bluuebunny/crossref_metadata_2025 huggingface dataset:
Towards a common carbon footprint assessment methodology for the water sector
Authors: Frijns Jos
Published: May 2011

This paper describes the carbon footprint methodology used in assessing the global warming potential of the Dutch water sector. The assessment includes CO2 emissions from energy consumption and methane and nitrous oxide emissions from water treatment processes. There is, however, debate on the amounts and mechanism of greenhouse gas (GHG) emissions, and a standardised approach is discussed. As a result of this approach, the contribution of GHG emissions to the total carbon footprint of the Dutch water sector appeared to be relatively high. Next to the lack of common emission factors for GHG and chemicals used, there is also no agreed‐upon approach related to the system boundaries and scope of carbon footprinting of the water cycle. For reasons of benchmarking and monitoring of climate change reduction targets, a common carbon footprint assessment methodology for the water sector will be required.
Analytical and Sensory Characterization of the Aroma of “Langhe D.O.C. Nebbiolo” Wines: Influence of the Prefermentative Cold Maceration with Dry Ice
Authors: Petrozziello Maurizio , Guaita Massimo , Motta Silvia , Panero Loretta , Bosso Antonella
Published: May 2011

Abstract:  The present work concerns the determination of the volatile compounds, as well as the description of the olfactory characteristics, of “Langhe DOC Nebbiolo” wines produced through the prefermentative cold maceration with dry ice, on full scale (30 quintals) in private cellars. The free volatile compounds and those obtained by enzymatic and chemical hydrolysis of the glycosilated precursors were analyzed by gas chromatography‐mass spectrometry. Among free volatile compounds, a loss of alcohols with 6 carbon atoms in the cryo‐macerated trials was observed. The cryo‐maceration technique, when correctly applied, increased the floral and fruity notes (blackberry, cherry, and plum). This increase is related to the extraction of polyphenolic compounds (maceration effect), but no volatile compounds, neither in free form, nor produced by enzymatic hydrolysis of the glycosilated precursors, can discriminate the wines according to their content of polyphenols. Only some compounds produced by chemical hydrolysis of the precursors can discriminate the wines (statistically significant differences): 3‐hydroxy‐β‐damascone, methylvanillate, and blumenol C. These compounds, however, form during aging and do not have any influence on young wines aroma.
Antioxidant Capacity Changes of 3 Cultivar Chinese Pomegranate (Punica granatum L.) Juices and Corresponding Wines
Authors: Zhuang Huiting , Du Jinhua , Wang Yongmei
Published: May 2011

Abstract:  This study investigated the antioxidant capacities that included Ferric reducing antioxidant power (FRAP), 1, 1‐diphenyl‐2‐picryl hydrazine (DPPH), ·OH and O2·−‐scavenging abilities, total polyphenols (TP) and total anthocyanins (TA) in pomegranate (Punica granatum L.) juice (PJ) and pomegranate wine (PW). The correlations among them were also analyzed. Both PJ and PW showed significantly high TP and antioxidant capacities, but some differences existed among these cultivars. There was 1596.67 mg/L TP found in sweet PJ, while sour PJ showed the highest titratable acidity of 35.90 g/L and lowest pH value at 2.56. Red PJ was found to have the highest TA (82.26 mg/L) in the 3 cultivars. Sweet PJ showed higher DPPH‐scavenging ability and higher FRAP than others. Both PJ and PW exhibited high and relatively stable ·OH‐scavenging abilities, in which sour PJ and sour PW had higher O2·− scavenging capacity than others. Significant positive correlations were observed among TP, DPPH, and FRAP in both PJ and PW. A high correlation between antioxidant capacities and TP indicated that phenolic compounds were major contributors to the high antioxidant activity of PJ and PW.

Practical Application:  The information presented might be valuable for developing functional foods made of Chinese pomegranate.
Potential Risks Resulting from Fruit/Vegetable–Drug Interactions: Effects on Drug‐Metabolizing Enzymes and Drug Transporters
Authors: Rodríguez‐Fragoso Lourdes , Martínez‐Arismendi José Luis , Orozco‐Bustos Danae , Reyes‐Esparza Jorge , Torres Eliseo , Burchiel Scott W.
Published: May 2011

Abstract:  It has been well established that complex mixtures of phytochemicals in fruits and vegetables can be beneficial for human health. Moreover, it is becoming increasingly apparent that phytochemicals can influence the pharmacological activity of drugs by modifying their absorption characteristics through interactions with drug transporters as well as drug‐metabolizing enzyme systems. Such effects are more likely to occur in the intestine and liver, where high concentrations of phytochemicals may occur. Alterations in cytochrome P450 and other enzyme activities may influence the fate of drugs subject to extensive first‐pass metabolism. Although numerous studies of nutrient–drug interactions have been published and systematic reviews and meta‐analyses of these studies are available, no generalizations on the effect of nutrient–drug interactions on drug bioavailability are currently available. Several publications have highlighted the unintended consequences of the combined use of nutrients and drugs. Many phytochemicals have been shown to have pharmacokinetic interactions with drugs. The present review is limited to commonly consumed fruits and vegetables with significant beneficial effects as nutrients and components in folk medicine. Here, we discuss the phytochemistry and pharmacokinetic interactions of the following fruit and vegetables: grapefruit, orange, tangerine, grapes, cranberry, pomegranate, mango, guava, black raspberry, black mulberry, apple, broccoli, cauliflower, watercress, spinach, tomato, carrot, and avocado. We conclude that our knowledge of the potential risk of nutrient–drug interactions is still limited. Therefore, efforts to elucidate potential risks resulting from food–drug interactions should be intensified in order to prevent undesired and harmful clinical consequences.
Pamela and Creative Cognition
Authors: JONES WENDY
Published: May 2011

