bluuebunny/crossref_metadata_2025 huggingface dataset:
Multinationals' Offshore Operations, Tax Avoidance, and Firm‐Specific Information Flows: International Evidence
Authors: Kim Jeong‐Bon , Li TieMei
Published: January 2014

Using a large sample of multinational enterprises (MNEs) over the period 1999–2009, this study investigates whether and how offshore operations via offshore financial centers (OFCs) impact the extent to which firm‐specific information is incorporated into stock price, relative to common information. Our analyses show that, irrespective of whether a firm is a Type I offshore firm (directly having headquarters registered in OFCs) or a Type II offshore firm (indirectly setting up subsidiaries in OFCs), the amount of firm‐specific information flowing into stock price is lower for offshore firms than for non‐offshore firms. We also find that as offshore firms become more aggressive in their tax avoidance strategies, their stock prices impound a lower amount of firm‐specific information relative to common information. Finally, we find that a strong offshore proclivity also deters firm‐specific information flows, thereby driving up stock price synchronicity. Our results suggest that the opaque and complex nature of business and financial transactions in OFCs, coupled with their institutional characteristics, that is, weak and flexible legal enforcement, zero or extremely low taxation, and low litigation risk, provide offshore firms with not only stronger incentives but also the opportunities and means to adopt opaque disclosure policies and aggressive earnings management.
The International Integrated Reporting Framework: Key Issues and Future Research Opportunities
Authors: Cheng Mandy , Green Wendy , Conradie Pieter , Konishi Noriyuki , Romi Andrea
Published: January 2014

This paper has three main aims. First, the paper introduces the concept of integrated reporting () as described by the International Integrated Reporting Council (IIRC). A background to the development of the  concept over the 4‐year period from the inception of the IIRC in 2010 is provided, culminating in the release by the IIRC of a Consultation Draft (CD) of the  framework in March 2013. Second, the paper discusses key issues currently being debated relating to the CD that the IIRC will need to resolve prior to the expected release of their  framework in late 2013. This discussion is based on issues identified and reported to the IIRC by a subcommittee of the International Association for Accounting Education and Research (IAAER) comprised of international accounting academics. Finally, the paper identifies a range of potential research issues relating to the development and implementation of .
Role of beta‐defensin 2 and interleukin‐4 receptor as stroke outcome biomarkers
Authors: García‐Berrocoso Teresa , Giralt Dolors , Bustamante Alejandro , Llombart Victor , Rubiera Marta , Penalba Anna , Boada Cristina , Espadaler Marc , Molina Carlos , Montaner Joan
Published: January 2014

Acute ischemic stroke is a complex disease with huge interindividual evolution variability that makes challenging the prediction of an adverse outcome. Our aim was to study the association of bloodstream signatures to early neurological outcome after stroke, by combining a subpooling of samples strategy with protein array discovery approach. Plasma samples from 36 acute stroke patients (< 4.5 h from onset) were equally pooled within outcome groups: worsening, stability, and improvement (n = 3 pools of four patients each, for each outcome group). These nine pools were screened using a 177 antibodies library, and 35 proteins were found altered regarding outcome classification (p < 0.1). Processes of inflammation, immune response, coagulation, and apoptosis were regulated by these proteins. Ten representative candidates, mainly cytokines and chemokines, were assayed for replication in individual baseline plasma samples from 80 new stroke patients: β‐defensin2, MIP‐3b, plasminogen activator inhibitor 1 active, β‐cell‐attracting chemokine 1, Exodus‐2, interleukin‐4 receptor (IL‐4R), IL‐12p40, leukemia inhibitor factor, MIP‐1b, and tumor necrosis factor‐related weak inducer of apoptosis. Multivariate logistic regression analysis showed β‐defensin 2 (ORadj 4.87 [1.13–20.91] p = 0.033) and IL‐4R (ORadj 3.52 [1.03–12.08] p = 0.045) as independent predictors of worsening at 24 h after adjustment by clinical variables. Both biomarkers improve the prediction by 19% as compared to clinical information, suggesting a potential role for risk stratification in acute thrombolyzed stroke patients.

image

Early neurological deterioration after stroke is not easily predictable. The use of blood biomarkers might help in decision‐making processes regarding this complication. By combining a sub‐pooling of samples strategy with protein array discovery approach, we have found two new biomarkers: beta‐defensin‐2 and interleukin‐4 receptor. Both biomarkers improve the prediction of poor‐outcome over clinical variables in the acute phase of stroke.
Advisory jurisdiction and the European Court of Human Rights: a magic bullet for dialogue and docket-control?
Authors: Dzehtsiarou Kanstantsin , O'Meara Noreen
Published: January 2014

Protocol 16 ECHR will provide for an extension of the advisory jurisdiction of the European Court of Human Rights (ECtHR), enabling highest national courts to request advisory opinions on questions of principle concerning the interpretation of the European Convention on Human Rights (ECHR) or its protocols. This extension of the ECtHR's advisory jurisdiction aims to achieve two goals: a reduction in the ECtHR's excessive docket, and the enhancement of dialogue between the ECtHR and (highest) national courts. While the aims of this reform initiative are laudable, we argue that Protocol 16 is likely to fail to achieve its objectives. Our analysis suggests that rather than facilitating the Court's adjudicatory function, extended advisory jurisdiction has the potential to impact on the Court's constitutionalist function in a manner that can be better achieved through the Court's contentious cases. The burden that this procedure will place on the Court's already overstretched resources would risk delays to contentious cases and potentially undermine judicial comity should requests for advisory opinions be declined. Furthermore, evidence of ‘constructive’ dialogue between highest national courts and the ECtHR is emerging in contentious cases without the need for a reformed advisory opinions mechanism. Rather than achieving its objectives, Protocol 16 risks exacerbating the Court's backlog and nullifying the positive effects of advisory opinions on dialogue.
Experimental evolution reveals habitat‐specific fitness dynamics among Wolbachia clades in Drosophila melanogaster
Authors: Versace Elisabetta , Nolte Viola , Pandey Ram Vinay , Tobler Ray , Schlötterer Christian
Published: January 2014

