bluuebunny/crossref_metadata_2025 huggingface dataset:
Enough is Enough
Authors: Whittaker‐Johns Barbara
Published: December 2003

Abstract.God, the sacred, is radically indwelling and immanent; therefore, nature is enough to satisfy our thirst for transcendence. The transcendent is the radically indwelling capacity of love to bring new life and a sense of being at home in the universe to us and to the people of the world. The epic of evolution, the scientific story of evolution understood as cosmogenesis, is the foundation of this position. However, my primary focus is on one particular practice of internalizing the transforming power of the scientific story of evolution—that of story. Here the practice of story is understood as framing moments of our encounters with matters of faith and love, creating narratives out of these moments, embracing these narratives as a kind of personal scripture, and telling these narratives as a source of inspiration and of guidance about how we are to be of use in the world.
On the Relationship between Science and Ethics
Authors: Pigliucci Massimo
Published: December 2003

Abstract.The relationship between ethics and science has been discussed within the framework of continuity versus discontinuity theories, each of which can take several forms. Continuity theorists claim that ethics is a science or at least that it has deep similarities with the modus operandi of science. Discontinuity theorists reject such equivalency, while at the same time many of them claim that ethics does deal with objective truths and universalizable statements, just not in the same sense as science does. I propose here a third view ofquasi‐continuity(or, equivalently,quasi‐discontinuity) that integrates ethics and science as equal partners toward the uncovering of new knowledge. In this third way, a program envisioned by William James but made practicable only by contemporary scientific advancement, science can and must inform ethics at a deep level, and ethical theory— while going beyond science—cannot do without it. In particular, I identify four areas of ethics‐science collaboration: neurobiological research into the basis of moral judgment, comparative anthropology, comparative evolutionary biology of primates, and game‐theoretical modeling. I provide examples within each of these fields to show how they link to ethical theories (including prescriptive work) and questions. The essay concludes with a brief discussion of the light that a scientifically informed ethics can shed on some classical problems in moral theory, such as the relationships between rationality and selfishness, egoism and altruism, as well as the concept of social contract. A joint research program involving both philosophers and scientists is called for if we wish to move ethical theory into the twenty‐first century.
Minding Animals, Minding Earth: Old Brains, New Bottlenecks
Authors: Bekoff Marc
Published: December 2003

Abstract. I emphasize the importance of broadening behavioral, ecological, and conservation science into a more integrative, interdisciplinary, socially responsible, compassionate, spiritual, and holistic endeavor. I stress the significance of studies of animal behavior, especially ethological research concerned with animal emotions in which individuals are named and recognized for their own personalities, for helping us to learn not only about the nonhuman animal beings with whom we share Earth but also about who we are and our place in nature. We are best understood in relationship with others. To this end I develop the notions of “minding animals” and “deep ethology.” Animals are sources of wisdom, a way of knowing.

We are all citizens of Earth, members of a global community in which intimate reciprocal and beneficent peaceful relationships are mandatory. A world without cruelty and with boundless compassion, respect, grace, humility, spirituality, and love would be a better world in which to live. We have compelling responsibilities for making Earth a better and more peaceful habitat for all beings. It is essential that we do better than our ancestors. We must reflect and step lightly as we “redecorate” nature. Time is not on our side.

I plead for the development of heartfelt and holistic science that allows for joy and play. Science need not be suspicious of things it cannot fully understand. We must not avert our eyes or other senses from the eyes and voices of other beings who urgently need our uncompromising and unconditional aid and love. We can do much more than we have done for animals and the Earth.
Religion, Science, and Evolution: Paul Tillich's Fourth Way
Authors: Grigg Richard
Published: December 2003

Abstract. In his book God After Darwin John Haught provides a useful categorization of theological approaches to evolution: some theologians actively oppose Darwinian evolution, another group maintains that science and religion have nothing to say to one another, and a third seeks to engage evolution. Haught wishes to pursue the third way. But many theological attempts to talk about divine action in the world, including divine involvement in the process of evolution, run afoul of the scientific principle of the conservation of matter‐energy. Haught's reliance on the now‐familiar notion that information can have causal efficacy does not in fact escape this difficulty. I suggest a fourth approach, represented by a constructive reading of Paul Tillich's theology. The central argument is that Tillich offers a way of taking Darwinian evolution up into one's ultimate concern without claiming that God has any causal relation to evolution. God provides no historical telos for evolution, but rather a “depth teleology” that springs from the manner in which God, as the depth of the structure of finite being, is the object of Christian faith.
Globalization, the new economy, and the commodification of language and identity
Authors: Heller Monica
Published: December 2003

The globalized new economy is bound up with transformations of language and identity in many different ways (cf., e.g. Bauman 1997; Castells 2000; Giddens 1990). These include emerging tensions between State‐based and corporate identities and language practices, between local, national and supra‐national identities and language practices, and between hybridity and uniformity. Ethnolinguistic minorities provide a particularly revealing window into these processes. In this paper, I explore ways in which the globalized new economy has resulted in the commodification of language and identity, sometimes separately, sometimes together. The paper is based on recent ethnographic, sociolinguistic research in francophone areas of Canada.
Global Englishes, Rip Slyme, and performativity
Authors: Pennycook Alastair
Published: December 2003