This article challenges the belief that the mind as a creative, generative entity belongs exclusively to a poetic rather than a novelistic tradition. Pamela offers a general model of creativity through Mr B.'s recreation of his character. This process accords with the findings of cognitive science that creativity has its source in emotion as well as cognition. This argument is embedded in a larger claim, that novels often give portrayals of human nature that are true to neuroscience's view of how the brain works, to suggest that a biocultural focus might enable literary studies to pursue important new directions.
Patterns of scintigraphic uptake in the fetlock joint of Thoroughbred racehorses and the effect of increased radiopharmaceutical uptake in the distal metacarpal/tarsal condyle on performance
Authors: TROPE G. D. , ANDERSON G. A. , WHITTON R. C.
Published: May 2011

Reasons for performing study:The effect of palmar osteochondral disease lesions on performance of Thoroughbred (TB) racehorses is unclear. There is a need to describe patterns of radiopharmaceutical uptake (IRU) in fetlock joints of TB racehorses and to evaluate post scintigraphy performance.

Hypotheses:IRU in the metacarpal (MC) and metatarsal (MT) condyles is more common than IRU in the parasagittal grooves and is associated with poorer post diagnosis performance than controls.

Methods:Location of IRU within the fetlock region was identified and graded subjectively in TB racehorses. Performance variables were determined from race records for horses with moderate/marked MC/MT condylar IRU (cases), other horses undergoing scintigraphy (scintigraphy controls) and age/sex matched controls from the last race in which a case participated (controls). Statistical analyses included quantile regression, Fisher's exact test, Kaplan‐Meier survival curves, log‐rank test and Cox regression.

Results:Metacarpal/MT condylar IRU was identified in 103/220 horses with only 3/220 having parasagittal IRU. Moderate/marked IRU was identified in the MC and MT condylar regions in 62 and 39 horses, respectively, with palmaromedial and plantarolateral IRU most common. Fore‐ and hindlimb cases had fewer starts, reduced earnings (P<0.001) and reduced earnings/start than controls respectively. Cases were more likely to return to racing later than controls following a rest period. Cases were older than scintigraphy controls.

Conclusions:In TB racehorses presenting for lameness MC/MT condylar IRU is the most common abnormality identified within the fetlock joint. Racehorses with moderate/marked condylar IRU have a shortened racing career, are less successful than age/sex matched controls and are older than other racehorses presented for scintigraphy.

Potential relevance:Overload of the MC/MT condyles is a common and significant problem in TB racehorses that is readily identified with scintigraphy. Scintigraphy of horses that are lame or performing poorly is less useful for screening for potential condylar fractures.
Transient Activation of the HOG MAPK Pathway Regulates Bimodal Gene Expression
Authors: Pelet Serge , Rudolf Fabian , Nadal-Ribelles Mariona , de Nadal Eulàlia , Posas Francesc , Peter Matthias
Published: May 2011

Bimodal expression of genes is activated in response to osmotic stress.
Relationship Between Clinical Signs and Transmission of an Infectious Disease and the Implications for Control
Authors: Charleston Bryan , Bankowski Bartlomies M. , Gubbins Simon , Chase-Topping Margo E. , Schley David , Howey Richard , Barnett Paul V. , Gibson Debi , Juleff Nicholas D. , Woolhouse Mark E. J.
Published: May 2011

Livestock experiments provide precise parameters for incubation and infectious periods for foot-and-mouth disease virus.
Designing the Next Generation of Chemical Separation Membranes
Authors: Gin Douglas L. , Noble Richard D.
Published: May 2011

New materials can be prepared as membranes that may allow their performance to beat long-standing limits.
Fitting Science and Screen
Lab Coats in Hollywood
Science, Scientists, and Cinema

by David A. Kirby

MIT Press, Cambridge, MA, 2011. 279 pp. $27.95, £20.95. ISBN 9780262014786.
Authors: MacIver Malcolm A.
Published: May 2011

Offering a fresh perspective on the intersection between science and cinema, Kirby focuses on the roles scientist-consultants play in attempts to depict science and researchers in movies more plausibly.
Twisted Disks
Authors: Salo Heikki
Published: May 2011

Ripple patterns in Saturn's and Jupiter's rings result from collisions with comets.
Indigenous Genomics
Authors: Hayes Vanessa
Published: May 2011

Studies of indigenous peoples are a crucial part of genomic research, not only to define the extent of human diversity but to provide medical benefit to all people. There are more than 370 million indigenous people living in almost half the countries of the world. Exploding interest in indigenous genomics and global population structure has raised debate about issues of informed consent and community benefit. As was evident in March 2011 during the African and Southern African Society of Human Genetics Meeting in Cape Town, South Africa, the inclusion of indigenous people in future genomic research is paramount, but ethical guidelines must address local concerns. Scientific practices and values must be integrated with indigenous governance so that such genomic research can continue, with the benefits fully realized by all.
Jürg Tschopp (1951–2011)
Authors: O'Neill Luke A.
Published: May 2011

A scientist's basic research into cell death and inflammation will continue to have a clinical impact.
EPA Proposal Would Exempt Some GMOs From Registry
Authors: Reardon Sara
Published: May 2011

The U.S. Environmental Protection Agency has proposed a draft rule that would exempt cisgenic organisms—which contain only genetic material from sexually compatible species—from the requirement to be registered with EPA before being field-tested or marketed.
Anonymous Alcoholic Bankrolls Trial of Controversial Therapy
Authors: Enserink Martin
Published: May 2011

An anonymous Dutch donor has given the University of Amsterdam a half-million euros to conduct a rigorous placebo-controlled study of baclofen to treat addiction.
Red in Tooth and Claw Among the Literati
Authors: Kean Sam
Published: May 2011

Upset by the isolation of their field, some critics are trying to bring Darwin's ideas and recent science to the study of literature. They haven't been popular.
Dating Duo Illuminates Modern Humans' Journey
Authors: Balter Michael
Published: May 2011

