bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative Immune Responses of Patients with Chronic Pulmonary Diseases during the 2-Year Period after Pneumococcal Vaccination
Authors: Chen Meng , Hisatomi Yuki , Furumoto Akitsugu , Kawakami Kenji , Masaki Hironori , Nagatake Tsuyoshi , Sueyasu Yoshiko , Iwanaga Tomoaki , Aizawa Hisamichi , Oishi Kazunori
Published: December 2006

Antibody responses to a 23-valent pneumococcal vaccine for
Streptococcus pneumoniae
serotypes 6B, 14, 19F, and 23F in 84 patients with chronic pulmonary diseases over a 2-year period after vaccination were examined by using a third-generation enzyme-linked immunosorbent assay. Of these patients, 28 (31%) were low responders who had developed increases of at least twofold in the levels of serotype-specific immunoglobulin G (IgG) in sera for none of the four serotypes at 1 month after vaccination. Although no specific clinical features of low responders were evident, their prevaccination levels of IgG for all serotypes were higher than those of responders. In responders, the levels of IgG specific for serotypes 14 and 23F in sera were greatly increased 1 month after vaccination and those specific for serotypes 6B and 19F were moderately increased. In contrast, no significant increases in the levels of IgG specific for serotypes 6B, 19F, and 23F in the low responders during the same period were found, but the levels of IgG specific for serotype 14 did increase. Although a rapid decline in the levels of IgG for all serotypes in responders between 1 month and 6 months after vaccination was found, the levels of IgG specific for serotypes 14 and 23F in sera remained higher than the prevaccination levels for at least 2 years after vaccination. These data suggest the need for the revaccination of responders but not low responders among patients with chronic pulmonary diseases. Revaccination as early as 3 years postvaccination is recommended for responders to increase the reduced levels of IgG in sera, especially those specific for the weak vaccine antigens.
Evaluation of NucliSens easyMAG for Automated Nucleic Acid Extraction from Various Clinical Specimens
Authors: Loens K. , Bergs K. , Ursi D. , Goossens H. , Ieven M.
Published: December 2006

The objectives of this study were to evaluate the performance of the NucliSens easyMAG platform for nucleic acid extraction from different clinical specimens compared to NucliSens miniMAG platform and manual QIAGEN extraction. The NucliSens easyMAG and the NucliSens miniMAG showed equal performance on 215 throat swabs since real-time nucleic acid sequence-based amplification scored the same samples positive for
Mycoplasma pneumoniae
(
n
= 9) and
Chlamydia pneumoniae
(
n
= 5) RNAs, although internal control RNA was slightly better detected with the NucliSens easyMAG (99.3% versus 96.8%). NucliSens easyMAG extracted nucleic acids more efficiently (higher recovery and/or fewer inhibitors) compared to QIAGEN extraction by showing, on average, lower Ct values in real-time LightCycler PCR, although 4 individual specimen out of 45 were found positive only with QIAGEN. For nine
M. pneumoniae
-positive throat swabs, the mean difference in Ct values between NucliSens easyMAG extraction and QIAGEN extraction was −2.26 (range, −5.77 to +0.60); for the detection of five
C. pneumoniae
-positive throat swabs, the average difference in Ct values between the two methods was −3.38 (range, −6.62 to −2.02); and for the detection of cytomegalovirus in 24 blood samples, the mean difference in Ct values between the two methods was −0.95 (range, −5.51 to +1.68). The NucliSens easyMAG is considerably easier to perform, efficiently extracts nucleic acids from throat swabs and whole blood, is automated, and has high throughput.
Geno- and Phenotypic Diversity of Avian Isolates ofStreptococcus gallolyticussubsp.gallolyticus(Streptococcus bovis) and Associated Diagnostic Problems
Authors: Chadfield M. S. , Christensen J. P. , Decostere A. , Christensen H. , Bisgaard M.
Published: December 2006

Recently, strains ofStreptococcus boviswere reclassified asStreptococcus gallolyticus. In the present study we describe for the first time an outbreak ofS. gallolyticusin a broiler flock. Mortality during the first week was normal (<1%), with a final total mortality at the end of production reaching 4.3%. Specific symptoms were not observed. Postmortem pathology demonstrated enlarged and light spleens and livers accompanied by multifocal irregular necroses surrounded by a hemorrhagic zone. In addition, these birds suffered from arthritis and osteomyelitis. Strains isolated from liver and spleen lesions showed clonality as demonstrated by pulsed-field gel electrophoresis. Compared to strains representing previously derived phylogeny, including theS. bovis-S. equinuscomplex, the 16S rRNA-derived phylogeny of the strains investigated in this study demonstrated a paraphyletic group (S. gallolyticus) well separated from two monophyletic groups: (i)S. equinus-S. bovisplusS. infantariusand (ii)S. alactolyticusplusS. intestinalis. According to information in GenBank, none of the strains included from the two monophyletic groups have been isolated from birds. Further biochemical analyses, including tannase activity, identified for the first time avian isolates belonging toS. gallolyticussubsp.gallolyticus. However, these investigations also demonstrated a clear heterogeneity with pigeon isolates.
Use of Pulsed-Field Gel Electrophoresis of Conserved XbaI Fragments for Identification of SwineSalmonellaSerotypes
Authors: Gaul Stephen B. , Wedel Stephanie , Erdman Matthew M. , Harris D. L. , Harris Isabel Turney , Ferris Kathleen E. , Hoffman Lorraine
Published: December 2006

