bluuebunny/crossref_metadata_2025 huggingface dataset:
Sublethal Staphylococcal Enterotoxin B Challenge Model in Pigs To Evaluate Protection following Immunization with a Soybean-Derived Vaccine
Authors: Hudson Laura C. , Seabolt Brynn S. , Odle Jack , Bost Kenneth L. , Stahl Chad H. , Piller Kenneth J.
Published: November 2012

In an effort to develop a sustainable platform for manufacturing protein-based vaccine candidates, we expressed a triple mutant of staphylococcal enterotoxin B carrying the L45R, Y89A, and Y94A modifications in transgenic soybean seeds (soy-mSEB). Soy-mSEB possessed no detectable superantigen activity
in vitro
. We found that this soybean-derived, nontoxic mutant of SEB could be stably expressed, stored in seeds for extended periods at room temperature without degradation, and easily purified from contaminating soy proteins. Vaccination of pigs with purified soy-mSEB, or the identical triple mutant expressed in
Escherichia coli
(
E. coli
-mSEB), resulted in high antibody titers against the native toxin in immunized animals. In fact, titers were indistinguishable regardless of the immunogen used, demonstrating the equivalence of soy-mSEB and
E. coli
-mSEB vaccinations. Antisera from either immunized group were able to block native SEB superantigen activity in an
in vitro
neutralization assay. Similar results were obtained when immunized animals were challenged with a sublethal dose of native toxin. Significant reductions in toxin-induced serum cytokine levels were observed in soy-mSEB- and
E. coli
-mSEB-immunized pigs compared to control animals. The reductions in SEB-induced cytokine responses were similar regardless of the immunogen used for vaccination. Surprisingly, however, some clinical symptoms, such as prostration, lethargy, emesis, and/or diarrhea, were still observed in all immunized animals. These studies demonstrate the potential for soybean-derived proteins as a platform technology for sustainable vaccine manufacturing and the usefulness of a sublethal challenge model in pigs for evaluating the efficacy of potential SEB vaccine candidates.
Immunization of Mice with Formalin-Inactivated Spores from Avirulent Bacillus cereus Strains Provides Significant Protection from Challenge with Bacillus anthracis Ames
Authors: Vergis James M. , Cote Christopher K. , Bozue Joel , Alem Farhang , Ventura Christy L. , Welkos Susan L. , O'Brien Alison D.
Published: November 2012

Bacillus anthracis
spores are the infectious form of the organism for humans and animals. However, the approved human vaccine in the United States is derived from a vegetative culture filtrate of a toxigenic, nonencapsulated
B. anthracis
strain that primarily contains protective antigen (PA). Immunization of mice with purified spore proteins and formalin-inactivated spores (FIS) from a nonencapsulated, nontoxigenic
B. anthracis
strain confers protection against
B. anthracis
challenge when PA is also administered. To investigate the capacity of the spore particle to act as a vaccine without PA, we immunized mice subcutaneously with FIS from nontoxigenic, nonencapsulated
B. cereus
strain G9241 pBCXO1
−
/pBC210
−
(dcG9241), dcG9241 Δ
bclA
, or 569-UM20 or with exosporium isolated from dcG9241. FIS vaccination provided significant protection of mice from intraperitoneal or intranasal challenge with spores of the virulent
B. anthracis
Ames or Ames Δ
bclA
strain. Immunization with dcG9241 Δ
bclA
FIS, which are devoid of the immunodominant spore protein BclA, provided greater protection from challenge with either Ames strain than did immunization with FIS from BclA-producing strains. In addition, we used prechallenge immune antisera to probe a panel of recombinant
B. anthracis
Sterne spore proteins to identify novel immunogenic vaccine candidates. The antisera were variably reactive with BclA and with 10 other proteins, four of which were previously tested as vaccine candidates. Overall our data show that immunization with FIS from nontoxigenic, nonencapsulated
B. cereus
strains provides moderate to high levels of protection of mice from
B. anthracis
Ames challenge and that neither PA nor BclA is required for this protection.
Antibody Responses to Vaccination among South African HIV-Exposed and Unexposed Uninfected Infants during the First 2 Years of Life
Authors: Reikie Brian A. , Naidoo Shalena , Ruck Candice E. , Slogrove Amy L. , de Beer Corena , la Grange Heleen , Adams Rozanne C. M. , Ho Kevin , Smolen Kinga , Speert David P. , Cotton Mark F. , Preiser Wolfgang , Esser Monika , Kollmann Tobias R.
Published: November 2012

HIV-exposed but uninfected (HEU) infants born to HIV-infected mothers from areas in the world with a high burden of infectious disease suffer higher infectious morbidity and mortality than their HIV unexposed uninfected (HUU) peers. Vaccination provides protection from infection. The possibility exists that altered response to vaccination contributes to the higher rate of infection in HEU than in HUU infants. While short-term, cross-sectional studies support this notion, it is unclear whether or not HEU infants develop long-term protective immune responses following the WHO extended program on immunization (EPI). Vaccine-specific antibody responses were compared between HEU and HUU infants from 2 weeks until 2 years of age in a longitudinal South African cohort. Total IgG and antibodies specific for
Bordetella pertussis
,
Haemophilus influenzae
type b (Hib), tetanus toxoid, hepatitis B virus (HepB), and measles virus were measured at multiple time points throughout the first 2 years of life. Prevaccine antibodies (maternal antibodies passively acquired) specific for tetanus were lower in HEU than in HUU infants, while prevaccine antibodies to HepB were higher in HEU than in HUU infants. Both groups responded similarly to tetanus, Hib, and HepB vaccination. HEU demonstrated stronger pertussis vaccine responses, developing protective titers 1 year earlier than HUU patients, and maintained higher anti-tetanus titers at 24 months of age. Vaccine-induced antibodies to measles virus were similar in both groups at all time points. Our results suggest that the current EPI vaccination program as practiced in South Africa leads to the development of vaccine-specific antibody responses that are equivalent in HEU and HUU infants. However, our data also suggest that a large fraction of both HEU and HUU South African infants have antibody titers for several infectious threats that remain below the level of protection for much of their first 2 years of life.
PspK of Streptococcus pneumoniae Increases Adherence to Epithelial Cells and Enhances Nasopharyngeal Colonization
Authors: Keller L. E. , Jones C. V. , Thornton J. A. , Sanders M. E. , Swiatlo E. , Nahm M. H. , Park I. H. , McDaniel L. S.
Published: November 2012