In this article I suggest that while recent sociolinguistic work focusing on crossing, styling the Other or language boundaries is raising significant questions concerning how we relate language, identity and popular culture, these insights have largely passed by the sociolinguistics of world Englishes. This latter work is still caught between arguments about homogeneity and heterogeneity, between arguments based on liberal accommodationism, linguistic imperialism or linguistic hybridity that do not allow for sufficiently complex understandings of what is currently happening with global Englishes. Focusing particularly on rap music, I suggest that we need, at the very least, a critical understanding of globalization, a focus on popular cultural flows, and a way of taking up performance and performativity in relationship to identity and culture.
Communicating a global reach: Inflight magazines as a globalizing genre in tourism
Authors: Thurlow Crispin , Jaworski Adam
Published: December 2003

Like most multinational or international corporations, marketing for airlines hinges on careful strategies of differentiation and promotion; an increasingly important part of the way this is achieved is for airlines to position themselves as ‘global’ and to promise their passengers the kudos of being global citizens and travellers. It is in this way that globalization can be seen to be not about just economic reordering, but also strategic, commercial rebranding; globalization is thus both cultural discourse and identity resource. This is exemplified by the 72 inflight magazines we examine here, and achieved by a series of discursive strategies such as: their cashing in on the global cachet (or ‘worldliness’) of English; their drawing on metonymic repertoires of global cities/destinations and celebrities; and their striking displays of global route‐maps regardless of actual network. Inflight magazines are both global medium (or genre) in themselves and carriers of global messages. Drawing on the discourses of nationality and globality, they epitomize a post‐modern tendency towards the ‘globalization of nationality’ as well as espousing a global lifestyle.
The Eurozone and financial integration: the employment relations issues
Authors: Grahl John , Teague Paul
Published: December 2003

Together the macroeconomic and financial attributes of monetary union are changing the rules governing the European economy. This paper assesses the consequences of these changes for employment relations in member states. The analysis focuses on three matters in particular. First, how labour market actors are responding to the fact that macroeconomic policies are not only out of their own control but may therefore fail to correspond to the needs of particular countries. Second, how employment relations are adapting to the new microeconomic challenges released by the move towards deeper financial integration inside the EU, which has its main impact on the structures of corporate finance and thus on corporate strategy. Finally the implications of both patterns of change for the EU employment policy are assessed to gauge the impact of monetary union on the European social model.
Employment relations in Central and Eastern Europe in 2002: towards EU accession
Authors: Martin Roderick , Cristescu‐Martin Anamaria
Published: December 2003

The difficult process of bringing CEE institutions and practices into line with those in the existing EU was accelerated at Copenhagen in December 2002 as accession negotiations were concluded. Elsewhere in the CEE, organised labour remained weak under high levels of unemployment and fewer workers entering the labour force as individual employers took advantage of favourable bargaining conditions and the absence of significant multi‐employer bargaining. Unions did, however, retain their political influence as seen in the revisions to Labour Codes in several countries in 2002. They also had expectations of greater post‐accession influence through their incorporation into the EU's social partnership framework.
Small cell lung cancer: from molecular biology to novel therapeutics
Authors: Pisick Evan , Jagadeesh Simha , Salgia Ravi
Published: December 2003

Small cell lung cancer (SCLC) is an aggressive tumor which metastasizes early. Patients with this disease have a poor prognosis even with immediate treatment. Because of the aggressive nature of this disease, all aspects of this tumor are studied extensively. This review will provide an update of the biology of SCLC at both the molecular and cellular levels. Cellular pathways and their relationship to cellular function will also be discussed. Treatment of both primary limited‐ and extensive‐stage diseases as well as recurrent disease will be discussed including chemotherapy, thoracic radiotherapy, and surgery. The role of novel therapeutics being investigated will also be addressed.
Laser‐assisted microdissection applied to frozen surgical pathologic specimens – methodological aspects on RT‐PCR
Authors: Saal Isabelle , Gustin Anne , Rombaut Katia , Pelaez Pilar , Rorive Sandrine , Remmelink Myriam , Salmon Isabelle
Published: December 2003

Laser‐assisted microdissection has become a unique technique for an accurate gene‐expression profiling analysis in human tissues. The introduction of this approach requires the development of a reliable, efficient, and reproducible procedure for tissue processing. We report a systematic evaluation of the different relevant steps required to obtain sufficient quantity and good quality RNA for reverse transcriptase‐polymerase chain reaction when using frozen surgical pathologic tissues as starting material. We propose an optimized and very efficient method with respect to time and effort that can easily be put into practice in research laboratory as well as in any pathology laboratory.
Decision Making with Uncertain Judgments: A Stochastic Formulation of the Analytic Hierarchy Process*
Authors: Hahn Eugene D.
Published: December 2003

In the analytic hierarchy process (AHP), priorities are derived via a deterministic method, the eigenvalue decomposition. However, judgments may be subject to error. A stochastic characterization of the pairwise comparison judgment task is provided and statistical models are introduced for deriving the underlying priorities. Specifically, a weighted hierarchical multinomial logit model is used to obtain the priorities. Inference is then conducted from the Bayesian viewpoint using Markov chain Monte Carlo methods. The stochastic methods are found to give results that are congruent with those of the eigenvector method in matrices of different sizes and different levels of inconsistency. Moreover, inferential statements can be made about the priorities when the stochastic approach is adopted, and these statements may be of considerable value to a decision maker. The methods described are fully compatible with judgments from the standard version of AHP and can be used to construct a stochastic formulation of it.
Does Knowledge Mediate the Effect of Context on Performance? Some Initial Evidence
Authors: Dröge Cornelia , Claycomb Cindy , Germain Richard
Published: December 2003

