bluuebunny/crossref_metadata_2025 huggingface dataset:
The Popularity of Jane Eyre in China
Authors: Wu Qinghong , Huang Lu
Published: August 2011

Charlotte Bronte’s Jane Eyre has gradually achieved a great popularity among Chinese readers from its first introduction into China in 1917 to the 1980s when China introduced an open door policy. Therefore, millions of copies of various versions of the novel have been sold in China in the past 30 years. Though the interpretations of the novel change under different political and social circumstances, it is always fascinating to readers and academic scholars in China, for it not only possesses a literary and aesthetic charm of its own, but also has rich thematic meanings and implied moral teachings somewhat similar to traditional Chinese ethics, and gives Chinese readers resonance to life. It is so well appreciated by Chinese readers that it is recommended by the Chinese national education department as one of the fundamental compulsory reading books for primary and middle school students. In fact, it seems to have melt into the Chinese culture and may have far‐reaching influence in China.
Chaucer and Italy: Contexts and/of Sources
Authors: Clarke K. P.
Published: August 2011

This critical overview of Chaucer and Italy examines the sources he encountered there, such as Boccaccio’sFilostrato,TeseidaandDecameronas well as Dante’sComedìa. It covers the most important and influential publications on the subject since the early 1980s, as well as highlighting new and forthcoming work. This new work emphasizes the material contexts of these sources, such as the commentaries frequently accompanying the early copies of theComedìa, and the influence of Boccaccio’s own early manuscripts on subsequent copies of his work. Work on Chaucer’s Italian sources remains vital and stimulating and offers Chaucerians a wealth of new reading contexts for his work.
Ishmael as Evolving Character in Melville’s Redburn, White‐Jacket, Mardi, and Moby‐Dick
Authors: Broek Michael
Published: August 2011

Most contemporary treatments of Herman Melville focus on issues related to cultural studies, biography, textual issues, performance, and New Media. Close textual readings, including those grounded in the study of character, have been largely out of vogue since the advent of Post‐colonial studies and French theory. However, these contemporary treatments may fail to capture the evolution of character and theme that is prevalent across many of the author’s early novels. Ishmael in particular, who is notoriously hard to grasp, may be usefully understood as developing across Redburn, White‐Jacket, and Mardi, before he emerges in the opening line of Moby‐Dick. Redburn uses language that clearly connects him to Ishmael, a young man going to sea in search of his father’s wisdom but instead discovering a cosmopolitan world of madness and deprivation, while in White‐Jacket, the title character attempts to shield himself from the horrors of a life aboard a ‘Man‐of‐War’ by insulating his ‘soul’ within the confines of a handmade white jacket. Failing in his attempt to isolate himself, he falls from the mainmast into the sea, his jacket caught in the arms of the rigging. Finally, in Mardi, Melville’s allegorical circumnavigation of the South Pacific, Babbalanja philosophizes in the same vein as Ishmael, trying to grasp some meaning out of the complexities of his experience. This article will analyze the characters of Redburn, White‐Jacket, Babbalanja, and Ishmael as representing the working out of the issues facing a single, evolving character, a character who adopts multiple perceptions in order to attempt integrating the many aspects of his self with the jarring and disassociating realities of the world he faces. Understood thus, Ishmael’s apparent strangeness within the narrative of Moby‐Dick may be more usefully understood as the ultimate manifestation of a complex, purposeful, and long‐running evolution.
Increasing impact of mental health presentations on New South Wales public hospital emergency departments 1999–2006
Authors: Tankel Alan S , Di Palma Matthew J , Kramer Kathy M , van der Zwan Rick
Published: August 2011

Objective: The percentage of mental health (MH) presentations to New South Wales (NSW) EDs in recent years has not been described at the statewide level. Evidence from other states suggests this burden might be increasing. The present study aims to evaluate recent trends in MH presentations to NSW EDs, including geographic variations and the spectrum of MH disorders encountered.

Methods: Data were sourced from the NSW Emergency Department Data Collection, which collates information from routine reporting undertaken in all NSW EDs. The database employs ICD‐9 diagnostic descriptors. The present study retrospectively analyses presentations from 1999 to 2006 for patients who received an MH primary discharge diagnosis, as defined by ICD‐9. In addition, rates of presentation for poisoning are considered. The percentage of MH presentations to the three largest hospital categories – principal referral, major metropolitan and rural base (i.e. major non‐metropolitan) – is examined.

Results: During 1999 and 2006, there were 9 013 357 ED presentations at the study sites. Of these, 3.22% received an MH primary discharge diagnosis. An additional 0.93% received a diagnosis of poisoning. In 2006, there were 15 262 more MH presentations than in 1999, a 49.78% increase. Over this period, total annual ED presentations increased by 201 763 (19.30%). General population growth was 6.31%. Across the state, 32.0% of MH presentations were for psychotic conditions and 68.0% for neurotic illnesses.

