bluuebunny/crossref_metadata_2025 huggingface dataset:
Validation Assessment of Risk Scores to Predict Postthrombolysis Intracerebral Haemorrhage
Authors: Cucchiara Brett , Kasner Scott , Tanne David , Levine Steven , Demchuk Andrew , Messe Steve , Sansing Lauren , Lees Kennedy , Lyden Patrick ,   for the SAINT Investigators
Published: December 2010

Background

Two clinical risk scores, the Haemorrhage After Thrombolysis and Multicentre Stroke Survey scores, have been proposed to predict the risk of intracerebral haemorrhage following thrombolysis in acute ischaemic stroke.

Aims

To validate Haemorrhage After Thrombolysis and Multicentre Stroke Survey scores as predictors of post-tissue plasminogen activator symptomatic intracerebral haemorrhage and asymptomatic intracerebral haemorrhage in an independent cohort.

Methods

Haemorrhage After Thrombolysis and Multicentre Stroke Survey scores were calculated for the cohort of tissue plasminogen activator-treated patients enrolled in the placebo arms of the SAINT-I and SAINT-II trials. The absolute risk of symptomatic intracerebral haemorrhage and asymptomatic intracerebral haemorrhage associated with each scoring system was determined. The overall predictive value was assessed using c-statistics.

Results

Symptomatic intracerebral haemorrhage occurred in 5·6% of 965 patients treated with tissue plasminogen activator in the SAINT cohorts. The risk of symptomatic intracerebral haemorrhage was modestly greater, with higher Haemorrhage After Thrombolysis scores (0: 4·1%, 1: 4·1%, 2: 8·8%, 3: 12·5%, 4: 0%, 5: no subjects). Similar results were seen with the Multicentre Stroke Survey score (0: 0%, 1: 4·8%, 2: 2·3%, 3: 7·3%, 4: 6·3%). In logistic regression, the Haemorrhage After Thrombolysis score was associated with the risk of symptomatic intracerebral haemorrhage (odds ratio=1·41 per point, 95% confidence interval: 1·05–1·89, P=0·021) and asymptomatic intracerebral haemorrhage (odds ratio=1·59 per point, 95% confidence interval: 1·33–1·92, P<0·001). The Multicentre Stroke Survey score was modestly associated with the risk of symptomatic intracerebral haemorrhage (odds ratio=1·43 per point, 95% confidence interval: 0·95–2·15, P=0·084) and asymptomatic intracerebral haemorrhage (odds ratio=1·63 per point, 95% confidence interval: 1·27–2·08, P<0·001). The c-statistic was 0·59 for predicting symptomatic intracerebral haemorrhage and 0·61 for asymptomatic intracerebral haemorrhage for both the Haemorrhage After Thrombolysis and the Multicentre Stroke Survey scores.

Conclusions

While both the Haemorrhage After Thrombolysis and Multicentre Stroke Survey scores were associated with a risk of symptomatic intracerebral haemorrhage, discriminatory ability was limited.
MULTI‐PHASE HORMUZ SALT DIAPIRISM IN THE SOUTHERN ZAGROS, SW IRAN
Authors: Motamedi H. , Sepehr M. , Sherkati S. , Pourkermani M.
Published: December 2010

In the Zagros orogenic belt in SW Iran, a 14 km thick sedimentary succession is decoupled from crystalline basement by the thick Infracambrian Hormuz complex. The complex consists of evaporites (“Hormuz salt”) together with marls, sandstones, carbonates and volcanic rocks and is known from emergent salt diapirs in the Fars and High Zagros areas. In this paper, we document discrete pulses of halokinetic activity using seismic profiles, well data and recent field studies. The earliest phase of mobilization of the Hormuz salt occurred in the pre‐Silurian. Later pulses of salt movement took place during opening of Neo‐Tethys in Permo‐Triassic times and during ophiolite obduction onto the NE Arabian plate margin in the Late Cretaceous. Halokinesis was intense during the Neogene Zagros orogeny when some salt plugs breached the surface.
MEMBRANE SEAL LEAKAGE IN NON‐FRACTURED CAPROCKS BY THE FORMATION OF OIL‐WET FLOW PATHS
Authors: Teige G.M.G. , Hermanrud C. , Rueslåtten H.G.
Published: December 2010

Leakage through non‐fractured shaly caprock sequences is often envisioned as hydrocarbon percolation through water‐wet pore networks. Leakage by this process requires that the buoyancy of the hydrocarbon column overcomes the capillary entry pressure of the caprock pores. If it does not, then leakage through caprocks that are not hydrofractured depends on diffusion of hydrocarbons, which is an extremely slow process. As a result, the hydrocarbon residence time would be almost endless.

We suggest a novel model for leakage through non‐fractured caprock shales. Two lines of laboratory‐based research motivate our suggestion. One of these addresses wettability alterations, while the other addresses pore‐scale fluid flow in the presence of capillary sealing. The latter approach has demonstrated that water can flow vertically through water‐wet oil reservoirs and further through caprocks, whereas the oil remains in the reservoir. This recognition is a consequence of the observation that residual water in the reservoir can be both continuous and mobile.

We suggest that such flow of residual water through reservoirs and into caprocks can lead to the establishment of oil‐wet flow paths in membrane seals. This process takes place in a sequence of events that leads to wettability alterations in the seals. Such wettability alterations allow seals to leak by multiphase Darcy flow, which would hardly happen if the seals were water wet and with sufficiently small pore throats.

