Biochemistry, Biophysics, and Structural Biology Commons^™

Stealth Carbon Nanotubes: Strategies To Coat Carbon Nanotubes To Prevent Opsonization And Improve Biodistribution, Nalinikanth Kotagiri

Graduate Theses and Dissertations

Carbon nanotubes (CNT) have recently been in the limelight for its potential role in disease diagnostics and therapeutics. Even before these medical applications can be realized, there is a need to address issues like opsonization, phagocytosis by macrophages and sequestration to liver and spleen for eventual elimination from the body. We believe coating CNT with biocompatible and opsonin resistant moieties will not only help CNT in traversing the blood stream to reach the target organ, but also improve biodistribution tremendously. We set out to achieve this by firstly identifying a new compound, mAmp, which is a fluorescent derivative of the …

The Role Of Albino3 And The Lipid Environment In Chloroplast Signal Recognition Particle Targeting, Nathan Lewis

Graduate Theses and Dissertations

Signal recognition particles (SRPs) in pro- and eukaryotes function in cotranslational targeting of nascent poplypeptides to an SRP receptor at the target membrane. A unique chloroplast SRP (cpSRP) functions post-translationally to direct light-harvesting chlorophyll-binding proteins (LHCPs) to the receptor cpFtsY at the thylakoid membrane for LHCP insertion in a process involving the integral membrane protein Albino3 (Alb3) and requiring GTP. Work here focuses on understanding cpSRP targeting events at the thylakoid membrane, specifically those involving Alb3 and the lipid environment.

We show an interaction between the novel cpSRP subunit cpSRP43 and the soluble, stromal-exposed C terminus of Albino3 (Alb3-Cterm). We …

Applications Of Site-Specific Recombination Systems In Transgene Expression And Marker Gene Removal, Mehmet Aydin Akbudak

Graduate Theses and Dissertations

Site Specific Recombination systems, such as FLP-FRT and Cre-lox, have been successfully used for site-specific gene integration and marker-gene deletion in plant systems. They are very useful tools in the integration of single-copy full-length transgene cassettes into the genome because the transgene integration via conventional methods often generate multi-copy locus. Such complex locus containing direct and inverted repeats of full-length and truncated copies of the transgene cassette generate aberrant RNA resulting in gene silencing. Therefore, for stable gene expression, a single copy transgene locus is preferred. However, even single copy locus sometimes succumbs to gene silencing. Although the mechanism is …

Tryptophan Anchored Peptides In Lipid Bilayer Membranes: Control Of Peptide Orientation And The Phase Behavior Of Cholesterol-Containing Ternary Lipid Mixtures, Johanna Maria Rankenberg

Graduate Theses and Dissertations

Model WALP peptides and "next generation" WALP-derived hydrophobic model peptides were employed to discover principles that govern protein-lipid interactions in biological membranes.

Ternary cholesterol-containing lipid mixtures were examined in the presence of WALP peptides of different lengths (acetyl-GWW(LA)_nLWWA-ethanolamide, with n between 3 and 8). Deuterium NMR spectra from labeled lipids reveal that WALP peptides may stabilize lipid ordered "raft" domains and therefore promote lipid phase separation, albeit to a minor extent. The results depend upon whether dioleolyl- or diphytanoyl-phosphatidylcholine is present as the fluid lipid component.

Several WALP peptides were modified to remove anchoring Trp residues from one …

Intraspecific Variation In Two Cosmopolitan Myxomycetes, Didymium Squamulosum And Didymium Difforme (Physarales: Didymiaceae), Katherine Elizabeth Winsett

Graduate Theses and Dissertations

The myxomycetes (plasmodial slime molds or myxogastrids) are one of three groups considered to be true slime molds (class Eumycetozoa sensu Olive 1975). Two vegetative states--amoebae and plasmodia--along with a spore-producing fruiting body characterize the life cycle of the myxomycetes. These organisms are associated with decaying plant material and are found in all terrestrial habitats worldwide. A number of species are considered cosmopolitan, being found worldwide, where they are associated with a diversity of microhabitats and substrates. A review of the literature, including molecular investigations in all three groups of slime molds, is presented, and this is followed by four …