This research examines the links among four firm context variables, knowledge, and financial performance. Adopting a knowledge‐based, contingency view of the firm and assuming that strategy's locus is knowledge creation and application, we hypothesize that knowledge completely mediates the effects of context on performance. The results from analyzing 208 manufacturers found a positive relationship between applied knowledge and financial performance, but none between knowledge creation and performance. As for context, production technology routineness and low demand unpredictability were positively related only to greater applied knowledge; high technological turbulence was positively related to both knowledge creation and knowledge application; and firm size had no effect on either knowledge construct. No direct context‐performance relationships were found (i.e., all effects were indirect): knowledge, in particular applied knowledge, completely mediated the impact of context on performance.
Confirmatory Factor Analysis of the End‐User Computing Satisfaction Instrument: Replication within an ERP Domain*
Authors: Somers Toni M. , Nelson Klara , Karimi Jahangir
Published: December 2003

Over the past decade, organizations have made significant investments in enterprise resource planning (ERP) systems. The realization of benefits from these investments depends on supporting effective use of information technology (IT) and satisfying IT users. User satisfaction with information systems is one of the most important determinants of the success of those systems. Drawing upon a sample of 407 end users of ERP systems and working within the framework of confirmatory factor analysis (CFA), this study examines the structure and dimensionality, and reliability and validity of the end‐user computing satisfaction (EUCS) instrument posited by Doll and Torkzadeh (1988). In response to Klenke's (1992) motion to cross‐validate management information system (MIS) instruments and to retest the end user computing satisfaction instrument using new data, this study's results, consistent with previous findings, confirm that the EUCS instrument maintains its psychometric stability when applied to users of enterprise resource planning application software. Implications of these results for practice and research are provided.
Multiple Conceptualizations of Small Business Web Use and Benefit*
Authors: Pflughoeft Kurt A. , Ramamurthy K. , Soofi Ehsan S. , Yasai‐Ardekani Masoud , Zahedi Fatemeh (Mariam)
Published: December 2003

Small businesses play an important role in the U.S. economy and there is anecdotal evidence that use of the Web is beneficial to such businesses. There is, however, little systematic analysis of the conditions that lead to successful use of and thereby benefits from the Web for small businesses. Based on the innovation adoption, organizations, and information systems (IS) implementation literature, we identify a set of variables that are related to adoption, use, and benefits of information technology (IT), with particular emphasis on small businesses. These variables are reflective of an organization's contextual characteristics, its IT infrastructure, Web use, and Web benefits. Since the extant research does not suggest a single theoretical model for Web use and benefits in the context of small businesses, we adopt a modeling approach and explore the relationships between “context‐IT‐use‐benefit” (CIUB) through three models—partial‐mediator, reduced partial‐mediator, and mediator. These models posit that the extent of Web use by small businesses and the associated benefits are driven by organizations’ contextual characteristics and their IT infrastructure. They differ in the endogeneity/exogeneity of the extent of IT sophistication, and in the direct/mediated effects of organizational context. We examine whether the relationships between variables identified in the literature hold within the context of these models using two samples of small businesses with national coverage, including various sizes, and representing several industry sectors. The results show that the evidence for patterns of relationships is similar across the two independent samples for two of these models. We highlight the relationships within the reduced partial‐mediator and mediator models for which conclusive evidence are given by both samples. Implications for small business managers and providers of Web‐based technologies are discussed.
Liquid secretion properties of airway submucosal glands
Authors: Ballard Stephen T. , Inglis Sarah K.
Published: December 2003

The tracheobronchial submucosal glands secrete liquid that is important for hydrating airway surfaces, supporting mucociliary transport, and serving as a fluid matrix for numerous secreted macromolecules including the gel‐forming mucins. This review details the essential structural elements of airway glands and summarizes what is currently known regarding the ion transport processes responsible for producing the liquid component of gland secretion. Liquid secretion most likely arises from serous cells and is principally under neural control with muscarinic agonists, substance P, and vasoactive intestinal peptide (VIP) functioning as effective secretogogues. Liquid secretion is driven by the active transepithelial secretion of both Cl− and HCO3− and at least a portion of this process is mediated by the cystic fibrosis transmembrane conductance regulator (CFTR), which is highly expressed in glands. The potential role of submucosal glands in cystic fibrosis lung disease is discussed.
Impact of mitochondrial inhibition on excitability and cytosolic Ca2+ levels in brainstem motoneurones from mouse
Authors: Bergmann Friederike , Keller Bernhard U.
Published: December 2003

