bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immune Responses and Therapeutic Antitumor Effects of an Experimental DNA Vaccine Encoding Human Papillomavirus Type 16 Oncoproteins Genetically Fused to Herpesvirus Glycoprotein D
Authors: Diniz Mariana O. , Lasaro Marcio O. , Ertl Hildegund C. , Ferreira Luís C. S. 
Published: August 2010

ABSTRACT

Recombinant adenovirus or DNA vaccines encoding herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) genetically fused to human papillomavirus type 16 (HPV-16) oncoproteins (E5, E6, and E7) induce antigen-specific CD8
+
T-cell responses and confer preventive resistance to transplantable murine tumor cells (TC-1 cells). In the present report, we characterized some previously uncovered aspects concerning the induction of CD8
+
T-cell responses and the therapeutic anticancer effects achieved in C57BL/6 mice immunized with pgD-E7E6E5 previously challenged with TC-1 cells. Concerning the characterization of the immune responses elicited in mice vaccinated with pgD-E7E6E5, we determined the effect of the CD4
+
T-cell requirement, longevity, and dose-dependent activation on the E7-specific CD8
+
T-cell responses. In addition, we determined the priming/boosting properties of pgD-E7E6E5 when used in combination with a recombinant serotype 68 adenovirus (AdC68) vector encoding the same chimeric antigen. Mice challenged with TC-1 cells and then immunized with three doses of pgD-E7E6E5 elicited CD8
+
T-cell responses, measured by intracellular gamma interferon (IFN-γ) and CD107a accumulation, to the three HPV-16 oncoproteins and displayed
in vivo
antigen-specific cytolytic activity, as demonstrated with carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled target cells pulsed with oligopeptides corresponding to the

H-2D
b

-restricted immunodominant epitopes of the E7, E6, or E5 oncoprotein. Up to 70% of the mice challenged with 5 × 10
5
TC-1 cells and immunized with pgD-E7E6E5 controlled tumor development even after 3 days of tumor cell challenge. In addition, coadministration of pgD-E7E6E5 with DNA vectors encoding pGM-CSF or interleukin-12 (IL-12) enhanced the therapeutic antitumor effects for all mice challenged with TC-1 cells. In conclusion, the present results expand our previous knowledge on the immune modulation properties of the pgD-E7E6E5 vector and demonstrate, for the first time, the strong antitumor effects of the DNA vaccine, raising promising perspectives regarding the development of immunotherapeutic reagents for the control of HPV-16-associated tumors.
Complication of Corticosteroid Treatment by Acute
Plasmodium malariae
Infection Confirmed by Small-Subunit rRNA Sequencing
Authors: To Kelvin K. W. , Teng Jade L. L. , Wong Samson S. Y. , Ngan Antonio H. Y. , Yuen Kwok-Yung , Woo Patrick C. Y. 
Published: August 2010

ABSTRACT

We report a case of acute
Plasmodium malariae
infection complicating corticosteroid treatment for membranoproliferative glomerulonephritis in a patient from an area where
P. malariae
infection is not endemic. A peripheral blood smear showed typical band-form trophozoites compatible with
P. malariae
or
Plasmodium knowlesi
. SSU rRNA sequencing confirmed the identity to be
P. malariae
.
Diagnostic Limitations to Accurate Diagnosis of Cholera
Authors: Alam Munirul , Hasan Nur A. , Sultana Marzia , Nair G. Balakrish , Sadique A. , Faruque A. S. G. , Endtz Hubert P. , Sack R. B. , Huq A. , Colwell R. R. , Izumiya Hidemasa , Morita Masatomo , Watanabe Haruo , Cravioto Alejandro 
Published: August 2010

