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2010

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Polyurea Elastomer Protects Utility Pole Crossarms From Damage By Pileated Woodpeckers, Shelagh Tupper, William Andelt, John Cummings, Charles Weisner, Richard Harness Jan 2010

Polyurea Elastomer Protects Utility Pole Crossarms From Damage By Pileated Woodpeckers, Shelagh Tupper, William Andelt, John Cummings, Charles Weisner, Richard Harness

United States Department of Agriculture Wildlife Services: Staff Publications

Woodpeckers cause severe damage to utility poles and crossarms, resulting in substantial economic losses to utility companies. We evaluated effectiveness of a polyurea elastomer coating material for reducing damage by captive pileated woodpeckers (Dryocopus pileatus) to utility pole crossarms. Because woodpeckers inflicted essentially no damage to the fully coated crossarms, we infer that the coating material holds substantial promise for protecting utility pole crossarms. Additional research should be conducted to evaluate the coating under field conditions.


Increased Risk Of Chronic Wasting Disease In Rocky Mountain Elk Associated With Decreased Magnesium And Increased Manganese In Brain Tissue, Stephen N. White, Katherine I. O’Rourke, Thomas Gidlewski, Kurt C. Vercauteren, Michelle R. Mousel, Gregory E. Phillips, Terry R. Spraker Jan 2010

Increased Risk Of Chronic Wasting Disease In Rocky Mountain Elk Associated With Decreased Magnesium And Increased Manganese In Brain Tissue, Stephen N. White, Katherine I. O’Rourke, Thomas Gidlewski, Kurt C. Vercauteren, Michelle R. Mousel, Gregory E. Phillips, Terry R. Spraker

United States Department of Agriculture Wildlife Services: Staff Publications

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) of Rocky Mountain elk in North America. Recent studies suggest that tissue and blood mineral levels may be valuable in assessing TSE infection in sheep and cattle. The objectives of this study were to examine baseline levels of copper, manganese, magnesium, zinc, selenium, and molybdenum in the brains of Rocky Mountain elk with differing prion genotypes and to assess the association of mineral levels with CWD infection. Elk with leucine at prion position 132 had significantly lower magnesium levels than elk with 2 copies of methionine. Chronic wasting disease-positive elk …


Review Of Issues Concerning The Use Of Reproductive Inhibitors, With Particular Emphasis On Resolving Human-Wildlife Conflicts In North America, Kathleen A. Fagerstone, Lowell A. Miller, Gary Killian, Christi A. Yoder Jan 2010

Review Of Issues Concerning The Use Of Reproductive Inhibitors, With Particular Emphasis On Resolving Human-Wildlife Conflicts In North America, Kathleen A. Fagerstone, Lowell A. Miller, Gary Killian, Christi A. Yoder

United States Department of Agriculture Wildlife Services: Staff Publications

This manuscript provides an overview of past wildlife contraception efforts and discusses the current state of research. Two fertility control agents, an avian reproductive inhibitor containing the active ingredient nicarbazin and an immunocontraceptive vaccine, have received regulatory approval with the Environmental Protection Agency and are commercially available in the USA. OvoControl G Contraceptive Bait for Canada Geese and Ovo Control for pigeons are delivered as oral baits. An injectable immunocontraceptive vaccine (GonaCon Immunocontraceptive Vaccine) was registered with the Environmental Protection Agency for use in female white-tailed deer in September 2009. An injectable product (GonaCon Immunocontraceptive Vaccine) is registered for use …


Experimental Infection Of Raccoons (Procyon Lotor) With West Nile Virus, J. Jeffrey Root, Kevin T. Bentler, Nicole M. Nemeth, Thomas Gidlewski, Terry R. Spraker, Alan B. Franklin Jan 2010

Experimental Infection Of Raccoons (Procyon Lotor) With West Nile Virus, J. Jeffrey Root, Kevin T. Bentler, Nicole M. Nemeth, Thomas Gidlewski, Terry R. Spraker, Alan B. Franklin

United States Department of Agriculture Wildlife Services: Staff Publications

To characterize the responses of raccoons to West Nile virus (WNV) infection, we subcutaneously exposed them to WNV. Moderately high viremia titers (≤ 104.6 plaque forming units [PFU]/mL of serum) were noted in select individuals; however, peak viremia titers were variable and viremia was detectable in some individuals as late as 10 days post-inoculation (DPI). In addition, fecal shedding was prolonged in some animals (e.g., between 6 and 13 DPI in one individual), with up to105.0 PFU/fecal swab detected. West Nile virus was not detected in tissues collected on 10 or 16 DPI, and no histologic lesions attributable …


