bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of Serum Humoral Responses Induced by Oral Immunization with the Hepatitis B Virus Core Antigen and the Cholera Toxin B Subunit
Authors: Broos Katleen , Janssens Michiel E. , De Goeyse Ine , Vanlandschoot Peter , Leroux-Roels Geert , Geysen Dirk , Guisez Yves 
Published: March 2008

ABSTRACTThe hepatitis B virus core (HBc) virus-like particle (VLP) is known as one of the most immunogenic antigens and carrier vehicles in different immunization strategies. Recent findings are suggesting the potential of the HBc VLPs as an oral immunogen. Here, we focus on the induction of serum humoral responses by oral administration of HBc VLPs in preparations substantially free of lipopolysaccharide and immunomodulating encapsidated RNA. The full-length HBc antigen was used, because the C-terminal arginine-rich tail may contribute to the immunogenicity of the antigen as the region is involved in cell surface heparan sulfate binding and internalization of the protein. Serum antibody levels and isotypes were determined following oral administration of the HBc VLPs with the perspective of using the HBc VLP as an immunostimulatory and carrier molecule for epitopes of blood-borne diseases in oral immunization vaccination strategies. Following oral administration of the HBc VLP preparations to mice, a strong serum humoral response was induced with mainly immunoglobulin G2a (IgG2a) antibodies, pointing toward a Th1 response which is essential in the control of intracellular pathogens. Intraperitoneal immunization with the HBc VLP induced a stronger, mixed Th1/Th2 response. Finally, a comparison was made with the induced serum humoral response following oral administration of the recombinant cholera toxin B pentamer, a commonly used oral immunogen. These immunizations, in contrast, induced predominantly antibodies of the IgG1 isotype, indicative of a Th2 response. These data suggest that the HBc VLP can be an interesting carrier molecule in oral vaccine development.
Enzyme-Linked Immunosorbent Assay for Detection of
Plasmodium falciparum
Histidine-Rich Protein 2 in Blood, Plasma, and Serum
Authors: Kifude Carolyne M. , Rajasekariah Halli G. , Sullivan David J. , Stewart V. Ann , Angov Evelina , Martin Samuel K. , Diggs Carter L. , Waitumbi John N. 
Published: March 2008

ABSTRACT

Microscopy, the gold standard for the detection and quantification of malaria parasites in blood, is in many aspects deficient for this purpose. The method is poorly reproducible and can be inaccurate because
Plasmodium falciparum
parasites sequester for a portion of each asexual cycle. Due to these deficiencies, biomarkers such as
P. falciparum
histidine-rich protein 2 (PfHRP2) are increasingly being used. In this study, we evaluated the use of a commercial PfHRP2 enzyme-linked immunosorbent assay (ELISA) kit with some procedural modifications. We determined the linear range of the assay, including the lower limits of detection and quantitation, using recombinant PfHRP2 (rPfHRP2). In 10 repeat experiments, the linear range of optical densities (ODs) at 450 to 650 nm was from 0.05 ± 0.002 to 2.28 ± 0.042, corresponding to 3.91 to 250 ng/ml of rPfHRP2. The coefficient of variation (CV) at each target concentration ranged from 1.93 to 8.07%. Using cultured parasites, we confirmed the linear range of ODs as well as the association between the PfHRP2 ELISA results and the microscopic parasite densities. For whole-blood samples spiked with cultured, washed, ring-stage-infected red blood cells (iRBCs), the linear range was 11.7 to 750 iRBCs/μl, with CVs of 0.29 to 7.56%. The same spiked samples evaluated by microscopists had similar sensitivities, but the CVs were unacceptably high (20.7 to 161.6%). Stock rPfHRP2 was stable through four freeze-thaw cycles (
P
< 0.05; paired
t
test). When different patient sample types at different concentrations within the linear range of the assay are compared, the recoveries of PfHRP2 from blood and serum were within ±20%, whereas the recoveries from plasma ranged between +35 and −41%. We conclude that PfHRP2 ELISA using whole-blood and serum samples is a suitable adjunct to microscopy and could ultimately benefit malaria intervention trials.
Rapid Detection of the Mosaic Structure of the
Neisseria gonorrhoeae penA
Gene, Which Is Associated with Decreased Susceptibilities to Oral Cephalosporins
Authors: Ochiai Susumu , Ishiko Hiroaki , Yasuda Mitsuru , Deguchi Takashi 
Published: March 2008

