DNA (mtDNA) were inserted into pGEM plasmid vectors following a series of modification and optimization experiments of currently available methods for PCR-cloning. …

Identification And Characterization Of Mitochondrial Dna Variants In Alzheimer's Disease, Natasha Singh Hamblet

Theses and Dissertations in Biomedical Sciences

Alzheimer's Disease (AD) is a complex neurodegenerative disorder that affects a significant portion of the human population regardless of ethnicity or gender. A mitochondrial hypothesis of AD has been proposed based on a number of studies which establish altered oxidative phosphorylation (OXPHOS) and ATP synthesis in AD tissue. ATP demand is most prevalent in the brain; damage to OXPHOS could severely impair brain metabolism, thereby leading to a decline in cognitive function. Four out of five complexes in the OXPHOS pathway are partly encoded by mitochondrial DNA (mtDNA); thus, this may be a crucial site of lesions that alter brain …

In Vitro Expression Of Human Zona Pellucida Protein 3, Chih-Wei Chen

Theses and Dissertations in Biomedical Sciences

Human zona pellucida protein 3 (hZP3) is the putative receptor on the zona pellucida of the mature oocyte that recognizes and binds sperm, and therefore plays a critical role in fertilization.

A full length cDNA of hZP3 (1278 bp) was amplified from the human ovary mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The hZP3 cDNA was subcloned into PSK and pREP4 expression vectors. The cDNA of hZP3 was further characterized by restriction mapping, PCR, auto-sequencing and Southern blot analysis by using an internal oligonucleotide probe, and found to be identical to the one reported by J. Dean. Using autosequencing, 289 …

Trophic Transfer Of Fatty Acids, Sterols, And A Triterpenoid Alcohol Between Bacteria, A Ciliate, And The Copepod Acartia Tonsa, Melissa C. Ederington, George B. Mcmanus, H. Rodger Harvey

OES Faculty Publications

The incorporation of lipids into the copepod Acartia tonsa and its eggs was measured when it was fed either a bacterivorous ciliate (Pleuronema sp.) or a diatom (Thalassiosira weissflogii). Egg production was 10-fold higher on the diatom diet, whereas hatch success of eggs was the same for algal and ciliate diets. Adult copepods fed diatoms contained more total fatty acid and sterols than copepods fed the ciliate diet, and individual lipids reflected the dietary source. Eggs from diatom-fed copepods had fewer fatty acids but more sterols than eggs from copepods on a ciliate diet. Ciliate-fed copepods and …