The diversity and infection dynamics of the endosymbiont Wolbachia can be influenced by many factors, such as transmission rate, cytoplasmic incompatibility, environment, selection and genetic drift. The interplay of these factors in natural populations can result in heterogeneous infection patterns with substantial differences between populations and strains. The causes of these heterogeneities are not yet understood, partly due to the complexity of natural environments. We present experimental evolution as a new approach to study Wolbachia infection dynamics in replicate populations exposed to a controlled environment. A natural Drosophila melanogaster population infected with strains of Wolbachia belonging to different clades evolved in two laboratory environments (hot and cold) for 1.5 years. In both treatments, the rate of Wolbachia infection increased until fixation. In the hot environment, the relative frequency of different Wolbachia clades remained stable over 37 generations. In the cold environment, however, we observed marked changes in the composition of the Wolbachia population: within 15 generations, one Wolbachia clade increased more than 50% in frequency, whereas the other two clades decreased in frequency, resulting in the loss of one clade. The frequency change was highly reproducible not only among replicates, but also when flies that evolved for 42 generations in the hot environment were transferred to the cold environment. These results document how environmental factors can affect the composition of Wolbachia in D. melanogaster. The high reproducibility of the pattern suggests that experimental evolution studies can efficiently determine the functional basis of habitat‐specific fitness among Wolbachia strains.
Genetic variation in regulatory DNA elements: the case of OCA2 transcriptional regulation
Authors: Visser Mijke , Kayser Manfred , Grosveld Frank , Palstra Robert‐Jan
Published: January 2014

Mutations within the OCA2 gene or the complete absence of the OCA2 protein leads to oculocutaneous albinism type 2. The OCA2 protein plays a central role in melanosome biogenesis, and it is a strong determinant of the eumelanin content in melanocytes. Transcript levels of the OCA2 gene are strongly correlated with pigmentation intensities. Recent studies demonstrated that the transcriptional level of OCA2 is to a large extent determined by the noncoding SNP rs12913832 located 21.5 kb upstream of the OCA2 gene promoter. In this review, we discuss current hypotheses and the available data on the mechanism of OCA2 transcriptional regulation and how this is influenced by genetic variation. Finally, we will explore how future epigenetic studies can be used to advance our insight into the functional biology that connects genetic variation to human pigmentation.
Early Pregnancy Changes in Bacterial Vaginosis‐Associated Bacteria and Preterm Delivery
Authors: Nelson Deborah B. , Hanlon Alexandra , Nachamkin Irving , Haggerty Catherine , Mastrogiannis Dimitrios S. , Liu Congzhou , Fredricks David N.
Published: January 2014

Background

We evaluated the importance of measuring early vaginal levels of eight bacterial vaginosis (BV)‐associated bacteria, at two points in pregnancy, and the risk of spontaneous preterm delivery (SPTD) among pregnant women and the subgroup of pregnant women with a history of preterm delivery (PTD).

Methods

This prospective cohort study enrolled women at five urban obstetric practices at Temple University Hospital in Philadelphia PA. Women with singleton pregnancies less than 16 weeks gestation self‐collected vaginal swabs at two points in pregnancy, prior to 16 weeks gestation and between 20–24 weeks gestation, to measure the presence and level of eight BV‐associated bacteria. Women were followed‐up for gestational age at delivery via medical records.

Results

Among women reporting a prior PTD, women with higher levels of Leptotrichia/Sneathia species, BVAB1 and Mobiluncus spp., prior to 16 weeks gestation, were significantly more likely to experience a SPTD. In addition, pregnant women with a prior PTD and increasing levels of Leptotrichia/Sneathia species (aOR: 9.1, 95% CI 1.9, 42.9), BVAB1 (aOR: 16.4, 95% CI 4.3, 62.7) or Megasphaera phylotype 1 (aOR: 6.2, 95% CI 1.9, 20.6), through 24 weeks gestation, were significantly more likely to experience an SPTD. Among the overall group of pregnant women, the levels of BV‐associated bacteria were not related to SPTD.