By improving a powerful dating technique, a professional and personal couple fills in the blanks of human evolution.
Ultrasensitive Detection of Drug-Resistant Pandemic 2009 (H1N1) Influenza A Virus by Rare-Variant-Sensitive High-Resolution Melting-Curve Analysis
Authors: Chen Neng , Pinsky Benjamin A. , Lee Betty P. , Lin Min , Schrijver Iris
Published: May 2011

Oseltamivir (Tamiflu), an oral neuraminidase inhibitor, has been widely used to treat pandemic 2009 (H1N1) influenza A. Although a majority of 2009 (H1N1) influenza A virus remains oseltamivir susceptible, the threat of resistance due to the His275Tyr mutation is highlighted by the limitations of alternative therapies and the potential for rapid, global fixation of this mutation in the circulating influenza A virus population. In order to better understand the emergence of resistance, we developed a rare-variant-sensitive high-resolution melting-curve analysis method (RVS-HRM) that is able to detect the His275Tyr oseltamivir resistance mutation to 0.5% in a background of susceptible virus. We applied RVS-HRM to clinical specimens from patients who developed oseltamivir resistance and demonstrated the ultrasensitive detection of influenza A virus N1 neuraminidase quasispecies. Interestingly, we were unable to detect the oseltamivir resistance mutation in pretreatment samples, suggesting that resistant virus does not reach even this very low detection threshold until exposed to selective drug pressure. Thus, patients naive to oseltamivir are most likely to be susceptible when this drug is used as a first-line treatment modality.
Combined Deletions of
pfhrp2
and
pfhrp3
Genes Result in Plasmodium falciparum Malaria False-Negative Rapid Diagnostic Test
Authors: Houzé Sandrine , Hubert Véronique , Le Pessec Gaëlle , Le Bras Jacques , Clain Jérôme
Published: May 2011

We report a case of misdiagnosis of
Plasmodium falciparum
malaria from Brazil with negative PfHRP2-based rapid diagnostic tests (RDTs), leading to inappropriate case management. Genetic tests showed the deletion of both
pfhrp2
and
pfhrp3
genes. The detection of two distinct
P. falciparum
target antigens is then advisable.
Quality Control Guidelines for Amphotericin B, Itraconazole, Posaconazole, and Voriconazole Disk Diffusion Susceptibility Tests with Nonsupplemented Mueller-Hinton Agar (CLSI M51-A Document) for Nondermatophyte Filamentous Fungi
Authors: Espinel-Ingroff A. , Canton E. , Fothergill A. , Ghannoum M. , Johnson E. , Jones R. N. , Ostrosky-Zeichner L. , Schell W. , Gibbs D. L. , Wang A. , Turnidge J.
Published: May 2011

Although Clinical and Laboratory Standards Institute (CLSI) disk diffusion assay standard conditions are available for susceptibility testing of filamentous fungi (molds) to antifungal agents, quality control (QC) disk diffusion zone diameter ranges have not been established. This multicenter study documented the reproducibility of tests for one isolate each of five molds (
Paecilomyces variotii
ATCC MYA-3630,
Aspergillus fumigatus
ATCC MYA-3626,
A. flavus
ATCC MYA-3631,
A. terreus
ATCC MYA-3633, and
Fusarium verticillioides
[
moniliforme
] ATCC MYA-3629) and
Candida krusei
ATCC 6258 by the CLSI disk diffusion method (M51-A document). The zone diameter ranges for selected QC isolates were as follows:
P. variotii
ATCC MYA-3630, amphotericin B (15 to 24 mm), itraconazole (20 to 31 mm), and posaconazole (33 to 43 mm);
A. fumigatus
ATCC MYA-3626, amphotericin B (18 to 25 mm), itraconazole (11 to 21 mm), posaconazole (28 to 35 mm), and voriconazole (25 to 33 mm); and
C. krusei
, amphotericin B (18 to 27 mm), itraconazole (18 to 26 mm), posaconazole (28 to 38 mm), and voriconazole (29 to 39 mm). Due to low testing reproducibility, zone diameter ranges were not proposed for the other three molds.
Point-Counterpoint: Should All Stools Be Screened for Shiga Toxin-Producing Escherichia coli?
Authors: Marcon Mario J. , Kiska Deanna L. , Riddell Scott W. , Gilligan Peter
Published: May 2011

In October 2009, the Centers for Disease Control and Prevention recommended that clinical laboratories test all stools submitted for the detection of enteric bacterial pathogens for the presence of Shiga toxin-producing
Escherichia coli
(STEC). In order to do this, it is recommended that all stools be cultured for
Escherichia coli
O157:H7 on selective medium as well as that testing for the presence of Shiga toxin be done by immunoassay to detect non-O157 STEC (3). There are a variety of products that are FDA approved for detection of Shiga toxin. Further, it is recommended that Shiga toxin detection be done by testing overnight enrichment broth cultures of stools rather than directly examining stools for this toxin. This recommendation was made approximately 18 months ago. We have asked Mario Marcon of Nationwide's Children Hospital in Columbus, OH, to explain the rationale for his decision to follow this recommendation, while we have asked Deanna Kiska and Scott Riddell of Upstate University Hospital in Syracuse, NY, why these guidelines have not been adopted by their laboratory.
Frequent Detection of Respiratory Viruses without Symptoms: Toward Defining Clinically Relevant Cutoff Values
Authors: Jansen Rogier R. , Wieringa Joanne , Koekkoek Sylvie M. , Visser Caroline E. , Pajkrt Dasja , Molenkamp Richard , de Jong Menno D. , Schinkel Janke
Published: May 2011