Motoneurones (MNs) are particularly affected by the inhibition of mitochondrial metabolism, which has been linked to their selective vulnerability during pathophysiological states like hypoxia and amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disorder. To elucidate underlying events, we used sodium cyanide (CN) as a pharmacological inhibitor of complex IV of the mitochondrial respiratory chain (‘chemical hypoxia’) and investigated the cellular response in vulnerable and resistant neurone types. Bath application of 2 mm CN activated TTX‐insensitive Na+ conductances in vulnerable hypoglossal MNs, which depolarized these MNs by 10.2 ± 1.1 mV and increased their action potential activity. This response was mimicked by sodium azide (2 mm) and largely prevented by preincubation with the antioxidants ascorbic acid (1 mm) and Trolox (750 μm), indicating an involvement of reactive oxygen species (ROS) in the activation mechanism. CN also elevated cytosolic [Ca2+] levels through (i) Ca2+ release from mitochondria‐controlled stores, (ii) significant retardation of cytosolic Ca2+ clearance rates, even when cytosolic ATP levels were held constant during whole‐cell recording, and (iii) secondary Ca2+ influx during elevated firing rates. Blocking mitochondrial ATP production additionally raised cytosolic Ca2+ levels and prolonged recovery of Ca2+ transients with a delay of 5–6 min. Comparative studies on hypoglossal MNs, facial MNs and dorsal vagal neurones suggested that CN responses were dominated by the activation of K+ conductances in resistant neurones, thus reducing excitability during mitochondrial inhibition. In summary, our observations therefore support a model where selective MN vulnerability results from a synergistic accumulation of risk factors, including low cytosolic Ca2+ buffering, strong mitochondrial impact on [Ca2+]i, and a mitochondria‐controlled increase in electrical excitability during metabolic disturbances.
Energy metabolism in heart failure
Authors: Ventura‐Clapier Renée , Garnier Anne , Veksler Vladimir
Published: December 2003

Heart failure (HF) is a syndrome resulting from the inability of the cardiac pump to meet the energy requirements of the body. Despite intensive work, the pathogenesis of the cardiac intracellular abnormalities that result from HF remains incompletely understood. Factors that lead to abnormal contraction and relaxation in the failing heart include metabolic pathway abnormalities that result in decreased energy production, energy transfer and energy utilization. Heart failure also affects the periphery. Patients suffering from heart failure always complain of early muscular fatigue and exercise intolerance. This is linked in part to intrinsic alterations of skeletal muscle, among which decreases in the mitochondrial ATP production and in the transfer of energy through the phosphotransfer kinases play an important role. Alterations in energy metabolism that affect both cardiac and skeletal muscles argue for a generalized metabolic myopathy in heart failure. Recent evidence shows that decreased expression of mitochondrial transcription factors and mitochondrial proteins are involved in mechanisms causing the energy starvation in heart failure. This review will focus on energy metabolism alterations in long‐term chronic heart failure with only a few references to compensated hypertrophy when necessary. It will briefly describe the energy metabolism of normal heart and skeletal muscles and their alterations in chronic heart failure. It is beyond the scope of this review to address the metabolic switches occurring in compensated hypertrophy; readers could refer to well‐documented reviews on this subject.
The scents of androstenone in humans
Authors: Araneda Ricardo C. , Firestein Stuart
Published: December 2003

Specific anosmia, the inability to detect a particular odour, has been well documented for decades in human and animal populations. Indeed, the existence of specific anosmias was a favoured argument used to support a receptor‐mediated mechanism of odourant detection prior to the molecular identification of a large family of olfactory G‐protein‐coupled receptors (GPCRs) in the early 1990s (Amoore, 1974; Buck & Axel, 1991). One well known anosmia in humans is the inability to sense 5‐α‐androst‐16‐en‐3‐one (androstenone). Androstenone is variously described as having an unpleasant (urine, sweat) or pleasant odour (sweet, floral), yet a fraction of the population cannot detect its presence. Moreover, androstenone is a pheromone in boars and is found in urine and axillary sweat in humans, making it a prospective candidate for odour‐mediated communication in humans. While a role for androstenone as a human pheromone is open to debate, a widely accepted finding is the ability of humans who are initially insensitive to androstenone to acquire sensitivity to it upon continued exposure (Wysocki et al. 1989). Since the 1989 anecdotal discovery of C.J. Wysocki, several other studies have shown that humans and other species can acquire sensitivity to androstenone as well as to other odourants (Wang et al. 1993; Pause et al. 1999; Dalton et al. 2002). However, the mechanism(s) of this increased sensitivity are poorly understood. In this issue of The Journal of Physiology
Wang et al. (2004) provide evidence for a mechanism of increased sensitivity in the olfactory epithelium of humans.
RNA Leaching of Transcription Factors Disrupts Transcription in Myotonic Dystrophy
Authors: Ebralidze A. , Wang Y. , Petkova V. , Ebralidse K. , Junghans R. P.
Published: December 2003

