bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Brain Functional MRI and DTI Visualization With Virtual Reality
Authors: Chen Bin , Moreland John , Zhang Jingyu
Published: March 2012

Magnetic resonance diffusion tensor imaging (DTI) and functional MRI (fMRI) are two active research areas in neuroimaging. DTI is sensitive to the anisotropic diffusion of water exerted by its macromolecular environment and has been shown useful in characterizing structures of ordered tissues such as the brain white matter, myocardium, and cartilage. The diffusion tensor provides two new types of information of water diffusion: the magnitude and the spatial orientation of water diffusivity inside the tissue. This information has been used for white matter fiber tracking to review physical neuronal pathways inside the brain. Functional MRI measures brain activations using the hemodynamic response. The statistically derived activation map corresponds to human brain functional activities caused by neuronal activities. The combination of these two methods provides a new way to understand human brain from the anatomical neuronal fiber connectivity to functional activities between different brain regions. In this study, virtual reality (VR) based MR DTI and fMRI visualization with high resolution anatomical image segmentation and registration, ROI definition and neuronal white matter fiber tractography visualization and fMRI activation map integration is proposed. Rationale and methods for producing and distributing stereoscopic videos are also discussed.
Topography of the Northern Hemisphere of Mercury from MESSENGER Laser Altimetry
Authors: Zuber Maria T. , Smith David E. , Phillips Roger J. , Solomon Sean C. , Neumann Gregory A. , Hauck Steven A. , Peale Stanton J. , Barnouin Olivier S. , Head James W. , Johnson Catherine L. , Lemoine Frank G. , Mazarico Erwan , Sun Xiaoli , Torrence Mark H. , Freed Andrew M. , Klimczak Christian , Margot Jean-Luc , Oberst Jürgen , Perry Mark E. , McNutt Ralph L. , Balcerski Jeffrey A. , Michel Nathalie , Talpe Matthieu J. , Yang Di
Published: March 2012

Mercury Inside and Out

The MESSENGER spacecraft orbiting Mercury has been in a ∼12-hour eccentric, near-polar orbit since 18 March 2011 (see the Perspective by

McKinnon

).

Smith
et al.

(p.
214
, published online 21 March) present the most recent determination of Mercury's gravity field, based on radio tracking of the MESSENGER spacecraft between 18 March and 23 August 2011. The results point to an interior structure that differs from those of the other terrestrial planets: the density of the planet's solid outer shell suggests the existence of a deep reservoir of high-density material, possibly an Fe-S layer.

Zuber
et al.

(p.
217
, published online 21 March) used data obtained by the MESSENGER laser altimeter through to 24 October 2011 to build a topographic map of Mercury's northern hemisphere. The map shows less variation in elevation, compared with Mars or the Moon, and its features add to the body of evidence that Mercury has sustained geophysical activity for much of its history.
Enantioselective C-H Crotylation of Primary Alcohols via Hydrohydroxyalkylation of Butadiene
Authors: Zbieg Jason R. , Yamaguchi Eiji , McInturff Emma L. , Krische Michael J.
Published: March 2012

Bonded at the Source

Asymmetric catalysis is a relatively mature field in the laboratory, with a diverse array of techniques available for the selective transformation of organic compounds. However, scaling up these techniques for industrial application remains challenging, in part because many catalysts act best on reagents that have been expensively modified, although this process often generates copious waste.

Zbieg
et al.

(p.
324
, published online 22 March) combat this challenge with a ruthenium-based catalyst that couples an unmodified bulk commodity feedstock (butadiene) with alcohols, forming carbon-carbon bonds to generate complex products with high selectivity.
Microbial Exposure During Early Life Has Persistent Effects on Natural Killer T Cell Function
Authors: Olszak Torsten , An Dingding , Zeissig Sebastian , Vera Miguel Pinilla , Richter Julia , Franke Andre , Glickman Jonathan N. , Siebert Reiner , Baron Rebecca M. , Kasper Dennis L. , Blumberg Richard S.
Published: March 2012

Microbes: Early and Often

Epidemiological studies have suggested that the increase in the incidence of asthma and other inflammatory diseases seen in many parts of the world may be due to a reduced exposure to microbes during early childhood.

Olszak
et al.

(p. 489, published online 22 March) now show that commensal microflora help to regulate the numbers and functions of natural killer T (NKT) cells in the colon and lung in mice. Germ-free mice had elevated numbers of NKT cells in these tissues and were more susceptible to chemically induced colitis and allergic asthma. Neonatal recolonization of germ-free mice with microflora prevented enhanced colitis and asthma sensitivity; however, exposure of adult mice to these conditions was not effective. Thus, early exposure to microbes has important, lasting effects on the immune system's sensitivity to inflammation.
Replicating Adenovirus-Simian Immunodeficiency Virus (SIV) Vectors Efficiently Prime SIV-Specific Systemic and Mucosal Immune Responses by Targeting Myeloid Dendritic Cells and Persisting in Rectal Macrophages, Regardless of Immunization Route
Authors: Patterson L. Jean , Kuate Seraphin , Daltabuit-Test Mara , Li Qingsheng , Xiao Peng , McKinnon Katherine , DiPasquale Janet , Cristillo Anthony , Venzon David , Haase Ashley , Robert-Guroff Marjorie
Published: March 2012

