bluuebunny/crossref_metadata_2025_split huggingface dataset:
Protection of Sheep against Rift Valley Fever Virus and Sheep Poxvirus with a Recombinant Capripoxvirus Vaccine
Authors: Soi Reuben K. , Rurangirwa Fred R. , McGuire Travis C. , Rwambo Paul M. , DeMartini James C. , Crawford Timothy B. 
Published: September 2010

ABSTRACT
Rift Valley fever (RVF) is an epizootic viral disease of sheep that can be transmitted from sheep to humans, particularly by contact with aborted fetuses. A capripoxvirus (CPV) recombinant virus (rKS1/RVFV) was developed, which expressed the Rift Valley fever virus (RVFV) Gn and Gc glycoproteins. These expressed glycoproteins had the correct size and reacted with monoclonal antibodies (MAb) to native glycoproteins. Mice vaccinated with rKS1/RVFV were protected against RVFV challenge. Sheep vaccinated with rKS1/RVFV twice developed neutralizing antibodies and were significantly protected against RVFV and sheep poxvirus challenge. These findings further document the value of CPV recombinants as ruminant vaccine vectors and support the inclusion of RVFV genes encoding glycoproteins in multivalent recombinant vaccines to be used where RVF occurs.
Identification, Characterization, and Application of a Recombinant Antigen for the Serological Investigation of Feline Hemotropic Mycoplasma Infections
Authors: Wolf-Jäckel Godelind A. , Jäckel Christian , Museux Kristina , Hoelzle Katharina , Tasker Séverine , Lutz Hans , Hofmann-Lehmann Regina 
Published: September 2010

ABSTRACTIn felids, three hemotropic mycoplasma species (hemoplasmas) have been described:Mycoplasma haemofelis, “CandidatusMycoplasma haemominutum,” and “CandidatusMycoplasma turicensis.” In particular,M. haemofelismay cause severe, potentially life-threatening hemolytic anemia. No routine serological assays for feline hemoplasma infections are available. Thus, the goal of our project was to identify and characterize anM. haemofelisantigen (DnaK) that subsequently could be applied as a recombinant antigen in a serological assay. The gene sequence of this protein was determined using consensus primers and blood samples from two naturallyM. haemofelis-infected Swiss pet cats, an experimentallyM. haemofelis-infected specific-pathogen-free cat, and a naturallyM. haemofelis-infected Iberian lynx (Lynx pardinus). TheM. haemofelisDnaK gene sequence showed the highest identity to an analogous protein of a porcine hemoplasma (72%).M. haemofelisDnaK was expressed recombinantly in anEscherichia coliDnaK knockout strain and purified using Ni affinity, size-exclusion, and anion-exchange chromatography. It then was biochemically and functionally characterized and showed characteristics typical for DnaKs (secondary structure profile, thermal denaturation, ATPase activity, and DnaK complementation). Moreover, its immunogenicity was assessed using serum samples from experimentally hemoplasma-infected cats. In Western blotting or enzyme-linked immunosorbent assays, it was recognized by sera from cats infected withM. haemofelis, “Ca.Mycoplasma haemominutum,” and “Ca.Mycoplasma turicensis,” respectively, but not from uninfected cats. This is the first description of a full-length purified recombinant feline hemoplasma antigen that can readily be applied in future pathogenesis studies and may have potential for application in a diagnostic serological test.
Phenotypic and Genotypic Characterization of Group B Streptococcal Isolates in Southern Brazil
Authors: Palmeiro Jussara K. , Dalla-Costa Libera M. , Fracalanzza Sérgio E. L. , Botelho Ana C. N. , da Silva Nogueira Keite , Scheffer Mara C. , de Almeida Torres Rosângela S. L. , de Carvalho Newton Sérgio , Cogo Laura Lúcia , Madeira Humberto M. F. 
Published: September 2010