Myotonic dystrophy type 1 (DM1) is caused by a CUG

n

expansion (
n
≈ 50 to 5000) in the 3′ untranslated region of the mRNA of the DM protein kinase gene. We show that mutant RNA binds and sequesters transcription factors (TFs), with up to 90% depletion of selected TFs from active chromatin. Diverse genes are consequently reduced in expression, including the ion transporter CIC-1, which has been implicated in myotonia. When TF specificity protein 1 (Sp1) was overexpressed in DM1-affected cells, low levels of messenger RNA for CIC-1 were restored to normal. Transcription factor leaching from chromatin by mutant RNA provides a potentially unifying pathomechanistic explanation for this disease.
Genome of
Geobacter sulfurreducens:
Metal Reduction in Subsurface Environments
Authors: Methé B. A. , Nelson K. E. , Eisen J. A. , Paulsen I. T. , Nelson W. , Heidelberg J. F. , Wu D. , Wu M. , Ward N. , Beanan M. J. , Dodson R. J. , Madupu R. , Brinkac L. M. , Daugherty S. C. , DeBoy R. T. , Durkin A. S. , Gwinn M. , Kolonay J. F. , Sullivan S. A. , Haft D. H. , Selengut J. , Davidsen T. M. , Zafar N. , White O. , Tran B. , Romero C. , Forberger H. A , Weidman J. , Khouri H. , Feldblyum T. V. , Utterback T. R. , Van Aken S. E. , Lovley D. R. , Fraser C. M.
Published: December 2003

The complete genome sequence of
Geobacter sulfurreducens
, a δ-proteobacterium, reveals unsuspected capabilities, including evidence of aerobic metabolism, one-carbon and complex carbon metabolism, motility, and chemotactic behavior. These characteristics, coupled with the possession of many two-component sensors and many c-type cytochromes, reveal an ability to create alternative, redundant, electron transport networks and offer insights into the process of metal ion reduction in subsurface environments. As well as playing roles in the global cycling of metals and carbon, this organism clearly has the potential for use in bioremediation of radioactive metals and in the generation of electricity.
Redistribution of Intracellular Oxygen in Hypoxia by Nitric Oxide: Effect on HIF1α
Authors: Hagen Thilo , Taylor Cormac T. , Lam Francis , Moncada Salvador
Published: December 2003

Cells exposed to low oxygen concentrations respond by initiating defense mechanisms, including the stabilization of hypoxia-inducible factor (HIF) 1α, a transcription factor that upregulates genes such as those involved in glycolysis and angiogenesis. Nitric oxide and other inhibitors of mitochondrial respiration prevent the stabilization of HIF1α during hypoxia. In studies of cultured cells, we show that this effect is a result of an increase in prolyl hydroxylase–dependent degradation of HIF1α. With the use of Renilla luciferase to detect intracellular oxygen concentrations, we also demonstrate that, upon inhibition of mitochondrial respiration in hypoxia, oxygen is redistributed toward nonrespiratory oxygen-dependent targets such as prolyl hydroxylases so that they do not register hypoxia. Thus, the signaling consequences of hypoxia may be profoundly modified by nitric oxide.
An Early Cretaceous Tribosphenic Mammal and Metatherian Evolution
Authors: Luo Zhe-Xi , Ji Qiang , Wible John R. , Yuan Chong-Xi
Published: December 2003

Derived features of a new boreosphenidan mammal from the Lower Cretaceous Yixian Formation of China suggest that it has a closer relationship to metatherians (including extant marsupials) than to eutherians (including extant placentals). This fossil dates to 125 million years ago and extends the record of marsupial relatives with skeletal remains by 50 million years. It also has many foot structures known only from climbing and tree-living extant mammals, suggesting that early crown therians exploited diverse niches. New data from this fossil support the view that Asia was likely the center for the diversification of the earliest metatherians and eutherians during the Early Cretaceous.
Doubly Ionized Carbon Observed in the Plasma Tail of Comet Kudo-Fujikawa
Authors: Povich Matthew S. , Raymond John C. , Jones Geraint H. , Uzzo Michael , Ko Yuan-Kuen , Feldman Paul D. , Smith Peter L. , Marsden Brian G. , Woods Thomas N.
Published: December 2003

Comet C/2002 X5 (Kudo-Fujikawa) was observed near its perihelion of 0.19 astronomical unit by the Ultraviolet Coronagraph Spectrometer aboard the Solar and Heliospheric Observatory spacecraft. Images of the comet reconstructed from high-resolution spectra reveal a quasi-spherical cloud of neutral hydrogen and a variable tail of C+and C2+that disconnects from the comet and subsequently regenerates. The high abundance of C2+and C+, at least 24% relative to water, cannot be explained by photodissociation of carbon monoxide and is instead attributed to the evaporation and subsequent photoionization of atomic carbon from organic refractory compounds present in the cometary dust grains.This result serves to strengthen the connection between comets and the material from which the Solar System formed.
Finite-Frequency Tomography Reveals a Variety of Plumes in the Mantle
Authors: Montelli Raffaella , Nolet Guust , Dahlen F. A. , Masters Guy , Engdahl E. Robert , Hung Shu-Huei
Published: December 2003

We present tomographic evidence for the existence of deep-mantle thermal convection plumes.
P
-wave velocity images show at least six well-resolved plumes that extend into the lowermost mantle: Ascension, Azores, Canary, Easter, Samoa, and Tahiti. Other less well-resolved plumes, including Hawaii, may also reach the lowermost mantle. We also see several plumes that are mostly confined to the upper mantle, suggesting that convection may be partially separated into two depth regimes. All of the observed plumes have diameters of several hundred kilometers, indicating that plumes convey a substantial fraction of the internal heat escaping from Earth.
The Baculovirus GP64 Protein Mediates Highly Stable Infectivity of a Human Respiratory Syncytial Virus Lacking Its Homologous Transmembrane Glycoproteins
Authors: Oomens A. G. P. , Wertz Gail W.
Published: December 2003