Response Of Captive Skunks To Microencapsulated Tetracycline, Brandon S. Schmit, Thomas M. Primus, Jerome C. Hurley, Dennis J. Kohler, Shawna F. Graves Jan 2010

Response Of Captive Skunks To Microencapsulated Tetracycline, Brandon S. Schmit, Thomas M. Primus, Jerome C. Hurley, Dennis J. Kohler, Shawna F. Graves

United States Department of Agriculture Wildlife Services: Staff Publications

A captive striped skunk (Mephitis mephitis) study was conducted between February and June 2004 at the United States Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services National Wildlife Research Center, Fort Collins, Colorado, USA. The main objective was to determine the percentage of adult striped skunks that were marked after consuming placebo oral rabies vaccine (ORV) baits containing 100 mg of an experimental microencapsulated (coated microparticle) tetracycline hydrochloride biomarker. Biomarkers were identified in the canine teeth and mandibles of five of five skunks that consumed an ORV bait. A second objective was to determine if …


New Evidence Suggests Southern China As A Common Source Of Multiple Clusters Of Highly Pathogenic H5n1 Avian Influenza Virus, Bin Wu, Chengmin Wang, Guoying Dong, Yunhai Guo, Dale L. Nolte, Thomas Jude Deliberto, Jianguo Xu, Mingxing Duan, Hongxuan He Jan 2010

New Evidence Suggests Southern China As A Common Source Of Multiple Clusters Of Highly Pathogenic H5n1 Avian Influenza Virus, Bin Wu, Chengmin Wang, Guoying Dong, Yunhai Guo, Dale L. Nolte, Thomas Jude Deliberto, Jianguo Xu, Mingxing Duan, Hongxuan He

United States Department of Agriculture Wildlife Services: Staff Publications

Highly pathogenic H5N1 avian influenza is considered an avian disease, although there is some evidence of limited human-to-human transmission of the virus. A global effort is underway to control or eradicate the highly pathogenic H5N1 avian influenza virus in poultry and prevent human exposure, both of which may also reduce the risk of pandemic emergence. Hemagglutinin gene sequences from 215 human H5N1 influenza viruses were used to trace the source and dispersal pattern of human H5N1 influenza viruses on a global scale. A mutation network and phylogenetic analyses of the hemagglutinin gene show that human H5N1 influenza viruses can be …


Spring Grazing Impacts On The Vegetation Of Reed Canarygrass–Invaded Wetlands, Heidi Hilhouse, Susan J. Tunnell, James L. Stubbendieck Jan 2010

Spring Grazing Impacts On The Vegetation Of Reed Canarygrass–Invaded Wetlands, Heidi Hilhouse, Susan J. Tunnell, James L. Stubbendieck

Department of Agronomy and Horticulture: Faculty Publications

The Rainwater Basin region in Nebraska is critically important stopover habitat for spring waterfowl migrations, but the ability of these sites to produce sufficient food for migrating waterfowl is endangered by the invasion of reed canarygrass (Phalaris arundinacea L.). This species produces thick litter layers and abundant aboveground biomass, reducing germination and seedling survival of the annual plant species responsible for much of the seed production in the area. Cattle grazing often is used as a management tool in the Rainwater Basin to slow or reverse reed canarygrass invasion and to improve growing conditions for more desirable plant species. …


Factors Affecting The Alkaline Cooking Performance Of Selected Corn And Sorghum Hybrids, Weston B. Johnson, Wajira S. Ratnayake, David S. Jackson, Kyung-Min Lee, Timothy J. Herrman, Scott R. Bean, Stephen Mason Jan 2010

Factors Affecting The Alkaline Cooking Performance Of Selected Corn And Sorghum Hybrids, Weston B. Johnson, Wajira S. Ratnayake, David S. Jackson, Kyung-Min Lee, Timothy J. Herrman, Scott R. Bean, Stephen Mason

Department of Agronomy and Horticulture: Faculty Publications

Dent com (Zea mays L.) and sorghum (Sorghum hieolor L. Moench) sample sets representative of commonly grown hybrids and diverse physical attributes were analyzed for alkaline cooking performance. The influence of kernel characteristics including hardness, density, starch properties (thermal. pasting, and crystallinity). starch content. protein content. and prolamin content on alkaline cooking performance was also determined. Com nixtamal moisture content was lower for hard, dense kernels with high protein contents; sorghum nixtamal moisture content was lower for kernels with low moisture contents and low starch relative crystallinities. Statistically significant (P < 0.05) regression equations showed that com nixtamal moisture content was influenced by TADD (tangential abrasive dehulling device) index. kernel moisture content. starch content. and protein content; sorghum nixtamal moisture content was influenced by starch relative crystallinity. kernel moisture content, and abrasive hardness index. Pericarp removal was not strongly correlated with kernel characterization tests. Location (environmental) and hybrid (genetic) factors influenced most kernel characteristics and nixtamalization processing variables.


