bluuebunny/crossref_metadata_2025 huggingface dataset:
Directed evolution of the substrate specificity of biotin ligase
Authors: Lu Wei‐Cheng , Levy Matthew , Kincaid Rodney , Ellington Andrew D.
Published: December 2013

We have developed selection scheme for directing the evolution of Escherichia coli biotin protein ligase (BPL) via in vitro compartmentalization, and have used this scheme to alter the substrate specificity of the ligase towards the utilization of the biotin analogue desthiobiotin. In this scheme, a peptide substrate (BAP) was conjugated to a DNA library encoding BirA, emulsified such that there was a single template per compartment, and protein variants were transcribed and translated in vitro. Those variants that could efficiently desthiobiotinylate their corresponding peptide:DNA conjugate were subsequently captured and amplified. Following just six rounds of selection and amplification several variants that demonstrated higher activity with desthiobiotin were identified. The best variants from Round 6, BirA6–40 and BirA6–47, showed 17‐fold and 10‐fold higher activity, respectively, their abilities to use desthiobiotin as a substrate. While selected enzymes contained a number of substitutions, a single mutation, M157T, proved sufficient to provide much greater activity with desthiobiotin. Further characterization of BirA6–40 and the single substitution variant BirAM157T revealed that they had twoto threefold higher kcat values for desthiobiotin. These variants had also lost much of their ability to utilize biotin, resulting in orthogonal enzymes that in conjunction with streptavidin variants that can utilize desthiobiotin may prove to be of great use in developing additional, robust conjugation handles for a variety of biological and biotechnological applications. Biotechnol. Bioeng. 2014;111: 1071–1081. © 2013 Wiley Periodicals, Inc.
Physiological and transcriptional characterization of Escherichia coli strains lacking interconversion of phosphoenolpyruvate and pyruvate when glucose and acetate are coutilized
Authors: Sabido Andrea , Sigala Juan Carlos , Hernández‐Chávez Georgina , Flores Noemí , Gosset Guillermo , Bolívar Francisco
Published: December 2013

Phosphoenolpyruvate (PEP) is a precursor involved in the biosynthesis of aromatics and other valuable compounds in Escherichia coli. The PEP:carbohydrate phosphotransferase system (PTS) is the major glucose transport system and the largest PEP consumer. To increase intracellular PEP availability for aromatics production purposes, mutant strains of E. coli JM101 devoid of the ptsHIcrr operon (PB11 strain) have been previously generated. In this derivative, transport and growth rate on glucose decreased significantly. A laboratory evolved strain derived from PB11 that partially recovered its growth capacity on glucose was named PB12. In the present study, we blocked carbon skeletons interchange between PEP and pyruvate (PYR) in these ptsHIcrr− strains by deleting the pykA, pykF, and ppsA genes. The PB11 pykAF− ppsA− strain exhibited no growth on glucose or acetate alone, but it was viable when both substrates were consumed simultaneously. In contrast, the PB12 pykAF− ppsA− strain displayed a low growth rate on glucose or acetate alone, but in the mixture, growth was significantly improved. RT‐qPCR expression analysis of PB11 pykAF− ppsA− growing with both carbon sources showed a downregulation of all central metabolic pathways compared with its parental PB11 strain. Under the same conditions, transcription of most of the genes in PB12 pykAF− ppsA− did not change, and few like aceBAK, sfcA, and poxB were overexpressed compared with PB12. We explored the aromatics production capabilities of both ptsHIcrr− pykAF− ppsA− strains and the engineered PB12 pykAF− ppsA− tyrR− pheAev2+/pJLBaroGfbrtktA enhanced the yield of aromatic compounds when coutilizing glucose and acetate compared with the control strain PB12 tyrR− pheAev2+/pJLBaroGfbrtktA. Biotechnol. Bioeng. 2014;111: 1150–1160. © 2013 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.
Drivers, Practices and Outcomes of Low‐carbon Operations: Approaches of German Automotive Suppliers to Cutting Carbon Emissions
Authors: Böttcher Christian Felix , Müller Martin
Published: December 2013

The aim of this paper is to investigate the determinants of measures used by German automotive suppliers to cut carbon emissions, and their impact on performance. To achieve this aim the concept of low‐carbon operations is developed based on the broader concept of sustainable operations. An integrated conceptual model to research the drivers, practices and outcome of low‐carbon operation is derived. The model is tested using variance‐based structural equations modelling with data collected from 159 German automotive suppliers. It is found that stakeholder pressure, and to some extent expectations about competitiveness, drive the adoption of low‐carbon operational practices. The relative and absolute effect of those drivers varies along the different practices. Regarding the outcomes, positive effects on carbon and indirect effects on economic performance are found. However, the results regarding both drivers and outcomes of the practices under consideration show significant differences with respect to firm size. Copyright © 2013 John Wiley & Sons, Ltd and ERP Environment
Business Strategies of Incumbents in the Market for Electric Vehicles: Opportunities and Incentives for Sustainable Innovation
Authors: Wesseling J. H. , Niesten E. M. M. I. , Faber J. , Hekkert M. P.
Published: December 2013