Conclusion: Mental health presentations to NSW EDs are increasing. There is a need to review emergency services to accommodate these changes. The reasons for this growth remain unclear and require formal prospective evaluation.
Microbiology of ‘Candidatus Accumulibacter’ in activated sludge
Authors: He Shaomei , McMahon Katherine D.
Published: August 2011

‘Candidatus Accumulibacter’ is a biotechnologically important bacterial group that can accumulate large amounts of intracellular polyphosphate, contributing to biological phosphorus removal in wastewater treatment. Since its first molecular identification more than a decade ago, this bacterial group has drawn significant research attention due to its high abundance in many biological phosphorus removal systems. In the past 6 years, our understanding of Accumulibacter microbiology and ecophysiology has advanced rapidly, largely owing to genomic information obtained through shotgun metagenomic sequencing efforts. In this review, we focus on the metabolism, physiology, fine‐scale population structure and ecological distribution of Accumulibacter, aiming to integrate the information learned so far and to present a more complete picture of the microbiology of this important bacterial group.
Comparison of Two Different α1‐Adrenoceptor Antagonists, Tamsulosin and Silodosin, in the Treatment of Male Lower Urinary Tract Symptoms Suggestive of Benign Prostatic Hyperplasia: A Prospective Randomized Crossover Study
Authors: YOKOYAMA Teruhiko , HARA Ryoei , FUJII Tomohiro , JO Yoshimasa , MIYAJI Yoshiyuki , NAGAI Atsushi
Published: August 2011

Objectives: We assessed the efficacy and safety of two α1‐adrenoceptor antagonists, tamsulosin and silodosin, in the treatment of male lower urinary tract symptoms.

Methods: Men aged 50 years or older who had a total International Prostate Symptom Score (IPSS) of 8 or higher were enrolled in this study. Forty‐six patients were randomized into two groups. Twenty‐three patients were initially prescribed tamsulosin 0.2 mg once daily for 3 months, followed by silodosin 4 mg twice daily for 3 months (group T); the other group of 23 patients were initially prescribed silodosin, followed by tamsulosin (group S). Patients then switched to the alternative treatment after a 1‐month clearance period. Evaluations included clinical determination of IPSS, quality‐of‐life index, maximum flow rate and postvoid residual urine volume before and after treatment.

Results: A total of 46 men, 23 in group T and 23 in group S, were treated and 41 (89.1%) completed the treatment. IPSS, quality‐of‐life index, maximum flow rate and postvoid residual urine volume were significantly improved in both groups after treatment. The changes in the total IPSS from baseline in groups S and T at 3 months were −6.6 and −7.5, respectively. There were no significant differences between the two groups. After taking both medications, 18 patients preferred silodosin, 11 preferred tamsulosin and others felt they had the same effects. Six and none patients experienced adverse events during silodosin and tamsulosin treatment, respectively.

Conclusion: Two types of α1‐adrenoceptor antagonists in the same individuals provide similar efficacy. Profiles and difference of each drug should be considered in making treatment choice.
Respiratory disease in the donkey
Authors: Thiemann A. K.
Published: August 2011

The donkey suffers from a similar range of respiratory diseases as the horse; however, there are a number of subtle variations, knowledge of which can influence the success of treatment. As an animal adapted to a semi‐arid terrain, there are variations in physiology, anatomy and disease susceptibility. The nonathletic nature of the donkey means that delayed presentation is common with many diseases and, while there may be enhanced resistance to some transboundary and parasitic diseases, there may be equal or increased severity of illness to some endemic diseases, e.g. equine influenza. Donkeys frequently live to geriatric ages and the clinician should be aware of the increased risk of conditions such as tracheal collapse, fibrosing pneumonia and neoplasia in this age group. As with any condition that causes stress and inappetance, respiratory disease in the donkey may be complicated by hyperlipaemia and good nursing care is an essential component of treatment.
Clinical, radiological and ultrasonographic features, treatment and outcome in 22 horses with caudal distal radial osteochondromata
Authors: WRIGHT I. M. , MINSHALL G. J.
Published: August 2011

Reasons for performing study:Although a well recognised clinical entity, only small numbers of osteochondromata on the caudal distal radius have previously been published and its occurrence in young racing Thoroughbreds has not previously been reported. Identification and management of associated lesions in the deep digital flexor tendon have received scant attention in the literature.

Hypothesis:Osteochondromata of the caudal distal radius occur commonly in young racing Thoroughbreds. They vary in size and location, sagittally and proximodistally, but the majority cause impingement damage to the deep digital flexor tendon.

Methods:Case records and diagnostic images of horses with osteochondromata of the caudal distal radius were reviewed retrospectively and follow‐up information obtained.

Results:Twenty‐five osteochondromata were identified in 22 horses, 19 of which were Thoroughbreds. All osteochondromata were metaphyseal. Twenty‐two were in the middle one‐third of the bone and laceration of the adjacent deep digital flexor tendon was identified in 21 limbs. Treatment in all cases consisted of removal of the osteochondroma with debridement of the deep digital flexor tendon when this was affected. All horses returned to work and none exhibited any evidence of recurrence.

Conclusion:Osteochondromata of the caudal distal radius occur in young racing Thoroughbreds but are also identified in other horses. They have a consistent metaphyseal location and most are found in the middle one‐third of the radius. Size varies, but most cause laceration of the adjacent deep digital flexor tendon. Treatment by removal of the mass and debridement of the tendon is associated with a good prognosis.

Potential relevance:Osteochondromata of the caudal distal radius are an important cause of tenosynovitis of the carpal sheath of the digital flexor tendons in young racing Thoroughbreds. When present in the most common location of the middle one‐third of the bone, they are likely to cause impingement damage to the deep digital flexor tendon. Tenoscopic surgery offers a good prognosis.
Cross‐sectional study of antimicrobial‐resistant bacteria in horses. Part 2: Risk factors for faecal carriage of antimicrobial‐resistant Escherichia coli in horses
Authors: MADDOX T. W. , PINCHBECK G. L. , CLEGG P. D. , WEDLEY A. L. , DAWSON S. , WILLIAMS N. J.
Published: August 2011

Reasons for performing study: The increasing prevalence of antimicrobial resistant bacteria such as antimicrobial‐resistant and extended spectrum β‐lactamase (ESBL)‐producing Escherichia coli represents a significant problem for human and veterinary medicine. Despite this, the risk factors for faecal carriage of such bacteria by horses in the UK, particularly those in the wider community, have not been well described.