One implication of this model is that capillary sealing will be effective only for a limited period of time. The model also predicts that residual oil saturation in the shales as a whole remains low, as the suggested flow paths only develop locally and develop narrow leakage pathways.
Common Health Policy Interests and the Shaping of Global Pharmaceutical Policies
Authors: Koivusalo Meri
Published: December 2010

In order to achieve more ethical global health outcomes, health policies must be driven by health priorities and should take into account broader health policy requirements, including the needs of specific national health systems. It is thus important to recognize that the division of interests in key policy areas are not necessarily between the priorities of rich and poor countries, but between (1) pharmaceutical industry interests and health policy interests, and (2) national industrial and trade policy interests and public health policies. In this article I will focus on two broad common interests for health policy officials. Both have become important in the context of current global negotiations relating to access to medicines; pandemic influenza; and public health, innovation, and intellectual property rights. These are (1) ensuring access, availability, and the safety of pharmaceuticals, and (2) ensuring that research-and-development efforts respond to public health needs. I argue that these issues are not solely the concern of developing countries because the diminishing national policy space for health in pharmaceutical policies presents a challenge to all governments, including rich ones.
Olympic Bidding, Multicultural Nationalism, Terror, and the Epistemological Violence of ‘Making Britain Proud’
Authors: Falcous Mark , Silk Michael L.
Published: December 2010

This paper excavates the entanglement of British nationalist identity politics with sport, terrorism, place re‐imagining, mega‐event bidding, and corporate neoliberalism. We focus on London's 2012 olympic bidding and the coalescence of corporate, state, civic, and sporting interests surrounding the national (re)imaginings that characterised the bid. We open with a critical reading of the bid narratives explicating how selective assertions of Britishness were envisioned through the motifs of harmonious multicultural unity, ‘youth’, and passion for sport. We focus on how these narratives offered up ‘idealised’ multicultural citizens and harmonious diversity as a reactionary form of nationalist ‘pride politics’ (Fortier 2005). We subsequently juxtapose these narratives with a critical reading of English press and political discourse in the aftermath of the 7 July 2005 bombings – the day after London was awarded the olympic games. This juxtaposition reveals the tensions and ambiguities between assertions of inclusive civic nationalism – that apparently transcends ethnic difference – and the geo‐politics of the ‘war on terror’ within Britain's post‐imperial self imaginings. Specifically, we tease out the place – and ambiguities – of the 2012 olympics within these imaginings reading the London games as an exemplar of a soft‐core ideological spectacle informing selective nationalist narratives within the context of unfolding neoliberal politics.
Tolerance to the Prophylactic Effects of Carbamazepine and Related Mood Stabilizers in the Treatment of Bipolar Disorders
Authors: Post Robert M. , Weiss Susan R.B.
Published: December 2010

Tolerance development after successful long‐term treatment of bipolar disorder is under recognized, as are ways to prevent or show its occurrence or reverse it once it has occurred. We review the clinical literature which suggests that tolerance can develop to most treatment approaches in bipolar illness and present an animal model of tolerance development to anticonvulsant effects of carbamazepine or lamotrigine on amgydala‐kindled seizures. In this model tolerance does not have a pharmacokinetic basis, but is contingent upon the drug being present in the brain at the time of amygdala stimulation. The occurrence of seizures in the absence of drug is sufficient to reverse tolerance and re‐establish anticonvulsant efficacy. Based on the model, we hypothesize that some episode‐induced compensatory adaptive changes in gene expression fail to occur in tolerant subjects and that episodes off medication re‐induce these changes and renew drug effectiveness. Approaches that slow or reverse tolerance development in the animal model are reviewed so that they can be tested for their applicability in the clinic. Criteria for assessing tolerance development are offered in the hope that this will facilitate a more systemic literature about its prevalence, prevention, and reversal. Careful longitudinal monitoring of episode occurrence is essential to understanding tolerance development in the affective disorder and its treatment.
Metastasis in the subcarinal lymph node with unknown primary tumor
Authors: Eckardt Jens , Olsen Karen Ege , Petersen Henrik
Published: December 2010

A 59‐year‐old man with previous anaplastic large cell T‐cell lymphoma stage 3A was admitted with an isolated positron emission tomography(PET)‐positive spot in a subcarinal lymph node. Diagnosis was achieved with endobronchial ultrasound‐guided fine‐needle aspiration demonstrating a well‐differentiated squamous cell carcinoma but no primary tumor was visible on PET‐computed tomography. Because of his previous lymphoma the patient was scheduled for mediastinoscopy where the diagnosis was confirmed. Subsequent gastroscopy was normal and a right‐sided thoracotomy showed no evidence of cancer elsewhere, only an inoperable metastasis in a subcarinal lymph node which infiltrated the trachea, esophagus and aorta. Such isolated squamous cell carcinoma in a subcarinal lymph node without a primary tumor despite invasive work‐up has not been reported before.
Energy Efficiency of Refrigeration Systems for High-Heat-Flux Microelectronics
Authors: Phelan P. E. , Gupta Y. , Tyagi H. , Prasher R. S. , Catano J. , Michna G. , Zhou R. , Wen J. , Jensen M. , Peles Y.
Published: December 2010

Increasingly, military and civilian applications of electronics require extremely high-heat fluxes on the order of 1000 W/cm2. Thermal management solutions for these severe operating conditions are subject to a number of constraints, including energy consumption, controllability, and the volume or size of the package. Calculations indicate that the only possible approach to meeting this heat flux condition, while maintaining the chip temperature below 65°C, is to utilize refrigeration. Here, we report an initial thermodynamic optimization of the refrigeration system design. In order to hold the outlet quality of the fluid leaving the evaporator to less than approximately 20%, in order to avoid reaching critical heat flux, the refrigeration system design is dramatically different from typical configurations for household applications. In short, a simple vapor-compression cycle will require excessive energy consumption, largely because of the additional heat required to return the refrigerant to its vapor state before the compressor inlet. A better design is determined to be a “two-loop” cycle, in which the vapor-compression loop is coupled thermally to a pumped loop that directly cools the high-heat-flux chip.
Numerical Analysis of Transient Temperature Distribution Inside a Current Transformer
Authors: Cui Jie , Nadkarni Mahesh , Mahajan Satish M. , Robalino Diego M.
Published: December 2010