Streptococcus pneumoniae
(the pneumococcus) colonizes the human nasopharynx and can cause invasive disease aided by the pneumococcal capsule. Group II nontypeable
S. pneumoniae
(NTSp) lacks a polysaccharide capsule, and a subgroup of NTSp carriage isolates has been found to have a novel gene, pneumococcal surface protein K (
pspK
), which replaces the capsule locus. A recent rise in the number of NTSp isolates colonizing the human nasopharynx has been observed, but the colonization factors of NTSp have not been well studied. PspK has been shown to play a role in mouse colonization. We therefore examined PspK-mediated immune evasion along with adherence to host cells and colonization. PspK bound human secretory immunoglobulin A (sIgA) but not the complement regulator factor H and did not decrease C3b deposition on the pneumococcal surface. PspK increased binding of pneumococci to epithelial cells and enhanced pneumococcal colonization independently of the genetic background. Understanding how NTSp colonizes and survives within the nasopharynx is important due to the increase in NTSp carriage. Our data suggest that PspK may aid in the persistence of NTSp within the nasopharynx but is not involved in invasion.
Effect of Angiotensin II Receptor Blocker on Experimental Periodontitis in a Mouse Model of Marfan Syndrome
Authors: Suda Naoto , Moriyama Keiji , Ganburged Ganjargal
Published: November 2012

Marfan syndrome is an autosomal dominant disease characterized by aneurysm and dilatation of the aortic root, tall stature, and ectopia lentis. These manifestations reflect excessive signaling of transforming growth factor beta (TGF-β). Moreover, cases are frequently associated with severe periodontitis, which is a chronic inflammation of the gingiva, periodontal ligament, and alveolar bone. Recently, angiotensin II receptor blockers (ARBs) were discovered to be an effective drug class that can prevent aortic aneurysm and dilation in Marfan syndrome by inhibiting TGF-β signaling. To investigate the effect of ARB on the progression of periodontitis, the application of a potent ARB, telmisartan, was examined in a mouse model of Marfan syndrome (MgΔ). Six-week-old male heterozygous MgΔ and wild-type mice were challenged with
Porphyromonas gingivalis
, which causes chronic periodontitis, with and without telmisartan application. After infection, alveolar bone resorption was measured by micro-computed tomography (μCT), and inflammatory cytokine levels were examined. Infection of
Porphyromonas gingivalis
induced alveolar bone resorption in both MgΔ and wild-type mice. The amount of resorption was significantly larger in the former than the latter. Immunoarray and enzyme-linked immunosorbent assay (ELISA) analyses demonstrated that interleukin-17 (IL-17) and tumor necrosis factor alpha (TNF-α) levels were significantly higher in infected MgΔ mice than infected wild-type mice. Telmisartan treatment significantly suppressed the alveolar bone resorption of infected MgΔ mice. Telmisartan also significantly decreased levels of TGF-β, IL-17, and TNF-α in infected MgΔ mice to levels seen in infected wild-type mice. This study suggests that ARB can prevent the severe periodontitis frequently seen in Marfan syndrome.
Transcriptional Modulation of Enterotoxigenic Escherichia coli Virulence Genes in Response to Epithelial Cell Interactions
Authors: Kansal Rita , Rasko David A. , Sahl Jason W. , Munson George P. , Roy Koushik , Luo Qingwei , Sheikh Alaullah , Kuhne Kurt J. , Fleckenstein James M.
Published: November 2012

EnterotoxigenicEscherichia coli(ETEC) strains are a leading cause of morbidity and mortality due to diarrheal illness in developing countries. There is currently no effective vaccine against these important pathogens. Because genes modulated by pathogen-host interactions potentially encode putative vaccine targets, we investigated changes in gene expression and surface morphology of ETEC upon interaction with intestinal epithelial cellsin vitro. Pan-genome microarrays, quantitative reverse transcriptase PCR (qRT-PCR), and transcriptional reporter fusions of selected promoters were used to study changes in ETEC transcriptomes. Flow cytometry, immunofluorescence microscopy, and scanning electron microscopy were used to investigate alterations in surface antigen expression and morphology following pathogen-host interactions. Following host cell contact, genes for motility, adhesion, toxin production, immunodominant peptides, and key regulatory molecules, including cyclic AMP (cAMP) receptor protein (CRP) and c-di-GMP, were substantially modulated. These changes were accompanied by visible changes in both ETEC architecture and the expression of surface antigens, including a novel highly conserved adhesin molecule, EaeH. The studies reported here suggest that pathogen-host interactions are finely orchestrated by ETEC and are characterized by coordinated responses involving the sequential deployment of multiple virulence molecules. Elucidation of the molecular details of these interactions could highlight novel strategies for development of vaccines for these important pathogens.
Francisella tularensis Live Vaccine Strain Folate Metabolism and Pseudouridine Synthase Gene Mutants Modulate Macrophage Caspase-1 Activation
Authors: Ulland Tyler K. , Janowski Ann M. , Buchan Blake W. , Faron Matthew , Cassel Suzanne L. , Jones Bradley D. , Sutterwala Fayyaz S.
Published: November 2012