Highly sensitive techniques, such as PCR, have greatly improved the detection of respiratory viruses. However, the sensitivity of PCR tests also complicates clinical interpretation, as the presence of small amounts of viral targets may not necessarily have clinical relevance. We performed a prospective case-control study in asymptomatic and symptomatic young children. PCR detection of 14 respiratory viruses was performed in nasal washes, and results were quantified in copies per milliliter. A total of 141 cases and 157 controls were included. In 72% of the cases and 28% of the controls, at least one virus was identified. When stratified for age, at least one virus was identified in 47% of the controls younger than 1 year old. Rhinovirus (RV) was frequently detected in both symptomatic and asymptomatic individuals. Receiver operating characteristic analysis for quantitative rhinovirus detection showed that cutoff values for clinical relevance are feasible for RV. In contrast to rhinovirus, respiratory syncytial virus (RSV) was rarely detected in controls, suggesting that a positive RSV test result is almost always of clinical relevance, independent of viral quantity. In conclusion, our study shows that asymptomatic carriage of a respiratory virus occurs frequently in young children. However, significant differences in the amount of virus present were observed between cases and controls. This suggests that defining cutoff levels should be feasible and represents the next necessary step for diagnosing viral respiratory infections using molecular tests.
Kaposi's Sarcoma-Associated Herpesvirus Infection of Endothelial Cells Inhibits Neutrophil Recruitment through an Interleukin-6-Dependent Mechanism: a New Paradigm for Viral Immune Evasion
Authors: Butler L. M. , Jeffery H. C. , Wheat R. L. , Rae P. C. , Townsend K. , Alkharsah K. R. , Schulz T. F. , Nash G. B. , Blackbourn D. J.
Published: May 2011

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of Kaposi's sarcoma (KS), an endothelial cell (EC) neoplasm characterized by dysregulated angiogenesis and inflammation. KSHV infection of EC causes production of proinflammatory mediators, regarded as possible initiators of the substantial mononuclear leukocyte recruitment seen in KS. Conversely, KSHV immune evasion strategies exist, such as degradation of EC leukocyte adhesion receptors by viral proteins. Here, we report the effects of KSHV infection of primary EC on recruitment of flowing leukocytes. Infection did not initiate adhesion of any leukocyte subsetper se. However, on cytokine-stimulated EC, KSHV specifically inhibited neutrophil, but not PBL or monocyte, transmigration, an observation consistent with the inflammatory cell profile found in KS lesionsin vivo. This inhibition could be recapitulated on uninfected EC using supernatant from infected cultures. These supernatants contained elevated levels of human interleukin 6 (hIL-6), and both the KSHV- and the supernatant-induced inhibitions of neutrophil transmigration were abrogated in the presence of a hIL-6 neutralizing antibody. Furthermore, preconditioning of EC with hIL-6 mimicked the effect of KSHV. Using RNA interference (RNAi), we show that upregulation of suppressor of cytokine signaling 3 (SOCS3) was necessary for this effect of hIL-6. These studies reveal a novel paracrine mode of KSHV immune evasion, resulting in reduced recruitment of neutrophils, a cell type whose antiviral and antitumor roles are becoming increasingly appreciated. Moreover, the findings have implications for our understanding of the contribution of hIL-6 to the pathogenesis of other inflammatory disorders and tumors in which this cytokine is abundant.
An HIV-1 gp120 Envelope Human Monoclonal Antibody That Recognizes a C1 Conformational Epitope Mediates Potent Antibody-Dependent Cellular Cytotoxicity (ADCC) Activity and Defines a Common ADCC Epitope in Human HIV-1 Serum
Authors: Ferrari Guido , Pollara Justin , Kozink Daniel , Harms Tiara , Drinker Mark , Freel Stephanie , Moody M. Anthony , Alam S. Munir , Tomaras Georgia D. , Ochsenbauer Christina , Kappes John C. , Shaw George M. , Hoxie James A. , Robinson James E. , Haynes Barton F.
Published: May 2011

Among nonneutralizing HIV-1 envelope antibodies (Abs), those capable of mediating antibody-dependent cellular cytotoxicity (ADCC) activity have been postulated to be important for control of HIV-1 infection. ADCC-mediating Ab must recognize HIV-1 antigens expressed on the membrane of infected cells and bind the Fcγ receptor (FcR) of the effector cell population. However, the precise targets of serum ADCC antibody are poorly characterized. The human monoclonal antibody (MAb) A32 is a nonneutralizing antibody isolated from an HIV-1 chronically infected person. We investigated the ability of MAb A32 to recognize HIV-1 envelope expressed on the surface of CD4+T cells infected with primary and laboratory-adapted strains of HIV-1, as well as its ability to mediate ADCC activity. The MAb A32 epitope was expressed on the surface of HIV-1-infected CD4+T cells earlier than the CD4-inducible (CD4i) epitope bound by MAb 17b and the gp120 carbohydrate epitope bound by MAb 2G12. Importantly, MAb A32 was a potent mediator of ADCC activity. Finally, an A32 Fab fragment blocked the majority of ADCC-mediating Ab activity in plasma of subjects chronically infected with HIV-1. These data demonstrate that the epitope defined by MAb A32 is a major target on gp120 for plasma ADCC activity.
Epstein-Barr Virus Zta-Induced Immunomodulators from Nasopharyngeal Carcinoma Cells Upregulate Interleukin-10 Production from Monocytes
Authors: Lee Chien-Hsun , Yeh Tzu-Hao , Lai Hsiao-Ching , Wu Shih-Yi , Su Ih-Jen , Takada Kenzo , Chang Yao
Published: May 2011

