Physical Sciences and Mathematics Commons^™

Antifungal Amphiphilic Aminoglycoside K20: Bioactivities And Mechanism Of Action, Sanjib K. Shrestha, Cheng-Wei Tom Chang, Nicole Meissner, John Oblad, Jaya P. Shrestha, Kevin N. Sorensen, Michelle M. Grilley, Jon Y. Takemoto

Chemistry and Biochemistry Faculty Publications

K20 is a novel amphiphilic antifungal aminoglycoside that is synthetically derived from the antibiotic kanamycin A. Reported here are investigations of K20′s antimicrobial activities, cytotoxicity, and fungicidal mechanism of action. In vitro growth inhibitory activities against a variety of human and plant pathogenic yeasts, filamentous fungi, and bacteria were determined using microbroth dilution assays and time-kill curve analyses, and hemolytic and animal cell cytotoxic activities were determined. Effects on Cryptococcus neoformans H-99 infectivity were determined with a preventive murine lung infection model. The antifungal mechanism of action was studied using intact fungal cells, yeast lipid mutants, and small unilamellar lipid …

Environmental Health Effects Of Multiple Exposures: Systemic Risks And The Detroit River International Crossing Study, Tor H. Oiamo

Electronic Thesis and Dissertation Repository

This thesis examines cumulative exposures to traffic noise and outdoor air pollution on environmental and health related quality of life in Windsor, Ontario, and provides a critical analysis of the environmental assessment process for the Detroit River International Crossing (DRIC) Study. The research utilizes a systemic risk framework to understand environmental health and stress effects of cumulative exposures. The significance of this research is based on a relative absence of literature on the systemic health risks of cumulative exposures and the need to elucidate environmental annoyance as a health outcome for risk assessment. The objectives of the research were to …

Inhibitors Of Nα-Acetyl-L-Ornithine Deacetylase: Synthesis, Characterization And Analysis Of Their Inhibitory Potency, J. Hlavacek, J. Picha, V. Vanek, J. Jiracek, J. Slaninova, V. Fucik, M. Budesinsky, Danuta Gillner, Richard Holz

Richard C. Holz

A series of N α-acyl (alkyl)- and N α-alkoxycarbonyl-derivatives of l- and d-ornithine were prepared, characterized, and analyzed for their potency toward the bacterial enzyme N α-acetyl-l-ornithine deacetylase (ArgE). ArgE catalyzes the conversion of N α-acetyl-l-ornithine to l-ornithine in the fifth step of the biosynthetic pathway for arginine, a necessary step for bacterial growth. Most of the compounds tested provided IC50 values in the μM range toward ArgE, indicating that they are moderately strong inhibitors. N α-chloroacetyl-l-ornithine (1g) was the best inhibitor tested toward ArgE providing an IC50 value of 85 μM while N α-trifluoroacetyl-l-ornithine (1f), N α-ethoxycarbonyl-l-ornithine (2b), and …

The Dape-Encoded N-Succinyl-L,L-Diaminopimelic Acid Desuccinylase From Haemophilus Influenzae Contains Two Active-Site Histidine Residues, Danuta M. Gillner, David L. Bienvenue, Boguslaw P. Nocek, Andzrej Joachimiak, Vincentos Zachary, Brian Bennett, Richard C. Holz

Richard C. Holz

The catalytic and structural properties of the H67A and H349A dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae were investigated. On the basis of sequence alignment with the carboxypeptidase from Pseudomonas sp. strain RS-16, both H67 and H349 were predicted to be Zn(II) ligands. The H67A DapE enzyme exhibited a decreased catalytic efficiency (180-fold) compared with wild-type (WT) DapE towards N-succinyldiaminopimelic acid. No catalytic activity was observed for H349A under the experimental conditions used. The electronic paramagnetic resonance (EPR) and electronic absorption data indicate that the Co(II) ion bound to H349A-DapE is analogous to that of WT DapE after the …

Mutation Of H63 And Its Catalytic Affect On The Methionine Aminopeptidase From Escherichia Coli, Sanghamitra Mitra, Brian Bennett, Richard C. Holz

Richard C. Holz

In order to gain insight into the mechanistic role of a flexible exterior loop near the active site, made up of Y62, H63, G64, and Y65, that has been proposed to play an important role in substrate binding and recognition in the methionyl aminopeptidase from Escherichia coli (EcMetAP-I), the H63A enzyme was prepared. Mutation of H63 to alanine does not affect the ability of the enzyme to bind divalent metal ions. The specific activity of H63A EcMetAP-I was determined using four different substrates of varying lengths, namely, l-Met-p-NA, MAS, MGMM and MSSHRWDW. For the smallest/shortest substrate (l-Met-p-NA) the specific activity …