Francisella tularensisis a Gram-negative bacterium and the causative agent of the disease tularemia. Escape ofF. tularensisfrom the phagosome into the cytosol of the macrophage triggers the activation of the AIM2 inflammasome through a mechanism that is not well understood. Activation of the AIM2 inflammasome results in autocatalytic cleavage of caspase-1, resulting in the processing and secretion of interleukin-1β (IL-1β) and IL-18, which play a crucial role in innate immune responses toF. tularensis. We have identified the5-formyltetrahydrofolate cycloligasegene (FTL_0724) as being important forF. tularensislive vaccine strain (LVS) virulence. Infection of micein vivowith aF. tularensisLVSFTL_0724mutant resulted in diminished mortality compared to infection of mice with wild-type LVS. TheFTL_0724mutant also induced increased inflammasome-dependent IL-1β and IL-18 secretion and cytotoxicity in macrophagesin vitro. In contrast, infection of macrophages with aF. tularensisLVSrluD pseudouridine synthase(FTL_0699) mutant resulted in diminished IL-1β and IL-18 secretion from macrophagesin vitrocompared to infection of macrophages with wild-type LVS. In addition, theFTL_0699mutant was not attenuatedin vivo. These findings further illustrate thatF. tularensisLVS possesses numerous genes that influence its ability to activate the inflammasome, which is a key host strategy to control infection with this pathogenin vivo.
Natural Competence Promotes Helicobacter pylori Chronic Infection
Authors: Dorer Marion S. , Cohen Ilana E. , Sessler Tate H. , Fero Jutta , Salama Nina R.
Published: November 2012

Animal models are important tools for studies of human disease, but developing these models is a particular challenge with regard to organisms with restricted host ranges, such as the human stomach pathogen
Helicobacter pylori
. In most cases,
H. pylori
infects the stomach for many decades before symptoms appear, distinguishing it from many bacterial pathogens that cause acute infection. To model chronic infection in the mouse, a human clinical isolate was selected for its ability to survive for 2 months in the mouse stomach, and the resulting strain, MSD132, colonized the mouse stomach for at least 28 weeks. During selection, the
cagY
component of the Cag type IV secretion system was mutated, disrupting a key interaction with host cells. Increases in both bacterial persistence and bacterial burden occurred prior to this mutation, and a mixed population of
cagY
+
and
cagY
mutant cells was isolated from a single mouse, suggesting that mutations accumulate during selection and that factors in addition to the Cag apparatus are important for murine adaptation. Diversity in both alleles and genes is common in
H. pylori
strains, and natural competence mediates a high rate of interstrain genetic exchange. Mutations of the Com apparatus, a membrane DNA transporter, and DprA, a cytosolic competence factor, resulted in reduced persistence, although initial colonization was normal. Thus, exchange of DNA between genetically heterogeneous
H. pylori
strains may improve chronic colonization. The strains and methods described here will be important tools for defining both the spectrum of mutations that promote murine adaptation and the genetic program of chronic infection.
Close Geographic Association of Human Neoehrlichiosis and Tick Populations Carrying “Candidatus Neoehrlichia mikurensis” in Eastern Switzerland
Authors: Maurer Florian P. , Keller Peter M. , Beuret Christian , Joha Cornelia , Achermann Yvonne , Gubler Jacques , Bircher Daniela , Karrer Urs , Fehr Jan , Zimmerli Lukas , Bloemberg Guido V.
Published: November 2012

Neoehrlichiosis caused by “
Candidatus
Neoehrlichia mikurensis” is an emerging zoonotic disease. In total, six patients have been described in Europe, with the first case detected in 2007. In addition, seven patients from China were described in a report published in October 2012. In 2009, we diagnosed the first human case of “
Ca
. Neoehrlichia mikurensis” infection in the Zurich area (Switzerland). Here, we report two additional human cases from the same region, which were identified by broad-range 16S rRNA gene PCR. Both patients were immunocompromised and presented with similar clinical syndromes, including fever, malaise, and weight loss. A diagnostic multiplex real-time PCR was developed for specific detection of “
Ca
. Neoehrlichia mikurensis” infections. The assay is based on the signature sequence of a 280-bp fragment of the “
Ca
. Neoehrlichia mikurensis” 16S rRNA gene and incorporates a “
Ca
. Neoehrlichia mikurensis” species, a “
Ca
. Neoehrlichia” genus, and an
Anaplasmataceae
family probe for simultaneous screening. The analytical sensitivity was determined to be below five copies of the “
Ca
. Neoehrlichia mikurensis” 16S rRNA gene. Our results show that the assay is suitable for the direct detection of “
Ca
. Neoehrlichia mikurensis” DNA in clinical samples from, for example, blood and bone marrow. In addition, it allows for monitoring treatment response during antibiotic therapy. Using the same assay, DNA extracts from 1,916 ticks collected in four forests in close proximity to the patients' residences (<3 km) were screened. At all sampling sites, the minimal prevalence of “
Ca
. Neoehrlichia mikurensis” was between 3.5 to 8% in pools of either nymphs, males, or females, showing a strong geographic association between the three patients and the assumed vector.
Environmental Contamination by Carbapenem-Resistant Enterobacteriaceae
Authors: Lerner A. , Adler A. , Abu-Hanna J. , Meitus I. , Navon-Venezia S. , Carmeli Y.
Published: November 2012