Evaluation Of The Intercom Model For Predicting Growth Of Forest Herbs, Zorica S. Popović, J. L. Lindquist Jan 2010

Evaluation Of The Intercom Model For Predicting Growth Of Forest Herbs, Zorica S. Popović, J. L. Lindquist

Department of Agronomy and Horticulture: Faculty Publications

The ecophysiological model INTERCOM was evaluated for its suitability in predicting the growth of perennial forest herbs. A field experiment was conducted to obtain data on photosynthesis and growth parameters of two spring flowering understorey geophytes. Results were used to parameterize the model and its performance was evaluated using the average normalized difference (AE) between predicted and observed biomass and the leaf area index. The model was assumed to provide accurate simulations if the AE was smaller than 0.4. Adjusting the photosynthetic intensity parameters in the model to reflect observed changes in photosynthesis throughout the growing period resulted in the …


Two And Three-Dimensional Gene Transfer From Enzymatically Degradable Hydrogel Scaffolds, Yuguo Lei, Quinn K.T. Ng, Tatiana Segura Jan 2010

Two And Three-Dimensional Gene Transfer From Enzymatically Degradable Hydrogel Scaffolds, Yuguo Lei, Quinn K.T. Ng, Tatiana Segura

Department of Chemical and Biomolecular Engineering: Faculty Publications

The ability to genetically modify mesenchymal stem cells (MSCs) seeded inside synthetic hydrogel scaffolds would offer an alternative approach to guide MSC differentiation. In this report, we explored gene transfer to MSCs seeded on top, or inside matrix metalloproteinase (MMP) degradable hydrogels that were loaded with DNA/poly(ethylene imine) (PEI) polyplexes. DNA/PEI polyplexes were encapsulated inside poly(ethylene glycol) (PEG) hydrogels crosslinked with MMP degradable peptides via Michael Addition chemistry. Gene transfer was visualized and quantified through using a vector encoding for green fluorescent protein and luciferase. We found that gene transfer to MSCs was possible for cells seeded both in two …


A Model Of Tuberculosis Transmission And Intervention Strategies In An Urban Residential Area, Elsje Pienaar, Aaron M. Fluitt, Scott E. Whitney, Alison G. Freifeld, Hendrik J. Viljoen Jan 2010

A Model Of Tuberculosis Transmission And Intervention Strategies In An Urban Residential Area, Elsje Pienaar, Aaron M. Fluitt, Scott E. Whitney, Alison G. Freifeld, Hendrik J. Viljoen

Department of Chemical and Biomolecular Engineering: Faculty Publications

The model herein aims to explore the dynamics of the spread of tuberculosis (TB) in an informal settlement or township. The population is divided into households of various sizes and also based on commuting status. The model dynamics distinguishes between three distinct social patterns: the exposure of commuters during travel, random diurnal interaction and familial exposure at night. Following the general SLIR models, the population is further segmented into susceptible (S), exposed/latently infected (L), active/infectious (I), and recovered (R) individuals. During the daytime, commuters travel on public transport, while non-commuters randomly interact in the community to mimic chance encounters with …


Fed-Batch Fermentation Of Gm-Csf-Producing Glycoengineered Pichia Pastoris Under Controlled Specific Growth Rate, Pieter P. Jacobs, Mehmet Inan, Nele Festjens, Jurgen Haustraete, Annelies Van Hecke, Roland Contreras, Michael M. Meagher, Nico Callewaert Jan 2010

Fed-Batch Fermentation Of Gm-Csf-Producing Glycoengineered Pichia Pastoris Under Controlled Specific Growth Rate, Pieter P. Jacobs, Mehmet Inan, Nele Festjens, Jurgen Haustraete, Annelies Van Hecke, Roland Contreras, Michael M. Meagher, Nico Callewaert

Papers in Biomolecular Engineering

Background: Yeast expression systems with altered N-glycosylation are now available to produce glycoproteins with homogenous, defined N-glycans. However, data on the behaviour of these strains in high cell density cultivation are scarce.