Conclusion

Among the group of women reporting a prior PTD, increasing levels of BVAB1, Leptotrichia/Sneathia species, and Megasphaera phylotype 1, through mid‐pregnancy were related to an increased risk of SPTD.
Restoring pistil‐side self‐incompatibility factors recapitulates an interspecific reproductive barrier between tomato species
Authors: Tovar‐Méndez Alejandro , Kumar Aruna , Kondo Katsuhiko , Ashford Amy , Baek You S. , Welch Lillian , Bedinger Patricia A. , McClure Bruce A.
Published: January 2014

Interspecific reproductive barriers are poorly understood, but are central to the biological species concept. The pre‐zygotic barriers between red‐ and green‐fruited species in the tomato clade of the genus Solanum provide a model to better understand these barriers in plants. Compatibility usually follows the SI x SC rule: pollen from self‐compatible (SC) red‐fruited species is rejected on pistils of the predominantly self‐incompatible (SI) green‐fruited species, but the reciprocal crosses are compatible. This suggests that the interspecific reproductive barrier may be linked to the intraspecific SI mechanism. However, pollen from the SC red‐fruited species is also rejected by SC accessions of green‐fruited species that lack S‐RNase, a key protein expressed in pistils of SI Solanum species. Thus, multiple mechanisms may contribute to the barrier between red‐ and green‐fruited species. We tested whether an S‐RNase‐dependent barrier is sufficient for rejection of pollen from red‐fruited species by introducing functional S‐RNase, HT‐A and HT‐B genes from SI species into Solanum lycopersicum (cultivated tomato). We found that expressing S‐RNase in combination with either HT‐A or HT‐B in the pistil is sufficient to cause rejection of pollen from all four red‐fruited species. Thus, redundant mechanisms must operate side by side to prevent crosses between red‐ and green‐fruited species in the clade, underlining the complexity of interspecific pollination barriers. Our results also have implications for mating system transitions. We suggest that these transitions must occur in a specific sequence, and that the transition from SI to SC also affects interspecific compatibility.
Trans‐corneal reduction of anterior lens luxation in dogs with lens instability: a retrospective study of 19 dogs (2010–2013)
Authors: Montgomery Keith W. , Labelle Amber L. , Gemensky‐Metzler Anne J.
Published: January 2014

Objective

To evaluate the success rate and outcome of trans‐corneal reduction of anterior lens luxation (TR‐ALL) in dogs with lens instability.

Animals Studied

Nineteen dogs with anterior lens luxation.

Procedures

Medical records of dogs with anterior lens luxation (ALL) were reviewed: dogs were included if TR‐ALL was performed followed by medical therapy in the form of topical 0.005% latanoprost ophthalmic solution. The duration of ALL, vision status, ophthalmic examination abnormalities, reason for performing TR‐ALL and details of TR‐ALL were recorded. Success rate for TR‐ALL as well as postprocedure complications were also assessed.

Results

Twenty eyes from 19 dogs met the inclusion criteria. Median age was 6.5 years (0.3–15 years) and 47% were terriers (9/19). Successful TR‐ALL was achieved in 85% of eyes (17/20); failure was attributed to posterior synechia or vitreous expansion. Short‐term complications included corneal ulceration (2/20) and anterior uveitis (3/20); recurrence of anterior luxation occurred in only one dog. Median follow‐up for visual eyes (11) following successful TR‐ALL was 353 days (range of 1 to 1182 days). Vision was retained in 54.5% (6/11) of eyes with a median time to vision loss of 12 months as a result of glaucoma or presumed retinal detachment.

Conclusions

Trans‐corneal reduction of ALL provides a non‐surgical alternative to intracapsular lens extraction (visual eyes) or enucleation (nonvisual eyes) in dogs. Long‐term visual outcome of TR‐ALL is comparable to intracapsular lens extraction for ALL.
Paradise Locked: The 1918 Influenza Pandemic in American Samoa
Authors: McLane John Ryan
Published: January 2014

The 1918 influenza pandemic killed roughly 5% of the global population. In Polynesia death rates were greater, reaching as high as a quarter of inhabitants in Western Samoa. Despite being less than 100km from the disaster in Western Samoa, and despite the close cultural links with the New Zealand governed colony, American Samoa successfully excluded the infection for years; becoming the largest known state to avoid any deaths from the pandemic. This success was facilitated by isolation, limited trade, a colonial government with absolute power but little oversight, and a working relationship between the US Navy and the traditional Samoan elites. While the crisis would fracture relations with Western Samoa and further isolate American Samoa, the successful quarantine would be seen as a benevolent act by Samoans under American rule and would contribute directly to ongoing American control of the territory.
Doctors and All Blacks: How depression and its treatment is framed in New Zealand GP-targeted advertising
Authors: Wardell Susan Edith
Published: January 2014

Depression, and the treatment of depression is a complex and multifaceted issue, and part of how it is understood and managed by doctors is shaped by print advertising with specific corporate sources and sponsors. This study uses a framing analysis, with attention to semiotics and discourse, to identify the way which depression, depression treatment, and the clinical encounter are framed in New Zealand GP-targeted media. It focuses on two pharmaceutical advertisements (Avanza & Efexor-XR) and one public health campaign (‘The Journal’) running concurrently in NZ Doctor magazine and MIMS New Ethicals, also paying attention to relevant editorial material. Competing frames are identified within these media publications around the biochemical versus psychosocial models of mental illness. The use of former All Black John Kirwan as spokesperson for The Journal contrasts with more typical western feminizations of depression, but nevertheless relates to other New-Zealand-specific cultural stereotypes by linking male mental health specifically with self-help rather than professional treatment. While mirroring the ‘Green Prescription’ movement and seeming to empower patients, The Journal also relies on and reinforces the discourse of the authority of the medical profession to legitimate the treatment process.
Pakeha Identity and Whiteness: What does it mean to be White?
Authors: Gray Claire , Jaber Nabila , Anglem Jim
Published: January 2014