In the last decade, the global emergence of carbapenem resistance in
Enterobacteriaceae
has posed great concern to public health. Data concerning the role of environmental contamination in the dissemination of carbapenem-resistant
Enterobacteriaceae
(CRE) are currently lacking. Here, we aimed to examine the extent of CRE contamination in various sites in the immediate surroundings of CRE carriers and to assess the effects of sampling time and cleaning regimens on the recovery rate. We evaluated the performance of two sampling methods, CHROMAgar KPC contact plate and eSwab, for the detection of environmental CRE. eSwab was followed either by direct plating or by broth enrichment. First, 14 sites in the close vicinity of the carrier were evaluated for environmental contamination, and 5, which were found to be contaminated, were further studied. The environmental contamination decreased with distance from the patient; the bed area was the most contaminated site. Additionally, we found that the sampling time and the cleaning regimen were critical factors affecting the prevalence of environmental CRE contamination. We found that the CHROMAgar KPC contact plate method was a more effective technique for detecting environmental CRE than were eSwab-based methods. In summary, our study demonstrated that the vicinity of patients colonized with CRE is often contaminated by these organisms. Using selective contact plates to detect environmental contamination may guide cleaning efficacy and assist with outbreak investigation in an effort to limit the spread of CRE.
Identification by Full-Genome Analysis of a Bovine Rotavirus Transmitted Directly to and Causing Diarrhea in a Human Child
Authors: Doan Yen Hai , Nakagomi Toyoko , Aboudy Yair , Silberstein Ilana , Behar-Novat Esther , Nakagomi Osamu , Shulman Lester M.
Published: November 2012

The genome of rotaviruses consists of 11 segments of double-stranded RNA, and each genome segment has multiple genotypes. Thus, the genotype constellation of an isolate is often indicative of its host species. Albeit rarely, interspecies transmission occurs either by virions with nonreassorted or reassorted genomic segments. A rotavirus with the G6P[1] genotype, Ro8059, was isolated from the stool of a 1-year-old child during routine characterization of diarrheal specimens from a sentinel clinic in Israel in 1995. Since genotype G6P[1] is generally associated with bovine rotaviruses, and the child developed diarrhea within days of his first contact with calves at an urban farm, the aim of this study was to characterize the whole genomic constellation of Ro8059 and four G6P[1] bovine strains, BRV101, BRV105, BRV106, and CR231/39, by RNA-RNA hybridization and full genome sequencing to determine whether some or all of the segments were of bovine origin. The genome constellations of all four bovine G6P[1] strains were G6-P[1]-I2-R2-C2-M2-A3-N2-T6-E2-H3 for VP7-VP4-VP6-VP1-VP2-VP3-NSP1-NSP2-NSP3-NSP4-NSP5, respectively. Ro8059 shared the same genotype constellation with these bovine strains, with high nucleotide sequence identities (95.84 to 100%) for each of the 11 segments indicating that Ro8059 represented a direct interspecies whole-genome transmission of a nonreassorted rotavirus from a calf to a human infant. We conclude that this was the earliest example with a complete epidemiological link in which an entirely bovine rotavirus directly infected a human child and caused a symptomatic diarrheal illness. Thus, not all bovine rotaviruses are always naturally attenuated to the human host.
Multilocus Sequence Typing for Characterization of Staphylococcus pseudintermedius
Authors: Solyman S. M. , Black C. C. , Duim B. , Perreten V. , van Duijkeren E. , Wagenaar J. A. , Eberlein L. C. , Sadeghi L. N. , Videla R. , Bemis D. A. , Kania S. A.
Published: November 2012

Staphylococcus pseudintermedius
is an opportunistic pathogen in dogs. Four housekeeping genes with allelic polymorphisms were identified and used to develop an expanded multilocus sequence typing (MLST) scheme. The new seven-locus technique shows
S. pseudintermedius
to have greater genetic diversity than previous methods and discriminates more isolates based upon host origin.
The Herpes Simplex Virus 1 Us11 Protein Inhibits Autophagy through Its Interaction with the Protein Kinase PKR
Authors: Lussignol Marion , Queval Christophe , Bernet-Camard Marie-Françoise , Cotte-Laffitte Jacqueline , Beau Isabelle , Codogno Patrice , Esclatine Audrey
Published: November 2012

Autophagy is now known to be an essential component of host innate and adaptive immunity. Several herpesviruses have developed various strategies to evade this antiviral host defense. Herpes simplex virus 1 (HSV-1) blocks autophagy in fibroblasts and in neurons, and the ICP34.5 protein is important for the resistance of HSV-1 to autophagy because of its interaction with the autophagy machinery protein Beclin 1. ICP34.5 also counteracts the shutoff of protein synthesis mediated by the double-stranded RNA (dsRNA)-dependent protein kinase PKR by inhibiting phosphorylation of the eukaryotic translation initiation factor 2α (eIF2α) in the PKR/eIF2α signaling pathway. Us11 is a late gene product of HSV-1, which is also able to preclude the host shutoff by direct inhibition of PKR. In the present study, we unveil a previously uncharacterized function of Us11 by demonstrating its antiautophagic activity. We show that the expression of Us11 is able to block autophagy and autophagosome formation in both HeLa cells and fibroblasts. Furthermore, immediate-early expression of Us11 by an ICP34.5 deletion mutant virus is sufficient to render the cells resistant to PKR-induced and virus-induced autophagy. PKR expression and the PKR binding domain of Us11 are required for the antiautophagic activity of Us11. However, unlike ICP34.5, Us11 did not interact with Beclin 1. We suggest that the inhibition of autophagy observed in cells infected with HSV-1 results from the activity of not only ICP34.5 on Beclin 1 but also Us11 by direct interaction with PKR.
Rotavirus Viroplasm Proteins Interact with the Cellular SUMOylation System: Implications for Viroplasm-Like Structure Formation
Authors: Campagna Michela , Marcos-Villar Laura , Arnoldi Francesca , de la Cruz-Herrera Carlos F. , Gallego Pedro , González-Santamaría José , González Dolores , Lopitz-Otsoa Fernando , Rodriguez Manuel S. , Burrone Oscar R. , Rivas Carmen
Published: November 2012