ABSTRACT

One-hundred sixty-eight group B streptococcal (GBS) isolates from a Brazilian hospital were phenotypically and genotypically characterized. Isolates were recovered from human sources from April 2006 to May 2008 and classified as either invasive, noninvasive, or colonizing isolates. Classical methods for serotyping and antibiotic resistance profiling were employed. Clonal groups were also defined by pulsed-field gel electrophoresis (PFGE). Results showed that susceptibility to beta-lactam antimicrobials was predominant among the isolates. Only 4.7% were resistant to erythromycin and clindamycin. The
erm
(B) gene was widely detected in our GBS isolates, according to our phenotypic results (constitutive macrolide-lincosamide-streptogramin B [cMLS
B
] resistance phenotype), and the
erm
(A) gene was also detected in some isolates. MLS
B
resistance was restricted to strains isolated from patients with noninvasive infections and carriers. Serotype Ia was predominant (38.1%), serotype IV isolates were found at a high frequency (13.1%), and few isolates of serotype III were identified (3%). Pulsed-field gel electrophoresis results revealed a variety of types, reflecting the substantial genetic diversity among GBS strains, although a great number of isolates could be clustered into two major groups with a high degree of genetic relatedness. Three main PFGE clonal groups were found, and isolates sharing the same PFGE type were grouped into different serotypes. Furthermore, in a few cases, isolates from the same patients and possessing the same PFGE type were of different serotypes. These findings could be related to the occurrence of capsular switching by horizontal transfer of capsular genes.
Candida bracarensis
Bloodstream Infection in an Immunocompromised Patient
Authors: Warren Thomas A. , McTaggart Lisa , Richardson Susan E. , Zhang Sean X. 
Published: September 2010

ABSTRACT

Candida bracarensis
is a recently described
Candida
species which is phenotypically similar to
Candida glabrata
. A case of
C. bracarensis
bloodstream infection in a bone marrow transplant patient is described and confirms this organism as an opportunistic human pathogen. The organism can be distinguished from
C. glabrata
by its white color on CHROMagar and by DNA sequence analysis using D1/D2 and internal transcribed spacer (ITS) primers.
Nonhealing Wound Due toRhodococcus equiin an Apparently Immunocompetent Patient, Revealing CD8+T-Lymphocyte Deficiency
Authors: Denes Eric , Peignon-Orsoni Dominique , Terrade François-Xavier 
Published: September 2010

ABSTRACTWe describe a case of a nonhealing wound due toRhodococcus equi. Failure of the wound to heal led to immunological investigations and the discovery of a previously unknown CD8+T-lymphocyte deficit responsible for the chronic infection. The infection was cured after a 3-month course of a combination of antibiotics.
Molecular Characterization of Clinical Methicillin-Resistant
Staphylococcus aureus
Isolates in South Africa
Authors: Moodley A. , Oosthuysen W. F. , Dusé A. G. , Marais E. 
Published: September 2010

ABSTRACT

Eighty-two percent of 320 clinical methicillin-resistant
Staphylococcus aureus
(MRSA) isolates from various infection sites collected throughout South Africa were separated into five major globally prevalent clusters by SmaI pulsed-field gel electrophoresis,
spa
, and SCC
mec
typing. Only one Panton-Valentine leukocidin-positive isolate was detected. This is the first detailed MRSA epidemiology study for the whole country.
gyrB
Multiplex PCR To Differentiate between
Acinetobacter calcoaceticus
and
Acinetobacter
Genomic Species 3
Authors: Higgins Paul G. , Lehmann Marlene , Wisplinghoff Hilmar , Seifert Harald 
Published: September 2010

ABSTRACT

A previously established multiplex PCR that identifies to the species level
Acinetobacter baumannii
and
Acinetobacter
genomic species 13TU (GS13TU) was expanded to include
Acinetobacter calcoaceticus
and
Acinetobacter
genomic species 3.
Interaction of Decay-Accelerating Factor with Echovirus 7
Authors: Plevka Pavel , Hafenstein Susan , Harris Katherine G. , Cifuente Javier O. , Zhang Ying , Bowman Valorie D. , Chipman Paul R. , Bator Carol M. , Lin Feng , Medof M. Edward , Rossmann Michael G. 
Published: September 2010

ABSTRACTEchovirus 7 (EV7) belongs to theEnterovirusgenus within the familyPicornaviridae. Many picornaviruses use IgG-like receptors that bind in the viral canyon and are required to initiate viral uncoating during infection. However, in addition, some of the enteroviruses use an alternative or additional receptor that binds outside the canyon. Decay-accelerating factor (DAF) has been identified as a cellular receptor for EV7. The crystal structure of EV7 has been determined to 3.1-Å resolution and used to interpret the 7.2-Å-resolution cryo-electron microscopy reconstruction of EV7 complexed with DAF. Each DAF binding site on EV7 is near a 2-fold icosahedral symmetry axis, which differs from the binding site of DAF on the surface of coxsackievirus B3, indicating that there are independent evolutionary processes by which DAF was selected as a picornavirus accessory receptor. This suggests that there is an advantage for these viruses to recognize DAF during the initial process of infection.
The Innate Immune Adaptor Molecule MyD88 Restricts West Nile Virus Replication and Spread in Neurons of the Central Nervous System
Authors: Szretter Kristy J. , Daffis Stephane , Patel Jigisha , Suthar Mehul S. , Klein Robyn S. , Gale Michael , Diamond Michael S. 
Published: September 2010