Current transformer (CT) is a device that transfers the electrical energy from one circuit to another through a shared magnetic field. In a CT, heat is generated in the core, tank wall, and primary and secondary windings. The performance of a CT is well indicated by the temperature distribution inside it. In this study, numerical analysis was performed to predict the temperature distribution inside the CT at every instant under different load conditions. It was found that the numerical results obtained were in good agreement with the experimental measurements. Thus, it was concluded that the numerical method can be a useful tool in CT design and performance monitoring.
Response to Comment on “The Incidence of Fire in Amazonian Forests with Implications for REDD”
Authors: Aragão Luiz E. O. C. , Shimabukuro Yosio E.
Published: December 2010

Balch
et al
. suggest that the increased fire frequency reported in our study is principally due to post-deforestation activities. We present a new analysis demonstrating that for the majority of grid cells with positive fire trends, there is a low likelihood that these trends have resulted exclusively from post-deforestation activities. We therefore confirm that fires pose a growing threat to reducing emissions from deforestation and degradation (REDD) policies.
New Genes in
Drosophila
Quickly Become Essential
Authors: Chen Sidi , Zhang Yong E. , Long Manyuan
Published: December 2010

Essential and New

Genes can be broadly grouped into two sets on the basis of their contribution to fitness: those that are essential to the life of an organism and those that can be dispensed with. However, the degree of essentiality in evolutionarily “new” genes—genes that have originated in the recent past—is unknown.

Chen
et al.

(p.
1682
) investigated the origination and evolution of new genes within 12
Drosophila
species and found, surprisingly, that over one-third of genes that have originated within the last 3.5 million years show essential function and that these functions are overrepresented during larval development. Approximately the same proportion of older genes was essential, although many of these genes also appear to show enrichment at later developmental stages. These findings challenge conventional wisdom that would claim that essential genes are ancient and conserved among animal taxa.
Fetal and Adult Hematopoietic Stem Cells Give Rise to Distinct T Cell Lineages in Humans
Authors: Mold Jeff E. , Venkatasubrahmanyam Shivkumar , Burt Trevor D. , Michaëlsson Jakob , Rivera Jose M. , Galkina Sofiya A. , Weinberg Kenneth , Stoddart Cheryl A. , McCune Joseph M.
Published: December 2010

Lymphocytes Layer It On

Cells of the immune system begin to develop from hematopoietic stem cells (HSCs) during fetal life. In the adult, HSCs continue to produce immune cells to replenish dying cells or in response to an infection. In mice and birds, immune cell development occurs in a “layered” manner, whereby distinct populations of HSCs that arise at different times during development generate distinct immune cell lineages. In contrast, development of human immune cells, and T lymphocytes in particular, is thought to be linear.

Mold
et al.

(p.
1695
; see the Perspective by

Betz

) now show that T lymphocyte development in humans is also “layered,” and strategically so. T cells that arise from fetal HSCs are enriched in regulatory T cells, which promote immune tolerance, rather than classical T cells, which readily respond to foreign antigen. By favoring the development of regulatory T cell populations during fetal life, the immune system is perhaps better able to keep responses to maternal antigens in check. The development of large numbers of classical T cells is delayed until after birth when infectious agents represent a more imminent threat.
The Substrate of Greatwall Kinase, Arpp19, Controls Mitosis by Inhibiting Protein Phosphatase 2A
Authors: Gharbi-Ayachi Aicha , Labbé Jean-Claude , Burgess Andrew , Vigneron Suzanne , Strub Jean-Marc , Brioudes Estelle , Van-Dorsselaer Alain , Castro Anna , Lorca Thierry
Published: December 2010

Beyond the Greatwall

Protein phosphorylation and dephosphorylation provide a central mechanism that controls the eukaryotic cell division cycle and entry of cells into mitosis. A form of protein phosphatase 2A (PP2A) has an important role inhibiting phosphorylation-dependent activation of cyclin-dependent kinase 1 (CDK1) itself and also dephosphorylating substrates of the active CDK1 that promote mitosis. PP2A activity is inhibited when another protein kinase, known as Greatwall, is activated (see the Perspective by

Virshup and Kaldis

).

Mochida
et al.

(p.
1670
) and

Gharbi-Ayachi
et al.

(p.
1673
) searched for substrates of Greatwall that might participate in the cell cycle regulatory machinery. When phosphorylated by Greatwall, a pair of small related proteins, Arpp19 and α-endosulfine, inhibited activity of PP2A. These effects were critical for regulation of mitosis in
Xenopus
egg extracts and in human cancer cells. Greatwall itself is phosphorylated and activated by CDK1—thus, apparently contributing to a feed-forward loop that contributes to the switchlike commitment of cells to mitosis.
Enforcing the Greatwall in Mitosis
Authors: Virshup David M. , Kaldis Philipp
Published: December 2010

Initiation of the cell division cycle involves a signaling pathway that blocks the action of a specific phosphatase.
Britton Chance (1913–2010)
Authors: Dutton P. Leslie
Published: December 2010

A pioneering scientist and gold medal Olympian had unflagging energy and broad interests that fueled his research.
Asia's Looming Social Challenge: Coping With the Elder Boom
Authors: Stone Richard
Published: December 2010

Asia is graying fast. Long-term population studies under way or about to begin will chart demographic, economic, and health transitions—and give governments worldwide clues about how to cope with the aging boom.
Shining a Light on the Genome's 'Dark Matter'
Authors: Pennisi Elizabeth
Published: December 2010