Posttranslational modification by SUMO provides functional flexibility to target proteins. Viruses interact extensively with the cellular SUMO modification system in order to improve their replication, and there are numerous examples of viral proteins that are SUMOylated. However, thus far the relevance of SUMOylation for rotavirus replication remains unexplored. In this study, we report that SUMOylation positively regulates rotavirus replication and viral protein production. We show that SUMO can be covalently conjugated to the viroplasm proteins VP1, VP2, NSP2, VP6, and NSP5. In addition, VP1, VP2, and NSP2 can also interact with SUMO in a noncovalent manner. We observed that an NSP5 SUMOylation mutant protein retains most of its activities, such as its interaction with VP1 and NSP2, the formation of viroplasm-like structures after the coexpression with NSP2, and the ability to complement in
trans
the lack of NSP5 in infected cells. However, this mutant is characterized by a high degree of phosphorylation and is impaired in the formation of viroplasm-like structures when coexpressed with VP2. These results reveal for the first time a positive role for SUMO modification in rotavirus replication, describe the SUMOylation of several viroplasm resident rotavirus proteins, and demonstrate a requirement for NSP5 SUMOylation in the production of viroplasm-like structures.
Inefficient Vaginal Transmission of Tenofovir-Resistant HIV-1
Authors: Chateau Morgan , Swanson Michael D. , Garcia J. Victor
Published: November 2012

Transmission of drug-resistant HIV has been postulated to be a threat to current first-line antiretroviral therapy (ART) regimens and the efficacy of several antiretroviral-based preexposure prophylaxis (PrEP) strategies being tested. Here we evaluated the effect of the common tenofovir (TFV) resistance mutation K65R on vaginal HIV transmission. Our results demonstrate that despite no overt loss of overall replication competencein vivo, this mutation results in significantly reduced mucosal transmission. When transmitted, the mutant virus eventually reverted to the wild type in 2 of 3 animals examined.
Macropinocytosis-Like HIV-1 Internalization in Macrophages Is CCR5 Dependent and Leads to Efficient but Delayed Degradation in Endosomal Compartments
Authors: Gobeil Lise-Andrée , Lodge Robert , Tremblay Michel J.
Published: November 2012

HIV-1 endocytosis by a macropinocytosis-like mechanism has been shown to lead to productive infection in macrophages. However, little is known of this pathway. In this study, we examined HIV-1 endocytosis using biochemical approaches and imaging techniques in order to better understand the mechanisms that allow for productive infection of these cells via the endosomal pathway. We show here that this macropinocytosis-like mechanism is not the sole pathway involved in HIV-1 endocytosis in macrophages. However, this pathway specifically requires CCR5 engagement at the cell surface, which in turn suggests that the virus and its coreceptor are present in the endosomal environment simultaneously. Furthermore, although we observed efficient viral degradation following endocytosis, analyses of HIV-1 transport through the endolysosomal pathway revealed that viral degradation is delayed following endosomal internalization, possibly allowing the virus to complete its fusion.
The Viral Interferon Regulatory Factors of Kaposi's Sarcoma-Associated Herpesvirus Differ in Their Inhibition of Interferon Activation Mediated by Toll-Like Receptor 3
Authors: Jacobs Sarah R. , Gregory Sean M. , West John A. , Wollish Amy C. , Bennett Christopher L. , Blackbourn David J. , Heise Mark T. , Damania Blossom
Published: November 2012

Kaposi's sarcoma-associated herpesvirus (KSHV) infection is correlated with three human malignancies and can establish lifelong latent infection in multiple cell types within its human host. In order to establish and maintain infection, KSHV utilizes multiple mechanisms to evade the host immune response. One such mechanism is the expression of a family of genes with homology to cellular interferon (IFN) regulatory factors (IRFs), known as viral IRFs (vIRFs). We demonstrate here that KSHV vIRF1, -2, and -3 have a differential ability to block type I interferon signaling mediated by Toll-like receptor 3 (TLR3), a receptor we have previously shown to be activated upon KSHV infection. vIRF1, -2, and -3 inhibited TLR3-driven activation of IFN transcription reporters. However, only vIRF1 and vIRF2 inhibited increases in both IFN-β message and protein levels following TLR3 activation. The expression of vIRF1 and vIRF2 also allowed for increased replication of a virus known to activate TLR3 signaling. Furthermore, vIRF1 and vIRF2 may block TLR3-mediated signaling via different mechanisms. Altogether, this report indicates that vIRFs are able to block IFN mediated by TLRs but that each vIRF has a unique function and mechanism for blocking antiviral IFN responses.
Fixation of Emerging Interviral Recombinants in Cucumber Mosaic Virus Populations
Authors: Pita Justin S. , Roossinck Marilyn J.
Published: November 2012

Interstrain recombinants were observed in the progenies of the
Cucumber mosaic virus
(CMV) reassortant L
1
L
2
F
3
containing RNAs 1 and 2 from LS-CMV and RNA 3 from Fny-CMV. We characterized these recombinants, and we found that their fixation was controlled by the nature of the replicating RNAs 1 and 2. We demonstrate that the 2b gene partially affects this fixation process, but only in the context of homologous RNAs 1 and 2.
Structure of the St. Louis Encephalitis Virus Postfusion Envelope Trimer
Authors: Luca Vincent C. , Nelson Christopher A. , Fremont Daved H.
Published: November 2012

