bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differential Accessibility of a Rotavirus VP6 Epitope in Trimers Comprising Type I, II, or III Channels as Revealed by Binding of a Human Rotavirus VP6-Specific Antibody
Authors: Aiyegbo Mohammed S. , Eli Ilyas M. , Spiller Benjamin W. , Williams Dewight R. , Kim Robert , Lee David E. , Liu Tong , Li Sheng , Stewart Phoebe L. , Crowe James E. 
Published: October 2013

ABSTRACT

Previous human antibody studies have shown that the human V
H
1–46 antibody variable gene segment encodes much of the naturally occurring human B cell response to rotavirus and is directed to virus protein 6 (VP6). It is currently unknown why some of the V
H
1-46-encoded human VP6 monoclonal antibodies inhibit viral transcription while others do not. In part, there are affinity differences between antibodies that likely affect inhibitory activity, but we also hypothesize that there are differing modes of binding to VP6 that affect the ability to block the transcriptional pore on double-layered particles. Here, we used a hybrid method approach for antibody epitope mapping, including single-particle cryo-electron microscopy (cryo-EM) and enhanced amide hydrogen-deuterium exchange mass spectrometry (DXMS) to determine the location and mode of binding of a V
H
1-46-encoded antibody, RV6-25. The structure of the RV6-25 antibody–double-layered particle (DLP) complex indicated a very complex binding pattern that revealed subtle differences in accessibility of the VP6 epitope depending on its position in the type I, II, or III channels. These subtle variations in the presentation or accessibility of the RV VP6 capsid layer led to position-specific differences in occupancy for binding of the RV6-25 antibody. The studies also showed that the location of binding of the noninhibitory antibody RV6-25 on the apical surface of RV VP6 head domain does not obstruct the transcription pore upon antibody binding, in contrast to binding of an inhibitory antibody, RV6-26, deeper in the transcriptional pore.
Human Plasmacytoid Dendritic Cells Sense Lymphocytic Choriomeningitis Virus-Infected Cells
In Vitro
Authors: Wieland S. F. , Takahashi K. , Boyd B. , Whitten-Bauer C. , Ngo N. , de la Torre J.-C. , Chisari F. V. 
Published: October 2013

ABSTRACT
We previously reported that exosomal transfer of hepatitis C virus (HCV) positive-strand RNA from human Huh-7 hepatoma cells to human plasmacytoid dendritic cells (pDCs) triggers pDC alpha/beta interferon (IFN-α/β) production in a Toll-like receptor 7 (TLR7)-dependent, virus-independent manner. Here we show that human pDCs are also activated by a TLR7-dependent, virus-independent, exosomal RNA transfer mechanism by human and mouse hepatoma and nonhepatoma cells that replicate the negative-strand lymphocytic choriomeningitis virus (LCMV).
NF-κB Activation Coordinated by IKKβ and IKKε Enables Latent Infection of Kaposi's Sarcoma-Associated Herpesvirus
Authors: He Zhiheng , Zhao Jun , Zhang Junjie , Jung Jae U. , Feng Pinghui 
Published: October 2013

ABSTRACT

All herpesviruses share a remarkable propensity to establish latent infection. Human Kaposi's sarcoma-associated herpesvirus (KSHV) effectively enters latency after
de novo
infection, suggesting that KSHV has evolved with strategies to facilitate latent infection. NF-κB activation is imperative for latent infection of gammaherpesviruses. However, how NF-κB is activated during
de novo
herpesvirus infection is not fully understood. Here, we report that KSHV infection activates the inhibitor of κB kinase β (IKKβ) and the IKK-related kinase epsilon (IKKε) to enable host NF-κB activation and KSHV latent infection. Specifically, KSHV infection activated IKKβ and IKKε that were crucial for latent infection. Knockdown of IKKβ and IKKε caused aberrant lytic gene expression and impaired KSHV latent infection. Biochemical and genetic experiments identified RelA as a key player downstream of IKKβ and IKKε. Remarkably, IKKβ and IKKε were essential for phosphorylation of S
536
and S
468
of RelA, respectively. Phosphorylation of RelA S
536
was required for phosphorylation of S
468
, which activated NF-κB and promoted KSHV latent infection. Expression of the phosphorylation-resistant RelA S
536
A increased KSHV lytic gene expression and impaired latent infection. Our findings uncover a scheme wherein NF-κB activation is coordinated by IKKβ and IKKε, which sequentially phosphorylate RelA in a site-specific manner to enable latent infection after KSHV
de novo
infection.
Minimal Features of Efficient Incorporation of the Hemagglutinin-Neuraminidase Protein into Sendai Virus Particles
Authors: Essaidi-Laziosi Manel , Shevtsova Anastasia S. , Roux Laurent 
Published: October 2013