Since the publication of the human genome sequence in 2001, scientists have found that the so-called junk DNA that lies between genes actually carries out many important functions.
Cells Rewrite Their Own Destiny
Authors: Vogel Gretchen
Published: December 2010

By prompting a cell to overexpress a few genes, researchers have discovered in the past decade how to turn a skin or blood cell into a pluripotent cell: one that has regained the potential to become any number of cells in the body.
Strange New Tricks With Light
Authors: Service Robert F. , Cho Adrian
Published: December 2010

In the past decade, physicists and engineers pioneered new ways to guide and manipulate light, creating lenses that defy the fundamental limit on the resolution of an ordinary lens and even constructing "cloaks" that make an object invisible—sort of.
Evaluation of a Rapid and Completely Automated Real-Time Reverse Transcriptase PCR Assay for Diagnosis of Enteroviral Meningitis
Authors: Nolte Frederick S. , Rogers Beverly B. , Tang Yi-Wei , Oberste M. Steven , Robinson Christine C. , Kehl K. Sue , Rand Kenneth A. , Rotbart Harley A. , Romero Jose R. , Nyquist Ann-Christine , Persing David H.
Published: December 2010

Nucleic acid amplification tests (NAATs) for enterovirus RNA in cerebrospinal fluid (CSF) have emerged as the new gold standard for diagnosis of enteroviral meningitis, and their use can improve the management and decrease the costs for caring for children with enteroviral meningitis. The Xpert EV assay (Cepheid, Sunnyvale, CA) is a rapid, fully automated real-time PCR test for the detection of enterovirus RNA that was approved by the U.S. Food and Drug Administration for
in vitro
diagnostic use in March 2007. In this multicenter trial we established the clinical performance characteristics of the Xpert EV assay in patients presenting with meningitis symptoms relative to clinical truth. Clinical truth for enteroviral meningitis was defined as clinical evidence of meningitis, the absence of another detectable pathogen in CSF, and detection of enterovirus in CSF either by two reference NAATs or by viral culture. A total of 199 prospectively and 235 retrospectively collected specimens were eligible for inclusion in this study. The overall prevalence of enteroviral meningitis was 26.04%. The Xpert EV assay had a sensitivity of 94.69% (90% confidence interval [CI] = 89.79 to 97.66%), specificity of 100% (90% CI = 99.07 to 100%), positive predictive value of 100%, negative predictive value of 98.17, and an accuracy of 98.62% relative to clinical truth. The Xpert EV assay demonstrated a high degree of accuracy for diagnosis of enteroviral meningitis. The simplicity and on-demand capability of the Xpert EV assay should prove to be a valuable adjunct to the evaluation of suspected meningitis cases.
High Proportion of Wrongly Identified Methicillin-Resistant Staphylococcus aureus Carriers by Use of a Rapid Commercial PCR Assay Due to Presence of Staphylococcal Cassette Chromosome Element Lacking the
mecA
Gene
Authors: Blanc Dominique S. , Basset Patrick , Nahimana-Tessemo Immaculée , Jaton Katia , Greub Gilbert , Zanetti Giorgio
Published: December 2010

During a 9-month period, 217 patients were newly diagnosed as methicillin-resistant
Staphylococcus aureus
(MRSA) carriers by using a commercial rapid PCR-based test (GeneXpert). However, no MRSA was recovered by culturing the second swab in 61 of these patients. Further analyses showed that 28 (12.9%) of the patients harbored
S. aureus
isolates with a staphylococcal cassette chromosome element lacking the
mecA
gene and were thus incorrectly determined to be MRSA carriers.
Wild-Type MIC Distributions and Epidemiological Cutoff Values for Posaconazole and Voriconazole and Candida spp. as Determined by 24-Hour CLSI Broth Microdilution
Authors: Pfaller M. A. , Boyken L. , Hollis R. J. , Kroeger J. , Messer S. A. , Tendolkar S. , Diekema D. J.
Published: December 2010

We tested 16,191 strains of
Candida
against posaconazole and voriconazole, using the CLSI M27-A3 broth microdilution (BMD) method (24-h incubation), in order to define wild-type (WT) populations and epidemiological cutoff values (ECVs). From 2001 to 2009, 8,619 isolates of
Candida albicans
, 2,415 isolates of
C. glabrata
, 2,278 isolates of
C. parapsilosis
, 1,895 isolates of
C. tropicalis
, 508 isolates of
C. krusei
, 205 isolates of
C. lusitaniae
, 177 isolates of
C. guilliermondii
, and 93 isolates of
C. kefyr
were obtained from over 100 centers worldwide. The modal MICs (μg/ml) for posaconazole and voriconazole, respectively, were as follows: for
C. albicans
, 0.016 and 0.007; for
C. glabrata
, 0.5 and 0.06; for
C. parapsilosis
, 0.06 and 0.007; for
C. tropicalis
, 0.03 and 0.015; for
C. krusei
, 0.25 and 0.12; for
C. lusitaniae
, 0.03 and 0.007; for
C. guilliermondii
, 0.12 and 0.03; and for
C. kefyr
, 0.06 and 0.007. The ECVs (μg/ml [% of isolates that had MICs equal to or less than the ECV]) for posaconazole and voriconazole, respectively, were as follows: 0.06 (98.5) and 0.03 (98.9) for
C. albicans
, 2 (96.2) and 0.5 (90.4%) for
C. glabrata
, 0.25 (99.3) and 0.12 (97.9) for
C. parapsilosis
, 0.12 (97.6) and 0.06 (97.2) for
C. tropicalis
, 0.5 (99.8) and 0.5 (99.4) for
C. krusei
, 0.12 (95.6) and 0.03 (96.6) for
C. lusitaniae
, 0.5 (98.9) and 0.25 (98.3) for
C. guilliermondii
, and 0.25 (100.0) and 0.015 (100.0) for
C. kefyr
. In the absence of clinical breakpoints (CBPs) for posaconazole, these WT distributions and ECVs will be useful in surveillance for emergence of reduced susceptibility to posaconazole among
Candida
spp. Whereas a CBP for susceptibility of ≤1 μg/ml has been established for voriconazole and all species of
Candida
, it is notable that ECVs for this agent range from 10- to >100-fold lower than the CBP, depending on the species of
Candida
. The CBP is inadequate in detecting the emergence of voriconazole resistance among most
Candida
species encountered clinically. The CBPs for voriconazole should be reassessed, with consideration for development of species-specific CBPs.
Inhibition of Early Stages of HIV-1 Assembly by INI1/hSNF5 Transdominant Negative Mutant S6
Authors: Cano Jennifer , Kalpana Ganjam V.
Published: December 2010