SwineSalmonellaisolates (n= 674) from various locations throughout the United States and Canada were analyzed via pulsed-field gel electrophoresis (PFGE) with XbaI. PFGE subtypes were analyzed by cluster analysis and compared to conventional serotyping results. The analysis showed a correlation of serotype to PFGE subtype. In addition, conserved fragments were identified within the restriction patterns that were unique to each serotype. PFGE using XbaI restriction provided a possible alternative method for screening and identifying swineSalmonellaserotypes.
Necrotizing Fasciitis Due to a Methicillin-Sensitive
Staphylococcus aureus
Isolate Harboring an Enterotoxin Gene Cluster
Authors: Morgan William R. , Caldwell Michael D. , Brady Jennifer M. , Stemper Mary E. , Reed Kurt D. , Shukla Sanjay K.
Published: December 2006

Benign papular eruption on the left leg of a 72-year-old diabetic man developed into rapidly spreading necrotizing fasciitis despite antimicrobial therapy and surgical debridements. This led to eventual amputation to control the infection. The etiological agent was a
Staphylococcus aureus
isolate harboring the enterotoxin gene cluster
seg
,
sei
,
sem
,
sen
, and
seo
but lacked all common toxin genes, including Panton-Valentine leukocidin.
Use of Hydrogen Peroxide Vapor for Deactivation of
Mycobacterium tuberculosis
in a Biological Safety Cabinet and a Room
Authors: Hall Leslie , Otter Jonathan A. , Chewins John , Wengenack Nancy L.
Published: December 2006

Mycobacterium tuberculosis
is an important human pathogen that is routinely cultured in clinical and research laboratories.
M. tuberculosis
can contaminate surfaces and is highly resistant to disinfection. We investigated whether hydrogen peroxide vapor (HPV) is effective for the deactivation of
M. tuberculosis
on experimentally contaminated surfaces in a biological safety cabinet (BSC) and a room. Biological indicators (BIs) consisting of an ∼3-log
10
inoculum of
M. tuberculosis
on stainless steel discs and a 6-log
10
inoculum of
Geobacillus stearothermophilus
were exposed to HPV in BSC time course experiments and at 10 locations during room experiments. In three separate BSC experiments,
M. tuberculosis
BIs were transferred to growth media at 15-min intervals during a 180-min HPV exposure period. No
M. tuberculosis
BIs grew following 30 min of HPV exposure. In three separate room experiments,
M. tuberculosis
and
G. stearothermophilus
BIs were exposed to HPV for 90, 120, and 150 min, respectively. BIs for both microorganisms were deactivated in all 10 locations following 90 min of HPV exposure. HPV provides an alternative to traditional decontamination methods, such as formaldehyde fumigation, for laboratories and other areas contaminated with
M. tuberculosis
.
Evaluation of Dried Blood Spots for Human Immunodeficiency Virus Type 1 Drug Resistance Testing
Authors: McNulty Amanda , Jennings Cheryl , Bennett Diane , Fitzgibbon Joseph , Bremer James W. , Ussery Michael , Kalish Marcia L. , Heneine Walid , García-Lerma J. Gerardo
Published: December 2006

Dried blood spots (DBS) are simpler to prepare, store, and transport than plasma or serum and may represent a good alternative for drug resistance genotyping, particularly in resource-limited settings. However, the utility of DBS for drug resistance testing is unknown. We investigated the efficiency of amplification of large human immunodeficiency virus type 1 (HIV-1)polfragments (1,023 bp) from DBS stored at different temperatures, the type of amplified product(s) (RNA and/or DNA), and the similarity between plasma and DBS sequences. We evaluated two matched plasma/DBS panels stored for 5 to 6 years at several temperatures and 40 plasma/DBS specimens collected from untreated persons in Cameroon and stored for 2 to 3 years at −20°C. The amplification of HIV-1polwas done using an in-house reverse transcriptase-nested PCR assay. Reactions were done with and without reverse transcription to evaluate the contribution of HIV DNA topolsequences from DBS. Amplification was successful for the DBS samples stored for 5 to 6 years at −20°C or at −70°C but not for those stored at room temperature. Thirty-seven of the 40 (92.5%) DBS from Cameroon were amplifiable, including 8/11 (72.7%) with plasma virus loads of <10,000 RNA copies/ml and all 29 with plasma virus loads of >10,000. Proviral DNA contributed significantly to DBS sequences in 24 of the 37 (65%) specimens from Cameroon. The overall similarity between plasma and DBS sequences was 98.1%. Our results demonstrate the feasibility of DBS for drug resistance testing and indicate that −20°C is a suitable temperature for long-term storage of DBS. The amplification of proviral DNA from DBS highlights the need for a wider evaluation of the concordance of resistance genotypes between plasma and DBS.
Molecular Characterization of a New Variant of Rotavirus P[8]G9 Predominant in a Sentinel-Based Survey in Central Italy
Authors: Ansaldi Filippo , Pastorino Barbara , Valle Laura , Durando Paolo , Sticchi Laura , Tucci Pierluigi , Biasci Paolo , Lai Piero , Gasparini Roberto , Icardi Giancarlo ,   and the Paediatric Leghorn Group
Published: December 2006

Rotavirus P[8]G9 was recognized as the most widespread genotype during a sentinel-based survey in Italy; phylogenetic analysis of the VP7 and VP4 genes showed that Italian isolates constituted a closely related genetic cluster distinct from the other G9 strains recently isolated in other European countries, America, and Asia.
Usefulness of PCR-Restriction Fragment Length Polymorphism Typing of the Coagulase Gene To Discriminate Arbekacin-Resistant Methicillin-Resistant
Staphylococcus aureus
Strains
Authors: Ishino Keiko , Tsuchizaki Naofumi , Ishikawa Jun , Hotta Kunimoto
Published: December 2006