ABSTRACT

In
Neisseria gonorrhoeae
, the mosaic structure of the
penA
gene (encoding penicillin-binding protein 2 [PBP 2]), which is composed of fragments of the
penA
genes from
Neisseria cinerea
and
Neisseria perflava
, has been significantly associated with decreased susceptibility to cephalosporins, particularly oral cephalosporins. The aim of this study was to develop a rapid assay for the detection of mosaic PBP 2 of
N. gonorrhoeae
by real-time PCR. This assay successfully detected the mosaic
penA
gene of
N. gonorrhoeae
, and its sensitivity was ≥10
1
copies/reaction. Six hundred twenty-one clinical strains were examined by this assay for the presence of mosaic PBP 2, which was detected in 85 (39.4%) of 216 strains from 2002, 69 (40.6%) of 170 strains from 2003, 71 (44.4%) of 160 strains from 2004, and 31 (41.3%) of 75 strains from 2005. The MICs of cephalosporins for strains with the mosaic PBP 2 detected by the assay were statistically higher than those for strains without the mosaic PBP 2. One hundred sixty-six (64.8%) of 256 strains with the mosaic PBP 2 exhibited cefixime MICs of ≥0.5 μg/ml. The emergence and spread of strains with mosaic PBP 2 could be a threat to the cefixime treatment of gonorrhea. This real-time PCR assay for the detection of mosaic PBP 2 of
N. gonorrhoeae
is thus useful in the prediction of decreased susceptibilities to oral cephalosporins.
Ventriculoperitoneal Shunt Infection Caused by
Actinomyces neuii
subsp.
neuii
Authors: Watkins Richard R. , Anthony Kathy , Schroder Suzanne , Hall Gerri S. 
Published: March 2008

ABSTRACT

Actinomyces neuii
subsp.
neuii
is a rare isolate in clinical specimens. The organism was previously designated CDC coryneform group 1 and was renamed in 1994. A case of a ventriculoperitoneal shunt infection caused by this organism is described.
Development and Validation of a One-Step Real-Time PCR Assay for Simultaneous Detection of Subtype H5, H7, and H9 Avian Influenza Viruses
Authors: Monne Isabella , Ormelli Silvia , Salviato Annalisa , De Battisti Cristian , Bettini Francesca , Salomoni Angela , Drago Alessandra , Zecchin Bianca , Capua Ilaria , Cattoli Giovanni 
Published: March 2008

ABSTRACT

Among the different hemagglutinin (HA) subtypes of avian influenza (AI) viruses, H5, H7, and H9 are of major interest because of the serious consequences for the poultry industry and the increasing frequency of direct transmission of these viruses to humans. The availability of new tools to rapidly detect and subtype the influenza viruses can enable the immediate application of measures to prevent the widespread transmission of the infection. In this study, a novel one-step real-time reverse transcription-PCR (RRT-PCR) was developed to detect simultaneously the H5, H7, and H9 subtypes of AI viruses from clinical samples of avian origin. The sensitivity of the RRT-PCR assay was determined by using in vitro-transcribed RNA and 10-fold serial dilutions of titrated AI viruses. High sensitivity levels were obtained, with limits of detection ranging from 10
1
to 10
3
RNA copies and from 10
1
50% egg infectious dose (EID
50
)/100 μl to 10
2.74
EID
50
/100 μl with titrated viruses. Excellent results were achieved in the intra- and interassay variability tests. The comparison of the results with those obtained from the analysis of 725 avian samples by means of the reference method (virus isolation [VI]) showed a high level of agreement. To date, this is the first real-time PCR protocol available for the simultaneous detection of AI viruses belonging to subtypes H5, H7, and H9, and the results obtained indicate that this method is suitable as a routine laboratory test for the rapid detection and differentiation of the three most-important AI virus subtypes in samples of avian origin.
Mutations Affecting the MA Portion of the v-Abl Protein Reveal a Conserved Role of Gag in Abelson Murine Leukemia Virus (MLV) and Moloney MLV
Authors: Yi Chae-ryun , Rosenberg Naomi 
Published: March 2008