ABSTRACT

Type I interferons (IFN-α/β) control viral infection by triggering the expression of genes that restrict transcription, translation, replication, and assembly. Many viruses induce IFN responses after recognition by cytoplasmic or endosomal RNA sensors (RIG-I-like RNA helicases [RLR] and Toll-like receptors [TLR]), which signal through the cognate adaptor signaling molecules IPS-1, TRIF, and MyD88. Recent studies have demonstrated that IPS-1-dependent induction of IFN-α/β downstream of RLR recognition restricts West Nile virus (WNV) infection in many cell types, whereas TRIF-dependent TLR3 signaling limits WNV replication in neurons. Here, we examined the contribution of MyD88 signaling to the control of WNV by evaluating IFN induction and virus replication in genetically deficient cells and mice.
MyD88
−
/
−
mice showed increased lethality after WNV infection and elevated viral burden primarily in the brain, even though little effect on the systemic type I IFN response was observed. Intracranial inoculation studies corroborated these findings, as WNV spread more rapidly in the central nervous system of
MyD88
−
/
−
mice, and this phenotype preceded the recruitment of inflammatory leukocytes.
In vitro
, increased WNV replication was observed in
MyD88
−
/
−
macrophages and subsets of neurons but not in myeloid dendritic cells. MyD88 had an independent effect on recruitment of monocyte-derived macrophages and T cells into the brain that was associated with blunted induction of the chemokines that attract leukocytes. Our experiments suggest that MyD88 restricts WNV by inhibiting replication in subsets of cells and modulating expression of chemokines that regulate immune cell migration into the central nervous system.
Interleukin-1 Family Cytokines as Mucosal Vaccine Adjuvants for Induction of Protective Immunity against Influenza Virus
Authors: Kayamuro Hiroyuki , Yoshioka Yasuo , Abe Yasuhiro , Arita Shuhei , Katayama Kazufumi , Nomura Tetsuya , Yoshikawa Tomoaki , Kubota-Koketsu Ritsuko , Ikuta Kazuyoshi , Okamoto Shigefumi , Mori Yasuko , Kunisawa Jun , Kiyono Hiroshi , Itoh Norio , Nagano Kazuya , Kamada Haruhiko , Tsutsumi Yasuo , Tsunoda Shin-Ichi 
Published: September 2010

ABSTRACTA safe and potent adjuvant is needed for development of mucosal vaccines against etiological agents, such as influenza virus, that enter the host at mucosal surfaces. Cytokines are potential adjuvants for mucosal vaccines because they can enhance primary and memory immune responses enough to protect against some infectious agents. For this study, we tested 26 interleukin (IL) cytokines as mucosal vaccine adjuvants and compared their abilities to induce antigen (Ag)-specific immune responses against influenza virus. In mice intranasally immunized with recombinant influenza virus hemagglutinin (rHA) plus one of the IL cytokines, IL-1 family cytokines (i.e., IL-1α, IL-1β, IL-18, and IL-33) were found to increase Ag-specific immunoglobulin G (IgG) in plasma and IgA in mucosal secretions compared to those after immunization with rHA alone. In addition, high levels of both Th1- and Th2-type cytokines were observed in mice immunized with rHA plus an IL-1 family cytokine. Furthermore, mice intranasally immunized with rHA plus an IL-1 family cytokine had significant protection against a lethal influenza virus infection. Interestingly, the adjuvant effects of IL-18 and IL-33 were significantly decreased in mast cell-deficientW/Wvmice, indicating that mast cells have an important role in induction of Ag-specific mucosal immune responses induced by IL-1 family cytokines. In summary, our results demonstrate that IL-1 family cytokines are potential mucosal vaccine adjuvants and can induce Ag-specific immune responses for protection against pathogens like influenza virus.
RNA Structures Facilitate Recombination-Mediated Gene Swapping in HIV-1
Authors: Simon-Loriere Etienne , Martin Darren P. , Weeks Kevin M. , Negroni Matteo 
Published: September 2010