Results: Here, we report on cultivations under controlled specific growth rate of a GlycoSwitch-Man5 Pichia pastoris strain producing Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) at high levels (hundreds of milligrams per liter). We demonstrate that homogenous Man5GlcNAc2 N-glycosylation of the secreted proteins is achieved at all specific growth rates tested.

Conclusions: Together, these data illustrate that the GlycoSwitch-Man5 P. pastoris is a robust production …


Efficacy Of Singular And Stacked Brown Midrib 6 And 12 In The Modification Of Lignocellulose And Grain Chemistry, Scott E. Sattler, Deanna L. Funnell-Harris, Jeffrey F. Pedersen Jan 2010

Efficacy Of Singular And Stacked Brown Midrib 6 And 12 In The Modification Of Lignocellulose And Grain Chemistry, Scott E. Sattler, Deanna L. Funnell-Harris, Jeffrey F. Pedersen

Department of Agronomy and Horticulture: Faculty Publications

In sorghum, brown midrib (bmr) 6 and 12 impair the last two steps of monolignol synthesis. bmr genes were introduced into grain sorghum to improve the digestibility of lignocellulosic tissues for grazing or bioenergy uses following grain harvest. Near-isogenic grain sorghum hybrids (AWheatland X RTx430) were developed containing bmr6, bmr12, and the bmr6 bmr12 double mutant (stacked), and their impacts were assessed in a two-year field study. The bmr genes did not significantly impact grain or lignocellulosic tissue yield. Lignocellulosic tissue from bmr6, bmr12, and stacked hybrids had reduced lignin content and increased in …


Fine Mapping Of The Scn Resistance Locus Rhg1-B From Pi 88788, Myungsik Kim, David L. Hyten, Andrew F. Bent, Brian W. Diers Jan 2010

Fine Mapping Of The Scn Resistance Locus Rhg1-B From Pi 88788, Myungsik Kim, David L. Hyten, Andrew F. Bent, Brian W. Diers

Department of Agronomy and Horticulture: Faculty Publications

Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is the most economically damaging soybean [Glycine max (L.) Merr.] pest in the USA and genetic resistance is a key component for its control. Although SCN resistance is quantitative, the rhg1 locus on chromosome 18 (formerly known as Linkage Group G) confers a high level of resistance. The objective of this study was to fi ne-map the rhg1-b allele that is derived from plant introduction (PI) 88788. F2 and F3 plants and F3:4 lines from crosses between SCN resistant and susceptible genotypes were tested with genetic markers to …


Opto-Electrophoretic Detection Of Bio-Molecules Using Conducting Chalcogenide Glass Sensors, Zhiyong Yang, Megan K. Fah, Kelly A. Reynolds, Jonathan D. Sexton, Mark R. Riley, Marie-Laure Anne, Bruno Bureau, Pierre Lucas Jan 2010

Opto-Electrophoretic Detection Of Bio-Molecules Using Conducting Chalcogenide Glass Sensors, Zhiyong Yang, Megan K. Fah, Kelly A. Reynolds, Jonathan D. Sexton, Mark R. Riley, Marie-Laure Anne, Bruno Bureau, Pierre Lucas

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Novel telluride glasses with high electrical conductivity, wide infrared transparency and good resistance to crystallization are used to design an opto-electrophoretic sensor for detection and identification of hazardous microorganisms. The sensor is based on an attenuated total reflectance element made of Ge-As-Te glass that serves as both an optical sensing zone and an electrode for driving the migration of bio-molecules within the evanescent wave of the sensor. An electric field is applied between the optical element and a counter electrode in order to induce the migration of bio-molecules carrying surface charges. The effect of concentration and applied voltage is tested …


Udp-Glucose Dehydrogenase As A Novel Field-Specific Candidate Biomarker Of Prostate Cancer, Dali Huang, George P. Casale, Jun Tian, Subodh M. Lele, Vladimir M. Pisarev, Melanie A. Simpson, George P. Hemstreet Iii Jan 2010

Udp-Glucose Dehydrogenase As A Novel Field-Specific Candidate Biomarker Of Prostate Cancer, Dali Huang, George P. Casale, Jun Tian, Subodh M. Lele, Vladimir M. Pisarev, Melanie A. Simpson, George P. Hemstreet Iii