During lytic infection with Epstein-Barr virus (EBV), several viral lytic proteins function to evade immune recognition or to actively suppress immune cells. An EBV lytic transactivator, Zta, induces an immunosuppressive cytokine interleukin 10 (IL-10) in B cells, but whether it regulates IL-10 in the context of epithelial cells is unclear. In this study, we tested nasopharyngeal carcinoma (NPC) cell lines and found that Zta did not induce IL-10 in these epithelial cells. Interestingly, the conditioned medium of Zta-expressing NPC cells enhanced IL-10 production from monocytes. We further revealed that the IL-10-inducing effect involved at least two immunomodulators that were upregulated by Zta and secreted from NPC cells: granulocyte-macrophage colony-stimulating factor (GM-CSF) and prostaglandin E2(PGE2). Zta was recruited to and activated the GM-CSF promoter, thus upregulating GM-CSF expression. Zta also activated the promoter of cyclooxygenase-2 (COX-2), and Zta-induced COX-2 increased downstream PGE2production. Cotreatment with GM-CSF and PGE2synergistically induced IL-10 production from monocytes. The IL-10-inducing effect of the Zta-conditioned medium was reduced when GM-CSF or the COX-2/PGE2pathway was blocked. The conditioned medium of NPC cells with EBV lytic infection showed a similar increase of GM-CSF and PGE2levels as well as the IL-10-inducing effect on monocytes, and knockdown of Zta abolished all the effects. Therefore, through Zta-induced immunomodulators, EBV lytic infection in NPC cells can direct bystander monocytes to produce IL-10, which may be a novel way of EBV to promote local immunosuppression.
HIV-1 Nef Disrupts Intracellular Trafficking of Major Histocompatibility Complex Class I, CD4, CD8, and CD28 by Distinct Pathways That Share Common Elements
Authors: Leonard Jolie A. , Filzen Tracy , Carter Christoph C. , Schaefer Malinda , Collins Kathleen L.
Published: May 2011

The Nef protein is an important HIV virulence factor that promotes the degradation of host proteins to augment virus production and facilitate immune evasion. The best-characterized targets of Nef are major histocompatibility complex class I (MHC-I) and CD4, but Nef also has been reported to target several other proteins, including CD8β, CD28, CD80, CD86, and CD1d. To compare and contrast the effects of Nef on each protein, we constructed a panel of chimeric proteins in which the extracellular and transmembrane regions of the MHC-I allele HLA-A2 were fused to the cytoplasmic tails of CD4, CD28, CD8β, CD80, CD86, and CD1d. We found that Nef coprecipitated with and disrupted the expression of molecules with cytoplasmic tails from MHC-I HLA-A2, CD4, CD8β, and CD28, but Nef did not bind to or alter the expression of molecules with cytoplasmic tails from CD80, CD86, and CD1d. In addition, we used short interfering RNA (siRNA) knockdown and coprecipitation experiments to implicate AP-1 as a cellular cofactor for Nef in the downmodulation of both CD28 and CD8β. The interaction with AP-1 required for CD28 and CD8β differed from the AP-1 interaction required for MHC-I downmodulation in that it was mediated through the dileucine motif within Nef (LL164,165AA) and did not require the tyrosine binding pocket of the AP-1 μ subunit. In addition, we demonstrate a requirement for β-COP as a cellular cofactor for Nef that was necessary for the degradation of targeted molecules HLA-A2, CD4, and CD8. These studies provide important new information on the similarities and differences with which Nef affects intracellular trafficking and help focus future research on the best potential pharmaceutical targets.
Adaptation of Saffold Virus 2 for High-Titer Growth in Mammalian Cells
Authors: Hertzler Shannon , Liang Zhiguo , Treso Balint , Lipton Howard L.
Published: May 2011

Saffold viruses (SAFV) are a recently discovered group of human
Cardioviruses
closely related to Theiler's murine encephalomyelitis viruses (TMEV). Unlike TMEV and encephalomyocarditis virus, each of which is monotypic, SAFV are genetically diverse and include at least eight genotypes. To date, only Saffold virus 3 (SAFV-3) has been grown efficiently in mammalian cells
in vitro
. Here, we report the successful adaptation of SAFV-2 for efficient growth in HeLa cells after 13 passages in the alpha/beta interferon-deficient human glial cell line U118 MG. Nine amino acid changes were found in the adapted virus, with single mutations in VP2, VP3, and 2B, while 6 mutations arose in VP1. Most capsid mutations were in surface loops. Analysis of SAFV-2 revealed virus growth and cytopathic effect only in human cell lines, with large plaques forming in HeLa cells, with minimal cell association, and without using sialic acid to enter cells. Despite the limited growth of SAFV-2 in rodent cells
in vitro
, BALB/c mice inoculated with SAFV-2 showed antibody titers of >1:10
6
, and fluorescence-activated cell sorting (FACS) analysis revealed only minimal cross-reactivity with SFV-3. Intracerebral inoculation of 6-week-old FVB/n mice produced paralysis and acute neuropathological changes, including meningeal infiltrates, encephalitis, particularly of the limbic system, and spinal cord white matter inflammation.
NS4B Self-Interaction through Conserved C-Terminal Elements Is Required for the Establishment of Functional Hepatitis C Virus Replication Complexes
Authors: Paul David , Romero-Brey Inés , Gouttenoire Jérôme , Stoitsova Savina , Krijnse-Locker Jacomine , Moradpour Darius , Bartenschlager Ralf
Published: May 2011