This paper is concerned with a critical examination of whiteness among those who self identify as Pakeha within a New Zealand context. Through in-depth interviews with 15 men and women we explored the proposition that the adoption of a Pakeha identity may preclude an understanding of the ways that whiteness and privilege operate. Employing thematic and discourse analysis, four major themes were identified within the data; a Pakeha relationship with Maori, the reification of whiteness, a disengagement from privilege and a separation from other white people. The functionality and organisation of language were considered in order to examine participants’ detachment from dominant white culture. This article suggests that the assumption of a Pakeha self-identity may allow the bearer to discursively obscure both the cultural capital that whiteness provides and the privileges afforded by this capital. Ultimately, this research draws attention to the intersection of privilege and whiteness within New Zealand and offers an explanation for the persistence of white hegemony.
EUGENICS AND UTOPIA: SOCIAL IMAGINARIES OF TECHNOLOGIES FOR DEAFNESS
Authors: Best Alana , Howland Corinna , Snapp Jenny , Park Julie K
Published: January 2014

A community where everyone speaks Sign? A society where familial deafness condemns people to sterilisation or death? A world where sign languages are suppressed? All have been historically documented: Martha’s Vineyard from 17th-20th centuries; Germany in the 1930s-40s; internationally, for a century from 1880. These and other images comprise divergent social imaginaries which are a context for current and future technologies for deafness. These technologies include postnatal genetic and aural testing for deafness, and may in future include prenatal testing. Cochlear implants can enable profoundly deaf people to hear and newborn hearing screening has recently been introduced in New Zealand. Sign language is another technology whereby deaf people can communicate, create poetry and drama and tell jokes; yet its fortunes have fluctuated over time with oralism’s dominance. Our article draws on two small ethnographic studies in Auckland: one with two families with hereditary deafness; the other with two families and one young adult who had recently chosen cochlear implants, to suggest that individual and societal moral reasoning on the contested issues of technologies for deafness is embedded in different social imaginaries of normalcy.
KEREKERE AND INDIGENOUS SOCIAL ENTREPRENEURSHIP
Authors: Farrelly Trisia Angela , Vudiniabola Alisi Talatoka
Published: January 2014

Since the colonial period, kerekere as an indigenous Fijian mode of exchange has been blamed for stunting the economic development of indigenous Fijians. It has often been reduced to ‘begging’ and it has been used in connection with terms such as ‘corruption‘, and ‘dependency‘. This article strives for a more balanced and culturally complex account of kerekere. Business and vanua; modernity and tradition; moral economy and market economy are often imaged as dichotomous and irreconcilable by indigenous Fijians and others. However, this paper will show that these are false dichotomies, and yet the way they are imagined has a significant bearing on indigenous Fijian business discourse and practices and indigenous Fijian identity. After surveying politics of tradition and indigenous development literature, the authors appeal for integrated economic approaches in business discourse in Fiji which acknowledges kerekere as a potentially positive element in indigenous social entrepreneurship.
Reproduction, ethics and heritable deafness: three South Island families express their views.
Authors: Fitzgerald Ruth Patricia , Bathard Hayley , Broad Rosie , Legge Michael
Published: January 2014

Three consenting South Island (New Zealand) families with experience of living with a heritable type of deafness undertook a series of open ended interviews to ascertain their own views on reproductive decision making and genetic testing. All the families had undergone genetic testing however their orientation to testing in general was cautiously negative. They did not regard deafness as a suitable reason for considering the termination of a pregnancy, although some of the younger adults gave some hesitant consideration to the ‘right to choose’ for others, while rejecting this for themselves. The families were adamant that deafness was not a disability but rather an inconvenience and that only so because a disabling and audist society created certain obstacles in the pathway to achieving good schooling, employment and quality of life.
Left Behind By the Nation: ‘stranded Pakistanis’ in Bangladesh
Authors: Siddiqi Dina Mahnaz
Published: January 2014

To follow...
Adaptive adhesion by a beetle: Manipulation of liquid bridges and their breaking limits
Authors: Macner Ashley M. , Steen Paul H.
Published: January 2014

A drop brought into contact with a nearby substrate can wet and spread against the substrate, forming a liquid bridge that exerts a capillary force. This force due to surface tension can be used to “grab” the substrate, pulling it toward the drop. “Wet” adhesion results from the parallel action of an array of small liquid bridges. The Florida palm beetle, Hemisphaerota cyanea, uses wet adhesion to defend itself against attacking predators by adhering to the palm leaf using an array of about 120 000 μm-sized liquid bridges. The beetle's survival depends on the strength of adhesion which, in turn, depends on how liquid bridges break. Individual bridges break when they go unstable, according to their response curves. However, the ultimate strength of an individual bridge depends on the class of disturbances to which it is subjected, and it has been speculated that the beetle may have some control over this class. The authors experimentally study families of liquid bridge equilibria for their breaking limits when subjected to constant-length (L) and constant-force (F) disturbances. While to control constant-L disturbances is straightforward, to apply and control constant-F disturbances on a liquid bridge requires more ingenuity. The authors introduce an apparatus with a lever-arm and a ball-bearing slide. The authors then compare our experimentally measured bridge response curves to the force trace from experiments on the beetle (prior literature) to infer the mode of beetle detachment. Under normal loads, the beetle detaches as a constant-L instability for smaller loads and as a constant-F instability for larger loads. The beetle's ability to adjust the type and magnitude of loading in real time is not only crucial to its survival but has implications for the design of various engineering devices.
Dose-Dependent Effects of Lactobacillus rhamnosus on Serum Interleukin-17 Production and Intestinal T-Cell Responses in Pigs Challenged with Escherichia coli
Authors: Zhu Yao-Hong , Li Xiao-Qiong , Zhang Wei , Zhou Dong , Liu Hao-Yu , Wang Jiu-Feng
Published: January 2014