ABSTRACT

The treatment regimen for diarrhea depends greatly on correct diagnosis of its etiology. Recent diarrhea outbreaks in Bangladesh showed
Vibrio cholerae
to be the predominant cause, although more than 40% of the suspected cases failed to show cholera etiology by conventional culture methods (CMs). In the present study, suspected cholera stools collected from every 50th patient during an acute diarrheal outbreak were analyzed extensively using different microbiological and molecular tools to determine their etiology. Of 135 stools tested, 86 (64%) produced
V. cholerae
O1 by CMs, while 119 (88%) tested positive for
V. cholerae
O1 by rapid cholera dipstick (DS) assay; all but three samples positive for
V. cholerae
O1 by CMs were also positive for
V. cholerae
O1 by DS assay. Of 49 stools that lacked CM-based cholera etiology despite most being positive for
V. cholerae
O1 by DS assay, 25 (51%) had coccoid
V. cholerae
O1 cells as confirmed by direct fluorescent antibody (DFA) assay, 36 (73%) amplified primers for the genes
wbe
O1 and
ctxA
by multiplex-PCR (M-PCR), and 31 (63%) showed El Tor-specific lytic phage on plaque assay (PA). Each of these methods allowed the cholera etiology to be confirmed for 97% of the stool samples. The results suggest that suspected cholera stools that fail to show etiology by CMs during acute diarrhea outbreaks may be due to the inactivation of
V. cholerae
by
in vivo
vibriolytic action of the phage and/or nonculturability induced as a host response.
Improved Identification of Epidemiologically Related Strains of
Salmonella enterica
by use of a Fusion Algorithm Based on Pulsed-Field Gel Electrophoresis and Multiple-Locus Variable-Number Tandem-Repeat Analysis
Authors: Broschat S. L. , Call D. R. , Davis M. A. , Meng D. , Lockwood S. , Ahmed R. , Besser T. E. 
Published: August 2010

ABSTRACT

Pulsed-field gel electrophoresis (PFGE) and multiple-locus variable-number tandem-repeat analysis (MLVA) are used to assess genetic similarity between bacterial strains. There are cases, however, when neither of these methods quantifies genetic variation at a level of resolution that is well suited for studying the molecular epidemiology of bacterial pathogens. To improve estimates based on these methods, we propose a fusion algorithm that combines the information obtained from both PFGE and MLVA assays to assess epidemiological relationships. This involves generating distance matrices for PFGE data (Dice coefficients) and MLVA data (single-step stepwise-mutation model) and modifying the relative distances using the two different data types. We applied the algorithm to a set of
Salmonella enterica
serovar Typhimurium isolates collected from a wide range of sampling dates, locations, and host species. All three classification methods (PFGE only, MLVA only, and fusion) produced a similar pattern of clustering relative to groupings of common phage types, with the fusion results being slightly better. We then examined a group of serovar Newport isolates collected over a limited geographic and temporal scale and showed that the fusion of PFGE and MLVA data produced the best discrimination of isolates relative to a collection site (farm). Our analysis shows that the fusion of PFGE and MLVA data provides an improved ability to discriminate epidemiologically related isolates but provides only minor improvement in the discrimination of less related isolates.
Diagnosis of Bacteriuria and Leukocyturia by Automated Flow Cytometry Compared with Urine Culture
Authors: Pieretti Barbara , Brunati Pietro , Pini Beatrice , Colzani Carlo , Congedo Pierluigi , Rocchi Marco , Terramocci Riccardo 
Published: August 2010