Objectives: To characterise the risk factors for faecal carriage of antimicrobial‐resistant E. coli amongst horses in the mainland UK.

Methods: A cross‐sectional study of horses recruited by 65 randomly selected equine veterinary practices was conducted, with a faecal sample collected and self‐administered questionnaire completed by the horse owner. Faecal samples were cultured for antimicrobial‐resistant E. coli, with isolates confirmed as E. coli having their antimicrobial resistance profile determined. Multilevel, multivariable logistic regression models were used to investigate risk factors for the carriage of antimicrobial‐resistant E. coli in the sample population.

Results: Faecal samples and completed questionnaires were obtained for 627 horses located on 475 premises. Recent hospitalisation, contact with specific types of nonequid animals, the type of premises, the surrounding land use, the reason for veterinary treatment received in the last 6 months and antimicrobial treatment in the previous 10 days were identified as risk factors for many of the antimicrobial‐resistance outcomes considered. Being stabled on the same yard as a recently hospitalised horse was identified as a risk factor for increased risk of carriage of ESBL‐producing E. coli.

Conclusions and potential relevance: Increasing antimicrobial resistance may have significant health implications for the horse population of Great Britain. This form of epidemiological investigation highlights potential risk factors that may be controlled to limit the extent of the problem.
Pump Design for a Portable Renal Replacement System
Authors: Kang Jane , Scholz Tamera , Weaver Jason D. , Ku David N. , Rosen David W.
Published: August 2011

This work proposes a small, light, valveless pump design for a portable renal replacement system. By analyzing the working principle of the pump and exploring the design space using an analytical pump model, we developed a novel design for a cam-driven finger pump. Several cams sequentially compress fingers, which compress flexible tubes; thus eliminating valves. Changing the speed of the motor or size of the tube controls the flow rate. In vitro experiments conducted with whole blood using the pump measured Creatinine levels over time, and the results verify the design for the portable renal replacement system. The proposed pump design is smaller than 153 cm3 and consumes less than 1 W while providing a flow rate of more than 100 ml/min for both blood and dialysate flows. The smallest pump of a portable renal replacement system in the literature uses check valves, which considerably increase the overall manufacturing cost and possibility of blood clotting. Compared to that pump, the proposed pump design achieved reduction in size by 52% and savings in energy consumption by 89% with the removal of valves. This simple and reliable design substantially reduces the size requirements of a portable renal replacement system.
Biomechanical Property of a Natural Microneedle: The Caterpillar Spine
Authors: Ma G. J. , Shi L. T. , Wu C. W.
Published: August 2011

Recently, microneedles (or microneedle arrays) for transdermal drug delivery have received increasing attention because they can provide painless, minimal invasiveness and time-released drug delivery. However, it is very difficult to design such an eligible microneedle that meets all the requirements for mechanical strength, small insertion force, and good biocompatibility. In this paper, we investigate a biomicroneedle: caterpillar spine. It is found that this type of biomicroneedle can pierce mouse skin using a very small force (about 173 μN) without fracture and buckling failures. Such excellent properties are mainly a result of its optimal geometry evolved by Nature, the high hardness, and the reasonable high elastic modulus near the tip end. This finding may provide an inspiration for the development of improved transdermal drug delivery microneedles.
Design of a Multigrasp Transradial Prosthesis
Authors: Wiste Tuomas E. , Dalley Skyler A. , Atakan Varol H. , Goldfarb Michael
Published: August 2011

This paper describes the design and performance of a new prosthetic hand capable of multiple grasp configurations, and capable of fingertip forces and speeds comparable to those used by healthy subjects in typical activities of daily living. The hand incorporates four motor units within the palm, which together drive sixteen joints through tendon actuation. Each motor unit consists of a brushless motor that drives one or more tendons through a custom two-way clutch and pulley assembly. After presenting the design of the prosthesis, the paper presents a characterization of the hand’s performance. This includes its ability to provide eight grasp postures, as well as its ability to provide fingertip forces and finger speeds comparable to those described in the biomechanics literature corresponding to activities of daily living.
Factors for conversion between human and automatic read‐outs of CDMAM images
Authors: Figl Michael , Hoffmann Rainer , Kaar Marcus , Semturs Friedrich , Brasik Natasa , Birkfellner Wolfgang , Homolka Peter , Hummel Johann
Published: August 2011

Purpose:

According to the European protocol for the quality control of the physical and technical aspects of mammography screening (EPQCM) image quality of digital mammography devices has to be assessed using human evaluation of the CDMAM contrast‐detail phantom. This is accomplished by the determination of threshold thicknesses of gold disks with different diameters (0.08–2 mm) and revealed to be very time consuming. Therefore a software solution based on a nonprewhitening matched filter (NPW) model was developed at the University of Nijmegen. Factors for the conversion from automatic to human readouts have been determined by Younget al. [Proc. SPIE 614206, 1–13 (2006) and Proc. SPIE 6913, 69131C1 (2008)] using a huge amount of data of both human and automatic readouts. These factors depend on the observer groups and are purely phenomenological. The authors present an alternative approach to determine the factors by using the Rose observer model.