Although priming with replicating adenovirus type 5 host range mutant (Ad5hr)-human immunodeficiency virus (HIV)/simian immunodeficiency virus (SIV) recombinants, followed by HIV/SIV envelope boosting, has proven highly immunogenic, resulting in protection from SIV/simian-human immunodeficiency virus (SHIV) challenges, Ad5hr recombinant distribution, replication, and persistence have not been examined comprehensively in nonhuman primates. We utilized Ad5hr-green fluorescent protein and Ad5hr-SIV recombinants to track biodistribution and immunogenicity following mucosal priming of rhesus macaques by the intranasal/intratracheal, sublingual, vaginal, or rectal route. Ad recombinants administered by all routes initially targeted macrophages in bronchoalveolar lavage (BAL) fluid and rectal tissue, later extending to myeloid dendritic cells in BAL fluid with persistent expression in rectal mucosa 25 weeks after the last Ad immunization. Comparable SIV-specific immunity, including cellular responses, serum binding antibody, and mucosal secretory IgA, was elicited among all groups. The ability of the vector to replicate in multiple mucosal sites irrespective of delivery route, together with the targeting of macrophages and professional antigen-presenting cells, which provide potent immunogenicity at localized sites of virus entry, warrants continued use of replicating Ad vectors.
Programmed Death 1 Receptor ChangesEx Vivoin HIV-Infected Adults following Initiation of Highly Active Antiretroviral Therapy
Authors: Spitsin Sergei , Tustin Nancy B. , Riedel Eric , Tustin Richard , Murray Jennifer B. , Peck Lauren M. , Khan Mohammad , Quinn Joseph , Douglas Steven D.
Published: March 2012

This study investigates the short-term effects of highly active antiretroviral therapy (HAART) on programmed death 1 receptor (PD-1) expression and lymphocyte function. We compared lymphocytes from human immunodeficiency virus (HIV)-infected adults prior to the initiation of HAART with lymphocytes from the same subjects following 2 months of treatment. Short-term HAART resulted in a moderate increase in the expression of PD-1 on both CD4+and CD8+T cells; yet, there was still a significant reduction in viral load and recovery of CD4+T cells. After 2 months of HAART, lymphocytes from the subjects had a reduction in lymphoproliferative responses to phytohemagglutinin (PHA) and an increased response to theCandidarecall antigen and the HIV antigen p24 compared to pretreatment lymphocytes. PHA-stimulated peripheral blood mononuclear cells (PBMCs) from samples obtained 2 months after HAART produced higher levels of Th-1 cytokines (gamma interferon [IFN-γ] and tumor necrosis factor alpha[TNF-α]) than the levels observed for samples taken before treatment was initiated. There were no significant changes in the proinflammatory cytokine interleukin-2 (IL-2) or Th-2 cytokines (IL-4, IL-5, and IL-10) in the corresponding samples.Ex vivoPD-1 blockade significantly augmented PHA-induced lymphoproliferation as well as the levels of Th-1 cytokines and to a lesser extent the levels of Th-2 cytokines in PBMC cultures. The ability to downregulate PD-1 expression may be important in enhancing immune recovery in HIV infection.
Memory B Cell and Other Immune Responses in Children Receiving Two Doses of an Oral Killed Cholera Vaccine Compared to Responses following Natural Cholera Infection in Bangladesh
Authors: Leung Daniel T. , Rahman Mohammad Arif , Mohasin M. , Patel Sweta M. , Aktar Amena , Khanam Farhana , Uddin Taher , Riyadh M. Asrafuzzaman , Saha Amit , Alam Mohammad Murshid , Chowdhury Fahima , Khan Ashraful Islam , Charles Richelle , LaRocque Regina , Harris Jason B. , Calderwood Stephen B. , Qadri Firdausi , Ryan Edward T.
Published: March 2012

Current oral cholera vaccines induce lower protective efficacy and shorter duration of protection against cholera than wild-type infection provides, and this difference is most pronounced in young children. Despite this, there are limited data comparing immune responses in children following wild-type disease versus vaccination, especially with regard to memory responses associated with long-term immunity. Here, we report a comparison of immune responses in young children (2 to 5 years of age;n= 20) and older children (6 to 17 years of age;n= 20) given two doses of an oral killed cholera vaccine containing recombinant cholera toxin B subunit (CtxB) 14 days apart and compare these responses to those induced in similarly aged children recovering from infection withVibrio choleraeO1 Ogawa in Bangladesh. We found that the two vaccine groups had comparable vibriocidal and lipopolysaccharide (LPS)-specific plasma antibody responses. Vaccinees developed lower levels of IgG memory B cell (MBC) responses against CtxB but no significant MBC responses against LPS. In contrast, children recovering from natural cholera infection developed prominent LPS IgG and IgA MBC responses, as well as CtxB IgG MBC responses. Plasma LPS IgG, IgA, and IgM responses, as well as vibriocidal responses, were also significantly higher in children following disease than after vaccination. Our findings suggest that acute and memory immune responses following oral cholera vaccination in children are significantly lower than those observed following wild-type disease, especially responses targeting LPS. These findings may explain, in part, the lower efficacy of oral cholera vaccination in children.
Evaluation of Eight Serological Tests for Diagnosis of Imported Schistosomiasis
Authors: Kinkel Hans-Friedemann , Dittrich Sabine , Bäumer Britta , Weitzel Thomas
Published: March 2012

