bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structural and Nonstructural Protein Genome Regions of Eastern Equine Encephalitis Virus Are Determinants of Interferon Sensitivity and Murine Virulence
Authors: Aguilar Patricia V. , Adams A. Paige , Wang Eryu , Kang Wenli , Carrara Anne-Sophie , Anishchenko Michael , Frolov Ilya , Weaver Scott C. 
Published: March 2008

ABSTRACT
Eastern equine encephalitis virus (EEEV) causes sporadic epidemics of human and equine disease in North America, but South American strains have seldom been associated with human neurologic disease or mortality, despite serological evidence of infection. In mice, most North American and South American strains of EEEV produce neurologic disease that resembles that associated with human and equine infections. We identified a South American strain that is unable to replicate efficiently in the brain or cause fatal disease in mice yet produces 10-fold higher viremia than virulent EEEV strains. The avirulent South American strain was also sensitive to human interferon (IFN)-α, -β, and -γ, like most South American strains, in contrast to North American strains that were highly resistant. To identify genes associated with IFN sensitivity and virulence, infectious cDNA clones of a virulent North American strain and the avirulent South American strain were constructed. Two reciprocal chimeric viruses containing swapped structural and nonstructural protein gene regions of the North American and South American strains were also constructed and found to replicate efficiently in vitro. Both chimeras produced fatal disease in mice, similar to that caused by the virulent North American strain. Both chimeric viruses also exhibited intermediate sensitivity to human IFN-α, -β, and -γ compared to that of the North American and South American strains. Virulence 50% lethal dose assays and serial sacrifice experiments further demonstrated that both structural and nonstructural proteins are important contributors to neurovirulence and viral tissue tropism. Together, the results of this study emphasize the complex and important influences of structural and nonstructural protein gene regions on EEEV virulence.
Herpes Simplex Virus Type 2 UL56 Interacts with the Ubiquitin Ligase Nedd4 and Increases Its Ubiquitination
Authors: Ushijima Yoko , Koshizuka Tetsuo , Goshima Fumi , Kimura Hiroshi , Nishiyama Yukihiro 
Published: March 2008

ABSTRACT

The herpes simplex virus UL56 gene is conserved among most members of the
Alphaherpesvirinae
family and plays a critical role in viral pathogenicity in vivo. The HSV-2 UL56 protein (UL56) is a C-terminally anchored type II membrane protein that is predicted to be inserted into the virion envelope, leaving its N-terminal domain in the tegument. UL56 interacts with KIF1A and UL11. Here we report that UL56 also interacts with the ubiquitin ligase Nedd4 and increases its ubiquitination. Nedd4 was identified as a UL56-interacting protein by a yeast two-hybrid screen. UL56 bound to Nedd4 via its PY motifs. Nedd4 was phosphorylated and degraded in wild-type HSV-2-infected cells but not in cells infected with a UL56-deficient mutant. Ubiquitination assays revealed that UL56 increased ubiquitinated Nedd4, which was actively degraded in infected cells. UL56 also caused a decrease in Nedd4 protein levels and the increased ubiquitination in cotransfected cells. However, UL56 itself was not ubiquitinated, despite its interaction with Nedd4. Based on these findings, we propose that UL56 regulates Nedd4 in HSV-2-infected cells, although deletion of UL56 had no apparent effect on viral growth in vitro.
Recruitment of Antigen-Presenting Cells to the Site of Inoculation and Augmentation of Human Immunodeficiency Virus Type 1 DNA Vaccine Immunogenicity by In Vivo Electroporation
Authors: Liu Jinyan , Kjeken Rune , Mathiesen Iacob , Barouch Dan H. 
Published: March 2008