Hepatitis C virus (HCV) is an important human pathogen, persistently infecting more than 170 million individuals worldwide. Studies of the HCV life cycle have become possible with the development of cell culture systems supporting the replication of viral RNA and the production of infectious virus. However, the exact functions of individual proteins, especially of nonstructural protein 4B (NS4B), remain poorly understood. NS4B triggers the formation of specific, vesicular membrane rearrangements, referred to as membranous webs, which have been reported to represent sites of HCV RNA replication. However, the mechanism of vesicle induction is not known. In this study, a panel of 15 mutants carrying substitutions in the highly conserved NS4B C-terminal domain was generated. Five mutations had only a minor effect on replication, but two of them enhanced assembly and release of infectious virus. Ten mutants were replication defective and used for selection of pseudoreversions. Most of the pseudoreversions also localized to the highly conserved NS4B C-terminal domain and were found to restore replication competence upon insertion into the corresponding primary mutant. Importantly, pseudoreversions restoring replication competence also restored heterotypic NS4B self-interaction, which was disrupted by the primary mutation. Finally, electron microscopy analyses of membrane alterations induced by NS4B mutants revealed striking morphological abnormalities, which were restored to wild-type morphology by the corresponding pseudoreversion. These findings demonstrate the important role of the C-terminal domain in NS4B self-interaction and the formation of functional HCV replication complexes.
Neutralizing Monoclonal Antibodies against Hepatitis C Virus E2 Protein Bind Discontinuous Epitopes and Inhibit Infection at a Postattachment Step
Authors: Sabo Michelle C. , Luca Vincent C. , Prentoe Jannick , Hopcraft Sharon E. , Blight Keril J. , Yi MinKyung , Lemon Stanley M. , Ball Jonathan K. , Bukh Jens , Evans Matthew J. , Fremont Daved H. , Diamond Michael S.
Published: May 2011

The E2 glycoprotein of hepatitis C virus (HCV) mediates viral attachment and entry into target hepatocytes and elicits neutralizing antibodies in infected patients. To characterize the structural and functional basis of HCV neutralization, we generated a novel panel of 78 monoclonal antibodies (MAbs) against E2 proteins from genotype 1a and 2a HCV strains. Using high-throughput focus-forming reduction or luciferase-based neutralization assays with chimeric infectious HCV containing structural proteins from both genotypes, we defined eight MAbs that significantly inhibited infection of the homologous HCV strain in cell culture. Two of these bound E2 proteins from strains representative of HCV genotypes 1 to 6, and one of these MAbs, H77.39, neutralized infection of strains from five of these genotypes. The three most potent neutralizing MAbs in our panel, H77.16, H77.39, and J6.36, inhibited infection at an early postattachment step. Receptor binding studies demonstrated that H77.39 inhibited binding of soluble E2 protein to both CD81 and SR-B1, J6.36 blocked attachment to SR-B1 and modestly reduced binding to CD81, and H77.16 blocked attachment to SR-B1 only. Using yeast surface display, we localized epitopes for the neutralizing MAbs on the E2 protein. Two of the strongly inhibitory MAbs, H77.16 and J6.36, showed markedly reduced binding when amino acids within hypervariable region 1 (HVR1) and at sites ∼100 to 200 residues away were changed, suggesting binding to a discontinuous epitope. Collectively, these studies help to define the structural and functional complexity of antibodies against HCV E2 protein with neutralizing potential.
Arginine Methylation of the RGG Box Does Not Appear To Regulate ICP27 Import during Herpes Simplex Virus Infection
Authors: Souki Stuart K. , Hernandez Felicia P. , Sandri-Goldin Rozanne M.
Published: May 2011

Arginine methylation can regulate protein import and export and can modulate protein interactions. Herpes simplex virus 1 (HSV-1) ICP27 is a shuttling protein involved in viral mRNA export. We previously reported that ICP27 is methylated on three arginines within its RGG box and that arginine methylation regulates ICP27 export and its interaction with SRPK1 and Aly/REF. Here, we report that ICP27 was efficiently imported into the nucleus when hypomethylated as determined by
F
luorescence
R
ecovery
A
fter
P
hotobleaching (FRAP). Furthermore, coimmunoprecipitation of ICP27 with β-importin was not significantly affected by ICP27 hypomethylation. Thus, ICP27 import does not appear to be regulated by arginine methylation.
Viral Replication and Innate Host Responses in Primary Human Alveolar Epithelial Cells and Alveolar Macrophages Infected with Influenza H5N1 and H1N1 Viruses
Authors: Yu Wendy C. L. , Chan Renee W. Y. , Wang Jieru , Travanty Emily A. , Nicholls John M. , Peiris J. S. Malik , Mason Robert J. , Chan Michael C. W.
Published: May 2011

Highly pathogenic influenza H5N1 virus continues to pose a threat to public health. Although the mechanisms underlying the pathogenesis of the H5N1 virus have not been fully defined, it has been suggested that cytokine dysregulation plays an important role. As the human respiratory epithelium is the primary target cell for influenza viruses, elucidating the viral tropism and innate immune responses of influenza H5N1 virus in the alveolar epithelium may help us to understand the pathogenesis of the severe pneumonia associated with H5N1 disease. Here we used primary cultures of differentiated human alveolar type II cells, alveolar type I-like cells, and alveolar macrophages isolated from the same individual to investigate viral replication competence and host innate immune responses to influenza H5N1 (A/HK/483/97) and H1N1 (A/HK/54/98) virus infection. The viral replication kinetics and cytokine and chemokine responses were compared by quantitative PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA). We demonstrated that influenza H1N1 and H5N1 viruses replicated productively in type II cells and type I-like cells although with different kinetics. The H5N1 virus replicated productively in alveolar macrophages, whereas the H1N1 virus led to an abortive infection. The H5N1 virus was a more potent inducer of proinflammatory cytokines and chemokines than the H1N1 virus in all cell types. However, higher levels of cytokine expression were observed for peripheral blood monocyte-derived macrophages than for alveolar macrophages in response to H5N1 virus infection. Our findings provide important insights into the viral tropisms and host responses of different cell types found in the lung and are relevant to an understanding of the pathogenesis of severe human influenza disease.
NSs Protein of Rift Valley Fever Virus Promotes Posttranslational Downregulation of the TFIIH Subunit p62
Authors: Kalveram Birte , Lihoradova Olga , Ikegami Tetsuro
Published: May 2011