Identification Of An Active Site-Bound Nitrile Hydratase Intermediate Through Single Turnover Stopped-Flow Spectroscopy, Natalie Gumataotao, Misty L. Kuhn, Natalia Hajnas, Richard C. Holz

Richard C. Holz

Stopped-flow kinetic data were obtained for the iron-type nitrile hydratase from Rhodococcus equi TG328-2 (ReNHase) using methacrylonitrile as the substrate. Multiple turnover experiments suggest a three-step kinetic model that allows for the reversible binding of substrate, the presence of an intermediate, and the formation of product. Microscopic rate constants determined from these data are in good agreement with steady state data confirming that the stopped-flow method used was appropriate for the reaction. Single turnover stopped-flow experiments were used to identify catalytic intermediates. These data were globally fit confirming a three-step kinetic model. Independent absorption spectra acquired between 0.005 …

Immobilization Of The Aminopeptidase From Aeromonas Proteolytica On Mg2+/Al3+ Layered Double Hydroxide Particles, Steven Frey, Stephanie Guilmet, Richard Egan, Alyssa Bennett, Sarah Soltau, Richard Holz

Richard C. Holz

A novel biomaterial formed by the immobilization of the Aminopeptidase from Aeromonas proteolytica (AAP) on synthetic Mg2+ and Al3+ ion-containing layered double hydroxide (LDH) particles was prepared. Immobilization of AAP on the LDH particles in a buffered, aqueous mixture is rapid such that the maximum loading capacity, 1 × 10−9 moles of AAP/mg LDH, is achieved in a few minutes. X-ray powder diffraction of LDH samples before and after treatment with AAP indicates that the enzyme does not intercalate between the layers of LDH, but instead binds to the surface. Treatment of AAP/LDH with various amounts of salt in a …

Analyzing The Binding Of Co(Ii)-Specific Inhibitors To The Methionyl Aminopeptidases From Escherichia Coli And Pyrococcus Furiosus, Sanghamitra Mitra, George Sheppard, Jieyi Wang, Brian Bennett, Richard Holz

Richard C. Holz

Methionine aminopeptidases (MetAPs) represent a unique class of protease that is capable of the hydrolytic removal of an N-terminal methionine residue from nascent polypeptide chains. MetAPs are physiologically important enzymes; hence, there is considerable interest in developing inhibitors that can be used as antiangiogenic and antimicrobial agents. A detailed kinetic and spectroscopic study has been performed to probe the binding of a triazole-based inhibitor and a bestatin-based inhibitor to both Mn(II)- and Co(II)-loaded type-I (Escherichia coli) and type-II (Pyrococcus furiosus) MetAPs. Both inhibitors were found to be moderate competitive inhibitors. The triazole-type inhibitor was found to …

Direct Patterning Of Silanized-Biomolecules On Semiconductor Surfaces, Dorjderem Nyamjav, Richard Holz

Richard C. Holz

A novel approach to pattern silanized-biomolecules directly onto glass (SiO_x) substrates via Dip-Pen nanolithography (DPN) and microcontact printing (μCP) is presented. Subsequent hybridization reactions of DPN patterned silanized-DNA with its complementary strands provide “proof-of-concept” that the patterned oligonucleotides maintain their biological activities. The fabrication strategy does not require premodification of substrates and offers a cheap and robust way to immobilize molecules on electronically important semiconductor surfaces.

Direct Patterning Of A Cyclotriveratrylene Derivative For Directed Self-Assembly Of C60, Zachary Osner, Dorjderem Nyamjav, Richard Holz, Daniel Becker

Richard C. Holz

A novel apex-modified cyclotriveratrylene (CTV) derivative with an attached thiolane-containing lipoic acid linker was directly patterned onto gold substrates via dip-pen nanolithography (DPN). The addition of a dithiolane-containing linker to the apex of CTV provides a molecule that can adhere to a gold surface with its bowl-shaped cavity directed away from the surface, thereby providing a surface-bound CTV host that can be used for the directed assembly of guest molecules. Subsequent exposure of these CTV microarrays to C60 in toluene resulted in the directed assembly of predesigned, spatially controlled, high-density microarrays of C60. The molecular recognition capabilities of this CTV …

Structural Basis For Catalysis By The Mono- And Dimetalated Forms Of The Dape-Encoded N-Succinyl-L,L-Diaminopimelic Acid Desuccinylase, Boguslaw Nocek, Danuta Gillner, Yao Fan, Richard Holz, Andzrej Joachimiak