ABSTRACT

In vivo electroporation (EP) has been shown to augment the immunogenicity of plasmid DNA vaccines, but its mechanism of action has not been fully characterized. In this study, we show that in vivo EP augmented cellular and humoral immune responses to a human immunodeficiency virus type 1 Env DNA vaccine in mice and allowed a 10-fold reduction in vaccine dose. This enhancement was durable for over 6 months, and re-exposure to antigen resulted in anamnestic effector and central memory CD8
+
T-lymphocyte responses. Interestingly, in vivo EP also recruited large mixed cellular inflammatory infiltrates to the site of inoculation. These infiltrates contained 45-fold-increased numbers of macrophages and 77-fold-increased numbers of dendritic cells as well as 2- to 6-fold-increased numbers of B and T lymphocytes compared to infiltrates following DNA vaccination alone. These data suggest that recruiting inflammatory cells, including antigen-presenting cells (APCs), to the site of antigen production substantially improves the immunogenicity of DNA vaccines. Combining in vivo EP with plasmid chemokine adjuvants that similarly recruited APCs to the injection site, however, did not result in synergy.
Role of the Membrane-Spanning Domain of Human Immunodeficiency Virus Type 1 Envelope Glycoprotein in Cell-Cell Fusion and Virus Infection
Authors: Shang Liang , Yue Ling , Hunter Eric 
Published: March 2008

ABSTRACTThe membrane-spanning domain (MSD) of the human immunodeficiency virus type 1 (HIV-1) gp41 glycoprotein is critical for its biological activity. Previous C-terminal truncation studies have predicted an almost invariant core structure of 12 amino acid residues flanked by basic amino acids in the HIV-1 MSD that function to anchor the glycoprotein in the lipid bilayer. To further understand the role of specific amino acids within the MSD core, we initially replaced the core region with 12 leucine residues and then constructed recovery-of-function mutants in which specific amino acid residues (including a GGXXG motif) were reintroduced. We show here that conservation of the MSD core sequence is not required for normal expression, processing, intracellular transport, and incorporation into virions of the envelope glycoprotein (Env). However, the amino acid composition of the MSD core does influence the ability of Env to mediate cell-cell fusion and plays a critical role in the infectivity of HIV-1. Replacement of conserved amino acid residues with leucine blocked virus-to-cell fusion and subsequent viral entry into target cells. This restriction could not be released by C-terminal truncation of the gp41 glycoprotein. These studies imply that the highly conserved core residues of the HIV Env MSD, in addition to serving as a membrane anchor, play an important role in mediating membrane fusion during viral entry.
Ataxia Telangiectasia-Mutated Damage-Signaling Kinase- and Proteasome-Dependent Destruction of Mre11-Rad50-Nbs1 Subunits in Simian Virus 40-Infected Primate Cells
Authors: Zhao Xiaorong , Madden-Fuentes Ramiro J. , Lou Becky X. , Pipas James M. , Gerhardt Jeannine , Rigell Christopher J. , Fanning Ellen 
Published: March 2008

ABSTRACT
Although the mechanism of simian virus 40 (SV40) DNA replication has been extensively investigated with cell extracts, viral DNA replication in productively infected cells utilizes additional viral and host functions whose interplay remains poorly understood. We show here that in SV40-infected primate cells, the activated ataxia telangiectasia-mutated (ATM) damage-signaling kinase, γ-H2AX, and Mre11-Rad50-Nbs1 (MRN) assemble with T antigen and other viral DNA replication proteins in large nuclear foci. During infection, steady-state levels of MRN subunits decline, although the corresponding mRNA levels remain unchanged. A proteasome inhibitor stabilizes the MRN complex, suggesting that MRN may undergo proteasome-dependent degradation. Analysis of mutant T antigens with disrupted binding to the ubiquitin ligase CUL7 revealed that MRN subunits are stable in cells infected with mutant virus or transfected with mutant viral DNA, implicating CUL7 association with T antigen in MRN proteolysis. The mutant genomes produce fewer virus progeny than the wild type, suggesting that T antigen-CUL7-directed proteolysis facilitates virus propagation. Use of a specific ATM kinase inhibitor showed that ATM kinase signaling is a prerequisite for proteasome-dependent degradation of MRN subunits as well as for the localization of T antigen and damage-signaling proteins to viral replication foci and optimal viral DNA replication. Taken together, the results indicate that SV40 infection manipulates host DNA damage-signaling to reprogram the cell for viral replication, perhaps through mechanisms related to host recovery from DNA damage.
Simultaneous Tracking of Capsid, Tegument, and Envelope Protein Localization in Living Cells Infected with Triply Fluorescent Herpes Simplex Virus 1
Authors: Sugimoto Ken , Uema Masashi , Sagara Hiroshi , Tanaka Michiko , Sata Tetsutaro , Hashimoto Yasuhiro , Kawaguchi Yasushi 
Published: March 2008