Rift Valley fever virus (RVFV; family
Bunyaviridae
, genus
Phlebovirus
) is an important emerging pathogen of humans and ruminants. Its NSs protein has previously been identified as a major virulence factor that suppresses host defense through three distinct mechanisms: it directly inhibits beta interferon (IFN-β) promoter activity, it promotes the degradation of double-stranded RNA-dependent protein kinase (PKR), and it suppresses host transcription by disrupting the assembly of the basal transcription factor TFIIH through sequestration of its p44 subunit. Here, we report that in addition to PKR, NSs also promotes the degradation of the TFIIH subunit p62. Infection of cells with the RVFV MP-12 vaccine strain reduced p62 protein levels to below the detection limit early in the course of infection. This NSs-mediated downregulation of p62 was posttranslational, as it was unaffected by pharmacological inhibition of transcription or translation and MP-12 infection had no effect on p62 mRNA levels. Treatment of cells with proteasome inhibitors but not inhibition of lysosomal acidification or nuclear export resulted in a stabilization of p62 in the presence of NSs. Furthermore, p62 could be coprecipitated with NSs from lysates of infected cells. These data suggest that the RVFV NSs protein is able to interact with the TFIIH subunit p62 inside infected cells and promotes its degradation, which can occur directly in the nucleus.
Hepatitis C Virus Hijacks P-Body and Stress Granule Components around Lipid Droplets
Authors: Ariumi Yasuo , Kuroki Misao , Kushima Yukihiro , Osugi Kanae , Hijikata Makoto , Maki Masatoshi , Ikeda Masanori , Kato Nobuyuki
Published: May 2011

The microRNA miR-122 and DDX6/Rck/p54, a microRNA effector, have been implicated in hepatitis C virus (HCV) replication. In this study, we demonstrated for the first time that HCV-JFH1 infection disrupted processing (P)-body formation of the microRNA effectors DDX6, Lsm1, Xrn1, PATL1, and Ago2, but not the decapping enzyme DCP2, and dynamically redistributed these microRNA effectors to the HCV production factory around lipid droplets in HuH-7-derived RSc cells. Notably, HCV-JFH1 infection also redistributed the stress granule components GTPase-activating protein (SH3 domain)-binding protein 1 (G3BP1), ataxin-2 (ATX2), and poly(A)-binding protein 1 (PABP1) to the HCV production factory. In this regard, we found that the P-body formation of DDX6 began to be disrupted at 36 h postinfection. Consistently, G3BP1 transiently formed stress granules at 36 h postinfection. We then observed the ringlike formation of DDX6 or G3BP1 and colocalization with HCV core after 48 h postinfection, suggesting that the disruption of P-body formation and the hijacking of P-body and stress granule components occur at a late step of HCV infection. Furthermore, HCV infection could suppress stress granule formation in response to heat shock or treatment with arsenite. Importantly, we demonstrate that the accumulation of HCV RNA was significantly suppressed in DDX6, Lsm1, ATX2, and PABP1 knockdown cells after the inoculation of HCV-JFH1, suggesting that the P-body and the stress granule components are required for the HCV life cycle. Altogether, HCV seems to hijack the P-body and the stress granule components for HCV replication.
Respiratory and laryngeal symptoms secondary to gastro-oesophageal reflux
Authors: Rafferty G , Mainie I , McGarvey L P A
Published: May 2011

Gastro-oesophageal reflux may cause a range of laryngeal and respiratory symptoms. Mechanisms responsible include the proximal migration of gastric refluxate beyond the upper oesophageal sphincter causing direct irritation of the larynx and lower airway. Alternatively, refluxate entering the distal oesophagus alone may stimulate oesophageal sensory nerves and indirectly activate airway reflexes such as cough and bronchospasm. Recognising reflux as a cause for these extraoesophageal symptoms can be difficult as many patients do not have typical oesophageal symptoms (eg, heartburn) and clinical findings on laryngoscopy are not very specific. Acid suppression remains an effective treatment in the majority of patients but there is growing appreciation of the need to consider and treat non-acid and volume reflux. New opinions about the role of existing medical and surgical (laparoscopic techniques) treatment are emerging and a number of novel anti-reflux treatments are under development.
1-Hexyl n-cyanoacrylate compound (Neucrylate™ AN), a new berry aneurysm treatment. I. Theoretical basis and laboratory evaluation
Authors: Kerber Charles W , Pakbaz Ramin Sean
Published: May 2011

Purpose

To develop and optimize a system for treating cerebral berry aneurysm using a new polymer based liquid embolic agent.

Materials and methods

Using a variety of human corrosion casting aneurysm replicas placed in a circuit of pulsatile flowing blood, techniques were developed to optimize aneurysm filling and recreate good blood flow dynamics. As part of this effort, both experienced physicians and lay personnel were asked to treat these replicas to discover common failure modes. Additional surgically created aneurysms in dog carotid arteries were treated by experienced physicians.

Results

Physicians experienced in the use of microcatheters and endovascular techniques learned to use the device effectively after, on average, five aneurysm treatments. In the dog aneurysms, all were treated to complete filling.
COVARIANCE TRACKING WITH FORGETTING FACTOR AND RANDOM SAMPLING
Authors: ZHANG XUGUANG , LI XIAOLI , LIANG MING , WANG YANJIE
Published: May 2011

Covariance matching is an excellent algorithm of target tracking. In this paper, forgetting factor and random sampling methods are proposed to improve the robustness and efficiency of covariance tracking. First, a distance function between covariance matrixes is weighted by using a forgetting factor based on a fuzzy membership function to overcome the disturbances from similar targets. Then a random sampling method is applied to reduce the computing time in covariance matching and to facilitate real-time object tracking. Experiment results show that the algorithm proposed in this paper can effectively mitigate the clutter and occlusion problems at a high computing speed.
GROWTH AND CHARACTERIZATION OF Na0.5Bi0.5TiO3–K0.5Bi0.5TiO3 LEAD-FREE PIEZOELECTRIC CRYSTAL BY THE TSSG METHOD
Authors: SUN RENBING , LIN DI , ZHANG QINHUI , FANG BIJUN , ZHANG HAIWU , LI XIAOBING , WANG SHENG , WANG DONG , ZHAO XIANGYONG , LUO HAOSU
Published: May 2011