We compared the results of two typing methods for 678 strains of methicillin-resistant
Staphylococcus aureus
and methicillin-susceptible
S. aureus
. PCR-restriction fragment length polymorphism typing of the coagulase gene was a more reliable method than coagulase serotyping from the viewpoint of arbekacin resistance.
A Cautionary Tale: Lack of Consistency in Allele Sizes between Two Laboratories for a Published Multilocus Microsatellite Typing System
Authors: Pasqualotto Alessandro C. , Denning David W. , Anderson Michael J.
Published: December 2006

For species with low genetic diversity, typing using the differences in PCR fragment length resulting from variations in numbers of short tandem repeats has been shown to provide a high level of discrimination. This technique has been called multilocus microsatellite typing (MLMT) or multiple-locus variable-number tandem repeat analysis, and studies usually employ genetic or sequence analyzers to size PCR fragments to a high degree of precision. We set out to validate one such system that has been developed forAspergillus fumigatus(H. A. de Valk, J. F. G. M. Meis, I. M. Curfs, K. Muehlethaler, J. W. Mouton, and C. H. W. Klaassen, J. Clin. Microbiol. 43:4112-4120, 2005). The sizes of the alleles were compared both by sequencing and from two genotyping laboratories, where they used capillary electrophoresis (CE) for sizing. Size differences of up to 6 bases were found between the actual sizes reported by sequencing and the sizes reported by CE. In addition, because the two genotyping laboratories used different machines and running conditions, differences of up to 3 bases were identified between them. As the microsatellite markers used differ by repeat units of 3 or 4 bases, it was not possible to assign PCR fragments to the correct alleles without confirming the sizes of a range of alleles by direct sequencing. Lines of best fit were plotted for each CE machine against actual sizes and will therefore enable unsequenced PCR fragments to be assigned to the correct alleles. This study highlights the care required to ensure that an MLMT system undergoes a suitable correction procedure before data can be merged between different laboratories involved in the typing of individual species.
Evolutionary History and Global Spread of the Emerging G12 Human Rotaviruses
Authors: Rahman Mustafizur , Matthijnssens Jelle , Yang Xuelei , Delbeke Thomas , Arijs Ingrid , Taniguchi Koki , Iturriza-Gómara Miren , Iftekharuddin Nadia , Azim Tasnim , Van Ranst Marc
Published: December 2006

G12
rotaviruses were first detected in diarrheic children in the
Philippines in 1987, but no further cases were reported until 1998.
However, G12 rotaviruses have been detected all over the world in
recent years. Here, we report the worldwide variations of G12
rotaviruses to investigate the evolutionary mechanisms by which they
managed to spread globally in a short period of time. We sequenced the
complete genomes (11 segments) of nine G12 rotaviruses isolated in
Bangladesh, Belgium, Thailand, and the Philippines and compared them
with the genomes of other rotavirus strains. Our genetic analyses
revealed that after introduction of the VP7 gene of the rare G12
genotype into more common local strains through reassortment, a vast
genetic diversity was generated and several new variants with distinct
gene constellations emerged. These reassortment events most likely took
place in Southeast Asian countries and spread to other parts of the
world. The acquirement of gene segments from human-adapted rotaviruses
might allow G12 to better propagate in humans and hence to develop into
an important emerging human
pathogen.
Preferential Infection of Dendritic Cells during Human Immunodeficiency Virus Type 1 Infection of Blood Leukocytes
Authors: Cameron Paul U. , Handley Amanda J. , Baylis Dean C. , Solomon Ajantha E. , Bernard Nicholas , Purcell Damian F. J. , Lewin Sharon R.
Published: December 2006

Human immunodeficiency virus type 1 (HIV-1) transmission by the parenteral route is similar to mucosal transmission in the predominance of virus using the CCR5 coreceptor (R5 virus), but it is unclear whether blood dendritic cells (DCs), monocytes, or T cells are the cells initially infected. We used ex vivo HIV-1 infection of sorted blood mononuclear cells to model the in vivo infection of blood leukocytes. Using quantitative real-time PCR to detect full-length HIV-1 DNA, both sorted CD11c+myeloid and CD11c−plasmacytoid DCs were more frequently infected than other blood mononuclear cells, including CD16+or CD14+monocytes or resting CD4+T cells. There was a strong correlation between CCR5 coreceptor use and preferential DC infection across a range of HIV-1 isolates. After infection of unsorted blood mononuclear cells, HIV-1 was initially detected in the CD11c+DCs and later in other leukocytes, including clustering DCs and activated T cells. DC infection with R5 virus was productive, as shown by efficient transmission to CD4+T cells in coculture. Blood DCs infected with HIV-1 in vitro and cultured alone expressed only low levels of multiply spliced HIV-1 RNA unless cocultured with CD4+T cells. Early selective infection of immature blood DCs by R5 virus and upregulation of viral expression during DC-T-cell interaction and transmission provide a potential pathway for R5 selection following parenteral transmission.
Clade-Specific Differences between Human Immunodeficiency Virus Type 1 Clades B and C: Diversity and Correlations in C3-V4 Regions of gp120
Authors: Gnanakaran S. , Lang Dorothy , Daniels Marcus , Bhattacharya Tanmoy , Derdeyn Cynthia A. , Korber Bette
Published: December 2006

Current knowledge of human immunodeficiency virus type 1 envelope (Env) glycoprotein structure and function is based on studies of clade B viruses. We present evidence of sequence and structural differences in viral glycoprotein gp120 between clades B and C. In clade C, the C3 region α2-helix exhibits high sequence entropy at the polar face but maintains its amphipathicity, whereas in clade B it accommodates hydrophobic residues. The V4 hypervariable domain in clade C is shorter than that in clade B. Generally, shorter V4 loops are incompatible with a glycine occurring in the α2-helix in clade C, an intriguing association that could be exploited to inform Env immunogen design.
Downregulation of Gamma Interferon Receptor 1 by Kaposi's Sarcoma-Associated Herpesvirus K3 and K5
Authors: Li Qinglin , Means Robert , Lang Sabine , Jung Jae U.
Published: December 2006