The mechanism underlying the dose effect of probiotics on ameliorating diarrhea has not been fully elucidated. Here, low (1 × 10
9
CFU/ml) or high (1 × 10
11
CFU/ml) doses of
Lactobacillus rhamnosus
ATCC 7469 were administered orally to piglets for 1 week before F4 (K88)-positive enterotoxigenic
Escherichia coli
(F4
+
ETEC) challenge. Administration of a low, but not a high, dose of
L. rhamnosus
decreased the percentage of CD3
+
CD4
+
CD8
−
T cells in the peripheral blood. Notably, transiently increased serum concentrations of interleukin-17A (IL-17A) were observed after F4
+
ETEC challenge in pigs pretreated with a high dose of
L. rhamnosus
. Administration of
L. rhamnosus
increased the percentage of the small intestinal lamina propria CD3
+
CD4
+
CD8
−
cells and Peyer's patch CD3
+
CD4
−
CD8
−
and CD3
−
CD4
−
CD8
+
cells. The percentage of ileal intraepithelial CD3
+
CD4
−
CD8
+
cells increased only in the high-dose piglets. Administration of
L. rhamnosus
downregulated expression of ileal IL-17A after F4
+
ETEC challenge but had no effect on expression of gamma interferon (IFN-γ), IL-12, IL-4, and FOXP3 mRNA in the small intestine. Expression of jejunal IL-2, ileal transforming growth factor β1 (TGF-β1), and ileal IL-10 was upregulated in the low-dose piglets after F4
+
ETEC challenge. Our findings suggest that amelioration of infectious diarrhea in piglets by
L. rhamnosus
is associated with the generation of lamina propria CD3
+
CD4
+
CD8
−
T cells, the expansion of Peyer's patch CD3
+
CD4
−
CD8
−
and CD3
−
CD4
−
CD8
+
cells, and the attenuation of F4
+
ETEC-induced increase in CD3
+
CD4
+
CD8
+
T cells in the small intestine. However, consumption of high doses of
L. rhamnosus
may increase levels of serum IL-17A after F4
+
ETEC challenge, thus eliciting a strong proinflammatory response.
Immunogenicity and Safety of a Booster Dose of an Investigational Adjuvanted Polyprotein HIV-1 Vaccine in Healthy Adults and Effect of Administration of Chloroquine
Authors: Leroux-Roels Geert , Bourguignon Patricia , Willekens Julie , Janssens Michel , Clement Frédéric , Didierlaurent Arnaud M. , Fissette Laurence , Roman François , Boutriau Dominique
Published: January 2014

This phase II study evaluated the effect of chloroquine on the specific CD8
+
T-cell responses to and the safety of a booster dose of investigational human immunodeficiency virus type 1 (HIV-1) F4/AS01
B
vaccine containing 10 μg of recombinant fusion protein (F4) adjuvanted with the AS01
B
adjuvant system. Healthy adults aged 21 to 41 years, primed 3 years before with two F4/AS01
B
doses containing 10 or 30 μg of F4 (
ClinicalTrials.gov
registration number NCT00434512), were randomized (1:1) to receive the F4/AS01
B
booster administered alone or 2 days after chloroquine (300 mg). F4-specific CD8
+
/CD4
+
T-cell responses were characterized by intracellular cytokine staining and lymphoproliferation assays and anti-F4 antibodies by enzyme-linked immunosorbent assays (ELISAs). No effect of chloroquine on CD4
+
/CD8
+
T-cell and antibody responses and no vaccine effect on CD8
+
T-cell responses (cytokine secretion or proliferation) were detected following F4/AS01
B
booster administration.
In vitro
, chloroquine had a direct inhibitory effect on AS01
B
adjuvant properties; AS01-induced cytokine production decreased upon coincubation of cells with chloroquine. In the pooled group of participants primed with F4/AS01
B
containing 10 μg of F4, CD4
+
T-cell and antibody responses induced by primary vaccination persisted for at least 3 years. The F4/AS01
B
booster induced strong F4-specific CD4
+
T-cell responses, which persisted for at least 6 months with similar frequencies and polyfunctional phenotypes as following primary vaccination, and high anti-F4 antibody concentrations, reaching higher levels than those following primary vaccination. The F4/AS01
B
booster had a clinically acceptable safety and reactogenicity profile. An F4/AS01
B
booster dose, administered alone or after chloroquine, induced robust antibody and F4-specific CD4
+
T-cell responses but no significant CD8
+
T-cell responses (cytokine secretion or proliferation) in healthy adults. (This study has been registered at
ClinicalTrials.gov
under registration number NCT00972725).
Role of Phosphatidylserine in Phospholipid Flippase-Mediated Vesicle Transport in Saccharomyces cerevisiae
Authors: Takeda Miyoko , Yamagami Kanako , Tanaka Kazuma
Published: January 2014