ABSTRACT

Two transmembrane glycoproteins form spikes on the surface of Sendai virus, a member of the
Respirovirus
genus of the
Paramyxovirinae
subfamily of the
Paramyxoviridae
family: the hemagglutinin-neuraminidase (HN) and the fusion (F) proteins. HN, in contrast to F, is dispensable for viral particle production, as normal amounts of particles can be produced with highly reduced levels of HN. This HN reduction can result from mutation of an SYWST motif in its cytoplasmic tail to AFYKD. HN
AFYKD
accumulates at the infected cell surface but does not get incorporated into particles. In this work, we derived experimental tools to rescue HN
AFYKD
incorporation. We found that coexpression of a truncated HN harboring the wild-type cytoplasmic tail, the transmembrane domain, and at most 80 amino acids of the ectodomain was sufficient to complement defective HN
AFYKD
incorporation into particles. This relied on formation of disulfide-bound heterodimers carried out by the two cysteines present in the HN 80-amino-acid (aa) ectodomain. Finally, the replacement of the measles virus H cytoplasmic and transmembrane domains with the corresponding HN domains promoted measles virus H incorporation in Sendai virus particles.
Mobilization of Circulating Endothelial Progenitor Cells Correlates with the Clinical Course of Hantavirus Disease
Authors: Krautkrämer Ellen , Grouls Stephan , Hettwer David , Rafat Neysan , Tönshoff Burkhard , Zeier Martin 
Published: October 2013

ABSTRACT

Infections with hemorrhagic fever viruses are characterized by increased permeability leading to capillary leakage. Hantavirus infection is associated with endothelial dysfunction, and the clinical course is related to the degree of vascular injury. Circulating endothelial progenitor cells (cEPCs) play a pivotal role in the repair of the damaged endothelium. Therefore, we analyzed the number of cEPCs and their mobilizing growth factors in patients suffering from hantavirus disease induced by infection with
Puumala virus
. The numbers of EPCs of 36 hantavirus-infected patients and age- and gender-matched healthy controls were analyzed by flow cytometry. Concentrations of cEPC-mobilizing growth factors in plasma were determined by enzyme-linked immunosorbent assay. Laboratory parameters were correlated with the number of cEPCs. In patients infected with hantavirus, the number of cEPCs was significantly higher than that in healthy controls. Levels of mobilizing cytokines were upregulated in patients, and the mobilization of cEPCs is paralleled with the normalization of clinical parameters. Moreover, higher levels of cEPCs correlated with higher serum albumin levels and platelet concentrations. Our data indicate that cEPCs may play a role in the repair of hantavirus-induced endothelial damage, thereby influencing the clinical course and the severity of symptoms.
Nonmuscle Myosin Heavy Chain IIA Is a Critical Factor Contributing to the Efficiency of Early Infection of Severe Fever with Thrombocytopenia Syndrome Virus
Authors: Sun Yinyan , Qi Yonghe , Liu Chenxuan , Gao Wenqing , Chen Pan , Fu Liran , Peng Bo , Wang Haimin , Jing Zhiyi , Zhong Guocai , Li Wenhui 
Published: October 2013