Upon viral infection, the major defense mounted by the host immune system is activation of the interferon (IFN)-mediated antiviral pathway. In order to complete their life cycles, viruses must modulate the host IFN-mediated immune response. The K3 and K5 proteins of a human tumor-inducing herpesvirus, Kaposi's sarcoma-associated herpesvirus (KSHV), have been shown to downregulate the surface expression of host immune modulatory receptors by increasing their endocytosis rates, which leads to suppression of cell-mediated immunity. In this report, we demonstrate that K3 and K5 both specifically target gamma interferon receptor 1 (IFN-γR1) and induce its ubiquitination, endocytosis, and degradation, resulting in downregulation of IFN-γR1 surface expression and, thereby, inhibition of IFN-γ action. Mutational analysis indicated that K5 appeared to downregulate IFN-γR1 more strongly than K3 and that the amino-terminal ring finger motif and the carboxyl-terminal region of K5 were necessary for IFN-γR1 downregulation. These results suggest that KSHV K3 and K5 suppress both cytokine-mediated and cell-mediated immunity, which ensures efficient viral avoidance of host immune controls.
A Multicomponent RNA-Based Control System Regulates Subgenomic mRNA Transcription in a Tombusvirus
Authors: Lin Han-Xin , Xu Wei , White K. Andrew
Published: December 2006

During infections, positive-strand RNA tombusviruses transcribe two subgenomic (sg) mRNAs that allow for the expression of a subset of their genes. This process is thought to involve an unconventional mechanism involving the premature termination of the virally encoded RNA-dependent RNA polymerase while it is copying the virus genome. The 3′ truncated minus strands generated by termination are then used as templates for sg mRNA transcription. In addition to requiring an extensive network of long-distance RNA-RNA interactions (H.-X. Lin and K. A. White, EMBO J. 23:3365-3374, 2004), the transcription of tombusvirus sg mRNAs also involves several additional RNA structures. In vivo analysis of these diverse RNA elements revealed that they function at distinct steps in the process by facilitating the formation or stabilization of the long-distance interactions, modulating minus-strand template production, or promoting the initiation of sg mRNA transcription. All of the RNA elements characterized could be readily incorporated into a premature termination model for sg mRNA transcription. Overall, the analyses revealed a complex system that displays a high level of structural integration and functional coordination. This multicomponent RNA-based control system may serve as a useful paradigm for understanding related transcriptional processes in other positive-sense RNA viruses.
Degradation of Tyrosine Phosphatase PTPN3 (PTPH1) by Association with Oncogenic HumanPapillomavirus E6 Proteins
Authors: Jing Ming , Bohl Joanna , Brimer Nicole , Kinter Michael , Vande Pol Scott B.
Published: December 2006

Oncoproteins from DNA tumor viruses associate with critical cellular proteins to
regulate cell proliferation, survival, and differentiation.Human papillomavirus (HPV) E6 oncoproteins have been previously shown
to associate with a cellular HECT domain ubiquitin ligase termed E6AP
(UBE3A). Here we show that the E6-E6AP complex associates with and
targets the degradation of the protein tyrosine phosphatase PTPN3
(PTPH1) in vitro and in living cells. PTPN3 is a membrane-associated
tyrosine phosphatase with FERM, PDZ, and PTP domains previously
implicated in regulating tyrosine phosphorylation of growth factor
receptors and p97 VCP (valosin-containing protein, termed Cdc48 in
Saccharomyces cerevisiae
) and is mutated in a
subset of colon cancers. Degradation of PTPN3 by E6 requires E6AP, the
proteasome, and an interaction between the carboxy terminus of E6 and
the PDZ domain of PTPN3. In transduced keratinocytes, E6 confers
reduced growth factor requirements, a function that requires the PDZ
ligand of E6 and that can in part be replicated by inhibiting the
expression of PTPN3. This report demonstrates the potential of E6 to
regulate phosphotyrosine metabolism through the targeted degradation of
a tyrosine
phosphatase.
HLA-A2-Restricted Protection against Lethal Lymphocytic Choriomeningitis
Authors: Botten Jason , Whitton J. Lindsay , Barrowman Polly , Sidney John , Whitmire Jason K. , Alexander Jeff , Ting Joey P. C. , Bui Huynh-Hoa , Sette Alessandro , Buchmeier Michael J.
Published: December 2006

The consequences of human lymphocytic choriomeningitis virus (LCMV) infection can be severe, including aseptic meningitis in immunocompetent individuals, hydrocephalus or chorioretinitis in fetal infection, or a highly lethal outcome in immunosuppressed individuals. In murine models of LCMV infection, CD8+T cells play a primary role in providing protective immunity, and there is evidence that cellular immunity may also be important in related arenavirus infections in humans. For this reason, we sought to identify HLA-A2 supertype-restricted epitopes from the LCMV proteome and evaluate them as vaccine determinants in HLA transgenic mice. We identified four HLA-A*0201-restricted peptides—nucleoprotein NP69-77, glycoprotein precursor GPC10-18, GPC447-455, and zinc-binding protein Z49-58—that displayed high-affinity binding (≤275 nM) to HLA-A*0201, induced CD8+T-cell responses of high functional avidity in HLA-A*0201 transgenic mice, and were naturally processed from native LCMV antigens in HLA-restricted human antigen presenting cells. One of the epitopes (GPC447-455), after peptide immunization of HLA-A*0201 mice, induced CD8+T cells capable of killing peptide-pulsed HLA-A*0201-restricted target cells in vivo and protected mice against lethal intracranial challenge with LCMV.
Basolateral Entry and Release of Crimean-Congo Hemorrhagic Fever Virus in Polarized MDCK-1 Cells
Authors: Connolly-Andersen Anne-Marie , Magnusson Karl-Erik , Mirazimi Ali
Published: December 2006