Department of Biochemistry: Faculty Publications

Uridine diphosphate (UDP)-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose to yield UDP-glucuronic acid, a precursor for synthesis of glycosaminoglycans and proteoglycans that promote aggressive prostate cancer (PC) progression. The purpose of our study was to determine if the UGDH expression in normal appearing acini (NAA) from cancerous glands is a candidate biomarker for PC field disease/effect assayed by quantitative fluorescence imaging analysis (QFIA). A polyclonal antibody to UGDH was titrated to saturation binding and fluorescent microscopic images acquired from fixed, paraffin-embedded tissue slices were quantitatively analyzed. Specificity of the assay was confirmed by Western blot analysis and competitive inhibition …


N-Glycans On The Link Domain Of Human Hare/Stabilin-2 Are Needed For Hyaluronan Binding To Purified Ecto-Domain, But Not For Cellular Endocytosis Of Hyaluronan, Ed Harris, Simon Parry, Mark Sutton-Smith, Madhu S. Pandey, Maria Panico, Howard R. Morris, Stuart M. Haslam, Anne Dell, Paul H. Weigel Jan 2010

N-Glycans On The Link Domain Of Human Hare/Stabilin-2 Are Needed For Hyaluronan Binding To Purified Ecto-Domain, But Not For Cellular Endocytosis Of Hyaluronan, Ed Harris, Simon Parry, Mark Sutton-Smith, Madhu S. Pandey, Maria Panico, Howard R. Morris, Stuart M. Haslam, Anne Dell, Paul H. Weigel

Department of Biochemistry: Faculty Publications

The hyaluronic acid receptor for endocytosis (HARE)/Stabilin- 2 is the primary systemic scavenger receptor for 13 ligands including hyaluronan (HA), heparin and chondroitin sulfates. Most ligand-binding sites are within the 190 kDa isoform, which contains ~25 kDa of N-glycans and is the C-terminal half of the full-length 315 kDa HARE. Glycoproteomic analyses of purified recombinant human 190-HARE ecto-domain identified a diverse population of glycans at 10 of 17 consensus sites. The most diversity (and the only sialylated structures) occurred at N2280, within the HA-binding Link domain. To determine if these N-glycans are required for HA binding, …


In Vitro Stable Isotope Labeling For Discovery Of Novel Metabolites By Liquid Chromatography-Mass Spectrometry: Confirmation Of Γ-Tocopherol Metabolism In Human A549 Cell, Wen-Chu Yang, Fred E. Regnier, Qing Jiang, Jiri Adamec Jan 2010

In Vitro Stable Isotope Labeling For Discovery Of Novel Metabolites By Liquid Chromatography-Mass Spectrometry: Confirmation Of Γ-Tocopherol Metabolism In Human A549 Cell, Wen-Chu Yang, Fred E. Regnier, Qing Jiang, Jiri Adamec

Department of Biochemistry: Faculty Publications

A general approach for discovering novel catabolic metabolites from a parent biocompound was developed and validated on metabolism of γ-tocopherol in human A549 cell. Method is based on LC-MS analysis of in vitro stable isotope labeled metabolites and assumes that a parent compound and its metabolites share a common functional group that can be derivatized by well-documented reagents. In this method, two equal aliquots of extracted metabolites are separately derivatized with isotope-coded (heavy) and non-isotope-coded (light) form of derivatizing reagent, mixed at 1:1 ratio and analyzed using LC-MS. The metabolites with common functional group are then easily recognized by determination …


Regulation Of Sealing Ring Formation By L-Plastin And Cortactin In Osteoclasts, Tao Ma, Kavitha Sadashivalah, Nandakumar Madayiputhiya, Meenakshi A. Chellaia Jan 2010

Regulation Of Sealing Ring Formation By L-Plastin And Cortactin In Osteoclasts, Tao Ma, Kavitha Sadashivalah, Nandakumar Madayiputhiya, Meenakshi A. Chellaia

Department of Biochemistry: Faculty Publications

The aim of this study is to identify the exact mechanism(s) by which cytoskeletal structures are modulated during bone resorption. In this study, we have shown the possible role of different actin-binding and signaling proteins in the regulation of sealing ring formation. Our analyses have demonstrated a significant increase in cortactin and a corresponding decrease in L-plastin protein levels in osteoclasts subjected to bone resorption for 18 h in the presence of RANKL, M-CSF, and native bone particles. Time-dependent changes in the localization of L-plastin (in actin aggregates) and cortactin (in the sealing ring) suggest that these proteins may be …