INI1/hSNF5 is an HIV-1 integrase (IN) binding protein specifically incorporated into virions. A truncated mutant of INI1 (S6, amino acids 183 to 294) harboring the minimal IN binding Rpt1 domain potently inhibits HIV-1 particle production in a transdominant manner. The inhibition requires interaction of S6 with IN within Gag-Pol. While INI1 is a nuclear protein and harbors a masked nuclear export signal (NES), the transdominant negative mutant S6 is cytoplasmic, due to the unmasking of NES. Here, we examined the effects of subcellular localization of S6 on HIV-1 inhibition and further investigated the stages of assembly that are affected. We found that targeting a nuclear localization signal-containing S6 variant [NLS-S6(Rpt1)] to the nucleoplasm (but not to the nucleolus) resulted in complete reversal of inhibition of particle production. Electron microscopy indicated that although no electron-dense particles at any stage of assembly were seen in cells expressing S6, virions were produced in cells expressing the rescue mutant NLS-S6(Rpt1) to wild-type levels. Immunofluorescence analysis revealed that p24 exhibited a diffuse pattern of localization within the cytoplasm in cells expressing S6 in contrast to accumulation along the membrane in controls. Pulse-chase analysis indicated that in S6-expressing cells, although Gag(Pr55
gag
) protein translation was unaffected, processing and release of p24 were defective. Together, these results indicate that expression of S6 in the cytoplasm interferes with trafficking of Gag-Pol/Gag to the membrane and causes a defective processing leading to inhibition of assembly at an early stage prior to particle formation and budding.
Genetics Coupled to Quantitative Intact Proteomics Links Heritable Aphid and Endosymbiont Protein Expression to Circulative Polerovirus Transmission
Authors: Cilia M. , Tamborindeguy C. , Fish T. , Howe K. , Thannhauser T. W. , Gray S.
Published: December 2010

Yellow dwarf viruses in the family
Luteoviridae
, which are the causal agents of yellow dwarf disease in cereal crops, are each transmitted most efficiently by different species of aphids in a circulative manner that requires the virus to interact with a multitude of aphid proteins. Aphid proteins differentially expressed in F2
Schizaphis graminum
genotypes segregating for the ability to transmit
Cereal yellow dwarf virus-RPV
(CYDV-RPV) were identified using two-dimensional difference gel electrophoresis (DIGE) coupled to either matrix-assisted laser desorption ionization-tandem mass spectrometry or online nanoscale liquid chromatography coupled to electrospray tandem mass spectrometry. A total of 50 protein spots, containing aphid proteins and proteins from the aphid's obligate and maternally inherited bacterial endosymbiont,
Buchnera
, were identified as differentially expressed between transmission-competent and refractive aphids. Surprisingly, in virus transmission-competent F2 genotypes, the isoelectric points of the
Buchnera
proteins did not match those in the maternal
Buchnera
proteome as expected, but instead they aligned with the
Buchnera
proteome of the transmission-competent paternal parent. Among the aphid proteins identified, many were involved in energy metabolism, membrane trafficking, lipid signaling, and the cytoskeleton. At least eight aphid proteins were expressed as heritable, isoelectric point isoform pairs, one derived from each parental lineage. In the F2 genotypes, the expression of aphid protein isoforms derived from the competent parental lineage aligned with the virus transmission phenotype with high precision. Thus, these isoforms are candidate biomarkers for CYDV-RPV transmission in
S. graminum
. Our combined genetic and DIGE approach also made it possible to predict where several of the proteins may be expressed in refractive aphids with different barriers to transmission. Twelve proteins were predicted to act in the hindgut of the aphid, while six proteins were predicted to be associated with the accessory salivary glands or hemolymph. Knowledge of the proteins that regulate virus transmission and their predicted locations will aid in understanding the biochemical mechanisms regulating circulative virus transmission in aphids, as well as in identifying new targets to block transmission.
Basic Residues in the Nucleocapsid Domain of Gag Are Critical for Late Events of HIV-1 Budding
Authors: Dussupt Vincent , Sette Paola , Bello Nana F. , Javid Melodi P. , Nagashima Kunio , Bouamr Fadila
Published: December 2010