The diagnosis of schistosomiasis in individuals from countries where the disease is not endemic is challenging, and few data are available on the accuracy of serological diagnosis in those patients. We evaluated the performance of eight serological assays, including four commercial kits, in the diagnosis of imported schistosomiasis in individuals from areas where the disease is not endemic, including six enzyme-linked immunosorbent assays using three different antigens, an indirect hemagglutination assay, and an indirect immunofluorescent-antibody test. To analyze the assays, we used a total of 141 serum samples, with 121 derived from patients with various parasitic infections (among which were 37 cases of schistosomiasis) and 20 taken from healthy volunteers. The sensitivity values for detection of schistosomiasis cases ranged from 41% to 78% and were higher for
Schistosoma mansoni
than for
S. haematobium
infections. Specificity values ranged from 76% to 100%; false-positive results were most frequent for samples from patients with cestode infections. By combining two or more tests, sensitivity improved markedly and specificity decreased only moderately. Serological tests are useful instruments for diagnosing imported schistosomiasis in countries where the disease is not endemic, but due to limitations in test sensitivities, we recommend the use of two or more assays in parallel.
Improvement of Immunogenicity of Meningococcal Lipooligosaccharide by Coformulation with Lipidated Transferrin-Binding Protein B in Liposomes: Implications for Vaccine Development
Authors: Mistretta Noëlle , Guy Bruno , Bérard Yves , Dalençon François , Fratantonio Olivia , Grégoire Christophe , Lechevallier Aurélie , Lhéritier Philippe , Revet Laurent , Moreau Monique , Haensler Jean , Rokbi Bachra
Published: March 2012

Among various meningococcal antigens, lipooligosaccharide (LOS) and recombinant lipidated transferrin-binding protein B (rlip-TbpB) are considered to be putative vaccine candidates against group BNeisseria meningitidis. In the present work, we report the development of a new liposome-based vaccine formulation containing both rlip-TbpB and L8 LOS. The endotoxic activity of the liposomal LOS was evaluatedin vitrousing theLimulusAmebocyte Lysate assay and compared to the endotoxic activity of free LOS. Above a 250:1 lipid/LOS molar ratio, liposomes were shown to effectively detoxify the LOS as the endotoxic activity of the LOS was reduced by more than 99%. Immunogenicity studies in rabbits showed that the presence of rlip-TbpB dramatically increased the immunogenicity of the LOS. While the formulation raised a strong anti-TbpB response, it elicited a higher anti-LOS IgG level than the liposomal LOS alone. Sera from rabbits immunized with rlip-TbpB/liposomal LOS displayed increased ability to recognize LOS on live bacteria expressing the L8 immunotype and increased anti-LOS-specific bactericidal activity compared to sera from rabbits immunized with liposomal LOS alone. Measurement of interleukin-8 (IL-8) produced by HEK293 cells transfected with Toll-like receptor (TLR) after stimulation with rlip-TbpB showed that the protein is a TLR2 agonist, which is in accordance with the structure of its lipid. Furthermore, anin vivostudy demonstrated that the lipid moiety is not only required for its adjuvant effect but also has to be linked to the protein. Overall, the rlip-TbpB/LOS liposomal formulation was demonstrated to induce an effective anti-LOS response due to the adjuvant effect of rlip-TbpB on LOS.
Comparison of Seven Commercial Antigen and Antibody Enzyme-Linked Immunosorbent Assays for Detection of Acute Dengue Infection
Authors: Blacksell Stuart D. , Jarman Richard G. , Gibbons Robert V. , Tanganuchitcharnchai Ampai , Mammen Mammen P. , Nisalak Ananda , Kalayanarooj Siripen , Bailey Mark S. , Premaratna Ranjan , de Silva H. Janaka , Day Nicholas P. J. , Lalloo David G.
Published: March 2012

Seven commercial assays were evaluated to determine their suitability for the diagnosis of acute dengue infection: (i) the Panbio dengue virus Pan-E NS1 early enzyme-linked immunosorbent assay (ELISA), second generation (Alere, Australia); (ii) the Panbio dengue virus IgM capture ELISA (Alere, Australia); (iii) the Panbio dengue virus IgG capture ELISA (Alere, Australia); (iv) the Standard Diagnostics dengue virus NS1 antigen ELISA (Standard Diagnostics, South Korea); (v) the Standard Diagnostics dengue virus IgM ELISA (Standard Diagnostics, South Korea); (vi) the Standard Diagnostics dengue virus IgG ELISA (Standard Diagnostics, South Korea); and (vii) the Platelia NS1 antigen ELISA (Bio-Rad, France). Samples from 239 Thai patients confirmed to be dengue virus positive and 98 Sri Lankan patients negative for dengue virus infection were tested. The sensitivities and specificities of the NS1 antigen ELISAs ranged from 45 to 57% and 93 to 100% and those of the IgM antibody ELISAs ranged from 85 to 89% and 88 to 100%, respectively. Combining the NS1 antigen and IgM antibody results from the Standard Diagnostics ELISAs gave the best compromise between sensitivity and specificity (87 and 96%, respectively), as well as providing the best sensitivity for patients presenting at different times after fever onset. The Panbio IgG capture ELISA correctly classified 67% of secondary dengue infection cases. This study provides strong evidence of the value of combining dengue virus antigen- and antibody-based test results in the ELISA format for the diagnosis of acute dengue infection.
Towards Improved Accuracy of Bordetella pertussis Nucleic Acid Amplification Tests
Authors: Loeffelholz Michael
Published: March 2012

In many clinical microbiology laboratories, nucleic acid amplification tests such as PCR have become the routine methods for the diagnosis of pertussis. While PCR has greatly increased the ability of laboratories to detect
Bordetella pertussis
infections, it has also been associated with false-positive results that can, given the tendency of
B. pertussis
to cause outbreaks, result in unnecessary and costly control measures. The species specificity of
Bordetella
gene targets and their number of copies per genome greatly impact the performance characteristics of nucleic acid amplification tests for
B. pertussis
. It is crucial that laboratorians recognize these characteristics, to limit false-positive test results and prevent pseudo-outbreaks.
Concurrent Serotyping and Genotyping of Pneumococci by Use of PCR and Electrospray Ionization Mass Spectrometry
Authors: Massire Christian , Gertz Robert E. , Svoboda Pavel , Levert Keith , Reed Matthew S. , Pohl Jan , Kreft Rachel , Li Feng , White Neill , Ranken Ray , Blyn Larry B. , Ecker David J. , Sampath Rangarajan , Beall Bernard
Published: March 2012