Methods:

Their method uses the Rose theory which gives a relationship between threshold contrast, diameter of the object and number of incident photons. To estimate the conversion factors for the five diameters from 0.2 to 0.5 mm they exposed with five different current‐time products which resulted in 25 equations with five unknowns.

Results:

The theoretical conversion factors (in dependence of the diameters) amounted to be 1.61 ± 0.02 (0.2 mm diameter), 1.67 ± 0.02 (0.25 mm), 1.85 ± 0.02 (0.31 mm), 2.09 ± 0.02 (0.4 mm), and 2.28 ± 0.02 (0.5 mm). The corresponding phenomenological factors found in literature are 1.74 (0.2 mm), 1.78 (0.25 mm), 1.83 (0.31 mm), 1.88 (0.4 mm), and 1.93 (0.5 mm).

Conclusions:

They transferred the problem of determining the factors to a well known observer model which has been examined for many years and is also well established. This method reveals to be reproduceable and produces factors comparable to the phenomenological ones.
Race, Ethnicity, and NIH Research Awards
Authors: Ginther Donna K. , Schaffer Walter T. , Schnell Joshua , Masimore Beth , Liu Faye , Haak Laurel L. , Kington Raynard
Published: August 2011

NIH research project grants from 2000 to 2006 show evidence of racial/ethnic disparities in the probability of receiving an award.
Three Periods of Regulatory Innovation During Vertebrate Evolution
Authors: Lowe Craig B. , Kellis Manolis , Siepel Adam , Raney Brian J. , Clamp Michele , Salama Sofie R. , Kingsley David M. , Lindblad-Toh Kerstin , Haussler David
Published: August 2011

Patterns of vertebrate gene regulation have changed during the course of evolution.
Graduate Students’ Teaching Experiences Improve Their Methodological Research Skills
Authors: Feldon David F. , Peugh James , Timmerman Briana E. , Maher Michelle A. , Hurst Melissa , Strickland Denise , Gilmore Joanna A. , Stiegelmeyer Cindy
Published: August 2011

Teaching is not wasted time.
Interventions Shown to Aid Executive Function Development in Children 4 to 12 Years Old
Authors: Diamond Adele , Lee Kathleen
Published: August 2011

To be successful takes creativity, flexibility, self-control, and discipline. Central to all those are executive functions, including mentally playing with ideas, giving a considered rather than an impulsive response, and staying focused. Diverse activities have been shown to improve children’s executive functions: computerized training, noncomputerized games, aerobics, martial arts, yoga, mindfulness, and school curricula. All successful programs involve repeated practice and progressively increase the challenge to executive functions. Children with worse executive functions benefit most from these activities; thus, early executive-function training may avert widening achievement gaps later. To improve executive functions, focusing narrowly on them may not be as effective as also addressing emotional and social development (as do curricula that improve executive functions) and physical development (shown by positive effects of aerobics, martial arts, and yoga).
Effectiveness of Early Educational Intervention
Authors: Barnett W. S.
Published: August 2011

Early educational intervention has been proposed to partially offset the impacts of poverty and inadequate learning environments on child development and school success. A broad range of early educational interventions are found to produce meaningful, lasting effects on cognitive, social, and schooling outcomes. However, all interventions are not equally effective. Two major U.S. programs perform relatively poorly. Research provides some guidance regarding the features of highly effective programs, but much remains to be learned. New experimental studies of key program features would have a high payoff.
Partitioning Regulatory Mechanisms of Within-Host Malaria Dynamics Using the Effective Propagation Number
Authors: Metcalf C. J. E. , Graham A. L. , Huijben S. , Barclay V. C. , Long G. H. , Grenfell B. T. , Read A. F. , Bjørnstad O. N.
Published: August 2011

As malaria progresses, red blood cell availability and immune control change depending on the initial dose of parasites.
Detection of Emerging Sunspot Regions in the Solar Interior
Authors: Ilonidis Stathis , Zhao Junwei , Kosovichev Alexander
Published: August 2011

Helioseismic observations allow the detection of sunspots before they emerge at the surface of the Sun.
Aneuploidy Drives Genomic Instability in Yeast
Authors: Sheltzer Jason M. , Blank Heidi M. , Pfau Sarah J. , Tange Yoshie , George Benson M. , Humpton Timothy J. , Brito Ilana L. , Hiraoka Yasushi , Niwa Osami , Amon Angelika
Published: August 2011

Extra chromosomes cause general genomic instability in yeast.
Inhibitory Fcγ Receptor Engagement Drives Adjuvant and Anti-Tumor Activities of Agonistic CD40 Antibodies
Authors: Li Fubin , Ravetch Jeffrey V.
Published: August 2011

Engagement of an inhibitory receptor improves the anti-tumor efficacy of a monoclonal antibody.
Visualizing Individual Nitrogen Dopants in Monolayer Graphene
Authors: Zhao Liuyan , He Rui , Rim Kwang Taeg , Schiros Theanne , Kim Keun Soo , Zhou Hui , Gutiérrez Christopher , Chockalingam S. P. , Arguello Carlos J. , Pálová Lucia , Nordlund Dennis , Hybertsen Mark S. , Reichman David R. , Heinz Tony F. , Kim Philip , Pinczuk Aron , Flynn George W. , Pasupathy Abhay N.
Published: August 2011

Nitrogen atoms that replace carbon atoms in the graphene lattice strongly modify the local electronic structure.
A Light Account of Plumes
Feathers
The Evolution of a Natural Miracle

by Thor Hanson

Basic Books, New York, 2011. 352 pp. $25.99, C$30. £18.99. ISBN 9780465020133.
Authors: Stettenheim Peter
Published: August 2011