Structure Of The Proline Utilization A Proline Dehydrogenase Domain Inactivated By N-Propargylglycine Provides Insight Into Conformational Changes Induced By Substrate Binding And Flavin Reduction, Dhiraj Srivastava, Weidong Zhu, William H. Johnson Jr., Christian P. Whitman, Donald F. Becker, John J. Tanner Jan 2010

Structure Of The Proline Utilization A Proline Dehydrogenase Domain Inactivated By N-Propargylglycine Provides Insight Into Conformational Changes Induced By Substrate Binding And Flavin Reduction, Dhiraj Srivastava, Weidong Zhu, William H. Johnson Jr., Christian P. Whitman, Donald F. Becker, John J. Tanner

Department of Biochemistry: Faculty Publications

Proline utilization A (PutA) from Escherichia coli is a flavoprotein that has mutually exclusive roles as a transcriptional repressor of the put regulon and a membrane-associated enzyme that catalyzes the oxidation of proline to glutamate. Previous studies have shown that the binding of proline in the proline dehydrogenase (PRODH) active site and subsequent reduction of the FAD trigger global conformational changes that enhance PutA-membrane affinity. These events cause PutA to switch from its repressor to enzymatic role, but the mechanism by which this signal is propagated from the active site to the distal membrane-binding domain is largely unknown. Here, it …


Analysis Of Leigh Syndrome Mutations In The Yeast Surf1 Homolog Reveals A New Member Of The Cytochrome Oxidase Assembly Factor Family, Megan Bestwick, Mi-Young Jeong, Oleh Khalimonchuk, Hyung Kim, Dennis R. Winge Jan 2010

Analysis Of Leigh Syndrome Mutations In The Yeast Surf1 Homolog Reveals A New Member Of The Cytochrome Oxidase Assembly Factor Family, Megan Bestwick, Mi-Young Jeong, Oleh Khalimonchuk, Hyung Kim, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Three missense SURF1 mutations identified in patients with Leigh syndrome (LS) were evaluated in the yeast homolog Shy1 protein. Introduction of two of the Leigh mutations, F249T and Y344D, in Shy1 failed to significantly attenuate the function of Shy1 in cytochrome c oxidase (CcO) biogenesis as seen with the human mutations. In contrast, a G137E substitution in Shy1 results in a nonfunctional protein conferring a CcO deficiency. The G137E Shy1 mutant phenocopied shy1 Δ cells in impaired Cox1 hemylation and low mitochondrial copper. A genetic screen for allele-specific suppressors of the G137E …


Formation Of The Redox Cofactor Centers During Cox1 Maturation In Yeast Cytochrome Oxidase, Oleh Khalimonchuk, Megan Bestwick, Brigitte Meunier, Talina C. Watts, Dennis R. Winge Jan 2010

Formation Of The Redox Cofactor Centers During Cox1 Maturation In Yeast Cytochrome Oxidase, Oleh Khalimonchuk, Megan Bestwick, Brigitte Meunier, Talina C. Watts, Dennis R. Winge

Department of Biochemistry: Faculty Publications

The biogenesis of cytochrome c oxidase initiates with synthesis and maturation of the mitochondrionencoded Cox1 subunit prior to the addition of other subunits. Cox1 contains redox cofactors, including the low-spin heme a center and the heterobimetallic heme a3:CuB center. We sought to identify the step in the maturation of Cox1 in which the redox cofactor centers are assembled. Newly synthesized Cox1 is incorporated within one early assembly intermediate containing Mss51 in Saccharomyces cerevisiae. Subsequent Cox1 maturation involves the progression to downstream assembly intermediates involving Coa1 and Shy1. We show that the two heme a cofactor sites …


The Role Of Coa2 In Hemylation Of Yeast Cox1 Revealed By Its Genetic Interaction With Cox10, Megan Bestwick, Oleh Khalimonchuk, Fabien Pierrel, Dennis R. Winge Jan 2010