Crimean-Congo hemorrhagic fever virus (CCHFV) is an etiological agent of a disease with mortality rates in patients averaging 30%. The disease is characterized by fever, myalgia, and hemorrhage. Mechanisms underlying the hemorrhage have to our knowledge not been elucidated for CCHFV. Possibly, a direct or indirect viral effect on tight junctions (TJ) could cause the hemorrhage observed in patients, as TJ play a crucial role in vascular homeostasis and can cause leakage upon deregulation. Moreover, there is no knowledge regarding the site of entry and release of CCHFV in polarized epithelial cells. Such cells represent a barrier to virus dissemination within the host, and as a site of viral entry and release, they could play a key role in further spread. For the first time, we have shown preferential basolateral entry of CCHFV in Madin-Darby canine kidney 1 (MDCK-1) epithelial cells. Furthermore, we demonstrated basolateral release of CCHFV in polarized epithelial cells. Interestingly, by measuring transepithelial electrical resistance, we found no effect of CCHFV replication on the function of TJ in this study. Neither did we observe any difference in the localization of the TJ proteins ZO-1 and occludin in CCHFV-infected cells compared to mock-infected cells.
The Cytoplasmic Tail of the Severe Acute Respiratory Syndrome Coronavirus Spike Protein Contains a Novel Endoplasmic Reticulum Retrieval Signal That Binds COPI and Promotes Interaction with Membrane Protein
Authors: McBride Corrin E. , Li Jie , Machamer Carolyn E.
Published: December 2006

Like
other coronaviruses, severe acute respiratory syndrome coronavirus
(SARS CoV) assembles at and buds into the lumen of the endoplasmic
reticulum (ER)-Golgi intermediate compartment (ERGIC). Accumulation of
the viral envelope proteins at this compartment is a prerequisite for
virus assembly. Previously, we reported the identification of a dibasic
motif (KxHxx) in the cytoplasmic tail of the SARS CoV spike (S) protein
that was similar to a canonical dilysine ER retrieval signal. Here we
demonstrate that this motif is a novel and functional ER retrieval
signal which reduced the rate of traffic of the full-length S protein
through the Golgi complex. The KxHxx motif also partially retained two
different reporter proteins in the ERGIC region and reduced their rates
of trafficking, although the motif was less potent than the canonical
dilysine signal. The dibasic motif bound the coatomer complex I (COPI)
in an in vitro binding assay, suggesting that ER retrieval may
contribute to the accumulation of SARS CoV S protein near the virus
assembly site for interaction with other viral structural proteins. In
support of this, we found that the dibasic motif on the SARS S protein
was required for its localization to the ERGIC/Golgi region when
coexpressed with SARS membrane (M) protein. Thus, the cycling of SARS S
through the ER-Golgi system may be required for its incorporation into
assembling virions in the
ERGIC.
The Cellular Protein P58
IPK
Regulates Influenza Virus mRNA Translation and Replication through a PKR-Mediated Mechanism
Authors: Goodman Alan G. , Smith Jennifer A. , Balachandran Siddharth , Perwitasari Olivia , Proll Sean C. , Thomas Matthew J. , Korth Marcus J. , Barber Glen N. , Schiff Leslie A. , Katze Michael G.
Published: December 2006

We previously hypothesized that efficient translation of influenza virus mRNA requires the recruitment of P58
IPK
, the cellular inhibitor of PKR, an interferon-induced kinase that targets the eukaryotic translation initiation factor eIF2α. P58
IPK
also inhibits PERK, an eIF2α kinase that is localized in the endoplasmic reticulum (ER) and induced during ER stress. The ability of P58
IPK
to interact with and inhibit multiple eIF2α kinases suggests it is a critical regulator of both cellular and viral mRNA translation. In this study, we sought to definitively define the role of P58
IPK
during viral infection of mammalian cells. Using mouse embryo fibroblasts from P58
IPK−/−
mice, we demonstrated that the absence of P58
IPK
led to an increase in eIF2α phosphorylation and decreased influenza virus mRNA translation. The absence of P58
IPK
also resulted in decreased vesicular stomatitis virus replication but enhanced reovirus yields. In cells lacking the P58
IPK
target, PKR, the trends were reversed—eIF2α phosphorylation was decreased, and influenza virus mRNA translation was increased. Although P58
IPK
also inhibits PERK, the presence or absence of this kinase had little effect on influenza virus mRNA translation, despite reduced levels of eIF2α phosphorylation in cells lacking PERK. Finally, we showed that influenza virus protein synthesis and viral mRNA levels decrease in cells that express a constitutively active, nonphosphorylatable eIF2α. Taken together, our results support a model in which P58
IPK
regulates influenza virus mRNA translation and infection through a PKR-mediated mechanism which is independent of PERK.
Influenza Virion-Derived Viral Ribonucleoproteins Synthesize both mRNA and cRNA In Vitro
Authors: Vreede F. T. , Brownlee G. G.
Published: December 2006