Applications Of Variable Number Tandem Repeat Genotyping In The Validation Of An Animal Medical Model And Gene Flow Studies In Threatened Populations Of Reptiles, Candace D. Smith

Graduate Theses and Dissertations

We used variable number tandem repeats (VNTR) to validate the chicken as a human medical model for Pulmonary Arterial Hypertension. We identified seven regions on four chromosomes and interrogated for VNTR markers that significantly associate with Pulmonary Hypertension Syndrome/ascites. In those regions, we identified 7 candidate genes; AGTR1, ACE, p38MAPK, SST, 5HT2B, NET1, and CALM3 for further analysis as significantly contributing QTL for ascites/PHS. We also used variable number tandem repeats to measure gene flow and gather evidence for multiple paternity in a population of Timber rattlesnakes, Crotalus horridus. We were able to verify 1 VNTR that can be used …

The Role Of Multiple Ccaat-Binding Factors In Candida Albicans Gene Expression, Lashall Lynn Bates

Graduate Theses and Dissertations

The CCAAT-binding factor is a heterooligomeric transcription factor that is evolutionarily conserved in eukaryotes. In yeast, the DNA-binding component that interacts with the CCAAT consensus sequence in promoters consists of the subunits termed Hap2p, Hap3p and Hap5p. In yeast and fungi, a fourth subunit, Hap4p, is required for regulating gene expression. The goal of this research is to understand the function of the Candida albicans CCAAT-binding factor and how it relates to virulence and pathogenicity. C. albicans is a human opportunistic pathogen responsible for a variety of mucosal and systemic infections that result in significant morbidity and mortality, particularly in …

Characterization And Functional Regulation Of Bioactive Peptides In Avian Macrophages And Heterophils, Lakshmi Kannan

Graduate Theses and Dissertations

Oligopeptides and low molecular weight polypeptides play central roles as effectors and signal transducers acting as hormones, neurotransmitters, growth factors, toxins, and antimicrobial factors that are important for the survival of the organism. Owing to the ubiquitous involvement of peptides in many key regulatory processes, we have been interested to identify native peptides in different cells and tissues and understand their functions. To conduct our studies, we used avian macrophages and heterophils as models of specialized cells which constitute central components of innate immunity. These studies involved (a) qualitative identification and characterization of the peptides associated with high intensity mass …

The Membrane Interface Of Chloroplast Signal Recognition Particle-Dependent Protein Targeting, Naomi Jane Marty

Graduate Theses and Dissertations

A novel signal recognition particle (SRP) found in the chloroplast (cpSRP) works in combination with the cpSRP receptor, cpFtsY, to facilitate the post-translational targeting of a family of nuclear-encoded thylakoid proteins to the Alb3 translocase in thylakoid membranes. Work here focused on understanding events at the membrane that take place to ensure targeting of the cpSRP-dependent substrate to Alb3. Specifically, we sought to understand the structural and functional role of membrane binding by cpFtsY, a protein that exhibits the ability to partition between the membrane (thylakoid) and soluble (stroma) phase during protein targeting. We also sought to understand whether a …

Understanding The Role Of Histones During Chromosome Segregation: A Study Of The Effects Of Histone Gene Dosage In Saccharomyces Cerevisiae, Rebecca Leigh Danforth

Graduate Theses and Dissertations

Chromatin plays a role in all cellular functions that involve DNA. These include, but are not limited to replication, recombination, transcription, and chromosome segregation. Chromosome segregation is an extremely well conserved cellular process and is essential for maintaining the genetic integrity of a cell. There is very strong evidence indicating that chromatin structure is critical for maintaining the fidelity of chromosome transmission, but its specific role(s) in this process remains unclear. Chromatin is comprised of arrays of nucleosomes that serve to compact DNA. These nucleosomes consist of 146 bp of DNA that is wrapped around a histone octamer; two each …

Influence On Proline Upon The Folding And Geometry Of A Model Transmembrane Peptide, Rachel Thomas