Baculovirus GP64 is a low-pH-dependent membrane fusion protein required for virus entry and cell-to-cell transmission. Recently, GP64 has generated interest for practical applications in mammalian systems. Here we examined the membrane fusion function of GP64 from
Autographa californica multiple nucleopolyhedrovirus
(Ac
M
NPV) expressed in mammalian cells, as well as its capacity to functionally complement a mammalian virus, human respiratory syncytial virus (HRSV). Both authentic GP64 and GP
64/F
, a chimeric protein in which the GP64 cytoplasmic tail domain was replaced with the 12 C-terminal amino acids of the HRSV fusion (F) protein, induced low-pH-dependent cell-cell fusion when expressed transiently in HEp-2 (human) cells. Levels of surface expression and syncytium formation were substantially higher at 33°C than at 37°C. The open reading frames (ORFs) encoding GP64 or GP
64/F
, along with two marker ORFs encoding green fluorescent protein (GFP) and β-glucuronidase (GUS), were used to replace all three homologous transmembrane glycoprotein ORFs (small hydrophobic SH, attachment G, and F) in a cDNA of HRSV. Infectious viruses were recovered that lacked the HRSV SH, G, and F proteins and expressed instead the GP64 or GP
64/F
protein and the two marker proteins GFP and GUS. The properties of these viruses, designated RSΔ
sh,g,f
/GP64 or RSΔ
sh,g,f
/GP
64/F
, respectively, were compared to a previously described HRSV expressing GFP in place of SH but still containing the wild-type HRSV G and F proteins (RSΔ
sh
[A. G. Oomens, A. G. Megaw, and G. W. Wertz, J. Virol., 77:3785-3798, 2003]). By immunoelectron microscopy, the GP64 and GP
64/F
proteins were shown to incorporate into HRSV-induced filaments at the cell surface. Antibody neutralization, ammonium chloride inhibition, and replication levels in cell culture showed that both GP64 proteins efficiently mediated infectivity of the respective viruses in a temperature-sensitive, low-pH-dependent manner. Furthermore, RSΔ
sh,g,f
/GP64 and RSΔ
sh,g,f
/GP
64/F
replicated to higher levels and had significantly higher stability of infectivity than HRSVs containing the homologous HRSV G and F proteins. Thus, GP64 and a GP64/HRSV F chimeric protein were functional and efficiently complemented an unrelated human virus in mammalian cells, producing stable, infectious virus stocks. These results demonstrate the potential of GP64 for both practical applications requiring stable pseudotypes in mammalian systems and for studies of viral glycoprotein requirements in assembly and pathogenesis.
Evidence for an Essential Catalytic Role of the F10 Protein Kinase in Vaccinia Virus Morphogenesis
Authors: Szajner Patricia , Weisberg Andrea S. , Moss Bernard
Published: December 2003

Temperature-sensitive mutants of vaccinia virus, with genetic changes that map to the open reading frame encoding the F10 protein kinase, exhibit a defect at an early stage of viral morphogenesis. To further study the role of the enzyme, we constructed recombinant vaccinia virus vF10V5i, which expresses inducible V5 epitope-tagged F10 and is dependent on a chemical inducer for plaque formation and replication. In the absence of inducer, viral membrane formation was delayed and crescents and occasional immature forms were detected only late in infection. When the temperature was raised from 37 to 39°C, the block in membrane formation persisted throughout the infection. The increased stringency may be explained by a mild temperature sensitivity of the wild-type F10 kinase, which reduced the activity of the very small amount expressed in the absence of inducer, or by the thermolability of an unphosphorylated kinase substrate or uncomplexed F10-interacting protein. Further analyses demonstrated that tyrosine and threonine phosphorylation of the A17 membrane component was inhibited in the absence of inducer. The phosphorylation defect could be overcome by transfection of plasmids that express wild-type F10, but not by plasmids that express F10 with single amino acid substitutions that abolished catalytic activity. Although the mutated forms of F10 were stable and concentrated in viral factories, only the wild-type protein complemented the assembly and replication defects of vF10V5i in the absence of inducer. These studies provide evidence for an essential catalytic role of the F10 kinase in vaccinia virus morphogenesis.
Physical and Functional Interactions between Vaccinia Virus F10 Protein Kinase and Virion Assembly Proteins A30 and G7
Authors: Szajner Patricia , Weisberg Andrea S. , Moss Bernard
Published: December 2003