A pneumococcal serotyping/genotyping system (PSGS) was developed based upon targeted PCR, followed by electrospray ionization mass spectrometry and amplicon base composition analysis. Eight multiplex PCRs, 32 targeting serotype-determining capsular biosynthetic loci, and 8 targeting multilocus sequence typing (MLST) loci were employed for each of 229 highly diverse
Streptococcus pneumoniae
isolates. The most powerful aspect of the PSGS system was the identification of capsular serotypes accounting for the majority of invasive and carried pneumococcal strains. Altogether, 45 different serotypes or serogroups were correctly predicted among the 196 resolvable isolates, with only 2 unexpected negative results. All 33 isolates that represented 23 serotypes not included in the PSGS yielded negative serotyping results. A genotyping database was constructed using the base compositions of 65- to 100-bp sections of MLST alleles compiled within
http://www.mlst.net
. From this database, one or more MLST sequence types (STs) that comprised a PSGS genotype were identified. The end result of more PSGS genotypes (163) than conventional STs actually tested (155) was primarily due to amplification failures of 1 to 3 targets. In many instances, the PSGS genotype could provide resolution of single- and double-locus variants. This molecular serotyping/genotyping scheme is well suited to rapid characterization of large sets of pneumococcal isolates.
Molecular Characterization of Feline Infectious Peritonitis Virus Strain DF-2 and Studies of the Role of ORF3abc in Viral Cell Tropism
Authors: Bálint Ádám , Farsang Attila , Zádori Zoltán , Hornyák Ákos , Dencső László , Almazán Fernando , Enjuanes Luis , Belák Sándor
Published: March 2012

The full-length genome of the highly lethal feline infectious peritonitis virus (FIPV) strain DF-2 was sequenced and cloned into a bacterial artificial chromosome (BAC) to study the role of ORF3abc in the FIPV-feline enteric coronavirus (FECV) transition. The reverse genetic system allowed the replacement of the truncated ORF3abc of the original FIPV DF-2 genome with the intact ORF3abc of the canine coronavirus (CCoV) reference strain Elmo/02. The
in vitro
replication kinetics of these two viruses was studied in CrFK and FCWF-4 cell lines, as well as in feline peripheral blood monocytes. Both viruses showed similar replication kinetics in established cell lines. However, the strain with a full-length ORF3 showed markedly lower replication of more than 2 log
10
titers in feline peripheral blood monocytes. Our results suggest that the truncated ORF3abc plays an important role in the efficient macrophage/monocyte tropism of type II FIPV.
The PSAP Motif within the ORF3 Protein of an Avian Strain of the Hepatitis E Virus Is Not Critical for Viral InfectivityIn Vivobut Plays a Role in Virus Release
Authors: Kenney Scott P. , Pudupakam R. S. , Huang Yao-Wei , Pierson F. William , LeRoith Tanya , Meng Xiang-Jin
Published: March 2012

The ORF3 protein of hepatitis E virus (HEV) is a multifunctional protein important for virus replication. The ORF3 proteins from human, swine, and avian strains of HEV contain a conserved PXXP amino acid motif, resembling either Src homology 3 (SH3) cell signaling interaction motifs or “late domains” involved in host cell interactions aiding in particle release. Using an avian strain of HEV, we determined the roles of the conserved prolines within the PREPSAPP motif in HEV replication and infectivity in Leghorn male hepatoma (LMH) chicken liver cells and in chickens. Each proline was changed to alanine to produce 8 avian HEV mutants containing single mutations (P64, P67, P70, and P71 to A), double mutations (P64/67A, P64/70A, and P67/70A), and triple mutations (P64/67/70A). The results showed that avian HEV mutants are replication competentin vitro, and none of the prolines in the PXXPXXPP motif are essential for infectivityin vivo; however, the second and third prolines appear to aid in fecal virus shedding, suggesting that the PSAP motif, but not the PREP motif, is involved in virus release. We also showed that the PSAP motif interacts with the host protein tumor suppressor gene 101 (TSG101) and that altering any proline within the PSAP motif disrupts this interaction. However, we showed that the ORF2 protein expressed in LMH cells is efficiently released from the cells in the absence of ORF3 and that coexpression of ORF2 and ORF3 did not act synergistically in this release, suggesting that another factor(s) such as ORF1 or viral genomic RNA may be necessary for proper particle release.
The Double-Stranded RNA Bluetongue Virus Induces Type I Interferon in Plasmacytoid Dendritic Cells via a MYD88-Dependent TLR7/8-Independent Signaling Pathway
Authors: Ruscanu Suzana , Pascale Florentina , Bourge Mickael , Hemati Behzad , Elhmouzi-Younes Jamila , Urien Céline , Bonneau Michel , Takamatsu Haru , Hope Jayne , Mertens Peter , Meyer Gilles , Stewart Meredith , Roy Polly , Meurs Eliane F. , Dabo Stéphanie , Zientara Stéphan , Breard Emmanuel , Sailleau Corinne , Chauveau Emilie , Vitour Damien , Charley Bernard , Schwartz-Cornil Isabelle
Published: March 2012