ABSTRACT

Many viruses, including retroviruses, undergo frequent recombination, a process which can increase their rate of adaptive evolution. In the case of HIV, recombination has been responsible for the generation of numerous intersubtype recombinant variants with epidemiological importance in the AIDS pandemic. Although it is known that fragments of genetic material do not combine randomly during the generation of recombinant viruses, the mechanisms that lead to preferential recombination at specific sites are not fully understood. Here we reanalyze recent independent data defining (i) the structure of a complete HIV-1 RNA genome and (ii) favorable sites for recombination. We show that in the absence of selection acting on recombinant genomes, regions harboring RNA structures in the NL4-3 model strain are strongly predictive of recombination breakpoints in the HIV-1
env
genes of primary isolates. In addition, we found that breakpoints within recombinant HIV-1 genomes sampled from human populations, which have been acted upon extensively by natural selection, also colocalize with RNA structures. Critically, junctions between genes are enriched in structured RNA elements and are also preferred sites for generating functional recombinant forms. These data suggest that RNA structure-mediated recombination allows the virus to exchange intact genes rather than arbitrary subgene fragments, which is likely to increase the overall viability and replication success of the recombinant HIV progeny.
Intrinsic Cytoskeleton-Dependent Clustering of Influenza Virus M2 Protein with Hemagglutinin Assessed by FLIM-FRET
Authors: Thaa Bastian , Herrmann Andreas , Veit Michael 
Published: September 2010

ABSTRACT
The hemagglutinin (HA) of influenza virus organizes the virus bud zone, a domain of the plasma membrane enriched in raft lipids. Using fluorescence lifetime imaging microscopy-fluorescence resonance energy transfer (FLIM-FRET), a technique that detects close colocalization of fluorescent proteins in transfected cells, we show that the viral proton channel M2 clusters with HA but not with a marker for inner leaflet rafts. The FRET signal between M2 and HA depends on the raft-targeting signals in HA and on an intact actin cytoskeleton. We conclude that M2 contains an intrinsic signal that targets the protein to the viral bud zone, which is organized by raft-associated HA and by cortical actin.
Nucleoporin Phosphorylation Triggered by the Encephalomyocarditis Virus Leader Protein Is Mediated by Mitogen-Activated Protein Kinases
Authors: Porter Frederick W. , Brown Bradley , Palmenberg Ann C. 
Published: September 2010

ABSTRACT

Cardioviruses disrupt nucleocytoplasmic transport through the activity of their leader (L) protein. We have shown that hyperphosphorylation of nuclear pore proteins (nucleoporins or Nups), including Nup62, Nup153, and Nup214, is central to this L protein function and requires one or more cytosolic kinases. In this study, potential cellular enzymes involved in encephalomyocarditis virus (EMCV) L-directed Nup phosphorylation were screened with a panel of specific, cell-permeating kinase inhibitors. Extracellular signal-regulated receptor kinase (ERK) and p38 mitogen-activated protein kinase inhibitors (U0126 and SB203580) were sufficient to block Nup hyperphosphorylation in EMCV-infected or L-expressing cells. Recombinant L alone, in the absence of infection, triggered activation of ERK and p38, independent of their upstream signaling cascades. Conserved residues within the L zinc finger (Cys
19
) and acidic domain (Asp
48
,
51
,
52
,
55
) were essential for this activation and for the phosphorylation of Nups, suggesting that the phenomena are linked. Analysis of the hyperphosphorylated Nup species revealed only phosphoserine and phosphothreonine residues. The sizes of the tryptic phosphopeptides derived from Nup62 were compatible with sites in the Phe/Gly repeat domain which display common consensus sequences for ERK and p38 substrates. The results provide strong evidence that ERK and p38 are the probable effector kinases required for L-dependent inhibition of nuclear trafficking.
Impairment of Hepatitis B Virus Virion Secretion by Single-Amino-Acid Substitutions in the Small Envelope Protein and Rescue by a Novel Glycosylation Site
Authors: Ito Kiyoaki , Qin Yanli , Guarnieri Michael , Garcia Tamako , Kwei Karen , Mizokami Masashi , Zhang Jiming , Li Jisu , Wands Jack R. , Tong Shuping 
Published: September 2010