The p6 region of HIV-1 Gag contains two late (L) domains, PTAP and LYPXnL, that bind the cellular proteins Tsg101 and Alix, respectively. These interactions are thought to recruit members of the host fission machinery (ESCRT) to facilitate HIV-1 release. Here we report a new role for the p6-adjacent nucleocapsid (NC) domain in HIV-1 release. The mutation of basic residues in NC caused a pronounced decrease in virus release from 293T cells, although NC mutant Gag proteins retained the ability to interact with cellular membranes and RNAs. Remarkably, electron microscopy analyses of these mutants revealed arrested budding particles at the plasma membrane, analogous to those seen following the disruption of the PTAP motif. This result indicated that the basic residues in NC are important for virus budding. When analyzed in physiologically more relevant T-cell lines (Jurkat and CEM), NC mutant viruses remained tethered to the plasma membrane or to each other by a membranous stalk, suggesting membrane fission impairment. Remarkably, NC mutant release defects were alleviated by the coexpression of a Gag protein carrying a wild-type (WT) NC domain but devoid of all L domain motifs and by providing alternative access to the ESCRT pathway, through the in
trans
expression of the ubiquitin ligase Nedd4.2s. Since NC mutant Gag proteins retained the interaction with Tsg101, we concluded that NC mutant budding arrests might have resulted from the inability of Gag to recruit or utilize members of the host ESCRT machinery that act downstream of Tsg101. Together, these data support a model in which NC plays a critical role in HIV-1 budding.
Interplay between PKCδ and Sp1 on Histone Deacetylase Inhibitor-Mediated Epstein-Barr Virus Reactivation
Authors: Tsai Pei-Fang , Lin Sue-Jane , Weng Pei-Lun , Tsai Shu-Chun , Lin Jiun-Han , Chou Ya-Ching , Tsai Ching-Hwa
Published: December 2010

Epstein-Barr virus (EBV) undergoes latent and lytic replication cycles, and its reactivation from latency to lytic replication is initiated by expression of the two viral immediate-early transactivators, Zta and Rta.
In vitro
, reactivation of EBV can be induced by anti-immunoglobulin, tetradecanoyl phorbol acetate, and histone deacetylase inhibitor (HDACi). We have discovered that protein kinase C delta (PKCδ) is required specifically for EBV reactivation by HDACi. Overexpression of PKCδ is sufficient to induce the activity of the Zta promoter (Zp) but not of the Rta promoter (Rp). Deletion analysis revealed that the ZID element of Zp is important for PKCδ activation. Moreover, the Sp1 putative sequence on ZID is essential for PKCδ-induced Zp activity, and the physiological binding of Sp1 on ZID has been confirmed. After HDACi treatment, activated PKCδ can phosphorylate Sp1 at serine residues and might result in dissociation of the HDAC2 repressor from ZID. HDACi-mediated HDAC2-Sp1 dissociation can be inhibited by the PKCδ inhibitor, Rotterlin. Furthermore, overexpression of HDAC2 can suppress the HDACi-induced Zp activity. Consequently, we hypothesize that HDACi induces PKCδ activation, causing phosphorylation of Sp1, and that the interplay between PKCδ and Sp1 results in the release of HDAC2 repressor from Zp and initiation of Zta expression.
Peroxiredoxin 1 Is Required for Efficient Transcription and Replication of Measles Virus
Authors: Watanabe Akira , Yoneda Misako , Ikeda Fusako , Sugai Akihiro , Sato Hiroki , Kai Chieko
Published: December 2010

Measles is a highly contagious human disease caused by the measles virus (MeV). In this study, by proteomic analysis, we identified peroxiredoxin 1 (Prdx1) as a host factor that binds to the C-terminal region of the nucleoprotein (N; NTAIL) of MeV. GlutathioneS-transferase (GST) pulldown experiments showed that the Prdx1-binding site overlapped with the MeV phosphoprotein (P)-binding site on NTAILand that Prdx1 competed for the binding to NTAILwith the P protein, which is a component of RNA-dependent RNA polymerase (RdRp). Furthermore, RNA interference for Prdx1 resulted in a significant reduction in MeV growth in HEK293-SLAM cells. A minigenome assay indicated that Prdx1 suppression affected the viral RNA transcription and/or replication step. Relative quantification of viral RNA by real-time PCR (RT-PCR) showed that Prdx1 suppression not only reduced viral RNA transcription and replication but also enhanced polar attenuation in viral mRNA transcription. Surface plasmon resonance analysis showed that the binding affinity of Prdx1 to MeV-N was 40-fold lower than that of MeV-P to MeV-N, which suggested that Prdx1 might be involved in the early stage of MeV infection, when the expression level of Prdx1 was much higher than that of MeV-P. Since Prdx1 was expressed abundantly and constitutively in various cells, the results in this study indicate that Prdx1 is one of the inherent host factors implicated in MeV RNA synthesis.
Characterization of a
Xenopus tropicalis
Endogenous Retrovirus with Developmental and Stress-Dependent Expression
Authors: Sinzelle L. , Carradec Q. , Paillard E. , Bronchain O. J. , Pollet N.
Published: December 2010

We report on the identification and characterization of XTERV1, a full-length endogenous retrovirus (ERV) within the genome of the western clawed frog (
Xenopus tropicalis
). XTERV1 contains all the basic genetic elements common to ERVs, including the classical 5′-long terminal repeat (LTR)-
gag
-
pol
-
env
-3′-LTR architecture, as well as conserved functional motifs inherent to each retroviral protein. Using phylogenetic analysis, we show that XTERV1 is related to the
Epsilonretrovirus
genus. The
X. tropicalis
genome harbors a single full-length copy with intact
gag
and
pol
open reading frames that localizes to the centromeric region of chromosome 5. About 10 full-length defective copies of XTERV1 are found interspersed in the genome, and 2 of them could be assigned to chromosomes 1 and 3. We find that XTERV1 genes are zygotically transcribed in a regulated spatiotemporal manner during frog development, including metamorphosis. Moreover, XTERV1 transcription is upregulated under certain cellular stress conditions, including cytotoxic and metabolic stresses. Interestingly, XTERV1 Env is found to be homologous to FR47, a protein upregulated following cold exposure in the freeze-tolerant wood frog (
Rana sylvatica
). In addition, we find that
R. sylvatica
FR47 mRNA originated from a retroviral element. We discuss the potential role(s) of ERVs in physiological processes in vertebrates.
Targeting Hepatitis B Virus-Infected Cells with a T-Cell Receptor-Like Antibody
Authors: Sastry Konduru S. R. , Too Chien Tei , Kaur Kaval , Gehring Adam J. , Low Lionel , Javiad Alia , Pollicino Teresa , Li Li , Kennedy Patrick T. F. , Lopatin Uri , Macary Paul A. , Bertoletti Antonio
Published: December 2010