An early step in vaccinia virus morphogenesis, the association of crescent membranes with electron-dense granular material, is perturbed when expression of the viral protein encoded by the A30L or G7L open reading frame is repressed. Under these conditions, we found that phosphorylation of the A17 membrane protein, which is mediated by the F10 kinase, was severely reduced. Furthermore, A30 and G7 stimulated F10-dependent phosphorylation of A17 in the absence of other viral late proteins. Evidence for physical interactions between A30, G7, and F10 was obtained by their coimmunoprecipitation with antibody against A30 or F10. In addition, phosphorylation of A30 was dependent on the F10 kinase and autophosphorylation of F10 was stimulated by A30 and G7. Nevertheless, the association of A30, G7, and F10 occurred even with mutated, catalytically inactive forms of F10. Just as A30 and G7 are mutually dependent on each other for stability, F10 was nearly undetectable in the absence of A30 and G7. The reverse is not true, however, as repression of F10 did not diminish A30 or G7. Interaction of F10 with A30 and G7 presumably occurred within the virus factory areas of the cytoplasm, where each was concentrated. F10 localized predominantly in the cortical region of immature virions, beneath the membrane where A17 is located. F10 remained associated with the particulate core fraction of mature virions after treatment with a nonionic detergent and reducing agent. The formation of protein complexes such as the one involving A30, G7, and F10 may be a mechanism for the regulated packaging and processing of virion components.
The Kaposi's Sarcoma-Associated Herpesvirus Latency-Associated Nuclear Antigen 1 N Terminus Is Essential for Chromosome Association, DNA Replication, and Episome Persistence
Authors: Barbera Andrew J. , Ballestas Mary E. , Kaye Kenneth M.
Published: December 2003

To persist in latently infected, proliferating cells, Kaposi's sarcoma-associated herpesvirus (KSHV) episomes must replicate and efficiently segregate to progeny nuclei. Episome persistence in uninfected cells requires latency-associated nuclear antigen 1 (LANA1) in
trans
and
cis
-acting KSHV terminal repeat (TR) DNA. The LANA1 C terminus binds TR DNA, and LANA1 mediates TR-associated DNA replication in transient assays. LANA1 also concentrates at sites of KSHV TR DNA episomes along mitotic chromosomes, consistent with a tethering role to efficiently segregate episomes to progeny nuclei. LANA1 amino acids 5 to 22 constitute a chromosome association region (Piolot et al., J. Virol.
75:
3948-3959, 2001). We now investigate LANA1 residues 5 to 22 with scanning alanine substitutions. Mutations targeting LANA1
5
GMR
7
,
8
LRS
10
, and
11
GRS
13
eliminated chromosome association, DNA replication, and episome persistence. LANA1 mutated at
14
TG
15
retained the ability to associate with chromosomes but was partially deficient in DNA replication and episome persistence. These results provide genetic support for a key role of the LANA1 N terminus in chromosome association, LANA1-mediated DNA replication, and episome persistence.
The ORF2 Protein of Hepatitis E Virus Binds the 5′ Region of Viral RNA
Authors: Surjit Milan , Jameel Shahid , Lal Sunil K.
Published: December 2003

Hepatitis E virus (HEV) is a major human pathogen in much of the developing world. It is a plus-strand RNA virus with a 7.2-kb polyadenylated genome consisting of three open reading frames,
ORF1
,
ORF2
, and
ORF3
. Of these,
ORF2
encodes the major capsid protein of the virus and
ORF3
encodes a small protein of unknown function. Using the yeast three-hybrid system and traditional biochemical techniques, we have studied the RNA binding activities of ORF2 and ORF3, two proteins encoded in the 3′ structural part of the genome. Since the genomic RNA from HEV has been postulated to contain secondary structures at the 5′ and 3′ ends, we used these two terminal regions, besides other regions within the genome, in this study. Experiments were designed to test for interactions between the genomic RNA fusion constructs with ORF2 and ORF3 hybrid proteins in a yeast cellular environment. We show here that the ORF2 protein contains RNA binding activity. The ORF2 protein specifically bound the 5′ end of the HEV genome. Deletion analysis of this protein showed that its RNA binding activity was lost when deletions were made beyond the N-terminal 111 amino acids. Finer mapping of the interacting RNA revealed that a 76-nucleotide (nt) region at the 5′ end of the HEV genome was responsible for binding the ORF2 protein. This 76-nt region included the 51-nt HEV sequence, conserved across alphaviruses. Our results support the requirement of this conserved sequence for interaction with ORF2 and also indicate an increase in the strength of the RNA-protein interaction when an additional 44 bases downstream of this 76-nt region were included. Secondary-structure predictions and the location of the ORF2 binding region within the HEV genome indicate that this interaction may play a role in viral encapsidation.
Characterization of the Interaction between P143 and LEF-3 from Two Different Baculovirus Species:
Choristoneura fumiferana
Nucleopolyhedrovirus LEF-3 Can Complement
Autographa californica
Nucleopolyhedrovirus LEF-3 in Supporting DNA Replication
Authors: Chen Tricia , Sahri Daniela , Carstens Eric B.
Published: December 2003

The baculovirus protein P143 is essential for viral DNA replication in vivo, likely as a DNA helicase. We have demonstrated that another viral protein, LEF-3, first described as a single-stranded DNA binding protein, is required for transporting P143 into the nuclei of insect cells. Both of these proteins, along with several other early viral proteins, are also essential for DNA replication in transient assays. We now describe the identification, nucleotide sequences, and transcription patterns of the
Choristoneura fumiferana
nucleopolyhedrovirus (Cf
M
NPV) homologues of
p143
and
lef-3
and demonstrate that Cf
M
NPV LEF-3 is also responsible for P143 localization to the nucleus. We predicted that the interaction between P143 and LEF-3 might be critical for cross-species complementation of DNA replication. Support for this hypothesis was generated by substitution of heterologous P143 and LEF-3 between two different baculovirus species,
Autographa californica
nucleopolyhedrovirus and Cf
M
NPV, in transient DNA replication assays. The results suggest that the P143-LEF-3 complex is an important baculovirus replication factor.
Functional Analysis of Epstein-Barr Virus SM Protein: Identification of Amino Acids Essential for Structure, Transactivation, Splicing Inhibition, and Virion Production
Authors: Ruvolo Vivian , Sun Liang , Howard Karilynn , Sung Seung , Delecluse Henri-Jacques , Hammerschmidt Wolfgang , Swaminathan Sankar
Published: December 2003