Inquiry: The University of Arkansas Undergraduate Research Journal

The orientations, geometries and lipid interactions of designed transmembrane (TM) peptides have attracted significant experimental and theoretical interest. Because the amino acid proline will introduce a known discontinuity into an alpha-helix, it is important to measure the extent of helix kinking caused by a single proline within an isolated TM helical domain. For this purpose, acetyl-GWWLALALAP10ALALALWWA-ethanolamide was synthesized, and pairs of deuterated alanines were included by using 60-100% deuterated fluorenylmethoxycarbonyl-L-alanine ifmoc-d4-L-Ala) at selected sequence positions. Solid-state deuterium fH) magnetic resonance spectra from oriented, hydrated samples (1140, peptide/lipid; using several lipids) reveal signals from many of the alanine backbone Ca deuterons …

Evolution Of Gene Structure In Multicellular Eukaryotes, Maria Hester

Inquiry: The University of Arkansas Undergraduate Research Journal

We investigated the patterns of intron conservation in eukaryotes for five different genes. The genes examined were ribosomal proteins L8, S14 and S17, along with elongation factor 2B and triose phosphate isomerase. Intron conservation for S14, S17, and triose phosphate isomerase was determined for 32 species representing the major branches of multicellular eukaryotes. For 25 conserved introns 16 were phase 0, five were phase 1, and four were phase 2. Triose phosphate isomerase had five of nine conserved introns shared between plants and animals, where S14 had one of nine and S17 had one of seven. However, there were two …

Letter From The Dean, Lalit Verma

Discovery, The Student Journal of Dale Bumpers College of Agricultural, Food and Life Sciences

No abstract provided.

Computational Study Of A Novel Dinuclear Metal Complex, Dragos Seghete

Inquiry: The University of Arkansas Undergraduate Research Journal

The first compound containing an M-H-M was recently reported by Vicic et al. With recent availability of large computational resources, molecular modeling has become a reliable tool for confirming experimental results. The novel dinuclear Ni complex [( dippm)2Ni2Br2]( 0 -H) was investigated in this work from a theoretical perspective. Full geometry optimization was carried on the dinuclear Ni complex at the DFT/B3LYP with the 6-31G* basis set. The result verifies the linear Ni-H-Ni bond. Two different starting structures that converged to the same geometry confirm that a global minimum was reached. The computed structure differs from the experimentally determined one …

Mapping Of Ssdna Nicks Within Dsdna Genomes By Two-Dimensional Gel Electrophoresis, Wayne P. Wahls, K. Mark Dewall, Mari K. Davidson

Journal of the Arkansas Academy of Science

DNA molecules within chromosomes undergo constant, dynamic changes yet maintain the integrity of the primary DNA sequence. DNA replication, adjustment of helical density, resolution of catenenes, repair of DNA damage, and homologous recombination each involve breakage and religation of the phosphate backbone of the double helix. Although the analysis of dsDNA breaks is facile, the analysis of ssDNA nicks is not. The principal impediment is that conventional, one-dimensional electrophoresis methods cannot readily detect ssDNA nicks in the context of dsDNA breaks. We therefore developed a two dimensional (native/denaturing) gel electrophoresis approach to map the positions of ssDNA nicks. Analysis of …

Can Martian Life Exist Under Dry Conditions?, Michael Kendrick

Inquiry: The University of Arkansas Undergraduate Research Journal

Recently, we have begun to hear more and more about news coming from Mars. With the recent confirmation of evidence of liquid water in Mars' past, evidence indicating liquid water in Mars' present, and as many missions planned for this decade and the next than have successfully reached the planet in the last 40 years, life on Mars is beginning to become a more relevant issue in our study of the planet. Although there have been no indications of multicellular life on Mars, there may be a possibility of microbial life in the subsurface. Since it appears that all of …

Rec12 (Spo11) Recominase Of Fission Yeast Promotes A Backup, Distributive Pathway For Chromosome Segregation In Meiosis I, Wallace D. Sharif, Mari K. Davidson, Wayne P. Wahls

Journal of the Arkansas Academy of Science

We studied the relationship between recombination and segregation in the fission yeast Schizosaccharomyces pombe. In meiosis, chromosomes undergo two rounds of segregation to produce haploid meiotic products. Crossover meiotic ecombination (chiasmata) promotes chromosome segregation during meiosis I; achiasmatic chromosomes often suffer on disjunction in meiosis I. Recl2 protein is a topoisomerase IIortholog that introduces double-strand DNA breaks that nitiate recombination. The red2 null (deletion) and active site (Y98F) mutants lack recombination and crossovers, and onsequently suffer meiosis I nondisjunction. However, null mutants chromosomes segregate to opposite poles more frequently than predicted. Thus, fission yeast has a backup, "distributive segregation" pathway …