Writing for nonspecialists and lay readers, Hanson discusses aspects of the natural history of feathers, including their evolution, functions, anatomy, and development.
Quantifying Malaria Dynamics Within the Host
Authors: Day Karen P. , Fowkes Freya J. I.
Published: August 2011

A statistical method reveals the roles of the immune system and cell availability in regulating parasitemia.
NIH Uncovers Racial Disparity in Grant Awards
Authors: Kaiser Jocelyn
Published: August 2011

An in-depth analysis of grant data from the U.S. National Institutes of Health finds that black Ph.D. scientists were far less likely to receive NIH funding than a white scientist from a similar institution with the same research record.
Drug Developers Finally Take Aim at a Neglected Disease
Authors: Leslie Mitch
Published: August 2011

New and recycled compounds target the parasite responsible for Chagas disease, which afflicts millions in Latin America.
A Tropical Disease Hits the Road
Authors: Leslie Mitch
Published: August 2011

Generally considered one of the many neglected tropical diseases, Chagas disease has been drawing increased attention, which some attribute to its growth outside of Latin America.
Giving Children a Head Start Is Possible—But It's Not Easy
Authors: Mervis Jeffrey
Published: August 2011

Begun in 1965, the U.S. Head Start program is still trying to find the best way to prepare poor children for school.
A Passion for Early Education
Authors: Mervis Jeffrey
Published: August 2011

Joan Lombardi brings experience and energy to the Obama Administration's efforts to better coordinate federal children's health and education programs.
Cerebral Schistosomiasis Due to Schistosoma haematobium Confirmed by PCR Analysis of Brain Specimen
Authors: Imai Kentaro , Koibuchi Tomohiko , Kumagai Takashi , Maeda Takuya , Osada Yoshio , Ohta Nobuo , Koga Michiko , Nakamura Hitomi , Miura Toshiyuki , Iwamoto Aikichi , Fujii Takeshi
Published: August 2011

The case of a 25-year-old Japanese male who had cerebral schistosomiasis caused by
Schistosoma haematobium
is reported here. Although serum antibody tests showed a cross-reaction with other helminths and no ova were excreted in urine or feces, the existence of
Schistosoma haematobium
in the brain was confirmed by PCR analysis.
Epstein-Barr Virus Latent Membrane Protein 1 Induces Cancer Stem/Progenitor-Like Cells in Nasopharyngeal Epithelial Cell Lines
Authors: Kondo Satoru , Wakisaka Naohiro , Muramatsu Masamichi , Zen Yoh , Endo Kazuhira , Murono Shigeyuki , Sugimoto Hisashi , Yamaoka Shoji , Pagano Joseph S. , Yoshizaki Tomokazu
Published: August 2011

Recent studies suggest the existence of cancer stem cells (CSC) and cancer progenitor cells (CPC), although strict definitions of neither CSC nor CPC have been developed. We have produced evidence that the principal oncoprotein of Epstein-Barr virus (EBV), latent membrane protein 1 (LMP1), which is associated with human malignancies, especially nasopharyngeal carcinoma (NPC), promotes tumor cell invasion and metastasis, as well as the epithelial-mesenchymal transition (EMT). However, whether LMP1 is involved in the development of CSC/CPC is still unclear. This study investigates whether the expression of EBV-LMP1 is related to the development of CSC/CPC. Analysis of cancer stem cell markers reveals that LMP1 induces the CD44
high
CD24
low
CSC/CPC-like phenotype as well as self-renewal abilities in LMP1-expressing epithelial cell lines. In addition, we show here that LMP1 induction in epithelial cells causes high tumorigenicity and rapid cellular proliferation. Furthermore, we found that LMP1 expression increased the expression of several CPC markers as well as producing increased levels of EMT markers. Our findings indicate that LMP1 can induce a CPC-like rather than a CSC-like phenotype in epithelial cells and suggest that LMP1-induced phenotypic changes contribute to the development of NPC.
Replication and Transcription Activator (RTA) of Murine Gammaherpesvirus 68 Binds to an RTA-Responsive Element and Activates the Expression of ORF18
Authors: Hong Yun , Qi Jing , Gong Danyang , Han Chuanhui , Deng Hongyu
Published: August 2011

The replication and transcription activator (RTA), mainly encoded by open reading frame 50, is an immediate-early gene product that is conserved among all characterized gammaherpesviruses. Previous studies have demonstrated that RTA proteins of Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) can activate the promoter of many viral early lytic genes through direct or indirect mechanisms. Murine gammaherpesvirus 68 (MHV-68) is genetically related to KSHV and EBV, and the RTA homologue from MHV-68 also initiates the lytic cycle of gene expression. Although two RTA-dependent promoters had been identified in MHV-68, the mechanism of the interaction between RTA and the promoters was not characterized. In this study, we first identified an RTA-responsive promoter in the left origin of lytic replication region of MHV-68 through a reporter assay and mapped a 27-bp RTA-responsive element (RRE) through systematic deletions. Interestingly, sequence analysis identified a second RRE in this region. An electrophoretic mobility shift assay (EMSA) and a chromatin immunoprecipitation (ChIP) assay showed that RTA can bind directly to these two RREs
in vitro
or
in vivo
. Mutagenesis studies have further characterized the nucleotides important for mediating RTA binding by an EMSA. Moreover, we engineered RRE-deleted viruses and demonstrated in the context of the viral genome that one of the RREs mediates the RTA-dependent activation of an essential lytic gene, ORF18, during
de novo
infection. To our knowledge, this is the first time that RTA binding sites in MHV-68 have been identified. Since ORF18 regulates viral late gene expression, our study has also contributed to the delineation of the expression cascade of gammaherpesvirus lytic genes.
The Human Papillomavirus Type 16 E5 Oncoprotein Translocates Calpactin I to the Perinuclear Region
Authors: Krawczyk Ewa , Suprynowicz Frank A. , Hebert Jess D. , Kamonjoh Christopher M. , Schlegel Richard
Published: August 2011