Richard C. Holz

Biosynthesis of lysine and meso-diaminopimelic acid in bacteria provides essential components for protein synthesis and construction of the bacterial peptidoglycan cell wall. The dapE operon enzymes synthesize both meso-diaminopimelic acid and lysine and, therefore, represent potential targets for novel antibacterials. The dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase functions in a late step of the pathway and converts N-succinyl-l,l-diaminopimelic acid to l,l-diaminopimelic acid and succinate. Deletion of the dapE gene is lethal to Helicobacter pylori and Mycobacterium smegmatis, indicating that DapE's are essential for cell growth and proliferation. Since there are no similar pathways in humans, inhibitors …

Inhibitors Of Bacterial N-Succinyl-L,L-Diaminopimelic Acid Desuccinylase (Dape) And Demonstration Of In Vitro Antimicrobial Activity, Danuta Gillner, Nicola Armoush, Richard Holz, Daniel Becker

Richard C. Holz

The dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) is a critical bacterial enzyme for the construction of the bacterial cell wall. A screen biased toward compounds containing zinc-binding groups (ZBG’s) including thiols, carboxylic acids, boronic acids, phosphonates and hydroxamates has delivered a number of micromolar inhibitors of DapE from Haemophilus influenzae, including the low micromolar inhibitor L-captopril (IC₅₀ = 3.3 μM, K_i = 1.8 μM). In vitro antimicrobial activity was demonstrated for l-captopril against Escherichia coli.

Lysine Biosynthesis In Bacteria: A Metallodesuccinylase As A Potential Antimicrobial Target, Danuta Gillner, Daniel P. Becker, Richard C. Holz

Richard C. Holz

In this review, we summarize the recent literature on dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE) enzymes, with an emphasis on structure–function studies that provide insight into the catalytic mechanism. Crystallographic data have also provided insight into residues that might be involved in substrate and hence inhibitor recognition and binding. These data have led to the design and synthesis of several new DapE inhibitors, which are described along with what is known about how inhibitors interact with the active site of DapE enzymes, including the efficacy of a moderately strong DapE inhibitor.

The Fe-Type Nitrile Hydratase From Comamonas Testosteroni Ni1 Does Not Require An Activator Accessory Protein For Expression In Escherichia Coli, Misty Kuhn, Salette Martinez, Natalie Gumataotao, Uwe Bornscheuer, Dali Liu, Richard Holz

Richard C. Holz

We report herein the functional expression of an Fe-type nitrile hydratase (NHase) without the co-expression of an activator protein or the Escherichia coli chaperone proteins GroES/EL. Soluble protein was obtained when the α- and β-subunit genes of the Fe-type NHase Comamonas testosteroni Ni1 (CtNHase) were synthesized with optimized E. coli codon usage and co-expressed. As a control, the Fe-type NHase from Rhodococcus equi TG328–2 (ReNHase) was expressed with (ReNHase+Act) and without (ReNHase−Act) its activator protein, establishing that expression of a fully functional, metallated ReNHase enzyme requires the co-expression of its activator protein, similar to all other Fe-type NHase enzymes reported …

Immobilization Of Motile Bacterial Cells Via Dip-Pen Nanolithography, Dorjderem Nyamjav, Sergey Rozhok, Richard Holz

Richard C. Holz

A strategy to bind bacterial cells to surfaces in a directed fashion via dip-pen nanolithography (DPN) is presented. Cellular attachment to pre-designed DPN generated microarrays was found to be dependent on the shape and size of the surface feature. While this observation is likely due in part to a dense, well formed mercaptohexadecanoic acid (MHA) monolayer generated via DPN, it may also simply be due to the physical shape of the surface structure. Motile Pseudomonas aeruginosa bacterial cells were observed to bind to DPN generated mercaptohexadecanoic acid/poly-L-lysine (MHA/PLL) line patterns, 'blocks' made up of eight lines with 100 nm spacings, …

Structural Characterization Of Zn(Ii)-, Co(Ii)-, And Mn(Ii)-Loaded Forms Of The Arge-Encoded N-Acetyl-L-Ornithine Deacetylase From Escherichia Coli, Ye Tao, Jacob Shokes, Wade Mcgregor, Robert Scott, Richard Holz