ABSTRACT

We report here the construction of a triply fluorescent-tagged herpes simplex virus 1 (HSV-1) expressing capsid protein VP26, tegument protein VP22, and envelope protein gB as fusion proteins with monomeric yellow, red, and cyan fluorescent proteins, respectively. The recombinant virus enabled us to monitor the dynamics of these capsid, tegument, and envelope proteins simultaneously in the same live HSV-1-infected cells and to visualize single extracellular virions with three different fluorescent emissions. In Vero cells infected by the triply fluorescent virus, multiple cytoplasmic compartments were found to be induced close to the basal surfaces of the infected cells (the adhesion surfaces of the infected cells on the solid growth substrate). Major capsid, tegument, and envelope proteins accumulated and colocalized in the compartments, as did marker proteins for the
trans
-Golgi network (TGN) which has been implicated to be the site of HSV-1 secondary envelopment. Moreover, formation of the compartments was correlated with the dynamic redistribution of the TGN proteins induced by HSV-1 infection. These results suggest that HSV-1 infection causes redistribution of TGN membranes to form multiple cytoplasmic compartments, possibly for optimal secondary envelopment. This is the first real evidence for the assembly of all three types of herpesvirus proteins—capsid, tegument, and envelope membrane proteins—in TGN.
Cross-Recognition of Avian H5N1 Influenza Virus by Human Cytotoxic T-Lymphocyte Populations Directed to Human Influenza A Virus
Authors: Kreijtz J. H. C. M. , de Mutsert G. , van Baalen C. A. , Fouchier R. A. M. , Osterhaus A. D. M. E. , Rimmelzwaan G. F. 
Published: March 2008

ABSTRACTSince the number of human cases of infection with avian H5N1 influenza viruses is ever increasing, a pandemic outbreak caused by these viruses is feared. Therefore, in addition to virus-specific antibodies, there is considerable interest in immune correlates of protection against these viruses, which could be a target for the development of more universal vaccines. After infection with seasonal influenza A viruses of the H3N2 and H1N1 subtypes, individuals develop virus-specific cytotoxic T-lymphocyte responses, which are mainly directed against the relatively conserved internal proteins of the virus, like the nucleoprotein (NP). Virus-specific cytotoxic T lymphocytes (CTL) are known to contribute to protective immunity against infection, but knowledge about the extent of cross-reactivity with avian H5N1 influenza viruses is sparse. In the present study, we evaluated the cross-reactivity with H5N1 influenza viruses of polyclonal CTL obtained from a group of well-defined HLA-typed study subjects. To this end, the recognition of synthetic peptides representing H5N1 analogues of known CTL epitopes was studied. In addition, the ability of CTL specific for seasonal H3N2 influenza virus to recognize the NP of H5N1 influenza virus or H5N1 virus-infected cells was tested. It was concluded that, apart from some individual epitopes that displayed amino acid variation between H3N2 and H5N1 influenza viruses, considerable cross-reactivity exists with H5N1 viruses. This preexisting cross-reactive T-cell immunity in the human population may dampen the impact of a next pandemic.
Evidence of hexaploid karyotype in shortnose sturgeon
Authors: Fontana Francesco , Congiu Leonardo , Mudrak Vincent A. , Quattro Joseph M. , Smith Theodore I.J. , Ware Kent , Doroshov Serge I. 
Published: March 2008

A karyotype analysis by several staining techniques was carried out on triplicate samples of the shortnose sturgeon, Acipenser brevirostrum . The chromosome number was found to be 2n = 372 ± 6. A representative karyotype of 374 chromosomes was composed of 178 metacentrics/submetacentrics and 196 telocentrics/acrocentrics and microchromosomes. The signals of fluorescent in situ hybridization (FISH) with a HindIII satellite DNA probe were visible on 14 chromosomes. The signals obtained with a PstI satellite DNA probe appeared on 12 chromosomes. The FISH with a 5S rDNA probe revealed fluorescent signals on 6 chromosomes. These last results, compared with 2 signals in species with about 120 chromosomes and 4 in species with 240, support the hypothesis that A. brevirostrum is a hexaploid species, probably of hybrid origin. Based on these results, we propose a model explaining speciation events occurring in sturgeons by hybridization, genome duplication, and diploidization.
Development of a Brassica seed cDNA microarray
Authors: Xiang Daoquan , Datla Raju , Li Fengling , Cutler Adrian , Malik Meghna R. , Krochko Joan E. , Sharma Nirmala , Fobert Pierre , Georges Fawzy , Selvaraj Gopalan , Tsang Ed , Klassen Darrin , Koh Chushin , Deneault Jean-Sebastien , Nantel Andre , Nowak Jacek , Keller Wilf , Bekkaoui Faouzi 
Published: March 2008