The Epstein-Barr virus (EBV) SM protein is a posttranscriptional regulator of cellular and viral gene expression that binds and stabilizes target mRNAs and shuttles from nucleus to cytoplasm. SM enhances expression of several EBV genes required for lytic replication and is essential for virion production. SM increases accumulation of specific mRNAs but also inhibits expression of several intron-containing transcripts. The mechanism by which SM inhibits gene expression is poorly understood. The experiments described here had several aims: to determine whether specific domains of SM were responsible for activation or inhibition function; whether these functions could be separated; and whether one or more of these functions were essential for virion production. A mutational analysis of SM was performed, focusing on amino acids in SM that are evolutionarily conserved among SM homologs in other herpesviruses. Mutation of the carboxy-terminal region of SM revealed a region that is likely to be structurally important for SM protein conformation. In addition, several amino acids were identified that are critical for activation and inhibition function. A specific mutation of a highly conserved cysteine residue revealed that it was essential for gene inhibition but not for transactivation, indicating that these two functions operate through independent mechanisms. Furthermore, the ability of wild-type SM and the inability of the mutant to inhibit gene expression were shown to correlate with the ability to inhibit splicing of a human target gene and thereby prevent accumulation of its processed mRNA. Surprisingly, some mutations which preserved both activation and inhibition functions in vitro nevertheless abolished virion production, suggesting that other SM functions or protein-protein interactions are also required for lytic replication.
Spatial and Temporal Organization of Adeno-Associated Virus DNA Replication in Live Cells
Authors: Fraefel Cornel , Bittermann Anne Greet , Büeler Hansruedi , Heid Irma , Bächi Thomas , Ackermann Mathias
Published: December 2003

Upon cell entry, the genomes of herpes simplex virus type 1 (HSV-1) and adenovirus (Ad) associate with distinct nuclear structures termed ND10 or promyelocytic leukemia (PML) nuclear bodies (NBs). PML NB morphology is altered or disrupted by specific viral proteins as replication proceeds. We examined whether adeno-associated virus (AAV) replication compartments also associate with PML NBs, and whether modification or disruption of these by HSV-1 or Ad, both of which are helper viruses for AAV, is necessary at all. Furthermore, to add a fourth dimension to our present view of AAV replication, we established an assay that allows visualization of AAV replication in live cells. A recombinant AAV containing 40
lac
repressor binding sites between the AAV inverted terminal repeats was constructed. AAV Rep protein and helper virus-mediated replication of this recombinant AAV genome was visualized by binding of enhanced yellow fluorescent protein-
lac
repressor fusion protein to double-stranded AAV replication intermediates. We demonstrate in live cells that AAV DNA replication occurs in compartments which colocalize with AAV Rep. Early after infection, the replication compartments were small and varied in numbers from 2 to more than 40 per cell nucleus. Within 4 to 8 h, individual small replication compartments expanded and fused to larger structures which filled out much of the cell nucleus. We also show that AAV replication compartments can associate with modified PML NBs in Ad-infected cells. In wild-type HSV-1-infected cells, AAV replication compartments and PML NBs did not coexist, presumably because PML was completely disrupted by the HSV-1 ICP0 protein. However, alteration or disruption of PML appears not to be a prerequisite for AAV replication, as the formation of replication compartments was normal when the ICP0 mutants HSV-1 dl1403 and HSV-1 FXE, which do not affect PML NBs, were used as the helper viruses; under these conditions, AAV replication compartments did not associate with PML NBs.
Complete, Annotated Sequence of the Pseudorabies Virus Genome
Authors: Klupp Barbara G. , Hengartner Christoph J. , Mettenleiter Thomas C. , Enquist Lynn W.
Published: December 2003

We have obtained the complete DNA sequence of pseudorabies virus (PRV), an alphaherpesvirus also known as Aujeszky's disease virus or suid herpesvirus 1, using sequence fragments derived from six different strains (Kaplan, Becker, Rice, Indiana-Funkhauser, NIA-3, and TNL). The assembled PRV genome sequence comprises 143,461 nucleotides. As expected, it matches the predicted gene arrangement, genome size, and restriction enzyme digest patterns. More than 70 open reading frames were identified with homologs in related alphaherpesviruses; none were unique to PRV. RNA polymerase II transcriptional control elements in the PRV genome, including core promoters, splice sites, and polyadenylation sites, were identified with computer prediction programs. The correlation between predicted and experimentally determined transcription start and stop sites was excellent. The transcriptional control architecture is characterized by three key features: core transcription elements shared between genes, yielding divergent transcripts and a large number of coterminal transcripts; bifunctional transcriptional elements, yielding head-to-tail transcripts; and short repetitive sequences that could function as insulators against improperly terminated transcripts. Many of these features are conserved in the alphaherpesvirus subfamily and have important implications for gene array analyses.