ABSTRACTMutations in the S region of the hepatitis B virus (HBV) envelope gene are associated with immune escape, occult infection, and resistance to therapy. We previously identified naturally occurring mutations in the S gene that alter HBV virion secretion. Here we used transcomplementation assay to confirm that the I110M, G119E, and R169P mutations in the S domain of viral envelope proteins impair virion secretion and that an M133T mutation rescues virion secretion of the I110M and G119E mutants. The G119E mutation impaired detection of secreted hepatitis B surface antigen (HBsAg), suggesting immune escape. The R169P mutant protein is defective in HBsAg secretion as well and has a dominant negative effect when it is coexpressed with wild-type envelope proteins. Although the S domain is present in all three envelope proteins, the I110M, G119E, and R169P mutations impair virion secretion through the small envelope protein. Conversely, coexpression of just the small envelope protein of the M133T mutant could rescue virion secretion. The M133T mutation could also overcome the secretion defect caused by the G145R immune-escape mutation or mutation at N146, the site of N-linked glycosylation. In fact, the M133T mutation creates a novel N-linked glycosylation site (131NST133). Destroying this site by N131Q/T mutation or preventing glycosylation by tunicamycin treatment of transfected cells abrogated the effect of the M133T mutation. Our findings demonstrate that N-linked glycosylation of HBV envelope proteins is critical for virion secretion and that the secretion defect caused by mutations in the S protein can be rescued by an extra glycosylation site.
Genetic Identity and Biological Phenotype of a Transmitted/Founder Virus Representative of Nonpathogenic Simian Immunodeficiency Virus Infection in African Green Monkeys
Authors: Gnanadurai Clement Wesley , Pandrea Ivona , Parrish Nicholas F. , Kraus Matthias H. , Learn Gerald H. , Salazar Maria G. , Sauermann Ulrike , Töpfer Katharina , Gautam Rajeev , Münch Jan , Stahl-Hennig Christiane , Apetrei Christian , Hahn Beatrice H. , Kirchhoff Frank 
Published: September 2010

ABSTRACT

Understanding the lack of disease progression in nonpathogenic simian immunodeficiency virus (SIV) infections is essential for deciphering the immunopathogenesis of human AIDS. Yet,
in vivo
studies have been hampered by a paucity of infectious molecular clones (IMCs) of SIV suitable to dissect the viral and host factors responsible for the nonpathogenic phenotype. Here, we describe the identification, cloning, and biological analysis of the first transmitted/founder (T/F) virus representing a nonpathogenic SIV infection. Blood was collected at peak viremia from an acutely infected sabaeus monkey (
Chlorocebus sabaeus
) inoculated intravenously with an African green monkey SIV (SIVagm) strain (Sab92018) that had never been propagated
in vitro
. To generate IMCs, we first used conventional (bulk) PCR to amplify full-length viral genomes from peripheral blood mononuclear cell (PBMC) DNA. Although this yielded two intact SIVagmSab genomes, biological characterization revealed that both were replication defective. We then performed single-genome amplification (SGA) to generate partially overlapping 5′ (
n
= 10) and 3′ (
n
= 13) half genomes from plasma viral RNA. Analysis of these amplicons revealed clusters of nearly identical viral sequences representing the progeny of T/F viruses. Synthesis of the consensus sequence of one of these generated an IMC (Sab92018ivTF) that produced infectious CCR5-tropic virions and replicated to high titers in Molt-4 clone 8 cells and African green monkey PBMCs. Sab92018ivTF also initiated productive infection in sabaeus monkeys and faithfully recapitulated the replication kinetics and nonpathogenic phenotype of the parental Sab92018 strain. These results thus extend the T/F virus concept to nonpathogenic SIV infections and provide an important new tool to define viral determinants of disease nonprogression.
Portable Filter-Based Microdevice for Detection and Characterization of Circulating Tumor Cells
Authors: Lin Henry K. , Zheng Siyang , Williams Anthony J. , Balic Marija , Groshen Susan , Scher Howard I. , Fleisher Martin , Stadler Walter , Datar Ram H. , Tai Yu-Chong , Cote Richard J. 
Published: September 2010

Abstract
Purpose: Sensitive detection and characterization of circulating tumor cells (CTC) could revolutionize the approach to patients with early-stage and metastatic cancer. The current methodologies have significant limitations, including limited capture efficiency and ability to characterize captured cells. Here, we report the development of a novel parylene membrane filter-based portable microdevice for size-based isolation with high recovery rate and direct on-chip characterization of captured CTC from human peripheral blood.
Experimental Design: We evaluated the sensitivity and efficiency of CTC capture in a model system using blood samples from healthy donors spiked with tumor cell lines. Fifty-nine model system samples were tested to determine the recovery rate of the microdevice. Moreover, 10 model system samples and 57 blood samples from cancer patients were subjected to both membrane microfilter device and CellSearch platform enumeration for direct comparison.
Results: Using the model system, the microdevice achieved &gt;90% recovery with probability of 95% recovering at least one cell when five are seeded in 7.5 mL of blood. CTCs were identified in 51 of 57 patients using the microdevice, compared with only 26 patients with the CellSearch method. When CTCs were detected by both methods, greater numbers were recovered by the microfilter device in all but five patients.
Conclusions: This filter-based microdevice is both a capture and analysis platform, capable of multiplexed imaging and genetic analysis. The microdevice presented here has the potential to enable routine CTC analysis in the clinical setting for the effective management of cancer patients. Clin Cancer Res; 16(20); 5011–8. ©2010 AACR.