Brassica species represent several important crops including canola ( Brassica napus ). Understanding of genetic elements that contribute to seed-associated functions will impact future improvements in the canola crop. Brassica species share a very close taxonomic and molecular relationship with Arabidopsis thaliana. However, there are several subtle but distinct seed-associated agronomic characteristics that differ among the oil seed crop species. To address these, we have generated 67 535 ESTs predominately from Brassica seeds, analyzed these sequences, and identified 10 642 unigenes for the preparation of a targeted seed cDNA array. A set of 10 642 PCR primer pairs was designed and corresponding amplicons were produced for spotting, along with relevant controls. Critical quality control tests produced satisfactory results for use of this microarray in biological experiments. The microarray was also tested with specific RNA targets from embryos, germinating seeds, and leaf tissues. The hybridizations, signal intensities, and overall quality of these slides were consistent and reproducible. Additionally, there are 429 ESTs represented on the array that show no homology with any A. thaliana annotated gene or any gene in the Brassica genome databases or other plant databases; however, all of these probes hybridized to B. napus transcripts, indicating that the array also will be useful in defining expression patterns for genes so far unique to Brassica species.
The Open-Circuit Potential of a Polarizable and Reactive Electrode
Authors: Chen Zhan 
Published: March 2008

The open-circuit potential (OCP) of a polarizable and reactive electrode is modeled by the combination of the mixed potential theory and the Frumkin effect. The product of surface oxidation reaction alters the electrolyte double layer	structures at the interface, which conversely affects the surface reaction rate. Tribo-electrochemical experiments were carried out with a tungsten rotating disc electrode in a series of diluted KIO3 solutions at pH 2.00. The OCP change with time during the passivation of the electrode surface is fitted based on the model. All the fitting results can provide coherent and reasonable physical meanings, and the apparent surface reaction rate of 1.1*10^4 (mol/L)^-1*sec^-1 of the	tungsten electrode surface by KIO3 is thus obtained.
Inhibitive Effect of Seawater on the Corrosion of Particulate-Reinforced Aluminum-Matrix Composites
Authors: Ding Hongbo , Hawthorn George A. , Hihara Lloyd H. 
Published: March 2008

The corrosion rates of particulate 6092-T6 Al metal-matrix composites (MMCs) reinforced with 20 vol. % of B4C, SiC, and Al2O3 in ASTM seawater and real seawater were lower than those in 3.15 wt% NaCl or 0.5 M Na2SO4 solutions. Scanning electron microscopy revealed that precipitates consisting of agglomerated	whiskers formed on the surfaces of the MMCs after prolonged immersion in either ASTM seawater or real seawater. The precipitates were tentatively ascribed to be Al-Mg hydrotalcite-like compounds based on energy-dispersive X-ray and X-ray diffraction analyses. Accordingly, the precipitation was proposed to follow an induced hydrolysis process that requires a relatively low pH close to that of seawater. It was postulated	that the corrosion of the MMCs was inhibited both by precipitates that impede the diffusion of oxygen to cathodic sites and by the buffering effect of seawater, which attenuates pH fluctuations near the MMC surfaces.
Water Transport in Multilayer Organic Coatings
Authors: Allahar Kerry N. , Hinderliter Brian R. , Tallman Dennis , Bierwagen Gordon 
Published: March 2008