Actin Dynamics Regulate Myosin Assembly In Muscle Cells, John Dylan Cook

Inquiry: The University of Arkansas Undergraduate Research Journal

During muscle cell (i.e. myocyte )development, organization of actin and myosin into the contractile unit (i.e. sarcomere) is required for proper muscle contraction. By disrupting the processes of muscle cell development, I am able to see which steps play important roles in proper maturation of myocytes. Elucidation of the key pathways in muscle development could lead to a better understanding of human cardiac hypertrophies and muscle myopathies. Since actin filament formation precedes myosin organization, I am using actin assembly inhibitors to determine if actin filaments are a necessary prerequisite for myosin organization. It is hypothesized that disruption of actin will …

Properties Of Modified Tryptophans In A Membrane- Spanning Channel, Erin M. Scherer

Inquiry: The University of Arkansas Undergraduate Research Journal

An emerging concept in biology assigns the amino acid tryptophan specific roles at the membrane/water interface that help to determine the conformation and biological function of membrane-spanning proteins. Previous studies involving the antibiotic model system gramicidin A (gA) have illustrated the importance of the indole ring of tryptophan (Trp) in anchoring proteins to a bilayer membrane and promoting ionic currents. To further investigate these phenomena, derivatives of Trp that have lost hydrogen-bonding ability (1-methyl-Trp ), have an altered dipole moment (7-aza-Trp), or both (1-methyl-7-azaTrp) were chosen for incorporation into gA. Gramicidin analogues that incorporate these modified Trps were then analyzed …

Development Of A Yeast Biosensor Strain For The Identification Of Genotoxic Compounds, Robin M. Reed

Inquiry: The University of Arkansas Undergraduate Research Journal

Advances in combinatorial chemistry have provided the pharmaceutical industry with innumerable lead compounds that could potentially serve as therapeutic agents. One of the challenges in the further development of such compounds is to rapidly, yet inexpensively, distinguish those that have undesirable effects such as genotoxicity. Thus, a simple biological assay that would permit the identification of potential DNA mutagens, and be adaptable to high-throughput technologies would be cost effective in screening such lead compounds. The current methods use the Ames and SOS tests involving prokaryotic organisms, while systems that utilize mammalian cell culture and/or animal testing are time-consuming and expensive. …

Voltage Gating Of A Model Membrane Spanning Channel, Eric Miller

Inquiry: The University of Arkansas Undergraduate Research Journal

Channels in cell membranes are important for intercellular communication and especially for the function of the nervous system in higher vertebrates. These channels consist of proteins made from the 20 common amino acids. Channel proteins are embedded into the lipid bilayer membranes of living cells and function by allowing the specific passage of a positively charged material such as sodium or potassium ions across the membrane in response to an external signal. The external signaloeither a chemical signal or a voltageoregulates the opening and closing of channels. In an attempt to understand the voltage-dependent opening of channels (igatingi), we are …

Molecular Orbital Studies Of Nitrosyl Metalloporphyrin Complexes, John P. Graham, Genevive Brown

Journal of the Arkansas Academy of Science

Molecular orbital calculations are applied to the study of the structure and bonding of nitrosyl metalloporphyrin complexes. The Fenske-Hall approximate molecular orbital method and the Amsterdam Density Functional (ADF) method are used. The calculations provide qualitative and quantitative explanations for the observed structural differences between the nitrosyl porphyrin complexes ofiron, manganese, and cobalt. It is proposed that the energy of the highest occupied molecular orbital (HOMO) of these complexes is primarily responsible for the observed structural differences. The interaction between the nitrosyl ligand and metal dz2 orbital results in an antibonding orbital that is occupied in the complexes of Fe …

Melanocortin-1-Receptor (Mcr-1) Gene Polymorphisms Associated With The Chicken E Locus Alleles, Andrew Ellett