Richard C. Holz

The Zn, Co, and Mn K-edge extended X-ray absorption fine structure (EXAFS) spectra of the N-acetyl-l-ornithine deacetylase (ArgE) from Escherichia coli, loaded with one or two equivalents of divalent metal ions (i.e., [Zn(II)_(ArgE)], [Zn(II)Zn(II)(ArgE)], [Co(II)_(ArgE)], [Co(II)Co(II)(ArgE)], [Mn(II)_(ArgE)], and [Mn(II)Mn(II)(ArgE)]), were recorded. The Fourier transformed data (FT) for [Zn(II)_(ArgE)], [Zn(II)Zn(II)(ArgE)], [Co(II)_(ArgE)] and [Co(II)Co(II)(ArgE)] are dominated by a peak at 2.05 Å, that can be fit assuming five or six light atom (N,O) scatterers. Inclusion of multiple-scattering contributions from the outer-shell atoms of a histidine-imidazole ring resulted in reasonable Debye–Waller factors for these contributions and a slight reduction in the goodness-of-fit value …

Synthesis Of N-Succinyl-L,L-Diaminopimelic Acid Mimetics Via Selective Protection, V. Vanek, J. Picha, M. Budesinsky, M. Sanda, J. Jiracek, Richard Holz, J. Hlavacek

Richard C. Holz

The search for potential inhibitors that target so far unexplored bacterial enzyme mono-N-succinyl-L,Ldiaminopimelic acid desuccinylase (DapE) has stimulated a development of methodology for quick and efficient preparation of mono-N-acylated 2,6-diaminopimelic acid (DAP) derivatives bearing the different carboxyl groups or lipophilic moieties on their amino group.

The Fierce Green Fire: Vol 5 Issue 11, Wofford College Environmental Studies Program

The Fierce Green Fire

No abstract provided.

Analyzing Drone Footage To Assess Necessary Streetlight Placement: Where In New Washington Heights And Poe Mill Are Streetlights Most Necessary?, Savannah Jennings, Billy Tutt

Earth and Environmental Sciences Presentations

New Washington Heights and Poe Mill are low income neighborhoods with high crime rates and the people living there feel that more street lights would make the areas more safe at night. In our project, we aimed to answer the question: where are streetlights most appropriate in these neighborhoods? And can footage taken from the recent drone flyover help to improve our ability to accurately model streetlights in these neighborhoods? We will present our findings to the residents of Poe Mill and New Washington Heights in order to aid their efforts in making their streets safer. This question is relevant …

Farmers Markets In Urban Us Counties: The Relationship Between Income, Obesity, And Access, Mia El-Hamaki

Earth and Environmental Sciences Presentations

In the last decade there has been a convergence of factors that have contributed to an increase in food deserts, obesity, and income inequality in the US. Two-thirds of US adults are overweight or obese. This prevalence disproportionately affects low income areas with high minority populations (Ruelas et al., 2012). Simultaneously, research shows that grocery stores are tending to move away from low income, inner city communities and towards affluent urban and suburban areas. In order to combat the growing food deserts and obesity rates as a result of this shift, farmers markets have been proposed and utilized as a …

A Comparison Of Trout Streams: Examining The Reasons For Angler Visitation To Certain Counties Around North Carolina, Rob George

Earth and Environmental Sciences Presentations

Recreational fishing is a popular and profitable sport for states across the country (Ditton et al., 2002). Mountain trout angling provides as much as 174 million dollars annually to the state of North Carolina (NCWRC, 2009). This number shows that recreational trout fishing is an important economic factor for North Carolina. The goal of this project is to determine what variables correlate to the popularity of a county among trout anglers. Through personal experience, academic articles, and interviews with fly fishermen a list of variables was created in order to compare two streams in different counties. These variables include household …

Urban Garden Survival, Phoebe Ferguson, Rachel Martin

Biology Presentations

Urban gardens are community led plots designated for agricultural purposes in residential and urban areas. Greenville County has seen a recent growth in urban gardens with the assistance of non-profit groups like Gardening for Good. The current total in Greenville County stands at 79 with new gardens added every year. While the growth is encouraging, some gardens have failed. This study uses GIS to explore the social and ecological factors that correlate with urban garden survival in an effort to provide garden managers with information that will help them develop gardens that thrive and persist.

College Of Engineering Senior Design Competition Fall 2014, University Of Nevada, Las Vegas

Fred and Harriet Cox Senior Design Competition Projects

Part of every UNLV engineering student’s academic experience, the senior design project stimulates engineering innovation and entrepreneurship. Each student in their senior year chooses, plans, designs, and prototypes a product in this required element of the curriculum. A capstone to the student’s educational career, the senior design project encourages the student to use everything learned in the engineering program to create a practical, real world solution to an engineering challenge. The senior design competition helps focus the senior students in increasing the quality and potential for commercial application for their design projects. Judges from local industry evaluate the projects on …

Paris Talks: The Art Of Climate Communication, Phaedra Pezzullo

Sea Level Rise Collection

No abstract provided.