ABSTRACT

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a novel phlebovirus in the
Bunyaviridae
family. Most patients infected by SFTSV present with fever and thrombocytopenia, and up to 30% die due to multiple-organ dysfunction. The mechanisms by which SFTSV enters multiple cell types are unknown. SFTSV contains two species of envelope glycoproteins, Gn (44.2 kDa) and Gc (56 kDa), both of which are encoded by the M segment and are cleaved from a precursor polypeptide (about 116 kDa) in the endoplasmic reticulum (ER). Gn fused with an immunoglobulin Fc tag at its C terminus (Gn-Fc) bound to multiple cells susceptible to the infection of SFTSV and blocked viral infection of human umbilical vein endothelial cells (HUVECs). Immunoprecipitation assays following mass spectrometry analysis showed that Gn binds to nonmuscle myosin heavy chain IIA (NMMHC-IIA), a cellular protein with surface expression in multiple cell types. Small interfering RNA (siRNA) knockdown of NMMHC-IIA, but not the closely related NMMHC-IIB or NMMHC-IIC, reduced SFTSV infection, and NMMHC-IIA specific antibody blocked infection by SFTSV but not other control viruses. Overexpression of NMMHC-IIA in HeLa cells, which show limited susceptivity to SFTSV, markedly enhanced SFTSV infection of the cells. These results show that NMMHC-IIA is critical for the cellular entry of SFTSV. As NMMHC-IIA is essential for the normal functions of platelets and human vascular endothelial cells, it is conceivable that NMMHC-IIA directly contributes to the pathogenesis of SFTSV and may be a useful target for antiviral interventions against the viral infection.
A First-in-Class NAE Inhibitor, MLN4924, Blocks Lentiviral Infection in Myeloid Cells by Disrupting Neddylation-Dependent Vpx-Mediated SAMHD1 Degradation
Authors: Wei Wei , Guo Haoran , Liu Xianjun , Zhang Hong , Qian Lei , Luo Kun , Markham Richard B. , Yu Xiao-Fang 
Published: October 2013

ABSTRACT
MLN4924 is a first-in-class cancer drug that inhibits the Nedd8-activating enzyme (NAE). Herein, we report that MLN4924 inhibits Vpx/Vpr-induced SAMHD1 degradation by inhibiting the neddylation of E3 ubiquitin ligase and blocks macaque simian immunodeficiency virus (SIVmac) replication in myeloid cells. SAMHD1 is required for MLN4924-mediated SIVmac inhibition. Our findings indicate the potential efficacy of inhibiting neddylation as an antiretroviral strategy and identify the readily available anticancer drug MLN4924 as a candidate agent for that purpose.
Isolation and Characterization of the Herpes Simplex Virus 1 Terminase Complex
Authors: Heming Jason D. , Huffman Jamie B. , Jones Lisa M. , Homa Fred L. 
Published: October 2013

ABSTRACTDuring herpes simplex virus 1 (HSV-1) infection, empty procapsids are assembled and subsequently filled with the viral genome by means of a protein complex called the terminase, which is comprised of the HSV-1 UL15, UL28, and UL33 proteins. Biochemical studies of the terminase proteins have been hampered by the inability to purify the intact terminase complex. In this study, terminase complexes were isolated by tandem-affinity purification (TAP) using recombinant viruses expressing either a full-length NTAP-UL28 fusion protein (vFH476) or a C-terminally truncated NTAP-UL28 fusion protein (vFH499). TAP of the UL28 protein from vFH476-infected cells, followed by silver staining, Western blotting, and mass spectrometry, identified the UL15, UL28, and UL33 subunits, while TAP of vFH499-infected cells confirmed previous findings that the C terminus of UL28 is required for UL28 interaction with UL33 and UL15. Analysis of the oligomeric state of the purified complexes by sucrose density gradient ultracentrifugation revealed that the three proteins formed a complex with a molecular mass that is consistent with the formation of a UL15-UL28-UL33 heterotrimer. In order to assess the importance of conserved regions of the UL15 and UL28 proteins, recombinant NTAP-UL28 viruses with mutations of the putative UL28 metal-binding domain or within the UL15 nuclease domain were generated. TAP of UL28 complexes from cells infected with each domain mutant demonstrated that the conserved cysteine residues of the putative UL28 metal-binding domain and conserved amino acids within the UL15 nuclease domain are required for the cleavage and packaging functions of the viral terminase, but not for terminase complex assembly.
Genomewide Mapping and Screening of Kaposi's Sarcoma-Associated Herpesvirus (KSHV) 3′ Untranslated Regions Identify Bicistronic and Polycistronic Viral Transcripts as Frequent Targets of KSHV MicroRNAs
Authors: Bai Zhiqiang , Huang Yufei , Li Wan , Zhu Ying , Jung Jae U. , Lu Chun , Gao Shou-Jiang 
Published: October 2013