ABSTRACT

Abelson murine leukemia virus (Ab-MLV) arose from a recombination between
gag
sequences in Moloney MLV (Mo-MLV) and the c-
abl
proto-oncogene. The v-Abl oncoprotein encoded by Ab-MLV contains MA, p12, and a portion of CA sequences derived from the
gag
gene of Mo-MLV. Previous studies indicated that alteration of MA sequences affects the biology of Mo-MLV and Ab-MLV. To understand the role of these sequences in Ab-MLV transformation more fully, alanine substitution mutants that affect Mo-MLV replication were examined in the context of Ab-MLV. Mutations affecting Mo-MLV replication decreased transformation, while alanine mutations in residues dispensable for Mo-MLV replication did not. The altered v-Abl proteins displayed aberrant subcellular localization that correlated to transformation defects. Immunofluorescent analyses suggested that aberrant trafficking of the altered proteins and improper interaction with components of the cytoskeleton were involved in the phenotype. Similar defects in localization were observed when the Gag moiety containing these mutations was expressed in the absence of
abl
-derived sequences. These results indicate that MA sequences within the Gag moiety of the v-Abl protein contribute to proper localization by playing a dominant role in trafficking of the v-Abl molecule.
Comparison of Cellular Ribonucleoprotein Complexes Associated with the APOBEC3F and APOBEC3G Antiviral Proteins
Authors: Gallois-Montbrun Sarah , Holmes Rebecca K. , Swanson Chad M. , Fernández-Ocaña Mireia , Byers Helen L. , Ward Malcolm A. , Malim Michael H. 
Published: March 2008

ABSTRACT
The human apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3F (APOBEC3F [A3F]) and A3G proteins are effective inhibitors of infection by various retroelements and share ∼50% amino acid sequence identity. We therefore undertook comparative analyses of the protein and RNA compositions of A3F- and A3G-associated ribonucleoprotein complexes (RNPs). Like A3G, A3F is found associated with a complex array of cytoplasmic RNPs and can accumulate in RNA-rich cytoplasmic microdomains known as mRNA processing bodies or stress granules. While A3F RNPs display greater resistance to disruption by RNase digestion, the major protein difference is the absence of the Ro60 and La autoantigens. Consistent with this, A3F RNPs also lack a number of small polymerase III RNAs, including the RoRNP-associated Y RNAs, as well as 7SL RNA. Alu RNA is, however, present in A3F and A3G RNPs, and both proteins suppress Alu element retrotransposition. Thus, we define a number of subtle differences between the RNPs associated with A3F and A3G and speculate that these contribute to functional differences that have been described for these proteins.
Plasma Membrane Microdomains Containing Vesicular Stomatitis Virus M Protein Are Separate from Microdomains Containing G Protein and Nucleocapsids
Authors: Swinteck B. Dancho , Lyles Douglas S. 
Published: March 2008

ABSTRACT
Immunogold electron microscopy and analysis were used to determine the organization of the major structural proteins of vesicular stomatitis virus (VSV) during virus assembly. We determined that matrix protein (M protein) partitions into plasma membrane microdomains in VSV-infected cells as well as in transfected cells expressing M protein. The sizes of the M-protein-containing microdomains outside the virus budding sites (50 to 100 nm) were smaller than those at sites of virus budding (approximately 560 nm). Glycoprotein (G protein) and M protein microdomains were not colocalized in the plasma membrane outside the virus budding sites, nor was M protein colocalized with microdomains containing the host protein CD4, which efficiently forms pseudotypes with VSV envelopes. These results suggest that separate membrane microdomains containing either viral or host proteins cluster or merge to form virus budding sites. We also determined whether G protein or M protein was colocalized with VSV nucleocapsid protein (N protein) outside the budding sites. Viral nucleocapsids were observed to cluster in regions of the cytoplasm close to the plasma membrane. Membrane-associated N protein was colocalized with G protein in regions of plasma membrane of approximately 600 nm. In contrast to the case for G protein, M protein was not colocalized with these areas of nucleocapsid accumulation. These results suggest a new model of virus assembly in which an interaction of VSV nucleocapsids with G-protein-containing microdomains is a precursor to the formation of viral budding sites.
Upregulation of the TLR3 Pathway by Kaposi's Sarcoma-Associated Herpesvirus during Primary Infection
Authors: West John , Damania Blossom 
Published: March 2008