Protective multilayer coatings are designed with each layer providing distinctive features to reduce the transport of water, ions, and oxygen from the environment to the substrate.	The topcoat provides the barrier against the environment whereas the basecoat/primer provides adhesion to the substrate and additional protection such as galvanic or passivation. The change in the coating capacitance during the ingress or egress of water is associated with the change in the dielectric	property of the coating due to the large difference between the relative dielectrics of water and coating. The coating capacitance during wetting and drying can be monitored using the electrochemical impedance spectroscopy response of a high frequency signal applied to the coating. The evolution of water uptake during wetting and water loss during drying associated with a two-layer coating is presented. Embedded electrodes were used to access the primer and a hydrophil ic room temperature ionic liquid was used to simulate drying	while maintaining a non-aqueous electrical contact to the coating. The wetting and drying behaviors of epoxy, urethane, and epoxy primer/urethane topcoat coatings were experimentally measured. Mathematical models were developed based on Fick's second law for water transport in single-layer and two-layer coatings. Comparison of measured and simulated water uptake demonstrated that the assumptions used in the single-layer model were appropriate while the assumptions used for transport across the interface between the layers in the two- layer coating were inadequate.
Initial Electroless NiP Coating Behavior On Mg - Al - Zn Alloy AZ91
Authors: Anik Mustafa , Korpe Erhan 
Published: March 2008

Alloys represent the α and β phases of Alloy AZ91 were designed and the hypophosphite-reduced electroless nickel coating	characteristics on them in both uncoupled and coupled conditions were analyzed. Galvanic coupling of β phase to the	matrix phase in the coating bath was observed to decelerate the deposition rate on the matrix phase. This behavior was attributed to the effective	cathodic action of β phase in the coating bath as a result of the catalytic cathodic reactions on it.
Hot Corrosion Behavior of Newly Developed Nickel based Superalloy and Comparison with other Alloys
Authors: Injeti Gurrappa , Alasdair Wilson , Hailiang Du , Burnell-gray Jim , Datta Santu 
Published: March 2008

The current paper explains the hot corrosion characteristics of newly developed super alloy at two different temperatures, which corresponds to type I and type II hot corrosion. The	data was compared with other super alloys under similar experimental conditions in order to understand the nature of degradation mechanism of new super alloy. The results show that the new super alloy is highly susceptible to hot corrosion. The relevant reaction mechanisms that are responsible for its faster degradation under hot corrosion conditions were established and discussed. Based on the results obtained with different techniques, an electrochemical model was proposed to show that the hot corrosion of new super alloy is electrochemical in nature. Finally, it is stressed the necessity of application of high performance coatings for its protection against high temperature corrosion.
Long-term Epigenetic Therapy with Oral Zebularine Has Minimal Side Effects and Prevents Intestinal Tumors in Mice
Authors: Yoo Christine B. , Chuang Jody C. , Byun Hyang-Min , Egger Gerda , Yang Allen S. , Dubeau Louis , Long Tiffany , Laird Peter W. , Marquez Victor E. , Jones Peter A. 
Published: March 2008

Abstract
Recent successes in the application of epigenetic drugs for the treatment of myelodysplastic syndrome have raised questions on the safety of long-term administration of DNA methylation inhibitors. We treated preweaned cancer prone ApcMin/+ (Min) mice continuously with the DNA methylation inhibitor zebularine in their drinking water to determine the effects of the drug on normal mouse development as well as cancer prevention. Zebularine caused a tissue-specific reduction in DNA methylation at B1 short interspersed nucleotide elements in the small and large intestines of female Min mice but not in other organs examined after chronic oral treatment. No significant difference in the average weights of mice was observed during the treatment. In addition, analysis of global gene expression of colonic epithelial cells from the females indicated that only 3% to 6% of the genes were affected in their expression. We did not detect toxicity and abnormalities from the histopathologic analysis of liver and intestinal tissues. Lastly, we tested whether prevention of tumorigenesis can be achieved with chronic oral administration of zebularine in Min mice. The average number of polyps in Min females decreased from 58 to 1, whereas the average polyp number remained unaffected in Min males possibly due to differential activity of aldehyde oxidase. Taken together, our results show for the first time that long-term oral administration of zebularine causes a gender-specific abrogation of intestinal tumors while causing a tissue-specific DNA demethylation. Importantly, prolonged treatment of mice with epigenetic drugs resulted in only minor developmental and histologic changes.