The mechanisms regulating the synthesis of mRNA, cRNA, and viral genomic RNA (vRNA) by the influenza A virus RNA-dependent RNA polymerase are not fully understood. Early results suggested that the RNA polymerase “switched” from a transcriptase to a replicase during the viral life cycle in response to the expression of viral proteins. However, recently an alternative model suggesting that replication of influenza virus is regulated by stabilization of the replicative intermediates was proposed. According to this model, the virion-associated polymerase is capable of synthesizing both mRNA and cRNA. We now demonstrate that virion-derived viral ribonucleoproteins (vvRNPs) synthesize both mRNA and cRNA in vitro in the absence of non-virion-associated RNA polymerase or nucleoproteins. The authenticity of the in vitro-transcribed mRNA and cRNA was confirmed by terminal sequence analysis. The addition of non-virion-associated polymerase or NP had no effect on vvRNP activity. De novo synthesis of cRNA was found to be more sensitive than the capped primer-dependent synthesis of mRNA to the concentration of ATP, CTP, and GTP. We conclude that vvRNPs intrinsically possess both transcriptase and replicase activities and that there is no switch in the synthesis of mRNA to cRNA during the influenza virus life cycle.
Transmembrane Domains of Hepatitis C Virus Envelope Glycoproteins: Residues Involved in E1E2 Heterodimerization and Involvement of These Domains in Virus Entry
Authors: Ciczora Yann , Callens Nathalie , Penin François , Pécheur Eve-Isabelle , Dubuisson Jean
Published: December 2006

The transmembrane (TM) domains of hepatitis C virus (HCV) envelope glycoproteins E1 and E2 have been shown to play multiple roles during the biogenesis of the E1E2 heterodimer. By using alanine scanning insertion mutagenesis within the TM domains of HCV envelope glycoproteins, we have previously shown that the central regions of these domains as well as the N-terminal part of the TM domain of E1 are involved in heterodimerization. Here, we used a tryptophan replacement scan of these regions to identify individual residues that participate in those interactions. Our mutagenesis study identified at least four residues involved in heterodimerization: Gly 354, Gly 358, Lys 370, and Asp 728. Interestingly, Gly 354 and Gly 358 belong to a G
XXX
G oligomerization motif. Our tryptophan mutants were also used to generate retrovirus-based, HCV-pseudotyped particles (HCVpp) in order to analyze the effects of these mutations on virus entry. Surprisingly, two mutants consistently displayed higher infectivity compared to that of the wild type. In contrast, HCVpp infectivity was strongly affected for many mutants, despite normal E1E2 heterodimerization and normal levels of incorporation of HCV glycoproteins into HCVpp. The characterization of some of these HCVpp mutants in the recently developed in vitro fusion assay using fluorescent-labeled liposomes indicated that mutations reducing HCVpp infectivity without altering E1E2 heterodimerization affected the fusion properties of HCV envelope glycoproteins. In conclusion, this mutational analysis identified residues involved in E1E2 heterodimerization and revealed that the TM domains of HCV envelope glycoproteins play a major role in the fusion properties of these proteins.
Antiviral Protein APOBEC3G Localizes to Ribonucleoprotein Complexes Found in P Bodies and Stress Granules
Authors: Gallois-Montbrun Sarah , Kramer Beatrice , Swanson Chad M. , Byers Helen , Lynham Steven , Ward Malcolm , Malim Michael H.
Published: December 2006

Members of the APOBEC (apolipoproteinBmRNA-editing enzymecatalytic polypeptide 1-like) family of cytidine deaminases inhibit host cell genome invasion by exogenous retroviruses and endogenous retrotransposons. Because these enzymes can edit DNA or RNA and potentially mutate cellular targets, their activities are presumably regulated; for instance, APOBEC3G (A3G) recruitment into high-molecular-weight ribonucleoprotein (RNP) complexes has been shown to suppress its enzymatic activity. We used tandem affinity purification together with mass spectrometry (MS) to identify protein components within A3G-containing RNPs. We report that numerous cellular RNA-binding proteins with diverse roles in RNA function, metabolism, and fate determination are present in A3G RNPs but that most interactions with A3G are mediated via binding to shared RNAs. Confocal microscopy demonstrated that substantial quantities of A3G localize to cytoplasmic microdomains that are known as P bodies and stress granules (SGs) and are established sites of RNA storage and metabolism. Indeed, subjecting cells to stress induces the rapid redistribution of A3G and a number of P-body proteins to SGs. Among these proteins are Argonaute 1 (Ago1) and Argonaute 2 (Ago2), factors that are important for RNA silencing and whose interactions with A3G are resistant to RNase treatment. Together, these findings reveal that A3G associates with RNPs that are found throughout the cytosol as well as in discrete microdomains. We also speculate that the interplay between A3G, RNA-silencing pathways, and cellular sites of RNA metabolism may contribute to A3G's role as an inhibitor of retroelement mobility and as a possible regulator of cellular RNA function.
Vaccinia Virus 4c (A26L) Protein on Intracellular Mature Virus Binds to the Extracellular Cellular Matrix Laminin
Authors: Chiu Wen-Ling , Lin Chi-Long , Yang Min-Hsiang , Tzou Der-Lii M. , Chang Wen
Published: December 2006