Virus-specific CD8 T cells are activated when their T-cell receptors (TCRs) recognize the specific viral peptide/major histocompatibility complex (MHC) class I (pMHC) complexes present on the surface of infected cells. Antibodies able to recognize the specific pMHC can mimic TCR specificity and both represent a valuable biological tool to visualize pMHC complexes on infected cells and serve as a delivery system for highly targeted therapies. To evaluate these possibilities, we created a monoclonal antibody able to specifically recognize a hepatitis B virus (HBV) envelope epitope (Env at positions 183 to 91 [Env183-91]) presented by the HLA-A201 molecule, and we tested its ability to recognize HBV-infected hepatocytes and to deliver a cargo to a specific target. We demonstrate that this antibody detects and visualizes the processed product of HBV proteins produced in naturally HBV-infected cells, is not inhibited by soluble HBV proteins present in patient sera, and mediates the intracellular delivery of a fluorescent molecule to target cells. Additionally, compared to CD8 T cells specific for the same HBV epitope, the TCR-like antibody has both a superior sensitivity and a specificity focused on distinct amino acids within the epitope. These data demonstrate that a T-cell receptor-like antibody can be used to determine the quantitative relationship between HBV replication and specific antigen presentation to CD8 T cells and serves as a novel therapeutic delivery platform for personalized health care for HBV-infected patients.
Newcastle Disease Virus Expressing a Dendritic Cell-Targeted HIV Gag Protein Induces a Potent Gag-Specific Immune Response in Mice
Authors: Maamary Jad , Array Frida , Gao Qinshan , García-Sastre Adolfo , Steinman Ralph M. , Palese Peter , Nchinda Godwin
Published: December 2010

Viral vaccine vectors have emerged as an attractive strategy for the development of a human immunodeficiency virus (HIV) vaccine. Recombinant Newcastle disease virus (rNDV) stands out as a vaccine vector since it has a proven safety profile in humans, it is a potent inducer of both alpha interferon (IFN-α) and IFN-β) production, and it is a potent inducer of dendritic cell (DC) maturation. Our group has previously generated an rNDV vector expressing a codon-optimized HIV Gag protein and demonstrated its ability to induce a Gag-specific CD8
+
T cell response in mice. In this report we demonstrate that the Gag-specific immune response can be further enhanced by the targeting of the rNDV-encoded HIV Gag antigen to DCs. Targeting of the HIV Gag antigen was achieved by the addition of a single-chain Fv (scFv) antibody specific for the DC-restricted antigen uptake receptor DEC205 such that the DEC205 scFv-Gag molecule was encoded for expression as a fusion protein. The vaccination of mice with rNDV coding for the DC-targeted Gag antigen induced an enhanced Gag-specific CD8
+
T cell response and enhanced numbers of CD4
+
T cells and CD8
+
T cells in the spleen relative to vaccination with rNDV coding for a nontargeted Gag antigen. Importantly, mice vaccinated with the DEC205-targeted vaccine were better protected from challenge with a recombinant vaccinia virus expressing the HIV Gag protein. Here we demonstrate that the targeting of the HIV Gag antigen to DCs via the DEC205 receptor enhances the ability of an rNDV vector to induce a potent antigen-specific immune response.
Double-Stranded RNA Adenosine Deaminase ADAR-1-Induced Hypermutated Genomes among Inactivated Seasonal Influenza and Live Attenuated Measles Virus Vaccines
Authors: Suspène Rodolphe , Petit Vincent , Puyraimond-Zemmour David , Aynaud Marie-Ming , Henry Michel , Guétard Denise , Rusniok Christophe , Wain-Hobson Simon , Vartanian Jean-Pierre
Published: December 2010

We sought to examine ADAR-1 editing of measles and influenza virus genomes derived from inactivated seasonal influenza and live attenuated measles virus vaccines grown on chicken cells as the culture substrate. Using highly sensitive 3DI-PCR (R. Suspène et al., Nucleic Acids Res. 36:e72, 2008), it was possible to show that ADAR-1 could hyperdeaminate adenosine residues in both measles virus and influenza virus A genomes. Detailed analysis of the dinucleotide editing context showed preferences for 5′ArA and 5′UrA, which is typical of editing in mammalian cells. The hyperedited mutant frequency, including genomes and antigenomes, was a log greater for influenza virus compared to measles virus, suggesting a greater sensitivity to restriction by ADAR-1.
Production of Hepatitis C Virus Lacking the Envelope-Encoding Genes for Single-Cycle Infection by Providing Homologous Envelope Proteins or Vesicular Stomatitis Virus Glycoproteins in
trans
Authors: Li Rui , Qin Yan , He Ying , Tao Wanyin , Zhang Nan , Tsai Cheguo , Zhou Paul , Zhong Jin
Published: December 2010