Dendritic cells (DCs), especially plasmacytoid DCs (pDCs), produce large amounts of alpha/beta interferon (IFN-α/β) upon infection with DNA or RNA viruses, which has impacts on the physiopathology of the viral infections and on the quality of the adaptive immunity. However, little is known about the IFN-α/β production by DCs during infections by double-stranded RNA (dsRNA) viruses. We present here novel information about the production of IFN-α/β induced by bluetongue virus (BTV), a vector-borne dsRNA
Orbivirus
of ruminants, in sheep primary DCs. We found that BTV induced IFN-α/β in skin lymph and in blood
in vivo
. Although BTV replicated in a substantial fraction of the conventional DCs (cDCs) and pDCs
in vitro
, only pDCs responded to BTV by producing a significant amount of IFN-α/β. BTV replication in pDCs was not mandatory for IFN-α/β production since it was still induced by UV-inactivated BTV (UV-BTV). Other inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and IL-12p40, were also induced by UV-BTV in primary pDCs. The induction of IFN-α/β required endo-/lysosomal acidification and maturation. However, despite being an RNA virus, UV-BTV did not signal through Toll-like receptor 7 (TLR7) for IFN-α/β induction. In contrast, pathways involving the MyD88 adaptor and kinases dsRNA-activated protein kinase (PKR) and stress-activated protein kinase (SAPK)/Jun N-terminal protein kinase (JNK) were implicated. This work highlights the importance of pDCs for the production of innate immunity cytokines induced by a dsRNA virus, and it shows that a dsRNA virus can induce IFN-α/β in pDCs via a novel TLR-independent and Myd88-dependent pathway. These findings have implications for the design of efficient vaccines against dsRNA viruses.
The Second Receptor Binding Site of the Globular Head of the Newcastle Disease Virus Hemagglutinin-Neuraminidase Activates the Stalk of Multiple Paramyxovirus Receptor Binding Proteins To Trigger Fusion
Authors: Porotto Matteo , Salah Zuhair , DeVito Ilaria , Talekar Aparna , Palmer Samantha G. , Xu Rui , Wilson Ian A. , Moscona Anne
Published: March 2012

The hemagglutinin-neuraminidase (HN) protein of paramyxoviruses carries out three distinct activities contributing to the ability of HN to promote viral fusion and entry: receptor binding, receptor cleavage (neuraminidase), and activation of the fusion protein. The relationship between receptor binding and fusion triggering functions of HN are not fully understood. For Newcastle disease virus (NDV), one bifunctional site (site I) on HN′s globular head can mediate both receptor binding and neuraminidase activities, and a second site (site II) in the globular head is also capable of mediating receptor binding. The receptor analog, zanamivir, blocks receptor binding and cleavage activities of NDV HN′s site I while activating receptor binding by site II. Comparison of chimeric proteins in which the globular head of NDV HN is connected to the stalk region of either human parainfluenza virus type 3 (HPIV3) or Nipah virus receptor binding proteins indicates that receptor binding to NDV HN site II not only can activate its own fusion (F) protein but can also activate the heterotypic fusion proteins. We suggest a general model for paramyxovirus fusion activation in which receptor engagement at site II plays an active role in F activation.
Temporal- and Strain-Specific Host MicroRNA Molecular Signatures Associated with Swine-Origin H1N1 and Avian-Origin H7N7 Influenza A Virus Infection
Authors: Loveday Emma-Kate , Svinti Victoria , Diederich Sandra , Pasick John , Jean François
Published: March 2012

MicroRNAs (miRNAs) repress the expression levels of genes by binding to mRNA transcripts, acting as master regulators of cellular processes. Differential expression of miRNAs has been linked to virus-associated diseases involving members of theHepacivirus,Herpesvirus, andRetrovirusfamilies. In contrast, limited biological and molecular information has been reported on the potential role of cellular miRNAs in the life cycle of influenza A viruses (infA). In this study, we hypothesize that elucidating the miRNA expression signatures induced by low-pathogenicity swine-origin infA (S-OIV) pandemic H1N1 (2009) and highly pathogenic avian-origin infA (A-OIV) H7N7 (2003) infections could reveal temporal and strain-specific miRNA fingerprints during the viral life cycle, shedding important insights into the potential role of cellular miRNAs in host-infA interactions. Using a microfluidic microarray platform, we profiled cellular miRNA expression in human A549 cells infected with S- and A-OIVs at multiple time points during the viral life cycle, including global gene expression profiling during S-OIV infection. Using target prediction and pathway enrichment analyses, we identified the key cellular pathways associated with the differentially expressed miRNAs and predicted mRNA targets during infA infection, including the immune system, cell proliferation, apoptosis, cell cycle, and DNA replication and repair. By identifying the specific and dynamic molecular phenotypic changes (microRNAome) triggered by S- and A-OIV infection in human cells, we provide experimental evidence demonstrating a series of temporal and strain-specific host molecular responses involving different combinatorial contributions of multiple cellular miRNAs. Our results also identify novel potential exosomal miRNA biomarkers associated with pandemic S-OIV and deadly A-OIV-host infection.
Formation of Orthopoxvirus Cytoplasmic A-Type Inclusion Bodies and Embedding of Virions Are Dynamic Processes Requiring Microtubules
Authors: Howard Amanda R. , Moss Bernard
Published: March 2012