ABSTRACT
Urinary tract infection (UTI) is a widespread disease, and thus, the most common samples tested in diagnostic microbiology laboratories are urine samples. The “gold standard” for diagnosis is still bacterial culture, but a large proportion of samples are negative. Unnecessary culture can be reduced by an effective screening test. We evaluated the performance of a new urine cytometer, the Sysmex UF-1000i (Dasit), on 703 urine samples submitted to our laboratory for culture. We compared bacteria and leukocyte (WBC) counts performed with the Sysmex UF-1000i to CFU-per-milliliter quantification on CPS agar to assess the best cutoff values. Different cutoff values of bacteria/ml and WBC/ml were compared to give the best discrimination. On the basis of the results obtained in this study, we suggest that when the Sysmex UF-1000i analyzer is used as a screening test for UTI the cutoff values should be 65 bacteria/ml and 100 WBC/ml. Diagnostic performance in terms of sensitivity (98.2%), specificity (62.1%), negative predictive value (98.7%), positive predictive value (53.7%), and diagnostic accuracy (73.3%) were satisfactory. Screening with the Sysmex UF-1000i is acceptable for routine use. In our laboratory, we have reduced the number of bacterial cultures by 43%, speeded up their reporting, and decreased the inappropriate use of antibiotics.
Molecular Epidemiology of Multidrug-Resistant
Acinetobacter baumannii
in a Single Institution over a 10-Year Period
Authors: Runnegar Naomi , Sidjabat Hanna , Goh H. M. Sharon , Nimmo Graeme R. , Schembri Mark A. , Paterson David L. 
Published: August 2010

ABSTRACT

Multidrug-resistant
Acinetobacter baumannii
is a worldwide nosocomial menace. We sought to better understand its behavior through studying the molecular epidemiology of this organism at the Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia, over a 10-year period. Multilocus sequence typing (MLST), semiautomated repetitive sequence-based PCR (rep-PCR), and pulsed-field gel electrophoresis (PFGE) were performed on a selection of 31
A. baumannii
isolates collected over the 10-year period to determine their relationships to one another. MLST also allowed us to put this information in a global context. The presence or absence of
bla
OXA-23
was also established. The presence of
bla
OXA-23
closely correlated with carbapenem resistance in our collection. Sequence type 92 (ST92) was the dominant sequence type and was present in the hospital for 9 years. There was also evidence of the spread of ST69, ST73, and ST125 (novel) within the hospital, but this was not sustained over long periods. There were only single examples of the novel sequence types ST126 and ST127. The different typing methods clustered the isolates similarly; however, PFGE and rep-PCR were more discriminatory than MLST. Worldwide, ST92 and the associated clonal complex 92 represent the most sampled and widespread sequence type(s) and are also known as European clone 2 and worldwide clonal lineage 2. Antibiotic susceptibility within ST92 is variable, suggesting a role for mechanisms other than antibiotic resistance in its success.
Evaluation of BBL CHROMagar VanRE for Detection of Vancomycin-Resistant Enterococci in Rectal Swab Specimens
Authors: Stamper Paul D. , Shulder Stephanie , Bekalo Pearl , Manandhar Deepika , Ross Tracy L. , Speser Sharon , Kingery Julie , Carroll Karen C. 
Published: August 2010

ABSTRACT

A study was performed on 517 surveillance rectal swabs to evaluate a selective and differential chromogenic medium, the BBL CHROMagar VanRE (CVRE), which enables recovery and identification of VanA- and VanB-containing
Enterococcus faecium
(ENFM) and
Enterococcus faecalis
(ENFS) isolates. Compared to BBL Enterococcosel agar, a bile-esculin-azide-vancomycin (BEAV) agar, the initial overall sensitivity, specificity, and positive and negative predictive values of CVRE for the detection of vancomycin-resistant ENFM and ENFS were 99.1% and 94.8% and 84.2% and 99.7%, respectively. Among our patient population, more vancomycin-resistant enterococci (VRE) were recovered with CVRE than BEAV.
Efficiency of Fine-Needle Aspiration Compared with Other Sampling Techniques for Laboratory Diagnosis of Buruli Ulcer Disease
Authors: Herbinger Karl-Heinz , Beissner Marcus , Huber Kristina , Awua-Boateng Nana-Yaa , Nitschke Jörg , Thompson William , Klutse Erasmus , Agbenorku Pius , Assiobo Awovi , Piten Ebekalisai , Wiedemann Franz , Fleischmann Erna , Helfrich Kerstin , Adjei Ohene , Löscher Thomas , Bretzel Gisela 
Published: August 2010