St. Louis encephalitis virus (SLEV) is a mosquito-borne flavivirus responsible for several human encephalitis outbreaks over the last 80 years. Mature flavivirus virions are coated with dimeric envelope (E) proteins that mediate attachment and fusion with host cells. E is a class II fusion protein, the hallmark of which is a distinct dimer-to-trimer rearrangement that occurs upon endosomal acidification and insertion of hydrophobic fusion peptides into the endosomal membrane. Herein, we report the crystal structure of SLEV E in the posfusion trimer conformation. The structure revealed specific features that differentiate SLEV E from trimers of related flavi- and alphaviruses. SLEV E fusion loops have distinct intermediate spacing such that they are positioned further apart than previously observed in flaviviruses but closer together than Semliki Forest virus, an alphavirus. Domains II and III (DII and DIII) of SLEV E also adopt different angles relative to DI, which suggests that the DI-DII joint may accommodate spheroidal motions. However, trimer interfaces are well conserved among flaviviruses, so it is likely the differences observed represent structural features specific to SLEV function. Analysis of surface potentials revealed a basic platform underneath flavivirus fusion loops that may interact with the anionic lipid head groups found in membranes. Taken together, these results highlight variations in E structure and assembly that may direct virus-specific interactions with host determinants to influence pathogenesis.
SNX17 Facilitates Infection with Diverse Papillomavirus Types
Authors: Bergant Martina , Banks Lawrence
Published: November 2012

Previous studies have shown that the human papillomavirus type 16 (HPV-16) L2 capsid protein plays an essential role in viral infection, in part through its interaction with sorting nexin 17 (SNX17). We now show that this interaction between L2 and SNX17 is conserved across multiple PV types. Furthermore, we demonstrate that SNX17 is essential for infection with all PV types analyzed, indicating an evolutionarily highly conserved virus entry mechanism.
A Hepatitis C Virus NS5A Phosphorylation Site That Regulates RNA Replication
Authors: LeMay K. L. , Treadaway J. , Angulo I. , Tellinghuisen T. L.
Published: November 2012

The hepatitis C virus NS5A protein is essential for RNA replication and virion assembly. NS5A is phosphorylated on multiple residues during infections, but these sites remain uncharacterized. Here we identify serine 222 of genotype 2a NS5A as a phosphorylation site that functions as a negative regulator of RNA replication. This site is a component of the hyperphosphorylated form of NS5A, which is in good agreement with previous observations that hyperphosphorylation negatively affects replication.
Comparative Proteomics Reveal Fundamental Structural and Functional Differences between the Two Progeny Phenotypes of a Baculovirus
Authors: Hou Dianhai , Zhang Leike , Deng Fei , Fang Wei , Wang Ranran , Liu Xijia , Guo Lin , Rayner Simon , Chen Xinwen , Wang Hualin , Hu Zhihong
Published: November 2012

The replication of lepidopteran baculoviruses is characterized by the production of two progeny phenotypes: the occlusion-derived virus (ODV), which establishes infection in midgut cells, and the budded virus (BV), which disseminates infection to different tissues within a susceptible host. To understand the structural, and hence functional, differences between BV and ODV, we employed multiple proteomic methods to reveal the protein compositions and posttranslational modifications of the two phenotypes of
Helicoverpa armigera
nucleopolyhedrovirus. In addition, Western blotting and quantitative mass spectrometry were used to identify the localization of proteins in the envelope or nucleocapsid fractions. Comparative protein portfolios of BV and ODV showing the distribution of 54 proteins, encompassing the 21 proteins shared by BV and ODV, the 12 BV-specific proteins, and the 21 ODV-specific proteins, were obtained. Among the 11 ODV-specific envelope proteins, 8 either are essential for or contribute to oral infection. Twenty-three phosphorylated and 6
N
-glycosylated viral proteins were also identified. While the proteins that are shared by the two phenotypes appear to be important for nucleocapsid assembly and trafficking, the structural and functional differences between the two phenotypes are evidently characterized by the envelope proteins and posttranslational modifications. This comparative proteomics study provides new insight into how BV and ODV are formed and why they function differently.
Complete Genome Sequence of Duck Tembusu Virus, Isolated from Muscovy Ducks in Southern China
Authors: Zhu Wanjun , Chen Jidang , Wei Chunya , Wang Heng , Huang Zhen , Zhang Minze , Tang Fengfeng , Xie Jiexiong , Liang Huanbin , Zhang Guihong , Su Shuo
Published: November 2012

We report here the complete genomic sequence of the duck Tembusu virus (DTMUV) WJ-1 strain, isolated from Muscovy ducks. This is the first complete genome sequence of DTMUV reported in southern China. Compared with the other strains (TA, GH-2, YY5, and ZJ-407) that were previously found in eastern China, WJ-1 bears a few differences in the nucleotide and amino acid sequences. We found that there are 47 mutations of amino acids encoded by the whole open reading frame (ORF) among these five strains. The whole-genome sequence of DTMUV will help in understanding the epidemiology and molecular characteristics of duck Tembusu virus in southern China.
Complete Genome Sequence of a Coxsackievirus B3 Isolated from a Sichuan Snub-Nosed Monkey
Authors: He Wenqi , Lu Huijun , Zhao Kui , Song Deguang , Gai Xianying , Gao Feng
Published: November 2012