In cells infected with some orthopoxviruses, numerous mature virions (MVs) become embedded within large, cytoplasmic A-type inclusions (ATIs) that can protect infectivity after cell lysis. ATIs are composed of an abundant viral protein called ATIp, which is truncated in orthopoxviruses such as vaccinia virus (VACV) that do not form ATIs. To study ATI formation and occlusion of MVs within ATIs, we used recombinant VACVs that express the cowpox full-length ATIp or we transfected plasmids encoding ATIp into cells infected with VACV, enabling ATI formation. ATI enlargement and MV embedment required continued protein synthesis and an intact microtubular network. For live imaging of ATIs and MVs, plasmids expressing mCherry fluorescent protein fused to ATIp were transfected into cells infected with VACV expressing the viral core protein A4 fused to yellow fluorescent protein. ATIs appeared as dynamic, mobile bodies that enlarged by multiple coalescence events, which could be prevented by disrupting microtubules. Coalescence of ATIs was confirmed in cells infected with cowpox virus. MVs were predominantly at the periphery of ATIs early in infection. We determined that coalescence contributed to the distribution of MVs within ATIs and that microtubule-disrupting drugs abrogated coalescence-mediated MV embedment. In addition, MVs were shown to move from viral factories at speeds consistent with microtubular transport to the peripheries of ATIs, whereas disruption of microtubules prevented such trafficking. The data indicate an important role for microtubules in the coalescence of ATIs into larger structures, transport of MVs to ATIs, and embedment of MVs within the ATI matrix.
Nuclear Egress of Pseudorabies Virus Capsids Is Enhanced by a Subspecies of the Large Tegument Protein That Is Lost upon Cytoplasmic Maturation
Authors: Leelawong Mindy , Lee Joy I. , Smith Gregory A.
Published: March 2012

Herpesviruses morphogenesis occurs stepwise both temporally and spatially, beginning in the nucleus and concluding with the emergence of an extracellular virion. The mechanisms by which these viruses interact with and penetrate the nuclear envelope and subsequent compartments of the secretory pathway remain poorly defined. In this report, a conserved viral protein (VP1/2; pUL36) that directs cytoplasmic stages of egress is identified to have multiple isoforms. Of these, a novel truncated VP1/2 species translocates to the nucleus and assists the transfer of DNA-containing capsids to the cytoplasm. The capsids are handed off to full-length VP1/2, which replaces the nuclear isoform on the capsids and is required for the final cytoplasmic stages of viral particle maturation. These results document that distinct VP1/2 protein species serve as effectors of nuclear and cytoplasmic egress.
Cold-Adapted Pandemic 2009 H1N1 Influenza Virus Live Vaccine Elicits Cross-Reactive Immune Responses against Seasonal and H5 Influenza A Viruses
Authors: Jang Yo Han , Byun Young Ho , Lee Yoon Jae , Lee Yun Ha , Lee Kwang-Hee , Seong Baik Lin
Published: March 2012

The rapid transmission of the pandemic 2009 H1N1 influenza virus (pH1N1) among humans has raised the concern of a potential emergence of reassortment between pH1N1 and highly pathogenic influenza strains, especially the avian H5N1 influenza virus. Here, we report that the cold-adapted pH1N1 live attenuated vaccine (CApH1N1) elicits cross-reactive immunity to seasonal and H5 influenza A viruses in the mouse model. Immunization with CApH1N1 induced both systemic and mucosal antibodies with broad reactivity to seasonal and H5 strains, including HAPI H5N1 and the avian H5N2 virus, providing complete protection against heterologous and heterosubtypic lethal challenges. Our results not only accentuate the merit of using live attenuated influenza virus vaccines in view of cross-reactivity but also represent the potential of CApH1N1 live vaccine for mitigating the clinical severity of infections that arise from reassortments between pH1N1 and highly pathogenic H5 subtype viruses.
Functional Characterization of Residues Required for the Herpes Simplex Virus 1 E3 Ubiquitin Ligase ICP0 To Interact with the Cellular E2 Ubiquitin-Conjugating Enzyme UBE2D1 (UbcH5a)
Authors: Vanni Emilia , Gatherer Derek , Tong Lily , Everett Roger D. , Boutell Chris
Published: March 2012

The viral ubiquitin ligase ICP0 is required for efficient initiation of herpes simplex virus 1 (HSV-1) lytic infection and productive reactivation of viral genomes from latency. ICP0 has been shown to target a number of specific cellular proteins for proteasome-dependent degradation during lytic infection, including the promyelocytic leukemia protein (PML) and its small ubiquitin-like modified (SUMO) isoforms. We have shown previously that ICP0 can catalyze the formation of unanchored polyubiquitin chains and mediate the ubiquitination of specific substrate proteins
in vitro
in the presence of two E2 ubiquitin-conjugating enzymes, namely, UBE2D1 (UbcH5a) and UBE2E1 (UbcH6), in a RING finger-dependent manner. Using homology modeling in conjunction with site-directed mutagenesis, we identify specific residues required for the interaction between the RING finger domain of ICP0 and UBE2D1, and we report that point mutations at these residues compromise the ability of ICP0 to induce the colocalization of conjugated ubiquitin and the degradation of PML and its SUMO-modified isoforms. Furthermore, we show that RING finger mutants that are unable to interact with UBE2D1 fail not only to complement the plaque-forming defect of an ICP0-null mutant virus but also to mediate the derepression of quiescent HSV-1 genomes in cell culture. These data demonstrate that the ability of ICP0 to interact with cellular E2 ubiquitin-conjugating enzymes is fundamentally important for its biological functions during HSV-1 infection.
Dissection of the Role of the Stable Signal Peptide of the Arenavirus Envelope Glycoprotein in Membrane Fusion
Authors: Messina Emily L. , York Joanne , Nunberg Jack H.
Published: March 2012