ABSTRACTKaposi's sarcoma-associated herpesvirus (KSHV) is associated with several different human malignancies, including Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. KSHV establishes lifelong latency in the host and modulates the host immune response. Innate immunity is critical for controlling de novo viral infection. Toll-like receptors (TLRs) are key components of the innate immune system, and they serve as pathogen recognition receptors that stimulate the host antiviral response. In particular, TLR3 has been implicated in RNA virus recognition. Currently, there is no information regarding how KSHV infection modulates any TLR pathway. We report the first evidence that KSHV upregulates TLR3 expression in human monocytes during primary infection. This is also the first demonstration of a human DNA tumor virus upregulating TLR3, a TLR that thus far has been associated with the recognition of RNA viruses. We found that KSHV upregulates the TLR3 pathway and induces TLR3-specific cytokines and chemokines, including beta 1 interferon (IFN-β1) and CXCL10 (IP-10). Small interfering RNAs directed against TLR3 greatly reduced the ability of KSHV to upregulate IFN-β1 and CXCL10 upon infection.
Proteomics Analysis Unravels the Functional Repertoire of Coronavirus Nonstructural Protein 3
Authors: Neuman Benjamin W. , Joseph Jeremiah S. , Saikatendu Kumar S. , Serrano Pedro , Chatterjee Amarnath , Johnson Margaret A. , Liao Lujian , Klaus Joseph P. , Yates John R. , Wüthrich Kurt , Stevens Raymond C. , Buchmeier Michael J. , Kuhn Peter 
Published: March 2008

ABSTRACT

Severe acute respiratory syndrome (SARS) coronavirus infection and growth are dependent on initiating signaling and enzyme actions upon viral entry into the host cell. Proteins packaged during virus assembly may subsequently form the first line of attack and host manipulation upon infection. A complete characterization of virion components is therefore important to understanding the dynamics of early stages of infection. Mass spectrometry and kinase profiling techniques identified nearly 200 incorporated host and viral proteins. We used published interaction data to identify hubs of connectivity with potential significance for virion formation. Surprisingly, the hub with the most potential connections was not the viral M protein but the nonstructural protein 3 (nsp3), which is one of the novel virion components identified by mass spectrometry. Based on new experimental data and a bioinformatics analysis across the
Coronaviridae
, we propose a higher-resolution functional domain architecture for nsp3 that determines the interaction capacity of this protein. Using recombinant protein domains expressed in
Escherichia coli
, we identified two additional RNA-binding domains of nsp3. One of these domains is located within the previously described SARS-unique domain, and there is a nucleic acid chaperone-like domain located immediately downstream of the papain-like proteinase domain. We also identified a novel cysteine-coordinated metal ion-binding domain. Analyses of interdomain interactions and provisional functional annotation of the remaining, so-far-uncharacterized domains are presented. Overall, the ensemble of data surveyed here paint a more complete picture of nsp3 as a conserved component of the viral protein processing machinery, which is intimately associated with viral RNA in its role as a virion component.
Surface Loop Dynamics in Adeno-Associated Virus Capsid Assembly
Authors: DiPrimio Nina , Asokan Aravind , Govindasamy Lakshmanan , Agbandje-McKenna Mavis , Samulski R. Jude 
Published: March 2008