ABSTRACT
In accordance with recent WHO recommendations, this study evaluates the sensitivities of PCR and microscopy for fine-needle aspiration (FNA) versus techniques involving swabs and punch biopsy specimens and suggests that FNA can replace punch biopsies for nonulcerative lesions and may serve as an alternative for ulcerative lesions in cases where scarred edges prevent the collection of swabs.
Structural Evidence of Glycoprotein Assembly in Cellular Membrane Compartments prior to Alphavirus Budding
Authors: Soonsawad Pan , Xing Li , Milla Emerson , Espinoza Juan M. , Kawano Masaaki , Marko Michael , Hsieh Chyongere , Furukawa Hiromitsu , Kawasaki Masahiro , Weerachatyanukul Wattana , Srivastava Ranjana , Barnett Susan W. , Srivastava Indresh K. , Cheng R. Holland 
Published: August 2010

ABSTRACT

Membrane glycoproteins of alphavirus play a critical role in the assembly and budding of progeny virions. However, knowledge regarding transport of viral glycoproteins to the plasma membrane is obscure. In this study, we investigated the role of cytopathic vacuole type II (CPV-II) through
in situ
electron tomography of alphavirus-infected cells. The results revealed that CPV-II contains viral glycoproteins arranged in helical tubular arrays resembling the basic organization of glycoprotein trimers on the envelope of the mature virions. The location of CPV-II adjacent to the site of viral budding suggests a model for the transport of structural components to the site of budding. Thus, the structural characteristics of CPV-II can be used in evaluating the design of a packaging cell line for replicon production.
In Vitro
Analysis of Virus Particle Subpopulations in Candidate Live-Attenuated Influenza Vaccines Distinguishes Effective from Ineffective Vaccines
Authors: Marcus Philip I. , Ngunjiri John M. , Sekellick Margaret J. , Wang Leyi , Lee Chang-Won 
Published: August 2010

ABSTRACT

Two effective (
vac
+
) and two ineffective (
vac
−
) candidate live-attenuated influenza vaccines (LAIVs) derived from naturally selected genetically stable variants of A/TK/OR/71-delNS1[1-124] (H7N3) that differed only in the length and kind of amino acid residues at the C terminus of the nonstructural NS1 protein were analyzed for their content of particle subpopulations. These subpopulations included total physical particles (measured as hemagglutinating particles [HAPs]) with their subsumed biologically active particles of infectious virus (plaque-forming particles [PFPs]) and different classes of noninfectious virus, namely, interferon-inducing particles (IFPs), noninfectious cell-killing particles (niCKPs), and defective interfering particles (DIPs). The
vac
+
variants were distinguished from the
vac
−
variants on the basis of their content of viral subpopulations by (i) the capacity to induce higher quantum yields of interferon (IFN), (ii) the generation of an unusual type of IFN-induction dose-response curve, (iii) the presence of IFPs that induce IFN more efficiently, (iv) reduced sensitivity to IFN action, and (v) elevated rates of PFP replication that resulted in larger plaques and higher PFP and HAP titers. These
in vitro
analyses provide a benchmark for the screening of candidate LAIVs and their potential as effective vaccines. Vaccine design may be improved by enhancement of attributes that are dominant in the effective (
vac
+
) vaccines.
Open Reading Frame E3-10.9K of Subspecies B1 Human Adenoviruses Encodes a Family of Late Orthologous Proteins That Vary in Their Predicted Structural Features and Subcellular Localization
Authors: Frietze Kathryn M. , Campos Samuel K. , Kajon Adriana E. 
Published: August 2010