Vaccinia virus intracellular mature virus (IMV) binds to glycosaminoglycans (GAGs) on cells via three virion proteins, H3L, A27L, and D8L. In this study, we demonstrated that binding of IMV to BSC40 cells was competitively inhibited by soluble laminin but not by fibronectin or collagen V, suggesting that this cell surface extracellular matrix (ECM) protein may play a role in vaccinia virus entry. Moreover, IMV infection of GAG
−
sog9 cells was also inhibited by laminin, demonstrating that virion binding to laminin does not involve a prior interaction with GAGs. Furthermore, comparative envelope protein analyses of wild-type vaccinia virus strain Western Reserve, which binds to laminin, and of a mutant virus, IA27L, which does not, showed that the A26L open reading frame (ORF), encoding an envelope protein, was mutated in IA27L, resulting in A26L being absent from the IMV. Expression of the wild-type A26L ORF in IA27L resulted in laminin binding activity. Moreover, recombinant A26L protein bound to laminin in vitro with a high affinity, providing direct evidence that A26L is the laminin binding protein on IMV. In summary, these results reveal a novel role for the vaccinia viral envelope protein A26L in binding to the ECM protein laminin, an association that is proposed to facilitate IMV entry.
Positive and Negative Effects of Adenovirus Type 5 Helper Functions on Adeno-Associated Virus Type 5 (AAV5) Protein Accumulation Govern AAV5 Virus Production
Authors: Nayak Ramnath , Pintel David J.
Published: December 2006

Full replication of adeno-associated virus type 5 (AAV5) is sustained by adenovirus type 5 (Ad5) helper functions E1a, E1b, E2a, E4Orf6, and virus-associated (VA) RNA; however, their combined net enhancement of AAV5 replication was comprised of both positive and negative individual effects. Although Ad5 E4Orf6 was required for AAV5 genomic DNA replication, it also functioned together with E1b to degrade de novo-expressed, preassembled AAV5 capsid proteins and Rep52 in a proteosome-dependent manner. VA RNA enhanced accumulation of AAV5 protein, overcoming the degradative effects of E4Orf6, and was thus required to restore adequate amounts of AAV5 proteins necessary to achieve efficient virus production.
Public road transport crashes in a low income country
Authors: Dharmaratne S D , Stevenson M
Published: December 2006

Objective:To assess the safety of government versus non-government public road transportation.

Design:Descriptive study.

Setting:Kandy Municipality Area, Sri Lanka.

Subjects:All road traffic crashes reported to the Kandy Police from 1 October 1998 to 30 September 1999.

Main outcome:Involvement in a road traffic crash reported to the Kandy Police in which a government bus, private bus, or a three-wheeler was involved.

Results:During the study period, 132 government buses, 243 private buses, and 115 three wheelers were involved in 437 police reported road crashes. Of these crashes, eight (1.8%) were fatal and 132 (30.2%) were crashes resulting in injury requiring hospitalization. The majority of road crashes involved vehicle-vehicle interaction (63.4%) and vehicle-pedestrian interaction (17.8%), while the remainder consisted of vehicle-passenger and vehicle-road structure crashes. The research highlights an increased risk associated with travel on privately owned buses (RR = 2.0, 95% CI 1.6 to 2.5) and three wheelers (RR = 2.2, 95% CI 1.7 to 2.8) compared to travel on government buses. The disparity in crash rates between government and privately owned transportation modes can be explained, in part, by fewer safety requirements being imposed on the deregulated public transportation system.

Conclusions:Recommendations are made in order to address the differential in crash rates between public and private vehicle ownership used for public transportation in Sri Lanka.
Motor vehicle crash pedestrian deaths in New York City: the plight of the older pedestrian
Authors: Nicaj L , Wilt S , Henning K
Published: December 2006

Objective:To describe the epidemiology of pedestrian deaths due to motor vehicle crashes (MVCs) between age groups in New York City (NYC), with a comparison to national data.

Methods:Review of MVC pedestrian deaths in NYC and the US from 1998 to 2002. Data on deaths were obtained from the National Highway Traffic Safety Administration.

Results:Almost half (48%) of all MVC deaths in NYC were among pedestrians, compared with 12% nationally. Pedestrian death rates were highest among older age groups (⩾65 years). NYC’s older pedestrians were more likely than US older pedestrians to be killed at an intersection, during daytime and on weekdays.

Conclusion:Older people constitute a major proportion of MVC pedestrian deaths in NYC. Conditions (such as traffic exposure) surrounding pedestrian MVCs may differ by age group. The high burden of MVCs among older pedestrians in NYC highlights the importance of local-level analysis to guide public health planning.
Non-firearm weapon use and injury severity: priorities for prevention
Authors: Brennan I R , Moore S C , Shepherd J P
Published: December 2006

Objectives:To test the hypothesis that weapon-related violence (excluding firearms) results in more severe injury relative to the use of body parts (fists, feet and other body parts), and to rank order of injury severity by assault mechanism.

Design:Retrospective cohort study.

Participants:24 660 patients who were treated in a UK emergency department for violence-related injury.

Main outcome measure:Score on the Manchester Triage Scale.

Results:The use of a weapon resulted in significantly more serious injury (adjusted odds ratio (AOR) 1.13, 95% confidence interval (CI) 1.00 to 1.28). However, of all mechanisms of violent injury, the use of feet resulted in most severe injury (AOR 1.41, 95% CI 1.17 to 1.70), followed by blunt objects (AOR 1.35, 95% CI 1.14 to 1.58), other body parts (AOR 1.22, 95% CI 1.06 to 1.40) and sharp objects (AOR 1.09, 95% CI 0.91 to 1.5), compared with use of fists.

Conclusions:Use of weapons resulted in more severe injury than use only of body parts. The use of feet caused the most serious injuries, whereas the use of fists caused the least severe injuries. Injury severity varied by number of assailants and age of the patient—peaking at 47 years—but not by number of injuries. Preventing the use of feet in violence, and preventing group violence should be major priorities.
Prevalence of transportation safety measures portrayed in primetime US television programs and commercials
Authors: McGwin G , Modjarrad K , Reiland A , Tanner S , Rue L W
Published: December 2006

Objective:To determine the prevalence of transportation related safety behaviors, such as seatbelt and helmet use, in primetime US television programs and commercials.