Phospholipid flippases translocate phospholipids from the exoplasmic to the cytoplasmic leaflet of cell membranes to generate and maintain phospholipid asymmetry. The genome of budding yeast encodes four heteromeric flippases (Drs2p, Dnf1p, Dnf2p, and Dnf3p), which associate with the Cdc50 family noncatalytic subunit, and one monomeric flippase Neo1p. Flippases have been implicated in the formation of transport vesicles, but the underlying mechanisms are largely unknown. We show here that overexpression of the phosphatidylserine synthase gene
CHO1
suppresses defects in the endocytic recycling pathway in flippase mutants. This suppression seems to be mediated by increased cellular phosphatidylserine. Two models can be envisioned for the suppression mechanism: (i) phosphatidylserine in the cytoplasmic leaflet recruits proteins for vesicle formation with its negative charge, and (ii) phosphatidylserine flipping to the cytoplasmic leaflet induces membrane curvature that supports vesicle formation. In a mutant depleted for flippases, a phosphatidylserine probe GFP-Lact-C2 was still localized to endosomal membranes, suggesting that the mere presence of phosphatidylserine in the cytoplasmic leaflet is not enough for vesicle formation. The
CHO1
overexpression did not suppress the growth defect in a mutant depleted or mutated for all flippases, suggesting that the suppression was dependent on flippase-mediated phospholipid flipping. Endocytic recycling was not blocked in a mutant lacking phosphatidylserine or depleted in phosphatidylethanolamine, suggesting that a specific phospholipid is not required for vesicle formation. These results suggest that flippase-dependent vesicle formation is mediated by phospholipid flipping, not by flipped phospholipids.
Assessment of Microbiological Diagnostic Procedures for Respiratory Specimens from Cystic Fibrosis Patients in German Laboratories by Use of a Questionnaire
Authors: Häfner Laura , Peters Georg , Kahl Barbara C.
Published: January 2014

Respiratory specimens from cystic fibrosis (CF) patients challenge microbiological laboratories with their complexity of pathogens and atypical variants. We evaluated the diagnostic procedures in German laboratories by use of a questionnaire. Although most laboratories followed guidelines, some of them served only a small number of patients, while others did not use the recommended selective agars to culture the particular CF-relevant species.
Malsoor Virus, a Novel Bat Phlebovirus, Is Closely Related to Severe Fever with Thrombocytopenia Syndrome Virus and Heartland Virus
Authors: Mourya D. T. , Yadav P. D. , Basu A. , Shete A. , Patil D. Y. , Zawar D. , Majumdar T. D. , Kokate P. , Sarkale P. , Raut C. G. , Jadhav S. M.
Published: January 2014

During a survey in the year 2010, a novel phlebovirus was isolated from the
Rousettus leschenaultii
species of bats in western India. The virus was identified by electron microscopy from infected Vero E6 cells. Phylogenic analysis of the complete genome showed its close relation to severe fever with thrombocytopenia syndrome (SFTS) and Heartland viruses, which makes it imperative to further study its natural ecology and potential as a novel emerging zoonotic virus.
Enterovirus 2A
pro
Targets MDA5 and MAVS in Infected Cells
Authors: Feng Qian , Langereis Martijn A. , Lork Marie , Nguyen Mai , Hato Stanleyson V. , Lanke Kjerstin , Emdad Luni , Bhoopathi Praveen , Fisher Paul B. , Lloyd Richard E. , van Kuppeveld Frank J. M.
Published: January 2014

RIG-I-like receptors (RLRs) MDA5 and RIG-I are key players in the innate antiviral response. Upon recognition of viral RNA, they interact with MAVS, eventually inducing type I interferon production. The interferon induction pathway is commonly targeted by viruses. How enteroviruses suppress interferon production is incompletely understood. MDA5 has been suggested to undergo caspase- and proteasome-mediated degradation during poliovirus infection. Additionally, MAVS is reported to be cleaved during infection with coxsackievirus B3 (CVB3) by the CVB3 proteinase 3C
pro
, whereas MAVS cleavage by enterovirus 71 has been attributed to 2A
pro
. As yet, a detailed examination of the RLR pathway as a whole during any enterovirus infection is lacking. We performed a comprehensive analysis of crucial factors of the RLR pathway, including MDA5, RIG-I, LGP2, MAVS, TBK1, and IRF3, during infection of CVB3, a human enterovirus B (HEV-B) species member. We show that CVB3 inhibits the RLR pathway upstream of TBK1 activation, as demonstrated by limited phosphorylation of TBK1 and a lack of IRF3 phosphorylation. Furthermore, we show that MDA5, MAVS, and RIG-I all undergo proteolytic degradation in CVB3-infected cells through a caspase- and proteasome-independent manner. We convincingly show that MDA5 and MAVS cleavages are both mediated by CVB3 2A
pro
, while RIG-I is cleaved by 3C
pro
. Moreover, we show that proteinases 2A
pro
and 3C
pro
of poliovirus (HEV-C) and enterovirus 71 (HEV-A) exert the same functions. This study identifies a critical role of 2A
pro
by cleaving MDA5 and MAVS and shows that enteroviruses use a common strategy to counteract the interferon response in infected cells.

IMPORTANCE
Human enteroviruses (HEVs) are important pathogens that cause a variety of diseases in humans, including poliomyelitis, hand, foot, and mouth disease, viral meningitis, cardiomyopathy, and more. Like many other viruses, enteroviruses target the host immune pathways to gain replication advantage. The MDA5/MAVS pathway is responsible for recognizing enterovirus infections in the host cell and leads to expression of type I interferons (IFN-I), crucial antiviral signaling molecules. Here we show that three species of HEVs all employ the viral proteinase 2A (2A
pro
) to proteolytically target MDA5 and MAVS, leading to an efficient blockade upstream of IFN-I transcription. These observations suggest that MDA5/MAVS antagonization is an evolutionarily conserved and beneficial mechanism of enteroviruses. Understanding the molecular mechanisms of enterovirus immune evasion strategies will help to develop countermeasures to control infections with these viruses in the future.
The Bean Pod Mottle Virus RNA2-Encoded 58-Kilodalton Protein P58 Is Required in
cis
for RNA2 Accumulation
Authors: Lin Junyan , Guo Jiangbo , Finer John , Dorrance Anne E. , Redinbaugh Margaret G. , Qu Feng
Published: January 2014