ABSTRACT
Subspecies B1 human adenoviruses (HAdV-B1s) are important causative agents of acute respiratory disease, but the molecular bases of their distinct pathobiology are still poorly understood. Marked differences in genetic content between HAdV-B1s and the well-characterized HAdV-Cs that may contribute to distinct pathogenic properties map to the E3 region. Between the highly conserved E3-19K and E3-10.4K/RIDα open reading frames (ORFs), and in the same location as the HAdV-C ADP/E3-11.6K ORF, HAdV-B1s carry ORFs E3-20.1K and E3-20.5K and a polymorphic third ORF, designated E3-10.9K, that varies in the size of its predicted product among HAdV-B1 serotypes and genomic variants. As an initial effort to define the function of the E3-10.9K ORF, we carried out a biochemical characterization of E3-10.9K-encoded orthologous proteins and investigated their expression in infected cells. Sequence-based predictions suggested that E3-10.9K orthologs with a hydrophobic domain are integral membrane proteins. Ectopically expressed, C-terminally tagged (with enhanced green fluorescent protein [EGFP]) E3-10.9K and E3-9K localized primarily to the plasma membrane, while E3-7.7K localized primarily to a juxtanuclear compartment that could not be identified. EGFP fusion proteins with a hydrophobic domain were N and O glycosylated. EGFP-tagged E3-4.8K, which lacked the hydrophobic domain, displayed diffuse cellular localization similar to that of the EGFP control. E3-10.9K transcripts from the major late promoter were detected at late time points postinfection. A C-terminally hemagglutinin-tagged version of E3-9K was detected by immunoprecipitation at late times postinfection in the membrane fraction of mutant virus-infected cells. These data suggest a role for ORF E3-10.9K-encoded proteins at late stages of HAdV-B1 replication, with potentially important functional implications for the documented ORF polymorphism.
Partial Functional Rescue of
Helicoverpa armigera
Single Nucleocapsid Nucleopolyhedrovirus Infectivity by Replacement of F Protein with GP64 from
Autographa californica
Multicapsid Nucleopolyhedrovirus
Authors: Wang Manli , Yin Feifei , Shen Shu , Tan Ying , Deng Fei , Vlak Just M. , Hu Zhihong , Wang Hualin 
Published: August 2010

ABSTRACT

Two distinct envelope fusion proteins (EFPs) (GP64 and F) have been identified in members of the
Baculoviridae
family of viruses. F proteins are found in group II nucleopolyhedroviruses (NPVs) of alphabaculoviruses and in beta- and deltabaculoviruses, while GP64 occurs only in group I NPVs of alphabaculoviruses. It was proposed that an ancestral baculovirus acquired the
gp64
gene that conferred a selective advantage and allowed it to evolve into group I NPVs. The F protein is a functional analogue of GP64, as evidenced from the rescue of
gp64
-null
Autographa californica
multicapsid nucleopolyhedrovirus (MNPV) (AcMNPV) by F proteins from group II NPVs or from betabaculoviruses. However, GP64 failed to rescue an F-null
Spodoptera exigua
MNPV (SeMNPV) (group II NPV). Here, we report the successful generation of an infectious
gp64
-rescued group II NPV of
Helicoverpa armigera
(vHaBacΔF-gp64). Viral growth curve assays and quantitative real-time PCR (Q-PCR), however, showed substantially decreased infectivity of vHaBacΔF-gp64 compared to the HaF rescue control virus vHaBacΔF-HaF. Electron microscopy further showed that most vHaBacΔF-gp64 budded viruses (BV) in the cell culture supernatant lacked envelope components and contained morphologically aberrant nucleocapsids, suggesting the improper BV envelopment or budding of vHaBacΔF-gp64. Bioassays using pseudotyped viruses with a reintroduced
polyhedrin
gene showed that GP64-pseudotyped
Helicoverpa armigera
single nucleocapsid nucleopolyhedrovirus (HearNPV) significantly delayed the mortality of infected
H. armigera
larvae.
The Carboxy-Terminal Segment of the Human Cytomegalovirus DNA Polymerase Accessory Subunit UL44 Is Crucial for Viral Replication
Authors: Silva Laurie A. , Loregian Arianna , Pari Gregory S. , Strang Blair L. , Coen Donald M. 
Published: August 2010