The 0.96Na0.5Bi0.5TiO3–0.04K0.5Bi0.5TiO3 (NKBT96/4) lead-free single crystal was successfully grown by top-seeded solution growth method. The effective segregation coefficient for K was calculated to be 0.20. X-ray powder diffraction measurement showed that the as-grown crystal possesses rhombohedral perovskite structure at room temperature. X-ray rock curve measure indicated that the as-grown single crystal owned relatively higher quality. The NKBT96/4 single crystal exhibits apparent diffused dielectric behavior, where the diffusiveness index α is 1.79 for the [001]-oriented crystal planes. A notable thermal hysteresis, ΔT h , of 25°C was disclosed for the tetragonal antiferroelectric-tetragonal paraelectric phase transition in the dielectric property studies between heating and cooling. The highest remnant polarization, P r , 16.8 μC/cm2, was obtained at 100°C. The electromechanical coupling coefficient kt of the [001]-oriented crystal planes at room temperature was determined to be 38% using resonance–antiresonance technique. The room temperature piezoelectric constant d33 reaches 120 pC/N.
FERROELECTRIC SWITCHING PATH IN MONODOMAIN RHOMBOHEDRAL BiFeO3 CRYSTAL: A FIRST-PRINCIPLES STUDY
Authors: DING HANG-CHEN , SHI SI-QI , TANG WEI-HUA , DUAN CHUN-GANG
Published: May 2011

Based on density-functional calculations, we have studied possible ferroelectric switching path in monodomain single crystal of rhombohedral BiFeO3 , a prototypical multiferroic compound. By carefully studying the behaviors of FeO6 corner-sharing double-tetrahedrons, we find abrupt changes in total energy and oxygen atomic positions, and therefore polarizations, occur in the ferroelectric switching path of rhombohedral BiFeO3 . Detailed analyses suggest that such behavior might be caused by the frustrated magnetic ordering in the paraelectric phase of rhombohedral BiFeO3 , where three O atoms and the Bi atom are in the same plane perpendicular to the polarization direction. This is supported by the fact that the ferroelectric switching for paramagnetic BiFeO3 is smooth and has a much lower energy barrier than that of antiferromagnetic BiFeO3 .
The Spectroscopy and Micro-Structure of Eu3+ Ions Doped Double Perovskite Ba3Y2WO9
Authors: Huang Yanlin , Seo Hyo Jin
Published: May 2011

Eu3+-doped red-emitting phosphors of double perovskite Ba3Y2WO9 were synthesized by the solid-state reaction. The phosphors were characterized by X-ray powder diffraction. The luminescence properties, e.g., the excitation and emission spectra, luminescence decay times and CIE coordinates were investigated. The phosphors can be efficiently excited by near UV light to realize an intense red luminescence corresponding to the electric dipole transition 5D0 → 7F2 of Eu3+ ions. The double perovskite oxide was investigated by the site-selective emission spectra and the luminescence decay curves in the 5D0 → 7F0 region using a pulsed, tunable, narrowband dye laser. It is suggested that Eu3+ ions have three different crystallographic sites in this host. The site assignments of Eu3+ ions were discussed on the base of both optical spectroscopy and structural analysis. This is helpful to understand the occupation of rare earth ions doped in this perovskite structure.
An intramolecular transport metabolon: fusion of carbonic anhydrase II to the COOH terminus of the Cl−/HCO3− exchanger, AE1
Authors: Sowah Daniel , Casey Joseph R.
Published: May 2011

Anion exchanger 1 (AE1) is the plasma membrane Cl−/HCO3− exchanger of erythrocytes. Carbonic anhydrases (CA) provide substrate for AE1 by catalyzing the reaction, H2O + CO2 ↔ HCO3− + H+. The physical complex of CAII with AE1 has been proposed to maximize anion exchange activity. To examine the effect of CAII catalysis on AE1 transport rate, we fused either CAII-wild type or catalytically inactive CAII-V143Y to the cytoplasmic COOH terminus of AE1 to form AE1.CAII and AE1.CAII-V143Y, respectively. When expressed in transfected human embryonic kidney 293 cells, AE1.CAII had a similar Cl−/HCO3− exchange activity to AE1 alone, as assessed by the flux of H+ equivalents (87 ± 4% vs. AE1) or rate of change of intracellular Cl− concentration (93 ± 4% vs. AE1), suggesting that CAII does not activate AE1. In contrast, AE1.CAII-V143Y displayed transport rates for H+ equivalents and Cl− of 55 ± 2% and of 40 ± 2%, versus AE1. Fusion of CAII to AE1 therefore reduces anion transport activity, but this reduction is compensated for during Cl−/HCO3− exchange by the presence of catalytically active CAII. Overexpression of free CAII-V143Y acts in a dominant negative manner to reduce AE1-mediated HCO3− transport by displacement of endogenous CAII-wild type from its binding site on AE1. To examine whether AE1.CAII bound endogenous CAII, we coexpressed CAII-V143Y along with AE1 or AE1.CAII. The bicarbonate transport activity of AE1 was inhibited by CAII-V143Y, whereas the activity of AE1.CAII was unaffected by CAII-V143Y, suggesting impaired transport activity upon displacement of functional CAII from AE1 but not AE1.CAII. Taken together, these data suggest that association of functional CAII with AE1 increases Cl−/HCO3− exchange activity, consistent with the HCO3− transport metabolon model.