Bean pod mottle virus (BPMV) is a bipartite, positive-sense (+) RNA plant virus in the
Secoviridae
family. Its RNA1 encodes proteins required for genome replication, whereas RNA2 primarily encodes proteins needed for virion assembly and cell-to-cell movement. However, the function of a 58-kDa protein (P58) encoded by RNA2 has not been resolved. P58 and the movement protein (MP) of BPMV are two largely identical proteins differing only at their N termini, with P58 extending MP upstream by 102 amino acid residues. In this report, we unveil a unique role for P58. We show that BPMV RNA2 accumulation in infected cells was abolished when the start codon of P58 was eliminated. The role of P58 does not require the region shared by MP, as RNA2 accumulation in individual cells remained robust even when most of the MP coding sequence was removed. Importantly, the function of P58 required the P58 protein, rather than its coding RNA, as compensatory mutants could be isolated that restored RNA2 accumulation by acquiring new start codons upstream of the original one. Most strikingly, loss of P58 function could not be complemented by P58 provided in
trans
, suggesting that P58 functions in
cis
to selectively promote the accumulation of RNA2 copies that encode a functional P58 protein. Finally, we found that all RNA1-encoded proteins are
cis
-acting relative to RNA1. Together, our results suggest that P58 probably functions by recruiting the RNA1-encoded polyprotein to RNA2 to enable RNA2 reproduction.

IMPORTANCE
Bean pod mottle virus (BPMV) is one of the most important pathogens of the crop plant soybean, yet its replication mechanism is not well understood, hindering the development of knowledge-based control measures. The current study examined the replication strategy of BPMV RNA2, one of the two genomic RNA segments of this virus, and established an essential role for P58, one of the RNA2-encoded proteins, in the process of RNA2 replication. Our study demonstrates for the first time that P58 functions preferentially with the very RNA from which it is translated, thus greatly advancing our understanding of the replication mechanisms of this and related viruses. Furthermore, this study is important because it provides a potential target for BPMV-specific control, and hence could help to mitigate soybean production losses caused by this virus.
Toll-Like Receptor 7/8 (TLR7/8) and TLR9 Agonists Cooperate To Enhance HIV-1 Envelope Antibody Responses in Rhesus Macaques
Authors: Moody M. Anthony , Santra Sampa , Vandergrift Nathan A. , Sutherland Laura L. , Gurley Thaddeus C. , Drinker Mark S. , Allen Ashley A. , Xia Shi-Mao , Meyerhoff R. Ryan , Parks Robert , Lloyd Krissey E. , Easterhoff David , Alam S. Munir , Liao Hua-Xin , Ward Brandy M. , Ferrari Guido , Montefiori David C. , Tomaras Georgia D. , Seder Robert A. , Letvin Norman L. , Haynes Barton F.
Published: January 2014

The development of a vaccine that can induce high titers of functional antibodies against HIV-1 remains a high priority. We have developed an adjuvant based on an oil-in-water emulsion that incorporates Toll-like receptor (TLR) ligands to test whether triggering multiple pathogen-associated molecular pattern receptors could enhance immunogenicity. Compared to single TLR agonists or other pairwise combinations, TLR7/8 and TLR9 agonists combined were able to elicit the highest titers of binding, neutralizing, and antibody-dependent cellular cytotoxicity-mediating antibodies against the protein immunogen, transmitted/founder HIV-1 envelope gp140 (B.63521). We further found that the combination of TLR7/8 and TLR9 agonists was associated with the release of CXCL10 (IP-10), suggesting that this adjuvant formulation may have optimally stimulated innate and adaptive immunity to elicit high titers of antibodies.

IMPORTANCECombining TLR agonists in an adjuvant formulation resulted in higher antibody levels compared to an adjuvant without TLR agonists. Adjuvants that combine TLR agonists may be useful for enhancing antibody responses to HIV-1 vaccines.
HLA-B*57 Elite Suppressor and Chronic Progressor HIV-1 Isolates Replicate Vigorously and Cause CD4
+
T Cell Depletion in Humanized BLT Mice
Authors: Salgado Maria , Swanson Michael D. , Pohlmeyer Christopher W. , Buckheit Robert W. , Wu Jin , Archin Nancie M. , Williams Thomas M. , Margolis David M. , Siliciano Robert F. , Garcia J. Victor , Blankson Joel N.
Published: January 2014

Elite controllers or suppressors (ES) are HIV-1-infected patients who maintain undetectable viral loads without antiretroviral therapy. The mechanism of control remains unclear, but the HLA-B*57 allele is overrepresented in cohorts of these patients. However, many HLA-B*57 patients develop progressive disease, and some studies have suggested that infection with defective viruses may be the cause of the lack of high levels of virus replication and disease progression in ES. We therefore performed a comprehensive comparative
in vivo
and
in vitro
characterization of viruses isolated from well-defined ES. For this purpose, we first performed full-genome sequence analysis and
in vitro
fitness assays on replication-competent isolates from HLA-B*57 ES and HLA-B*57 chronic progressors (CPs). Under our experimental conditions, we found that isolates from ES and CPs can replicate
in vitro
. However, since inherently these assays involve the use of unnaturally
in vitro
-activated cells, we also investigated the replication competence and pathogenic potential of these HIV isolates
in vivo
using humanized BLT mice. The results from these analyses demonstrate that virus isolates from ES are fully replication competent
in vivo
and can induce peripheral and systemic CD4 T cell depletion. These results provide the first direct
in vivo
evidence that viral fitness does not likely determine clinical outcome in HLA-B*57 patients and that elite suppressors can control replication-competent, fully pathogenic viruses. A better understanding of the immunological bases of viral suppression in ES will serve to inform novel approaches to preventive and therapeutic HIV vaccine design.