ABSTRACT
Kaposi's sarcoma-associated herpesvirus (KSHV) encodes over 90 genes and 25 microRNAs (miRNAs). The KSHV life cycle is tightly regulated to ensure persistent infection in the host. In particular, miRNAs, which primarily exert their effects by binding to the 3′ untranslated regions (3′UTRs) of target transcripts, have recently emerged as key regulators of KSHV life cycle. Although studies with RNA cross-linking immunoprecipitation approach have identified numerous targets of KSHV miRNAs, few of these targets are of viral origin because most KSHV 3′UTRs have not been characterized. Thus, the extents of viral genes targeted by KSHV miRNAs remain elusive. Here, we report the mapping of the 3′UTRs of 74 KSHV genes and the effects of KSHV miRNAs on the control of these 3′UTR-mediated gene expressions. This analysis reveals new bicistronic and polycistronic transcripts of KSHV genes. Due to the 5′-distal open reading frames (ORFs), KSHV bicistronic or polycistronic transcripts have significantly longer 3′UTRs than do KSHV monocistronic transcripts. Furthermore, screening of the 3′UTR reporters has identified 28 potential new targets of KSHV miRNAs, of which 11 (39%) are bicistronic or polycistronic transcripts. Reporter mutagenesis demonstrates that miR-K3 specifically targets ORF31-33 transcripts at the lytic locus via two binding sites in the ORF33 coding region, whereas miR-K10a-3p and miR-K10b-3p and their variants target ORF71-73 transcripts at the latent locus through distinct binding sites in both 5′-distal ORFs and intergenic regions. Our results indicate that KSHV miRNAs frequently target the 5′-distal coding regions of bicistronic or polycistronic transcripts and highlight the unique features of KSHV miRNAs in regulating gene expression and life cycle.
Echovirus 7 Entry into Polarized Caco-2 Intestinal Epithelial Cells Involves Core Components of the Autophagy Machinery
Authors: Kim Chonsaeng , Bergelson Jeffrey M. 
Published: October 2013

ABSTRACT
Echovirus 7 enters polarized Caco-2 intestinal epithelial cells by a clathrin-mediated endocytic process and then moves through the endosomal system before releasing its genome into the cytoplasm. We examined the possible role in virus entry of core components of the autophagy machinery. We found that depletion of Beclin-1, Atg12, Atg14, Atg16, or LC3 with specific small interfering RNAs inhibited echovirus 7 infection upstream of uncoating but had little or no effect on virus attachment to the cell surface. These data indicate that multiple autophagy-related proteins are important for one or more events that occur after the virus has bound its receptor on the cell surface but before RNA is released from the virus capsid. Although we have not determined the mechanism by which each protein contributes to virus entry, we found that stable depletion of Atg16L1 interfered with virus internalization from the cell surface rather than with intracellular trafficking. Autophagy gene products may thus participate in the endocytic process that moves virus into polarized Caco-2 cells.
An Avian Retrovirus Uses Canonical Expression and Processing Mechanisms To Generate Viral MicroRNA
Authors: Yao Yongxiu , Smith Lorraine P. , Nair Venugopal , Watson Mick 
Published: October 2013

ABSTRACTTo date, the vast majority of known virus-encoded microRNAs (miRNAs) are derived from polymerase II transcripts encoded by DNA viruses. A recent demonstration that the bovine leukemia virus, a retrovirus, uses RNA polymerase III to directly transcribe the pre-miRNA hairpins to generate viral miRNAs further supports the common notion that the canonical pathway of miRNA biogenesis does not exist commonly among RNA viruses. Here, we show that an exogenous virus-specific region, termed the E element or XSR, of avian leukosis virus subgroup J (ALV-J), a member of avian retrovirus, encodes a novel miRNA, designated E (XSR) miRNA, using the canonical miRNA biogenesis pathway. Detection of novel microRNA species derived from the E (XSR) element, a 148-nucleotide noncoding RNA with hairpin structure, showed that the E (XSR) element has the potential to function as a microRNA primary transcript, demonstrating a hitherto unknown function with possible roles in myeloid leukosis associated with ALV-J.
SPORT AND EXERCISE MEDICINE (SEM) IN PRIMARY CARE: A NOVEL DUAL ARMED APPROACH TO IMPROVING PHYSICAL ACTIVITY (PA) AND MUSCULOSKELETAL (MSK) SERVICES IN A HERTFORDSHIRE GENERAL PRACTICE
Authors: Chew S , Noake J 
Published: October 2013