ABSTRACT
The amino-terminal 290 residues of UL44, the presumed processivity factor of human cytomegalovirus DNA polymerase, possess all of the established biochemical activities of the full-length protein, while the carboxy-terminal 143 residues contain a nuclear localization signal (NLS). We found that although the amino-terminal domain was sufficient for origin-dependent synthesis in a transient-transfection assay, the carboxy-terminal segment was crucial for virus replication and for the formation of DNA replication compartments in infected cells, even when this segment was replaced with a simian virus 40 NLS that ensured nuclear localization. Our results suggest a role for this segment in viral DNA synthesis.
HLA-Cw*03-Restricted CD8+T-Cell Responses Targeting the HIV-1 Gag Major Homology Region Drive Virus Immune Escape and Fitness Constraints Compensated for by Intracodon Variation
Authors: Honeyborne Isobella , Codoñer Francisco M. , Leslie Alasdair , Tudor-Williams Gareth , Luzzi Graz , Ndung'u Thumbi , Walker Bruce D. , Goulder Philip J. , Prado Julia G. 
Published: August 2010

ABSTRACTThe potential importance of HLA-C-restricted CD8+cytotoxic T lymphocytes (CTL) in HIV infection remains undetermined. We studied the dominant HLA-Cw*03-restricted CTL response to YVDRFFKTL296-304(YL9), within the conserved major homology region (MHR) of the Gag protein, in 80 HLA-Cw*03-positive individuals with chronic HIV infection to better define the efficacy of the YL9 HLA-C-restricted response. The HLA-Cw*03 allele is strongly associated with HIV sequence changes from Thr-303 to Val, Ile, or Ala at position 8 within the YL9 epitope (P= 1.62 × 10−10).In vitrostudies revealed that introduction of the changes T303I and T303A into the YL9 epitope both significantly reduced CTL recognition and substantially reduced the viral replicative capacity. However, subsequent selection of the Val-303 variant, via intracodon variation from Ile-303 (I303V) or Ala-303 (A303V), restored both viral fitness and CTL recognition, as supported by ourin vivodata. These results illustrate that HLA-C-restricted CTL responses are capable of driving viral immune escape within Gag, but in contrast to what was previously described for HLA-B-restricted Gag escape mutants, the common Cw*03-Gag-303V variant selected resulted in no detectable benefit to the host.
Novel Nipah Virus Immune-Antagonism Strategy Revealed by Experimental and Computational Study
Authors: Seto Jeremy , Qiao Liang , Guenzel Carolin A. , Xiao Sa , Shaw Megan L. , Hayot Fernand , Sealfon Stuart C. 
Published: August 2010

ABSTRACTNipah virus is an emerging pathogen that causes severe disease in humans. It expresses several antagonist proteins that subvert the immune response and that may contribute to its pathogenicity. Studies of its biology are difficult due to its high pathogenicity and requirement for biosafety level 4 containment. We integrated experimental and computational methods to elucidate the effects of Nipah virus immune antagonists. Individual Nipah virus immune antagonists (phosphoprotein and V and W proteins) were expressed from recombinant Newcastle disease viruses, and the responses of infected human monocyte-derived dendritic cells were determined. We developed an ordinary differential equation model of the infectious process that that produced results with a high degree of correlation with these experimental results. In order to simulate the effects of wild-type virus, the model was extended to incorporate published experimental data on the time trajectories of immune-antagonist production. These data showed that the RNA-editing mechanism utilized by the wild-type Nipah virus to produce immune antagonists leads to a delay in the production of the most effective immune antagonists, V and W. Model simulations indicated that this delay caused a disconnection between attenuation of the antiviral response and suppression of inflammation. While the antiviral cytokines were efficiently suppressed at early time points, some early inflammatory cytokine production occurred, which would be expected to increase vascular permeability and promote virus spread and pathogenesis. These results suggest that Nipah virus has evolved a unique immune-antagonist strategy that benefits from controlled expression of multiple antagonist proteins with various potencies.