IMPORTANCE
Elite suppressors are HIV-1-infected patients who have undetectable levels of viremia despite not being on antiviral drugs. One of the most fundamental questions about this phenomenon involves the mechanism of control. To address this question, we isolated virus from elite suppressors and from HIV-1-infected patients who have the usual progressive disease course. We compared how well the isolates from the two groups of patients replicated in culture and in humanized mice. Our results suggest that elite suppressors are capable of controlling HIV-1 due to the possession of unique host factors rather than infection with defective virus.
Feline Immunodeficiency Virus Envelope Glycoproteins Antagonize Tetherin through a Distinctive Mechanism That Requires Virion Incorporation
Authors: Morrison James H. , Guevara Rebekah B. , Marcano Adriana C. , Saenz Dyana T. , Fadel Hind J. , Rogstad Daniel K. , Poeschla Eric M.
Published: January 2014

BST2/tetherin inhibits the release of enveloped viruses from cells. Primate lentiviruses have evolved specific antagonists (Vpu, Nef, and Env). Here we characterized tetherin proteins of species representing both branches of the order Carnivora. Comparison of tiger and cat (Feliformia) to dog and ferret (Caniformia) genes demonstrated that the tiger and cat share a start codon mutation that truncated most of the tetherin cytoplasmic tail early in the Feliformia lineage (19 of 27 amino acids, including the dual tyrosine motif). Alpha interferon (IFN-α) induced tetherin and blocked feline immunodeficiency virus (FIV) replication in lymphoid and nonlymphoid feline cells. Budding of bald FIV and HIV particles was blocked by carnivore tetherins. However, infectious FIV particles were resistant, and spreading FIV replication was uninhibited. Antagonism mapped to the envelope glycoprotein (Env), which rescued FIV from carnivore tetherin restriction when expressed in
trans
but, in contrast to known antagonists, did not rescue noncognate particles. Also unlike the primate lentiviral antagonists, but similar to the Ebola virus glycoprotein, FIV Env did not reduce intracellular or cell surface tetherin levels. Furthermore, FIV-enveloped FIV particles actually required tetherin for optimal release from cells. The results show that FIV Envs mediate a distinctive tetherin evasion. Well adapted to a phylogenetically ancient tetherin tail truncation in the Felidae, it requires functional virion incorporation of Env, and it shields the budding particle without downregulating plasma membrane tetherin. Moreover, FIV has evolved dependence on this protein: particles containing FIV Env need tetherin for optimal release from the cell, while Env
−
particles do not.

IMPORTANCE
HIV-1 antagonizes the restriction factor tetherin with the accessory protein Vpu, while HIV-2 and the filovirus Ebola use their envelope (Env) glycoproteins for this purpose. It turns out that the FIV tetherin antagonist is also its Env protein, but the mechanism is distinctive. Unlike other tetherin antagonists, FIV Env cannot act in
trans
to rescue
vpu
-deficient HIV-1. It must be incorporated specifically into FIV virions to be active. Also unlike other retroviral antagonists, but similar to Ebola virus Env, it does not act by downregulating or degrading tetherin. FIV Env might exclude tetherin locally or direct assembly to tetherin-negative membrane domains. Other distinctive features are apparent, including evidence that this virus evolved an equilibrium in which tetherin is both restriction factor and cofactor, as FIV requires tetherin for optimal particle release.
Influence of Different Durations of Electroacupuncture Stimulation on Skin Blood Flow and Muscle Blood Volume
Authors: Mori Hidetoshi , Kuge Hiroshi , Tanaka Tim Hideaki , Taniwaki Eiichi
Published: January 2014

Objective

To determine whether skin blood flow (SBF) and muscle blood volume (MBV) responses differ according to the duration of electroacupuncture (EA) stimulation.

Methods

Two experiments were conducted. In Experiment 1, healthy male students (n=20) were randomly divided into two groups (an EA group and a no-stimulation group). In Experiment 2, healthy male students (n=37) were randomly divided into three groups with different stimulation durations of EA (5, 10 and 15 min). EA stimulation on the quadriceps femoris muscle was administered with a frequency of 1 Hz. SBF and MBV were estimated before, during and after stimulation.

Results

In Experiment 1 there was a significant difference between the groups (p=0.001). In the EA stimulation group, MBV increased immediately following EA (p=0.045) and 5 min (p=0.005) and 10 min (p=0.002) after EA compared with baseline. No significant change was observed in the no-stimulation group. In Experiment 2 there was no significant difference between the groups. All three stimulation durations (5, 10 and 15 min) yielded increases in MBV after stimulation compared with baseline. SBF following stimulation for 5 and 10 min was increased compared with baseline.

Conclusions

Based on these findings, we consider that 10 min EA stimulation could be a sufficient duration to induce a blood flow response in a clinical situation when increasing the blood flow is desirable.