Measurement Of The Pp→Zzproduction Cross Section And Constraints On Anomalous Triple Gauge Couplings In Four-Lepton Final States At √S=8tev, Cms Collaboration, Cern, Switzerland, V. Gaultney, Samantha Hewamanage, Stephan Linn, Pete E. Markowitz, German Martinez, Jorge Luis Rodriguez

Department of Physics

A measurement of the inclusive ZZproduction cross section and constraints on anomalous triple gauge couplings in proton–proton collisions at √s=8TeVare presented. The analysis is based on a data sample, corresponding to an integrated luminosity of 19.6fb−1, collected with the CMS experiment at the LHC. The measurements are performed in the leptonic decay modes ZZ →, where=e, μand =e, μ, τ. The measured total cross section σ(pp →ZZ) =7.7 ±0.5(stat)+0.5−0.4(syst)±0.4 (theo) ±0.2(lumi)pb, for both Zbosons produced in the mass range 60

Molecular Dynamics Study Of The Opening Mechanism For Dna Polymerase I, Carol A. Parish, Bill R. Miller Iii, Eugene Y. Wu

Chemistry Faculty Publications

During DNA replication, DNA polymerases follow an induced fit mechanism in order to rapidly distinguish between correct and incorrect dNTP substrates. The dynamics of this process are crucial to the overall effectiveness of catalysis. Although Xray crystal structures of DNA polymerase I with substrate dNTPs have revealed key structural states along the catalytic pathway, solution fluorescence studies indicate that those key states are populated in the absence of substrate. Herein, we report the first atomistic simulations showing the conformational changes between the closed, open, and ajar conformations of DNA polymerase I in the binary (enzyme:DNA) state to better understand its …

Analysis Of Drug Interactions With Lipoproteins By High Performance Affinity Chromatography, Matthew R. Sobansky

Department of Chemistry: Dissertations, Theses, and Student Research

High density lipoprotein (HDL), low density lipoprotein (LDL), and very low density lipoprotein (VLDL) are lipoproteins previously shown to bind many basic and neutral hydrophobic drugs in serum. These interactions impact the distribution, delivery, metabolism, and excretion of drugs and are important in determining drug activity, pharmacokinetics, and toxicity in the human body. Information about drug-lipoprotein interactions and the strength of these interactions can be useful in determining the distribution of drugs following administration.

The research presented in this dissertation uses high performance affinity chromatography (HPAC) and packed columns to study binding of the drug propranolol to immobilized lipoproteins such …

Spartanburg Humane Society Data Analysis: Hotspots And Resource Management, Leigh Robertson

Earth and Environmental Sciences Presentations

A data set from the Spartanburg Humane Society was provided with the hope that the use of GIS could show patterns and hotspots of dog and cat pickup locations. With a better understanding of where the majority of the animals come from, the Humane Society can better distribute their man power and resources to help as many homeless animals as possible. According to Beck (1973) the main reasons for stray and abandoned animals is negligence causing pet releases or escapes, and also breeding. Therefore, finding and targeting areas where there are many strays (potentially as a result of a release, …

Wing And Fin Outfitters: Using Gis To Determine The Ideal Location For A Tackle Shop In Charleston, Sc, Matt Giordano

Earth and Environmental Sciences Presentations

A few years down the road I hope to open a tackle shop in Charleston, South Carolina with my current college roommate. I have been fishing my entire life and since coming down to school I have spent many breaks down in Charleston. My interest in the area really blossomed during the summer of 2013 which I spent in Charleston fishing and working around the water. During my time in Charleston I have realized that many local shops are in good locations but they do not provide excellent service. Shops seem to get by on reputation rather than actual service. …

Kelvin Probe Force Microscopy On Graphene Thin Films For Solar Cell And Biosensing Applications, Faranak Sharifi

Electronic Thesis and Dissertation Repository

Graphene, a one atom thick planer sheet of carbon atoms, has attracted much attention in scientific and technological communities due to its remarkable electronic and physical properties. Graphene has been widely studied as an alternative to transparent conducting Indium Tin Oxide (ITO) electrodes in organic photovoltaics fabrication. Graphene platforms have also attracted interest in biological applications. However, large area graphene films are not yet widely commercialized because the fabrication techniques needed to prepare high quality graphene are expensive and non-scalable. More importantly, most of the low cost fabrication techniques require using toxic materials, which are not biocompatible for using graphene …