The Role Of Coa2 In Hemylation Of Yeast Cox1 Revealed By Its Genetic Interaction With Cox10, Megan Bestwick, Oleh Khalimonchuk, Fabien Pierrel, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Saccharomyces cerevisiae cells lacking the cytochrome c oxidase (CcO) assembly factor Coa2 are impaired in Cox1 maturation and exhibit a rapid degradation of newly synthesized Cox1. The respiratory deficiency of coa2 Δ cells is suppressed either by the presence of a mutant allele of the Cox10 farnesyl transferase involved in heme a biosynthesis or through impaired proteolysis by the disruption of the mitochondrial Oma1 protease. Cox10 with an N196K substitution functions as a robust gain-of-function suppressor of the respiratory deficiency of coa2 Δ cells but lacks suppressor activity for two other CcO assembly mutant strains, the coa1 Δ and shy1 …


Quantitative Nuclear Proteomics Identifies Mtor Regulation Of Dna Damage Response, Sricharan Bandhakavi, Young-Mi Kim, Seung-Hyun Ro, Hongwei Xie, Getiria Onsongo, Chang-Bong Jun, Do-Hyung Kim, Timothy J. Griffin Jan 2010

Quantitative Nuclear Proteomics Identifies Mtor Regulation Of Dna Damage Response, Sricharan Bandhakavi, Young-Mi Kim, Seung-Hyun Ro, Hongwei Xie, Getiria Onsongo, Chang-Bong Jun, Do-Hyung Kim, Timothy J. Griffin

Department of Biochemistry: Faculty Publications

Cellular nutritional and energy status regulates a wide range of nuclear processes important for cell growth, survival, and metabolic homeostasis. Mammalian target of rapamycin (mTOR) plays a key role in the cellular responses to nutrients. However, the nuclear processes governed by mTOR have not been clearly defined. Using isobaric peptide tagging coupled with linear ion trap mass spectrometry, we performed quantitative proteomics analysis to identify nuclear processes in human cells under control of mTOR. Within 3 h of inhibiting mTOR with rapamycin in HeLa cells, we observed downregulation of nuclear abundance of many proteins involved in translation and RNA modification. …


Phosphatidylinositol 5-Phosphate Links Dehydration Stress To The Activity Of Arabidopsis Trithorax-Like Factor Atx1, Ivan Ndamukong, David R. Jones, Hanna Lapko, Nullin Divecha, Zoya V. Avramova Jan 2010

Phosphatidylinositol 5-Phosphate Links Dehydration Stress To The Activity Of Arabidopsis Trithorax-Like Factor Atx1, Ivan Ndamukong, David R. Jones, Hanna Lapko, Nullin Divecha, Zoya V. Avramova

School of Biological Sciences: Faculty Publications

Background: Changes in gene expression enable organisms to respond to environmental stress. Levels of cellular lipid second messengers, such as the phosphoinositide PtdIns5P, change in response to a variety of stresses and can modulate the localization, conformation and activity of a number of intracellular proteins. The plant trithorax factor (ATX1) trimethylates the lysine 4 residue of histone H3 (H3K4me3) at gene coding sequences, which positively correlates with gene transcription. Microarray analysis has identified a target gene (WRKY70) that is regulated by both ATX1 and by the exogenous addition of PtdIns5P in Arabidopsis. Interestingly, ATX1 …


Evolution Of The Kdo2-Lipid A Biosynthesis In Bacteria, Stephen O. Opiyo, Rosevelt L. Pardy, Hideaki Moriyama, Etsuko N. Moriyama Jan 2010

Evolution Of The Kdo2-Lipid A Biosynthesis In Bacteria, Stephen O. Opiyo, Rosevelt L. Pardy, Hideaki Moriyama, Etsuko N. Moriyama

School of Biological Sciences: Faculty Publications

Background: Lipid A is the highly immunoreactive endotoxic center of lipopolysaccharide (LPS). It anchors the LPS into the outer membrane of most Gram-negative bacteria. Lipid A can be recognized by animal cells, triggers defense-related responses, and causes Gram-negative sepsis. The biosynthesis of Kdo2-lipid A, the LPS substructure, involves with nine enzymatic steps.

Results: In order to elucidate the evolutionary pathway of Kdo2-lipid A biosynthesis, we examined the distribution of genes encoding the nine enzymes across bacteria. We found that not all Gram-negative bacteria have all nine enzymes. Some Gram-negative bacteria have no genes encoding …


Agmatidine, A Modified Cytidine In The Anticodon Of Archaeal TrnaIle, Base Pairs With Adenosine But Not With Guanosine, Debabrata Mandal, Caroline Köhrer, Dan Su, Susan P. Russell, Kady Krivos, Colette M. Castleberry, Paul H. Blum, Patrick A. Limbach, Dieter Söll, Uttam L. Rajbhandary Jan 2010