Coxsackievirus B3 (CVB3) is an enterovirus in the family
Picornaviridae
that is significant to human health, being associated with myocarditis, aseptic meningitis, and pancreatitis, among other conditions. In addition to humans, Sichuan snub-nosed monkeys can be infected and killed by CVB3. Here, we report the first complete genome sequence of a novel coxsackievirus B3 strain, SSM-CVB3, which was isolated from a deceased Sichuan snub-nosed monkey with severe myocarditis. Our findings may aid in understanding the evolutionary characteristics and molecular pathogenesis of this virus.
Complete Genome Sequence of the Novel Escherichia coli Phage phAPEC8
Authors: Tsonos J. , Adriaenssens E. M. , Klumpp J. , Hernalsteens J.-P. , Lavigne R. , De Greve H.
Published: November 2012

Bacteriophage phAPEC8 is an
Escherichia coli
-infecting myovirus, isolated on an avian pathogenic
Escherichia coli
(APEC) strain. APEC strains cause colibacillosis in poultry, resulting in high mortality levels and important economic losses. Genomic analysis of the 147,737-bp double-stranded DNA phAPEC8 genome revealed that 53% of the 269 encoded proteins are unique to this phage. Its closest relatives include the
Salmonella
phage PVP-SE1 and the coliphage rv5, with 19% and 18% similar proteins, respectively. As such, phAPEC8 represents a novel, phylogenetically distinct clade within the
Myoviridae
, with molecular properties suitable for phage therapy applications.
Complete Genome Sequence of a Newly Identified Porcine Astrovirus Genotype 3 Strain US-MO123
Authors: Xiao Chao-Ting , Halbur Patrick G. , Opriessnig Tanja
Published: November 2012

Astrovirus (AstV) infections are among the most common causes of gastroenteritis and are also associated with extraintestinal manifestations in humans and many animals. Herein, for the first time, the complete genome sequence of newly identified porcine astrovirus genotype 3 (PAstV3) strain US-MO123 was determined. Sequence comparison and phylogenetic analysis showed that PAstV3 has the closest relationship with mink AstV and the human AstV strains VA1, VA2, and SG, indicating the same ancestral origin and zoonotic potential of the virus.
Pseudoplusia includens Densovirus Genome Organization and Expression Strategy
Authors: Huynh Oanh T. H. , Pham Hanh T. , Yu Qian , Tijssen Peter
Published: November 2012

The genome of a densovirus of a major phytophagous pest,
Pseudoplusia includens
, was analyzed. It contained 5,990 nucleotides (nt) and included inverted terminal repeats of 540 nt with terminal Y-shaped hairpins of 120 nt. Its DNA sequence and ambisense organization with 4 typical open reading frames demonstrated that it belonged to the genus
Densovirus
in the subfamily
Densovirinae
of the family
Parvoviridae
.
Complete Genomic Sequence of a Reovirus Isolate from Pekin Ducklings in China
Authors: Ma Guoming , Wang Dan , Shi Jiajian , Jiang Tiantian , Yuan Yuan , Zhang Dabing
Published: November 2012

The complete genomic sequence of a Pekin duck origin reovirus (DRV) from China was determined. The genome comprises 23,419 bp, with segments ranging from 1,191 bp (S4) to 3,959 bp (L1). Pairwise comparisons and phylogenetic analysis indicate that the Pekin duck origin reovirus is more closely related to the new type of Muscovy duck origin reovirus (N-MDRV) identified recently than to the chicken origin avian orthoreovirus (ARV) and the originally described Muscovy duck origin reovirus (ARV-Md).
Complete Genome of a Novel Endornavirus Assembled from Next-Generation Sequence Data
Authors: Espach Yolandi , Maree Hans J. , Burger Johan T.
Published: November 2012

Endornaviruses have large double-stranded RNA (dsRNA) genomes that carry a single open reading frame (ORF). Here we report the complete genome of a novel endornavirus, assembled from next-generation sequence data generated from
Vitis vinifera
-extracted dsRNA. Two different fungal hosts have been identified for this virus, suggesting that horizontal transmission of the virus is possible.
Evaluation of Acupuncture for Cancer Symptoms in a Cancer Institute in Brazil
Authors: D'Alessandro Eduardo , de Brito Christina , Cecatto Rebeca , Saul Maira , Atta Jose Antonio , Lin Chin An
Published: November 2012

Introduction

Acupuncture has been progressively included in the practice of mainstream medicine in recent decades. The State of Sao Paulo Cancer Institute is a public hospital established in 2008 and its acupuncture service follows the experience and model of several oncology centres in the USA, aiming to optimise the treatment of symptoms such as postoperative pain, oncological pain, neuropathic pain, nausea, vomiting, xerostomia and fatigue induced by chemotherapy. This paper describes the population given acupuncture treatment and the effects of the intervention on symptom management.

Methods

One hundred and eighty-three patients from our service were enrolled in the study. Baseline and final symptom intensity was recorded using a visual analogue score (VAS) ranging from 0 to 10 cm, with a higher score meaning higher symptom intensity.

Results

Fifty-four (29.50%) were receiving active treatment with chemotherapy and/or radiotherapy, 29 (15.85%) were receiving hormone therapy and 100 (54.65%) were considered to be in remission. The main symptoms were: oncological pain in 44 (24.04%), chemotoxicity in 34 (18.6%), lumbar pain in 53 (28.96%) and chronic postoperative pain in 54 (28.4%). The mean (SD) initial symptom score was 7.04 (1.8), which was reduced to 2.56 (2.75) after treatment (p<0.001), an improvement of 63.6% in control of the symptoms. Further analysis of the data showed that the effect was similar in different indications for acupuncture treatment.