BackgroundWith the advent of SEM as a recognised medical specialty in the United Kingdom, a number of service models have become apparent.1Despite government initiatives to increase for example PA promotion in the primary care setting, uptake, promotion and utilisation of these services has been poor or lacked longevity.2Schopwick General Practice has a patient population of over 12,000 and is a standard primary care service provider.AimTo develop a holistic reproducible dual armed model of SEM services in the primary care setting that is economically viable and sustainable.MethodsTwo SEM registrars in conjunction with the practice partners and manager, proposed, implemented and piloted a SEM service providing both musculoskeletal and physical activity interventions. Care provision included “one-stop” MSK clinics with capabilities for “see and treat” injection therapy, rehabilitation prescription & allied priority physiotherapy appointments. Novel PA prescription templates to assist practitioners were embedded in consultation software, & electronic ‘flag’ systems developed to highlight priority intervention patients. GPPAQ was made a mandatory component of the “new patient” registration procedure. Over a 12 month period audit of parameters including orthopaedic referral rates, frequency of physical activity interventions according to electronic note records, and patient service questionnaires were used to assess outcome measures of the service.ResultsDocumented PA advice increased fourfold over a 9 month period. Use of GPPAQ Pre and Post January 2011 increased significantly. All use Vision Exercise Related Read Codes Pre and Post January 2011 increased fivefold. Orthopaedic referral rates were reduced by approximately 50%, MRI scan requests were reduced by over half and in-house physiotherapy waiting times were reduced from 8 to 6 weeks despite the introduction of service specific appointments.ConclusionsNovel service models marrying both musculoskeletal and physical activity promotion services can make sound economical and patient satisfaction positive outcomes within primary care settings. We present a reproducible model that has recently been commissioned as a service by the local clinical commissioning group.
INFLUENCE OF THE STATIC IMAGES THAT EVOKE ILLUSORY MOTION PERCEPTION ON THE EYE MOVEMENTS AND THE BODY SWAY
Authors: Inoue Y 
Published: October 2013

Previous studies suggested that eye movements might be one of the factors that static images evoke illusory motion perception. The influence that visual information gives to posture control is also known to be big. However, the relationship between the posture control and the illusory motion perceptions has not been clear well. Therefore we examined how static illusory motion images affected the body sway and the eye movements. Seven participants (20±2 years old) took part in the experiment. Wavy image (W condition), two types of parallel images (P1, P2 condition), and expanded image (E condition) were used as four conditions that evoke illusory motion perception. One plain image was used as the control condition (CON condition) that does not evoke illusory motion perception. Participant was instructed to stand on a pressure sensor plate and viewed the images by a 46 inches monitor. Each image was presented for 2 minutes. Participants' postural movements were measured with centre of plantar pressure (COP) by a pressure sensor plate. The eye movements were measured with horizontal electro-oculogram. Comparing four illusory conditions, many participants reported that more illusory motion perception was evoked with W and E1 conditions. The number of horizontal eye movements was larger with W condition (p<0.01) and E1 condition (p<0.05) than with CON condition. Although no significant differences between the five conditions were observed on the total movement distance of the COP, the movements to swing of the COP of W condition tended to be different from that of CON condition. Although the COP of CON condition tended to sway slow and big, the COP of W condition tended to sway very smaller and fast. Our results suggested that the static images that evoke illusory motion perception more would induce faster eye movements and that these quick eye movements might induce small and fast body sway.
An application of the learning curve–cumulative summation test to evaluate training for endotracheal intubation in emergency medicine
Authors: Je Sangmo , Cho Youngsuk , Choi Hyuk Joong , Kang Boseung , Lim Taeho , Kang Hyunggoo 
Published: October 2013

ObjectiveThe learning curve–cumulative summation (LC-CUSUM) test allows for quantitative and individual assessments of the learning process. In this study, we evaluated the process of skill acquisition for performing endotracheal intubation (ETI) in three emergency medicine (EM) residents over a 2 year period in their first 2 years of their EM residency.MethodsWe evaluated 342 ETI cases performed by three EM residents using the LC-CUSUM test according to their rate of success or failure of ETI. A 90% success rate (SR) was chosen to define adequate performance and an SR of 80% was considered inadequate. After the learning phase, the standard CUSUM test was applied to ensure that performance was maintained.ResultsThe mean number of ETI cases required to reach the predefined level of performance was 74.7 (95% CI 62.0 to 87.3). CUSUM tests confirmed that performance was maintained after the learning phase.ConclusionsBy using the LC-CUSUM test, we were able to quantitatively monitor the acquisition of the skill of ETI by EM residents. The LC-CUSUM could be useful for monitoring the learning process for the training of airway management in the practice of EM.