Agmatidine, A Modified Cytidine In The Anticodon Of Archaeal TrnaIle, Base Pairs With Adenosine But Not With Guanosine, Debabrata Mandal, Caroline Köhrer, Dan Su, Susan P. Russell, Kady Krivos, Colette M. Castleberry, Paul H. Blum, Patrick A. Limbach, Dieter Söll, Uttam L. Rajbhandary

School of Biological Sciences: Faculty Publications

Modification of the cytidine in the first anticodon position of the AUA decoding tRNAIle (tRNAIle 2 ) of bacteria and archaea is essential for this tRNA to read the isoleucine codon AUA and to differentiate between AUA and the methionine codon AUG. To identify the modified cytidine in archaea, we have purified this tRNA species from Haloarcula marismortui, established its codon reading properties, used liquid chromatography–mass spectrometry (LC-MS) to map RNase A and T1 digestion products onto the tRNA, and used LC-MS/MS to sequence the oligonucleotides in RNase A digests. These analyses revealed that the modification of …


Isolation And Characterization Of Bacteria Resistant To Metallic Copper Surfaces, Christophe Espírito Santo, Paula Vasconcelos Morais, Gregor Grass Jan 2010

Isolation And Characterization Of Bacteria Resistant To Metallic Copper Surfaces, Christophe Espírito Santo, Paula Vasconcelos Morais, Gregor Grass

School of Biological Sciences: Faculty Publications

Metallic copper alloys have recently attracted attention as a new antimicrobial weapon for areas where surface hygiene is paramount. Currently it is not understood on a molecular level how metallic copper kills microbes, but previous studies have demonstrated that a wide variety of bacteria, including Escherichia coli, Staphylococcus aureus, and Clostridium difficile, are inactivated within minutes or a few hours of exposure. In this study, we show that bacteria isolated from copper alloy coins comprise strains that are especially resistant against the toxic properties exerted by dry metallic copper surfaces. The most resistant of 294 isolates were …


Nm23-H1 Can Induce Cell Cycle Arrest And Apoptosis In B Cells, Tathagata Choudhuri, Masanao Murakami, Rajeev Kaul, Sushil K. Sahu, Suchitra Mohanty, Subhash C. Verma, Pankaj Kumar, Erle S. Robertson Jan 2010

Nm23-H1 Can Induce Cell Cycle Arrest And Apoptosis In B Cells, Tathagata Choudhuri, Masanao Murakami, Rajeev Kaul, Sushil K. Sahu, Suchitra Mohanty, Subhash C. Verma, Pankaj Kumar, Erle S. Robertson

School of Biological Sciences: Faculty Publications

Nm23-H1 is a well-known tumor metastasis suppressor, which functions as a nucleoside-diphosphate kinase converting nucleoside diphosphates to nucleoside triphosphates with an expense of ATP. It regulates a variety of cellular activities, including proliferation, development, migration and differentiation known to be modulated by a series of complex signaling pathway. Few studies have addressed the mechanistic action of Nm23-H1 in the context of these cellular processes. To determine the downstream pathways modulated by Nm23-H1, we expressed Nm23-H1 in a Burkitt lymphoma derived B-cell line BJAB and performed pathway specific microarray analysis. The genes with significant changes in expression patterns were clustered in …


Increased Dendrite Branching In Aβpp/Ps1 Mice And Elongation Of Dendrite Arbors By Fasudil Administration, Brian A. Couch, George J. Demarco, Shannon L. Gourley, Anthony J. Koleske Jan 2010

Increased Dendrite Branching In Aβpp/Ps1 Mice And Elongation Of Dendrite Arbors By Fasudil Administration, Brian A. Couch, George J. Demarco, Shannon L. Gourley, Anthony J. Koleske

School of Biological Sciences: Faculty Publications

Amyloid-β (Aβ) overproduction and dendrite arbor atrophy are hallmarks of Alzheimer’s disease. The RhoA GTPase (Rho) signals through Rho kinase (ROCK) to control cytoskeletal dynamics and regulate neuron structure. Hyperactive Rho signaling destabilizes neurons leading to dendritic regression that can be rescued by genetic or pharmacological reduction of ROCK signaling. To understand what effect reduced ROCK signaling has on the dendrite arbors of mice that overproduce Aβ, we administered the ROCK inhibitor fasudil to AβPP/PS1 transgenic mice. We report that increased dendrite branching occurs in AβPP/PS1 mice and that fasudil promotes …