Design:Cross sectional study.

Setting:Top rated television programs and associated commercials from four major US television networks were reviewed for the prevalence of transportation safety related behaviors during a one month period in 2005. Programs were categorized according to the time and network of airing, program type, program rating, and—for commercials—type of product being advertised

Subjects:Occupants of automobiles, motorcycles, or bicycles in 507 instances in which a transportation scene was aired.

Results:Seatbelt use was depicted in 62% and 86% of individuals in television program and commercial automobile scenes, respectively. The prevalence of motorcycle helmet use was 47% in television programs and 100% in commercials. Bicycle helmets were used in 9% of television programs and 84% of commercials. The frequency of seatbelt use in programs and commercials varied by television rating and genre but did not differ by network, time of airing, or age of character portrayed.

Conclusions:The prevalence of safety related behaviors aired on major US networks during primetime slots is higher than previous reports but still much lower than national averages. Commercials, in contrast, portray transportation safety measures with a frequency that exceeds that of US television programs or most national surveys.
An exploration of human services system contacts prior to suicide in South Carolina: an expansion of the South Carolina Violent Death Reporting System
Authors: Weis M A , Bradberry C , Carter L P , Ferguson J , Kozareva D
Published: December 2006

Objective:To link South Carolina Violent Death Reporting System (SCVDRS) data with state government human services databases, enabling expanded analysis of suicide in South Carolina and providing a model for other jurisdictions.

Design:The SCVDRS database compiles data from vital statistics, coroner reports, and law enforcement incident and supplemental reports. The Office of Research and Statistics, South Carolina Budget and Control Board (ORS) created a “Data Warehouse”, to which a variety of state agencies and healthcare providers submit data on a regular basis. A unique identifier was used to link SCVDRS data to the Data Warehouse so that data may be analyzed on aggregate and case-specific levels. Year 2004 suicide data from SCVDRS were linked to South Carolina Uniform Billing codes from hospital in-patient and emergency room billing records, State Department of Mental Health service records, and criminal justice databases.

Results:SCVDRS year 2004 suicide data are augmented by hospitalization and emergency room visit data and diagnoses; State Department of Mental Health service provision; and criminal involvement. Of the 491 suicides occurring in 2004, 282 linked with hospitalization and emergency room data, 196 linked with criminal history databases, and 91 had previous contact with the State Department of Mental Health.

Conclusions:Linking SCVDRS data to additional human services databases enables greater examination of factors surrounding suicide. Results show the positive benefits of partnerships created through SCVDRS, illustrate how SCVDRS and human service databases may augment each other, and suggest practitioners should explore implementation of prevention programs in specific settings.
Seat belt use among Hispanic ethnic subgroups of national origin
Authors: Briggs N C , Schlundt D G , Levine R S , Goldzweig I A , Stinson N , Warren R C
Published: December 2006

Objective:Findings from over a dozen studies of Hispanic/white disparities in seat belt use have been inconsistent, variably revealing that seat belt use prevalence among Hispanics is higher, lower, or comparable to use among non-Hispanics. In contrast to previous studies, this study investigates disparities in seat belt use by Hispanic subgroups of national origin.

Methods:Data from the US Fatality Analysis Reporting System were used to compare seat belt use among 60 758 non-Hispanic whites and 6879 Hispanics (Mexican American (MA), n = 5175; Central American/South American (CASA), n = 876; Puerto Rican (PR), n = 412; Cuban (CU), n = 416) killed in crashes from 1999–2003. Logistic regression was used to adjust for age, gender, seat belt law, seat position, urban/rural region, and income.

Results:Overall adjusted odds ratios for seat belt use among Hispanic subgroups, relative to non-Hispanic whites, were 1.04 (95% confidence interval (CI) 0.85 to 1.28) for CUs, 1.17 (95% CI 0.95 to 1.44) for PRs, 1.33 (95% CI 1.25 to 1.42) for MAs, and 1.66 (95% CI 1.44 to 1.91) for CASAs. Relative to their non-Hispanic white counterparts, odds ratios among MA and CASA Hispanics were highest for men, younger age groups, drivers, primary law states, rural areas, and lower income quartiles.

Conclusion:Among all Hispanic subgroups, seat belt use was at least as prevalent as among non-Hispanic whites. In the CASA and MA subgroups, which have the most rapidly growing subpopulations of immigrants, seat belt use was significantly more common than among whites.
Using NVDRS data for suicide prevention: promising practices in seven states
Authors: Powell V , Barber C W , Hedegaard H , Hempstead K , Hull-Jilly D , Shen X , Thorpe G E , Weis M A
Published: December 2006

Objectives:This article describes how seven states participating in a new public health surveillance system for violent death in the US, the National Violent Death Reporting System (NVDRS), have used data to support local suicide prevention activities.

Setting:The NVDRS is unique in that it augments death certificate data with event and circumstance information from death investigation reports filed by coroners, medical examiners, and law enforcement. These data illuminate why the victim ended his or her life, fatal injury patterns, and toxicological findings at death.

Results:Current suicide prevention efforts using these data fall into three categories: describing the problem of suicide and identifying opportunities for intervention; collaborating on statewide suicide prevention plans; and forming new partnerships for targeted prevention initiatives. Taken together, these three areas show early promise for state suicide prevention efforts.

Conclusions:In each of the states, NVDRS data analyses are being shared with injury prevention colleagues, suicide prevention planning groups and policymakers, and adapted to respond to unique state and local suicide problems. A powerful surveillance tool, the NVDRS is bringing new clarity and direction to these state-based efforts. The NVDRS can serve as a model for other countries looking to establish timely suicide surveillance systems and data driven prevention strategies.