The arenavirus envelope glycoprotein (GPC) retains a stable signal peptide (SSP) as an essential subunit in the mature complex. The 58-amino-acid residue SSP comprises two membrane-spanning hydrophobic regions separated by a short ectodomain loop that interacts with the G2 fusion subunit to promote pH-dependent membrane fusion. Small-molecule compounds that target this unique SSP-G2 interaction prevent arenavirus entry and infection. The interaction between SSP and G2 is sensitive to the phylogenetic distance between New World (Junín) and Old World (Lassa) arenaviruses. For example, heterotypic GPC complexes are unable to support virion entry. In this report, we demonstrate that the hybrid GPC complexes are properly assembled, proteolytically cleaved, and transported to the cell surface but are specifically defective in their membrane fusion activity. Chimeric SSP constructs reveal that this incompatibility is localized to the first transmembrane segment of SSP (TM1). Genetic changes in TM1 also affect sensitivity to small-molecule fusion inhibitors, generating resistance in some cases and inhibitor dependence in others. Our studies suggest that interactions of SSP TM1 with the transmembrane domain of G2 may be important for GPC-mediated membrane fusion and its inhibition.
Trends in sudden cardiac death and its risk factors in Japan from 1981 to 2005: the Circulatory Risk in Communities Study (CIRCS)
Authors: Maruyama Minako , Ohira Tetsuya , Imano Hironori , Kitamura Akihiko , Kiyama Masahiko , Okada Takeo , Maeda Kenji , Yamagishi Kazumasa , Noda Hiroyuki , Ishikawa Yoshinori , Shimamoto Takashi , Iso Hiroyasu
Published: March 2012

Objective

There is little evidence whether sudden cardiac death (SCD) is increasing in Asia, although the incidence of coronary heart disease among urban middle-aged Japanese men has increased recently. We examined trends in the incidence of SCD and its risk factors in the Circulatory Risk in Communities Study.

Design and setting

This was a population-based longitudinal study. Surveillance of men and women for SCD incidence and risk factors was conducted from 1981 to 2005.

Subjects

The surveyed population was all men and women aged 30–84 years who lived in three rural communities and one urban community in Japan.

Main outcome measures

Trends in SCD incidence and its risk factors.

Results

Age-adjusted and sex-adjusted incidence of SCD decreased from 1981–1985 to 1991–1995, and plateaued thereafter. The annual incidence per 100 000 person-years was 76.0 in 1981–1985, 57.9 in 1986–1990, 39.3 in 1991–1995, 31.6 in 1996–2000 and 36.8 in 2001–2005. The prevalence of hypertension decreased from 1981–1985 to 1991–1995, and plateaued thereafter for men and women. The age-adjusted prevalence of current smoking for men decreased while that of diabetes mellitus increased for both sexes from 1981–1985 to 2001–2005.

Conclusions

The incidence of SCD decreased from 1981 to 1995 but was unchanged from 1996 to 2005. Continuous surveillance is necessary to clarify future trends in SCD in Japan because of an increasing incidence of diabetes mellitus.
Diabetes-induced osteoarthritis: from a new paradigm to a new phenotype
Authors: Berenbaum Francis
Published: March 2012

Several epidemiological and experimental data support the hypothesis that diabetes could be an independent risk factor for osteoarthritis (OA), at least in some patients, leading to the concept of a diabetes-induced OA phenotype. If confirmed, this new paradigm will have a dramatic impact on prevention of OA initiation and progression.
Republished review: Next generation sequencing for clinical diagnostics and personalised medicine: implications for the next generation cardiologist
Authors: Ware James S , Roberts Angharad M , Cook Stuart A
Published: March 2012

The fast moving field of genomic medicine is already impacting on clinical care and cardiologists are fortunate to be in a position to benefit early from the transformative advances in genomics. However, the challenges associated with genomics in the clinic in general, and with next generation sequencing technologies in particular, are significant and cardiologists need to be prepared if they wish to surf the wave of genomic opportunity. This paper presents an overview of the implications of next generation sequencing for clinical diagnostics and personalised medicine in the cardiology clinic.
Coronary artery spasm and ventricular arrhythmias
Authors: Looi Khang Li , Grace Andrew , Agarwal Sharad
Published: March 2012

Coronary artery spasm (CAS) is characterised by chest pain at rest and transient ST segment elevation on the ECG. The natural history of variant angina is not fully understood. Patients with CAS are younger, mostly female subjects and usually do not have traditional cardiovascular risk factors other than cigarette smoking. Cardiac arrhythmias are known to be associated with CAS. Ventricular arrhythmia is a well-recognised complication and sudden cardiac death has also been documented. The most important diagnostic tool in CAS is coronary angiography. 24 h ECG Holter monitoring can be very useful in the diagnosis of ventricular arrhythmias caused by CAS. The mainstay therapy for CAS is calcium channel blockers and nitrates. The use of β-blockers, especially the non-selective group, can promote attacks or prolong vasospastic state. The indication for implantable cardioverter defibrillator (ICD) implantation in a patient with CAS is still not clearly established. The role of primary prevention with the use of ICD is controversial; however, ICD implantation should be considered in high risk patients despite optimal medical treatment.
Sex work and the public health impacts of the 2010 Olympic Games: Table 1
Authors: Deering Kathleen N , Chettiar Jill , Chan Keith , Taylor Matthew , Montaner Julio SG , Shannon Kate
Published: March 2012

Objectives

The authors examined the impact of exposure to the 2010 Winter Olympics time period on outcomes measuring disruption of local sex work environments in Vancouver, Canada.

Methods

The authors conducted a before-and-after study, using multivariable logistic regression, to assess the relationship between exposure to the Olympics period (19 January–14 March 2010) versus the post-Olympics period (1 April–1 July 2010) and outcomes.