Conclusions

Use of acupuncture may have improved symptom control in patients enrolled in this study.
Chronic asymptomatic dislocation of the bearing in an Oxford unicompartmental knee replacement
Authors: Woodacre Timothy , Marshall Morwena , Awad Ashraf , Isaac David
Published: November 2012

Unicompartmental knee replacement is a common procedure for the treatment of osteoarthritis confined to one compartment of the knee. Dislocation of the mobile bearing is a recognised complication. We present a patient with a 6-year asymptomatic bearing dislocation, missed on original presentation.
Digital gangrene as the initial presentation of systemic lupus erythematosus
Authors: Adelowo Olufemi , Olaosebikan Hakeem , Ajani Wahab , Omosebi D T
Published: November 2012

Systemic lupus erythematosus (SLE) has rarely been reported among African blacks. However, recent reports may indicate otherwise. Digital gangrene in lupus patients has been reported in different populations and has mostly been seen in patients with long-standing lupus. Digital gangrene as the initial presentation of SLE has rarely been reported, even among African blacks. Two cases of digital gangrene as the initial presentation in Nigerian lupus patients are hereby presented. The medical and surgical managements, as well as relevant literature are hereby discussed.
A staged management of prolonged chylothorax in a patient with yellow nail syndrome
Authors: Tidder James , Pang Chun Lap
Published: November 2012

Spontaneous chylothorax remains an unusual condition with diverse aetiologies and non-unified management strategies. Owing to the rarity of the condition, the evidence of management remains from case reports and limited retrospective studies. This case represents a difficult-to-manage chylothorax secondary to yellow nail syndrome with initial failure of both surgical and conservative treatment methods. Pleurovenous shunting represents a surgical management approach allowing the patient to live with and control an ongoing chyle leak.
Pregnancy in Shwachman-Diamond syndrome: a novel genetic mutation with minimal consequence
Authors: Horne Gillian A , Chevassut Timothy
Published: November 2012

Shwachman-Diamond syndrome (SDS) is a rare inherited bone marrow failure syndrome, characterised by neutropenia, exocrine pancreatic dysfunction and often skeletal abnormalities. To date, and to our knowledge, we report a novel genetic mutation in SDS that, we believe, is associated with minimal consequence,and report the fertility and pregnancy in this individual.
Fetal warfarin syndrome
Authors: Starling Luke D , Sinha Ashutosh , Boyd Duncan , Furck Anke
Published: November 2012

A case of a baby born preterm with an antenatal diagnosis of aortic coarctation for which prostin was electively started at birth. The baby was found to be profoundly anaemic with no clear obstetric cause. Features consistent with antenatal intracerebral haemorrhage were noted on cranial ultrasonography in the context of severe coagulopathy, prompting investigations which confirmed fetal–maternal haemorrhage. It transpired that, following aortic and mitral valve replacements, the mother was anticoagulated with warfarin at conception, having misunderstood her cardiologist's advice that: ‘you cannot get pregnant whilst on warfarin’. Following conversion to low molecular weight heparin, she suffered a stroke, thus warfarin was restarted, with an international normalised ratio of 3–4.7 during pregnancy. Following transfer to the paediatric intensive care unit, fetal warfarin syndrome was diagnosed. The coagulopathy and anaemia were corrected and aortic coarctation was excluded. The baby returned to the neonatal intensive care unit for ongoing care and was discharged home in good condition around his due date. At the present time, there is no clinically overt neurological deficit.
Medical services at the first Winter Youth Olympic Games 2012 in Innsbruck/Austria
Authors: Blank Cornelia , Schamasch Patrick , Engebretsen Lars , Haslinger Simon , Ruedl Gerhard , Fink Christian , Schobersberger Wolfgang
Published: November 2012

Background

The Youth Olympic Games (YOG) are a new format designed by the International Olympic Committee. So far no reference data are available regarding the organisation or implementation of the medical services that were needed for the Winter Youth Olympic Games that took place for the first time in Innsbruck 9–24 January 2012.

Objectives

(1) To provide insight into what is needed to prepare for such a complex high level sporting event from a medical perspective, (2) to provide data on medical services for future organising committees and (3) to provide information on different National Olympic Committee (NOC) delegation structures and the consequences of registering a National Olympic Committee Team Physician.

Methods

A medical information system in the form of a patient data-management system was developed with all involved parties to standardise data collection. All medical encounters occurring at any IYOGOC medical service centre (including physiotherapy and psychology facilities) were tracked and collected in daily reports. Data evaluation was prepared based on different interest groups (Athletes, National Olympic Committees, Workforce, International Olympic Committee and Media) and analysed.

Results

327 medical encounters (42.8% athletes; out of these, 57.9% were accounted to athletes with own NOC team physician) were seen during the YOG 2012. The total number of hospital transports was 27.3%, of which 8.9% were hospitalised with an average length of 1.9 nights. Physiotherapy usage was low with only 19 medical encounters resulting in a referral to physiotherapy accounting for 67 treatments during the entire YOG. Psychological care service was not used at all. The main reason for illnesses was disorders of the respiratory system (28.8%), injuries mostly affected upper extremities (49.6%) and were mostly diagnosed with lacerations and contusions (26.2%). Injury (70.7%) and illness (29.3%) incidences in athletes were slightly lower than previous studies showed. 40.0% of NOC delegations registered their own team physicians, which led to a significant difference in usage frequency of medical service (–3%, p=0.012).

Conclusions

Medical service coverage at the first Winter Youth Olympic Games seemed to be appropriate. No disaster or epidemic disease challenged the medical service plan. Future organising committees could use the provided data as a reference for their planning efforts.