ABSTRACT
The HI loop is a prominent domain on the adeno-associated virus (AAV) capsid surface that extends from each viral protein (VP) subunit overlapping the neighboring fivefold VP. Despite the highly conserved nature of the residues at the fivefold pore, the HI loops surrounding this critical region vary significantly in amino acid sequence between the AAV serotypes. In order to understand the role of this unique capsid domain, we ablated side chain interactions between the HI loop and the underlying EF loop in the neighboring VP subunit by generating a collection of deletion, insertion, and substitution mutants. A mutant lacking the HI loop was unable to assemble particles, while a substitution mutant (10 glycine residues) assembled particles but was unable to package viral genomes. Substitution mutants carrying corresponding regions from AAV1, AAV4, AAV5, and AAV8 yielded (i) particles with titers and infectivity identical to those of AAV2 (AAV2 HI1 and HI8), (ii) particles with a decreased virus titer (1 log) but normal infectivity (HI4), and (iii) particles that synthesized VPs but were unable to assemble into intact capsids (HI5). AAV5 HI is shorter than all other HI loops by one amino acid. Replacing the missing residue (threonine) in AAV2 HI5 resulted in a moderate particle assembly rescue. In addition, we replaced the HI loop with peptides varying in length and amino acid sequence. This region tolerated seven-amino-acid peptide substitutions unless they spanned a conserved phenylalanine at amino acid position 661. Mutation of this highly conserved phenylalanine to a glycine resulted in a modest decrease in virus titer but a substantial decrease (1 log order) in infectivity. Subsequently, confocal studies revealed that AAV2 F661G is incapable of efficiently completing a key step in the infectious pathway nuclear entry, hinting at a possible perturbation of VP1 phospholipase activity. Molecular modeling studies with the F661G mutant suggest that disruption of interactions between F661 and an underlying P373 residue in the EF loop of the neighboring subunit might adversely affect incorporation of the VP1 subunit at the fivefold axis. Western blot analysis confirmed inefficient incorporation of VP1, as well as a proteolytically processed VP1 subunit that could account for the markedly reduced infectivity. In summary, our studies show that the HI loop, while flexible in amino acid sequence, is critical for AAV capsid assembly, proper VP1 subunit incorporation, and viral genome packaging, all of which implies a potential role for this unique surface domain in viral infectivity.
Contribution of CD8+T Cells to Containment of Viral Replication and Emergence of Mutations inMamu-A*01-Restricted Epitopes in Simian Immunodeficiency Virus-Infected Rhesus Monkeys
Authors: Kim Eun-Young , Veazey Ronald S. , Zahn Roland , McEvers Kimberly J. , Baumeister Susanne H. C. , Foster Gabriel J. , Rett Melisa D. , Newberg Michael H. , Kuroda Marcelo J. , Rieber E. Peter , Piatak Michael , Lifson Jeffrey D. , Letvin Norman L. , Wolinsky Steven M. , Schmitz Jörn E. 
Published: March 2008

ABSTRACTHere, we investigated the containment of virus replication in simian immunodeficiency virus (SIV) infection by CD8+lymphocytes. Escape mutations inMamu-A*01epitopes appeared first in SIV Tat TL8 and then in SIV Gag p11C. The appearance of escape mutations in SIV Gag p11C was coincident with compensatory changes outside of the epitope. Eliminating CD8+lymphocytes from rhesus monkeys during primary infection resulted in more rapid disease progression that was associated with preservation of canonical epitopes. These results confirm the importance of cytotoxic T cells in controlling viremia and the constraint on epitope sequences that require compensatory changes to go to fixation.
Very rare aetiology of giant popliteal pseudoaneurysm: anterior cruciate ligament surgery
Authors: Kanko M , Buluc L , Yavuz S , Muezzinoglu S , Berki T 
Published: March 2008

Abstract
There are various causes of the formation of arterial pseudoaneurysms, including trauma, surgical procedures, infection and iatrogenic injuries. A popliteal aneurysm was detected in a patient with pain and discomfort in his leg. The patient had a history of knee surgery. The aneurysm was treated surgically. Aneurysms following penetrating arterial injury resulting from surgical intervention requiring the use of surgical devices is one of the possible traumatic causes.
Severe asthma: approach and management
Authors: Reddy R C 
Published: March 2008

Abstract
Management of severe asthma remains a significant challenge. Patients with this condition do not respond adequately to inhaled corticosteroids and bronchodilators, forcing a search for alternative strategies. The clinician's initial priority is to firmly establish the diagnosis of severe asthma, as many conditions can mimic and/or aggravate this disease. Once the diagnosis is confirmed and confounding variables addressed, a variety of pharmacological and non-pharmacological approaches must be considered. Continuous use of oral corticosteroids carries a risk of significant adverse effects. Leukotriene modifiers and antibodies to IgE are effective for some patients but not for many others. Alternative anti-inflammatory drugs and novel or unconventional modalities may also be used. Although severe asthma remains a clinical dilemma, a rational diagnostic and therapeutic strategy can be used to improve patient outcomes.
An update on cardioembolic stroke
Authors: Weir N U 
Published: March 2008

Abstract
About one-fifth of all ischaemic strokes are cardioembolic. Recent years have seen considerable progress in our understanding of atrial fibrillation, the most important cause of cardioembolism, and evidence-based treatment strategies have emerged. Progress in relation to other cardioembolic disorders has been more limited and here stroke prevention strategies remain less certain and subject to debate. This article briefly reviews the methods currently used to identify and investigate cardioembolic stroke and then provides an update on stroke prevention in relation to atrial fibrillation, valvular heart disease, patent foramen ovale and left ventricular dysfunction (after myocardial infarction and chronic failure).