Inquiry: The University of Arkansas Undergraduate Research Journal

The melanocortin 1-receptor (MCl-R) gene has been associated with E locus phenotypes in chickens. Variant alleles of the E locus are important for accurate down color sexing and also for the inhibition of unwanted tissue pigmentation in broilers. PCR (polymerase chain reaction) based tests for various replacement substitutions found in the published E allele sequences gave unexpected results when tested against known alleles of the E locus. To resolve these issues and gain a better understanding of how replacement substitutions in the MCl-R gene are affecting E locus phenotypes, a number of accessions of the e+, eb, wheaten ( ewh …

Analysis Of Phytoestrogens By High Performance Liquid Chromatography, Anne A. Grippo, Yan Xie, Benjamin L. Rougeau, William V. Wyatt

Journal of the Arkansas Academy of Science

Phytoestrogens are biochemicals synthesized in plants which mimic steroidal estrogen activity in mammals. Analysis of these compounds in the legumes which produce them and in body fluids is important to the study of their physiological effects. High pressure liquid chromatography (HPLC) has been found to be an efficient and sensitive method of identification and quantitation of isoflavonoids, one class of phytoestrogen. Here we report the separation of three isoflavonoids, biochanin A, genistein and daidzein using an HPLC system with a Cg reverse phase column and a linear gradient mobile phase containing acetonitrile and acetic acid/water (10/90, v/v) over 60 minutes. …

Electron Shock Waves, Frances Terry, Mostafa Hemmati

Journal of the Arkansas Academy of Science

No abstract provided.

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 …

Multisite Microprobes For Electrochemical Recordings In Biological Dynamics, G. Sreenivas, S. S. Ang, R. M. Ranade, A. S. Salian, W. D. Brown

Journal of the Arkansas Academy of Science

For over 30 years, techniques have been developed that allow for the microscale (10-30 /mum) measurement of chemical signals with high temporal resolution (1-200 Hz). Such measurements, called in vivo electrochemical recordings, allow for the direct determination of neurotransmitter molecules and related compounds in biological systems. Multiple recordings, simultaneously performed at different, closely spaced, well defined locations throughout a three-dimensional tissue volume in the brain, are of interest in neuroscience. Developments in microelectronic techniques enable the fabrication of multi-electrode microprobes for recording extracellular action potentials generated by individual neurons simultaneously. A high-yield microfabrication process has been successfully developed for the …

Effective, Reliable, Inexpensive Cryofixation Device, Lawrence A. Mink, Roger A. Buchanan

Journal of the Arkansas Academy of Science

No abstract provided.

Thermal Decomposition Studies Of Selected Transition Metal Polysulfide Complexes. Ii. Effect Of Atmosphere On Decomposition, Benjamin Rougeau, Mark Draganjac

Journal of the Arkansas Academy of Science

Initial studies involved the thermal decomposition profile of five polysulfide complexes in air up to 550 °C. Since our first report to the Academy in 1990, we have obtained the capability to run samples up to 1500°C under various gases. Thermal Gravimetric Analysis (TGA) and Differential Thermal Analysis (DTA)of a series of transition metal polysulfide complexes are presented. Compounds analyzed included Cp2TiS5 ,MoS9 2",MoOS8 2", Zn(S x ) 2", Cd(S x ) 2-, Fe2S122 - and NiS82.

Conformational And Circular Dichroism Studies On N-Acetyl-L-Prolyl-D-Alanyl-Methylamide, S. Ramaprasad

Journal of the Arkansas Academy of Science

The compact ordered conformations of the molecule N-Acteyl-L-Prolyl-D-Alanyl-Methylamide have been studied by semiempirical energy calculations in vacuum and circular dichroism (CD) in solution. The presence of ordered structure has been observed in hydrogen bond promoting solvents like trifluoroethanol by CD studies. In hydrogen bond breaking solvents, like trifluoroacetic acid (TFA), significant fraction of the ordered conformers probably assume extended conformation without intramolecular hydrogen bonds and perhaps are in equilibrium with the fraction of compact ordered structures. The trend observed in going from nonpolar to polar solvent is also compatible with the previous NMR studies in solution. The semiempirical energy calculations …