The human papillomavirus type 16 (HPV-16) E5 oncoprotein is embedded in membranes of the endoplasmic reticulum and nuclear envelope with its C terminus exposed to the cytoplasm. Among other activities, E5 cooperates with the HPV E6 oncoprotein to induce koilocytosis in human cervical cells and keratinocytes
in vitro
. The effect of E5 on infected cells may rely on its interactions with various cellular proteins. In this study we identify calpactin I, a heterotetrameric, Ca
2+
- and phospholipid-binding protein complex that regulates membrane fusion, as a new cellular target for E5. Both the annexin A2 and p11 subunits of calpactin I coimmunoprecipitate with E5 in COS cells and in human epithelial cell lines, and an intact E5 C terminus is required for binding. Moreover, E5-expressing cells exhibit a perinuclear redistribution of annexin A2 and p11 and show increased fusion of perinuclear membrane vesicles. The C terminus of E5 is required for both the perinuclear redistribution of calpactin I and increased formation of perinuclear vacuoles. These results support the hypothesis that the E5-induced relocalization of calpactin I to the perinuclear region promotes perinuclear membrane fusion, which may underlie the development of koilocytotic vacuoles.
Y-Box-Binding Protein 1 Interacts with Hepatitis C Virus NS3/4A and Influences the Equilibrium between Viral RNA Replication and Infectious Particle Production
Authors: Chatel-Chaix Laurent , Melançon Pierre , Racine Marie-Ève , Baril Martin , Lamarre Daniel
Published: August 2011

The hepatitis C virus (HCV) NS3/4A protein has several essential roles in the virus life cycle, most probably through dynamic interactions with host factors. To discover cellular cofactors that are co-opted by HCV for its replication, we elucidated the NS3/4A interactome using mass spectrometry and identified Y-box-binding protein 1 (YB-1) as an interacting partner of NS3/4A protein and HCV genomic RNA. Importantly, silencing YB-1 expression decreased viral RNA replication and severely impaired the propagation of the infectious HCV molecular clone JFH-1. Immunofluorescence studies further revealed a drastic HCV-dependent redistribution of YB-1 to the surface of the lipid droplets, an important organelle for HCV assembly. Core and NS3 protein-dependent polyprotein maturation were shown to be required for YB-1 relocalization. Unexpectedly, YB-1 knockdown cells showed the increased production of viral infectious particles while HCV RNA replication was impaired. Our data support that HCV hijacks YB-1-containing ribonucleoparticles and that YB-1–NS3/4A–HCV RNA complexes regulate the equilibrium between HCV RNA replication and viral particle production.
Nuclear Receptor Signaling Inhibits HIV-1 Replication in Macrophages through Multipletrans-Repression Mechanisms
Authors: Hanley Timothy M. , Viglianti Gregory A.
Published: August 2011

Sexually transmitted pathogens activate HIV-1 replication and inflammatory gene expression in macrophages through engagement of Toll-like receptors (TLRs). Ligand-activated nuclear receptor (NR) transcription factors, including glucocorticoid receptor (GR), peroxisome proliferator-activated receptor gamma (PPARγ), and liver X receptor (LXR), are potent inhibitors of TLR-induced inflammatory gene expression. We therefore hypothesized that ligand-activated NRs repress both basal and pathogen-enhanced HIV-1 replication in macrophages by directly repressing HIV-1 transcription and by ameliorating the local proinflammatory response to pathogens. We show that the TLR2 ligand PAM3CSK4 activated virus transcription in macrophages and that NR signaling repressed both basal and TLR-induced HIV-1 transcription. NR ligand treatment repressed HIV-1 expression when added concurrently with TLR ligands and in the presence of cycloheximide, demonstrating that they act independently of new cellular gene expression. We found that treatment with NR ligands inhibited the association of AP-1 and NF-κB subunits, as well as the coactivator CBP, with the long terminal repeat (LTR). We show for the first time that the nuclear corepressor NCoR is bound to HIV-1 LTR in unstimulated macrophages and is released from the LTR after TLR engagement. Treatment with PPARγ and LXR ligands, but not GR ligands, prevented this TLR-induced clearance of NCoR from the LTR. Our data demonstrate that both classical and nonclassicaltrans-repression mechanisms account for NR-mediated HIV-1 repression. Finally, NR ligand treatment inhibited the potent proinflammatory response induced by PAM3CSK4 that would otherwise activate HIV-1 expression in infected cells. Our findings provide a rationale for studying ligand-activated NRs as modulators of basal and inflammation-induced HIV-1 replication.
De Novo
Infection of B Cells during Murine Gammaherpesvirus 68 Latency
Authors: Freeman Michael L. , Burkum Claire E. , Yager Eric J. , Woodland David L. , Blackman Marcia A.
Published: August 2011