Hepatitis C virus (HCV) infection is a major worldwide health problem. The envelope glycoproteins are the major components of viral particles. Here we developed a
trans
-complementation system that allows the production of infectious HCV particles in whose genome the regions encoding envelope proteins are deleted (HCVΔE). The lack of envelope proteins could be efficiently complemented by the expression of homologous envelope proteins in
trans
. HCVΔE production could be enhanced significantly by previously described adaptive mutations in NS3 and NS5A. Moreover, HCVΔE could be propagated and passaged in packaging cells stably expressing HCV envelope proteins, resulting in only single-round infection in wild-type cells. Interestingly, we found that vesicular stomatitis virus (VSV) glycoproteins could efficiently rescue the production of HCV lacking endogenous envelope proteins, which no longer required apolipoprotein E for virus production. VSV glycoprotein-mediated viral entry could allow for the bypass of the natural HCV entry process and the delivery of HCV replicon RNA into HCV receptor-deficient cells. Our development provides a new tool for the production of single-cycle infectious HCV particles, which should be useful for studying individual steps of the HCV life cycle and may also provide a new strategy for HCV vaccine development.
Spread of Herpes Simplex Virus to the Spinal Cord Is Independent of Spread to Dorsal Root Ganglia
Authors: Ohashi Masahiro , Bertke Andrea S. , Patel Amita , Krause Philip R.
Published: December 2010

Levels of herpes simplex virus 1 (HSV-1) and HSV-2 DNA in dorsal root ganglia (DRG) and spinal cord (SC) were quantified after inoculation of guinea pig genitals and footpads. In genital infection, viral DNA reached SC and DRG simultaneously (at 2 to 3 days after inoculation) but was more abundant in SC than in DRG. After inoculation of footpads, which lack parasympathetic innervation, the viruses spread more efficiently to DRG than to SC. These results show important differences between genital and footpad infections, including independence of spread to DRG and SC, and imply that autonomic neurons may play an important role in the pathogenesis of viral latency after genital inoculation.
Inverse relationship between percentage body weight change and finishing time in 643 forty-two-kilometre marathon runners
Authors: Zouhal Hassane , Groussard Carole , Minter Guenolé , Vincent Sophie , Cretual Armel , Gratas-Delamarche Arlette , Delamarche Paul , Noakes Timothy David
Published: December 2010

Objective

The purpose of this study was to determine the relationship between athletic performance and the change in body weight (BW) during a 42 km marathon in a large cohort of runners.

Methods

The study took place during the 2009 Mont Saint-Michel Marathon (France). 643 marathon finishers (560 males and 83 females) were studied. The change in BW during the race was calculated from measurements of each runner's BW immediately before and after the race.

Results

BW loss was 2.3±2.2% (mean±SEM) (p<0.01). BW loss was −3.1±1.9% for runners finishing the marathon in less than 3 h; −2.5±2.1% for runners finishing between 3 and 4 h; and −1.8±2.4% for runners who required more than 4 h to complete the marathon. The degree of BW loss was linearly related to 42 km race finishing time (p<0.0000001). Neither age nor gender influenced BW loss during the race.

Conclusions

BW loss during the marathon was inversely related to race finishing time in 643 marathon runners and was >3% in runners completing the race in less than 3 h. These data are not compatible with laboratory-derived data suggesting that BW loss greater than 2% during exercise impairs athletic performance. They match an extensive body of evidence showing that the most successful athletes in marathon and ultra-marathon running and triathlon events are frequently those who lose substantially more than 3–4% BW during competition.
20-Hydroxyvitamin D2 is a noncalcemic analog of vitamin D with potent antiproliferative and prodifferentiation activities in normal and malignant cells
Authors: Slominski Andrzej T. , Kim Tae-Kang , Janjetovic Zorica , Tuckey Robert C. , Bieniek Radoslaw , Yue Junming , Li Wei , Chen Jianjun , Nguyen Minh N. , Tang Edith K. Y. , Miller Duane , Chen Tai C. , Holick Michael
Published: December 2010

20-hydroxyvitamin D2 [20(OH)D2] inhibits DNA synthesis in epidermal keratinocytes, melanocytes, and melanoma cells in a dose- and time-dependent manner. This inhibition is dependent on cell type, with keratinocytes and melanoma cells being more sensitive than normal melanocytes. The antiproliferative activity of 20(OH)D2 is similar to that of 1,25(OH)2D3 and of newly synthesized 1,20(OH)2D2 but significantly higher than that of 25(OH)D3. 20(OH)D2 also displays tumorostatic effects. In keratinocytes 20(OH)D2 inhibits expression of cyclins and stimulates involucrin expression. It also stimulates CYP24 expression, however, to a significantly lower degree than that by 1,25(OH)2D3 or 25(OH)D3. 20(OH)D2 is a poor substrate for CYP27B1 with overall catalytic efficiency being 24- and 41-fold lower than for 25(OH)D3 with the mouse and human enzymes, respectively. No conversion of 20(OH)D2 to 1,20(OH)2D2 was detected in intact HaCaT keratinocytes. 20(OH)D2 also demonstrates anti-leukemic activity but with lower potency than 1,25(OH)2D3. The phenotypic effects of 20(OH)D2 are mediated through interaction with the vitamin D receptor (VDR) as documented by attenuation of cell proliferation after silencing of VDR, by enhancement of the inhibitory effect through stable overexpression of VDR and by the demonstration that 20(OH)D2 induces time-dependent translocation of VDR from the cytoplasm to the nucleus at a comparable rate to that for 1,25(OH)2D3. In vivo tests show that while 1,25(OH)2D3 at doses as low as 0.8 μg/kg induces calcium deposits in the kidney and heart, 20(OH)D2 is devoid of such activity even at doses as high as 4 μg/kg. Silencing of CY27B1 in human keratinocytes showed that 20(OH)D2 does not require its transformation to 1,20(OH)2D2 for its biological activity. Thus 20(OH)D2 shows cell-type dependent antiproliferative and prodifferentiation activities through activation of VDR, while having no detectable toxic calcemic activity, and is a poor substrate for CYP27B1.