Results

This study included 207 youth (14+ years) and adult sex workers (SWs) (Olympics: n=107; post-Olympics: n=100). SWs in the two periods were statistically similar, with an overall median age of 33 years (IQR: 28–40), and 106 (51.2%) of indigenous ancestry or ethnic minority. In separate multivariable logistic regression models, the Olympics period remained statistically significantly associated with perceived heightened police harassment of SWs without arrest (adjusted ORs (AOR): 3.95, 95% CIs 1.92 to 8.14), decreased availability of clients (AOR: 1.97, 95% CIs 1.11 to 3.48) and increased difficulty meeting clients due to road closures/construction (AOR: 7.68, 95% CIs 2.46 to 23.98). There were no significantly increased odds in perceived reports of new (0.999), youth (0.536) or trafficked SWs (zero reports) in the Olympic period.

Conclusions

To reduce potential adverse public health impacts of enhanced police harassment and displacement of local sex work markets, results suggest that evidence-based public health strategies need to be adopted for host cities of future events (eg, the London 2012 Olympic Games), such as the removal of criminal sanctions targeting sex work and the piloting and rigorous evaluation of safer indoor work spaces.
Access to and use of unauthorised online HIV self-tests by internet-using French-speaking men who have sex with men
Authors: Greacen Tim , Friboulet David , Fugon Lionel , Hefez Serge , Lorente Nicolas , Spire Bruno
Published: March 2012

Objectives

Although not authorised in France, HIV self-tests are easily available online. The authors questioned French-speaking internet-using men who have sex with men (MSM) concerning their access to and use of such tests.

Methods

Online questionnaire on sex websites and gay and HIV community websites. A multivariate logistic regression for rare events was used to identify factors associated with accessing self-tests.

Results

9169 MSM completed the questionnaire. 2748 (30%) were aware of online self-tests, but only 82 of those aware and not already HIV positive (3.5%) declared having accessed one and only 69 using it. In the multivariate model, living one's sex-life with men in absolute secrecy (odds ratio (OR)=3.90, 95% CI 1.73 to 8.76), knowing of the tests via a sex partner (OR=3.42, CI 1.39 to 8.41) or an internet search engine (OR=2.18, CI 1.26 to 3.74) but not through the general (OR=0.21, CI 0.11 to 0.42) or the gay press (OR=0.34, CI 0.16 to 0.73), having unprotected anal intercourse with a casual partner in the preceding year (OR=1.90, CI 1.17 to 3.06), using self-tests for other diseases (OR=2.66, CI 1.43 to 4.90), using poppers (OR=2.23, CI 1.35 to 3.67) and doping products (OR=3.53, CI 1.55 to 8) in the preceding year, having done a traditional HIV test but not in the preceding year (OR=1.93, CI 1.14 to 3.32), never having had suicidal thoughts (OR=0.54, CI 0.32 to 0.90) and living in the provinces (OR=1.80, CI 1.01 to 3.25) were all independently associated with accessing self-tests.

Conclusions

Although readiness to admit use of unauthorised medical products may have influenced results, few MSM aware of the existence of self-tests actually declared having accessed one. Accessing the self-test was independently associated with both living one's sex-life with men in total secrecy and having had unprotected anal intercourse in the last 12 months, indicating that autonomous self-testing may reduce barriers to testing for this vulnerable population.
Pregnancy and contraceptive use among women participating in an HIV prevention trial in Tanzania
Authors: Odutola Aderonke , Baisley Kathy , Hayes Richard J , Rusizoka Mary , Tanton Clare , Weiss Helen A , Changalucha John , Ross David A , Watson-Jones Deborah
Published: March 2012

Objectives

Information on pregnancy rates and factors associated with pregnancy and contraceptive use is important for clinical trials in women in sub-Saharan Africa where withdrawal of investigational products may be required in the event of pregnancy with a consequent effect on sample size and trial power.

Methods

A prospective cohort analysis of pregnancy and contraceptive use was conducted in Tanzanian women enrolled in a randomised placebo-controlled trial of herpes simplex virus-suppressive therapy with acyclovir to measure the effect on HIV incidence in HIV-negative women and on genital and plasma HIV viral load in HIV-positive women. The cohort was followed every 3 months for 12–30 months. Women at each visit were categorised into users or non-users of contraception. Pregnancy rates and factors associated with pregnancy incidence and contraceptive use were measured.

Results

Overall 254 of 1305 enrolled women became pregnant at least once during follow-up (pregnancy rate: 12.0/100 person-years). Younger age, being unmarried, higher baseline parity and changes in contraceptive method during follow-up were independently associated with pregnancy. Having paid sex and being HIV positive were associated with lower risk of pregnancy. Uptake of contraception was associated with young age, being unmarried, occupation, parity and the number and type of sexual partners.

Conclusions

Data on use of contraceptive methods and risk factors for pregnancy can help to guide decisions on trial eligibility and the need for additional counselling. Mandatory reliable contraceptive use in study participants may be required to reduce pregnancy rates in studies where pregnancy is contraindicated.
A GEOMETRIC APPROACH TO CORRELATIONS AND QUARK NUMBER SUSCEPTIBILITIES
Authors: BELLUCCI STEFANO , CHANDRA VINOD , TIWARI BHUPENDRA NATH
Published: March 2012

We study the thermodynamic geometry arising from the free energy for the two- and three-flavor finite temperature hot QCD near the critical temperature. We develop a geometric notion for QCD thermodynamics, relating it with the existing microscopic quantities, e.g. quark-number susceptibility, which appears naturally within an approximately self-consistent resummation of perturbative QCD. We further incorporate thermal fluctuations in the free energy, thus yielding the geometric properties of local and global chemical correlations. These investigations are perturbative in nature. Nevertheless, one could apply the same line of thought for the geometric realization of underlying quark susceptibilities, either in the fabric of lattice QCD or in that of nonperturbative QCD.