The mechanisms by which gammaherpesviruses maintain latency are unclear. Here we used a murine gammaherpesvirus model to show that previously uninfected B cells in immunocompetent mice can acquire virus during latency.
In vivo
depletion of T cells allowed viral reactivation, as measured by increased viral loads, but not enhanced transfer of virus to new cells. In the absence of both immune T cells and antibody following the transfer of latently infected cells into naïve animals, there was robust infection of new B cells. These data confirm that both T cells and antibody contribute to the control of gammaherpesvirus latency, reactivation, and spread.
Newcastle Disease Virus Expressing Human Immunodeficiency Virus Type 1 Envelope Glycoprotein Induces Strong Mucosal and Serum Antibody Responses in Guinea Pigs
Authors: Khattar Sunil K. , Samal Sweety , DeVico Anthony L. , Collins Peter L. , Samal Siba K.
Published: August 2011

Human immunodeficiency virus type 1 (HIV-1) is transmitted mainly through mucosal sites. Optimum strategies to elicit both systemic and mucosal immunity are critical for the development of vaccines against HIV-1. We therefore sought to evaluate the induction of systemic and mucosal immune responses by the use of Newcastle disease virus (NDV) as a vaccine vector. We generated a recombinant NDV, designated rLaSota/gp160, expressing the gp160 envelope (Env) protein of HIV-1 from an added gene. The gp160 protein expressed by rLaSota/gp160 virus was detected on an infected cell surface and was incorporated into the NDV virion. Biochemical studies showed that gp160 present in infected cells and in the virion formed a higher-order oligomer that retained recognition by conformationally sensitive monoclonal antibodies. Expression of gp160 did not increase the virulence of recombinant NDV (rNDV) strain LaSota. Guinea pigs were administered rLaSota/gp160 via the intranasal (i.n.) or intramuscular (i.m.) route in different prime-boost combinations. Systemic and mucosal antibody responses specific to the HIV-1 envelope protein were assessed in serum and vaginal washes, respectively. Two or three immunizations via the i.n. or i.m. route induced a more potent systemic and mucosal immune response than a single immunization by either route. Priming by the i.n. route was more immunogenic than by the i.m. route, and the same was true for the boosts. Furthermore, immunization with rLaSota/gp160 by any route or combination of routes induced a Th1-type response, as reflected by the induction of stronger antigen-specific IgG2a than IgG1 antibody responses. Additionally, i.n. immunization elicited a stronger neutralizing serum antibody response to laboratory-adapted HIV-1 strain MN.3. These data illustrate that it is feasible to use NDV as a vaccine vector to elicit potent humoral and mucosal responses to the HIV-1 envelope protein.
In SituCleavage of Baculovirus Occlusion-Derived Virus Receptor Binding Protein P74 in the Peroral Infectivity Complex
Authors: Peng Ke , van Lent Jan W. M. , Vlak Just M. , Hu Zhihong , van Oers Monique M.
Published: August 2011

Proteolytic processing of viral membrane proteins is common among enveloped viruses and facilitates virus entry. TheAutographa californicamulticapsid nucleopolyhedrovirus (AcMNPV) occlusion-derived virus (ODV) protein P74 is part of a complex of essential peroral infectivity factors (PIFs). Here we report that P74 is efficiently cleaved into two fragments of about equal size by an occlusion body (OB) endogenous alkaline protease during ODV release when AcMNPV OBs are derived from larvae. The cleavage is specific for P74, since the other known peroral infectivity factors in the same complex (PIF1, PIF2, and PIF3) were not cleaved under the same conditions. P74 cleavage was not observed in OBs produced in three different insect cell lines, suggesting a larval host origin of the responsible protease. P74 in OBs produced in larvae of two different host species was cleaved into fragments with the same apparent molecular mass, indicating that the virus incorporates a similar alkaline protease from different hosts. Coimmunoprecipitation analysis revealed that the two P74 subunit fragments remain associated with the recently discovered PIF complex. We propose that underin vivoODV infection conditions, P74 undergoes two sequential cleavage events, the first one being performed by an ODV-associated host alkaline protease and the second carried out by trypsin in the host midgut.
Determining the Frequency and Mechanisms of HIV-1 and HIV-2 RNA Copackaging by Single-Virion Analysis
Authors: Dilley Kari A. , Ni Na , Nikolaitchik Olga A. , Chen Jianbo , Galli Andrea , Hu Wei-Shau
Published: August 2011

HIV-1 and HIV-2 are derived from two distinct primate viruses and share only limited sequence identity. Despite this, HIV-1 and HIV-2 Gag polyproteins can coassemble into the same particle and their genomes can undergo recombination, albeit at an extremely low frequency, implying that HIV-1 and HIV-2 RNA can be copackaged into the same particle. To determine the frequency of HIV-1 and HIV-2 RNA copackaging and to dissect the mechanisms that allow the heterologous RNA copackaging, we directly visualized the RNA content of each particle by using RNA-binding proteins tagged with fluorescent proteins to label the viral genomes. We found that when HIV-1 and HIV-2 RNA are present in viral particles at similar ratios, ∼10% of the viral particles encapsidate both HIV-1 and HIV-2 RNAs. Furthermore, heterologous RNA copackaging can be promoted by mutating the 6-nucleotide (6-nt) dimer initiation signal (DIS) to discourage RNA homodimerization or to encourage RNA heterodimerization, indicating that HIV-1 and HIV-2 RNA can heterodimerize prior to packaging using the DIS sequences. We also observed that the coassembly of HIV-1 and HIV-2 Gag proteins is not required for the heterologous RNA copackaging; HIV-1 Gag proteins are capable of mediating HIV-1 and HIV-2 RNA copackaging. These results define the
cis-
and
trans
-acting elements required for and affecting the heterologous RNA copackaging, a prerequisite for the generation of chimeric viruses by recombination, and also shed light on the mechanisms of RNA-Gag recognition essential for RNA encapsidation.
High-Risk Human Papillomaviruses Repress Constitutive Kappa Interferon Transcription via E6 To Prevent Pathogen Recognition Receptor and Antiviral-Gene Expression
Authors: Reiser Jeanette , Hurst José , Voges Maike , Krauss Peter , Münch Peter , Iftner Thomas , Stubenrauch Frank
Published: August 2011

Persistent infections with human papillomavirus type 16 (HPV16), HPV18, or HPV31 are necessary for the development of cervical cancer, implying that HPVs have evolved immunoevasive mechanisms. Recent global transcriptome analyses indicated that these HPV types downregulate the constitutive expression of interferon (IFN)-stimulated genes (ISGs), but the underlying mechanism is not well understood. Comparative analyses of ISG transcription in keratinocytes with complete HPV16, -18, and -31 genomes revealed that antiviral genes (IFIT1 and MX1), genes involved in IFN signaling (STAT1), proapoptotic genes (TRAIL and XAF1), and pathogen recognition receptors (TLR3, RIG-I, and MDA5) are inhibited to similar extents by HPV16, -18, and -31. The lower expression of pathogen receptors in HPV-positive cells correlated with a greatly impaired induction of IFN-β and also of IFN-λ1, -2, and -3 upon receptor stimulation. IFN-κ is constitutively expressed in normal keratinocytes and is strongly repressed by HPV16, -18, and -31. ISGs downregulated in HPV-positive cells can be reactivated by IFN-κ expression. The viral E6 and E7 oncogenes are sufficient for IFN-κ repression, with E6 being mainly responsible. E6 inhibits IFN-κ transcription independently from binding to PDZ proteins. IFN-κ expression can be activated in only one cell line by E6AP knockdown but can be activated in all tested HPV-positive cells by addition of a DNA methyltransferase inhibitor, suggesting that HPVs modulate DNA methylation. Taken together, these results suggest that carcinogenic HPVs target IFN-κ by different pathways in keratinocytes to inhibit both antiviral ISGs and pathogen recognition receptors, which in turn reduces the expression of inducible IFNs.
Distal Leucines Are Key Functional Determinants of Alix-Binding Simian Immunodeficiency Virus SIV
smE543
and SIV
mac239
Type 3 L Domains
Authors: Bello Nana F. , Wu Fan , Sette Paola , Dussupt Vincent , Hirsch Vanessa M. , Bouamr Fadila
Published: August 2011

In addition to PTAP L domains, primate lentiviruses carry Alix-binding motifs that include the recently described type 3 SREK
P
Y
KE

V

TED

L

LHLNSLF sequence. We examined the requirements for the type 3 sequence motif in simian immunodeficiency virus SIV
smE543
and identified the
499

L

NS

L

F
503
sequence as a key functional determinant. Mutation of distal leucines
499
L and
502
L (LL mutant) caused an inhibitory effect on Alix-dependent SIV
smE543
release that was quantitatively similar to that observed following disruption of the type 3 L domain or RNA interference (RNAi) depletion of Alix. Similar results were obtained with the SIV
mac239
LL mutant. Thus, distal leucines are key determinants of SIV
smE543
and SIV
mac239
type 3 L domains.
Canine Distemper Virus Infects Canine Keratinocytes and Immune Cells by Using Overlapping and Distinct Regions Located on One Side of the Attachment Protein
Authors: Langedijk Johannes P. M. , Janda Jozef , Origgi Francesco C. , Örvell Claes , Vandevelde Marc , Zurbriggen Andreas , Plattet Philippe
Published: August 2011

The morbilliviruses measles virus (MeV) and canine distemper virus (CDV) both rely on two surface glycoproteins, the attachment (H) and fusion proteins, to promote fusion activity for viral cell entry. Growing evidence suggests that morbilliviruses infect multiple cell types by binding to distinct host cell surface receptors. Currently, the only known
in vivo
receptor used by morbilliviruses is CD150/SLAM, a molecule expressed in certain immune cells. Here we investigated the usage of multiple receptors by the highly virulent and demyelinating CDV strain A75/17. We based our study on the assumption that CDV-H may interact with receptors similar to those for MeV, and we conducted systematic alanine-scanning mutagenesis on CDV-H throughout one side of the β-propeller documented in MeV-H to contain multiple receptor-binding sites. Functional and biochemical assays performed with SLAM-expressing cells and primary canine epithelial keratinocytes identified 11 residues mutation of which selectively abrogated fusion in keratinocytes. Among these, four were identical to amino acids identified in MeV-H as residues contacting a putative receptor expressed in polarized epithelial cells. Strikingly, when mapped on a CDV-H structural model, all residues clustered in or around a recessed groove located on one side of CDV-H. In contrast, reported CDV-H mutants with SLAM-dependent fusion deficiencies were characterized by additional impairments to the promotion of fusion in keratinocytes. Furthermore, upon transfer of residues that selectively impaired fusion induction in keratinocytes into the CDV-H of the vaccine strain, fusion remained largely unaltered. Taken together, our results suggest that a restricted region on one side of CDV-H contains distinct and overlapping sites that control functional interaction with multiple receptors.