This paper focuses on the relation between large car manufacturers’ incentive and opportunity to innovate and their electric vehicle (EV) business strategies. We analyze how environmental regulation and the firm's incentive (measured by net income) and opportunity to innovate (measured by EV asset position, determined from a combination of patent, partnership and prototype data) affected EV sales over the period 1990–2011. During the EV's R&D period in the 1990s, large car manufacturers that were regulated by the full zero emission vehicle mandate developed a significantly stronger EV asset position, but did not sell significantly more EVs than their rivals. During the EV's commercialization period (2007–2011), large car manufacturers with both a strong incentive and a strong opportunity to innovate sold significantly more EVs. Based on these results, the paper offers a typology of business strategies, several managerial implications, and recommendations for policy makers to stimulate sustainable development. Copyright © 2013 John Wiley & Sons, Ltd and ERP Environment
Exploring the Relationship between Sustainability Reporting and Sustainability Management Practices
Authors: Pérez‐López Diego , Moreno‐Romero Ana , Barkemeyer Ralf
Published: December 2013

There has been a dramatic uptake of sustainability reporting (SR) and sustainability management practices over the last two decades, in turn raising questions about the relationship between external disclosure and internal practice of corporate sustainability. Previous literature has emphasized the role of external pressures in driving SR adoption. However, as recent practitioner surveys also identify the existence of internal motives for SR, a more comprehensive analysis of this relationship is needed.

In order to address this issue, we develop a framework accounting for four company‐level SR configurations, resulting from different levels of importance of external and internal motives for SR. Then, four exploratory cases serve to illustrate these configurations and to describe the respective roles of SR at the company level. As considerable diversity among these SR practices is found, our findings point to the relevance of both external and internal motives in understanding SR contribution to corporate sustainability. Copyright © 2013 John Wiley & Sons, Ltd and ERP Environment
Biological reduction of graphene oxide using plant leaf extracts
Authors: Lee Geummi , Kim Beom Soo
Published: December 2013

Two‐dimensional graphene has attracted significant attention due to its unique mechanical, electrical, thermal, and optical properties. Most commonly employed methods to chemically reduce graphene oxide to graphene use hydrazine or its derivatives as the reducing agent. However, they are highly hazardous and explosive. Various phytochemicals obtained from different natural sources such as leaves and peels of a plant are used as reducing agents in the preparation of different gold, silver, copper, and platinum nanoparticles. In this study, seven plant leaf extracts (Cherry, Magnolia, Platanus, Persimmon, Pine, Maple, and Ginkgo) were compared for their abilities to reduce graphene oxide. The optimized reaction conditions for the reduction of graphene oxide were determined as follows. Type of plant: Cherry (Prunus serrulata), reaction time: 12 h, composition of the reaction mixture: 16.7% v/v of plant leaf extract in total suspension, and temperature: 95°C. The degree of reduction caused by Cherry leaf extract was analyzed by elemental analysis and X‐ray photoelectron spectroscopy. The reduction of graphene oxide was also confirmed by ultraviolet‐visible spectroscopy, Fourier transform‐infrared spectroscopy, Raman spectroscopy, X‐ray diffraction, transmission electron microscopy, and thermogravimetric analysis. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:463–469, 2014
Correlation analysis of the taxane core functional group modification, enzyme expression, and metabolite accumulation profiles under methyl jasmonate treatment
Authors: Song Guang Hao , Zhao Chun Fang , Zhang Meng , Fu Chun Hua , Zhang Hua , Yu Long Jiang
Published: December 2013

Metabolomic and transcriptomic profiling data were obtained and integrated to elucidate the crucial network controls on taxol and its precursor biosynthesis during the taxane core functionalization within methyl jasmonate (MJ)‐induced Taxus chinensis cells. Twelve metabolites were identified using liquid chromatography‐electrospray ionization‐mass spectrometry. These metabolites contain taxol (paclitaxel), baccatin III (B‐III) and its analogs, a group structurally bearing multiple free hydroxyls (TAX), and another group of multiple acyl taxanes (MAT), including taxuyunnanine C (TC) and its analogs. The metabolomic profile showed a higher increase in TAX than in MAT. Particularly, the ratio of B‐III and taxol to the total taxane content increased more significantly in TAX than in MAT. The MAT proportion did not significantly change, although they are predominant components in cell cultures compared with TAX. Quantitative real‐time polymerase chain reaction (qRT‐RCR) was used to determine the transcription level of 20 genes, among which 11 were reported responsible for taxol biosynthesis and 9 were obtained from our previous transcriptomic data. The total expression levels of hydroxylase after 24 h and 6 days were higher than those of acylase. The principal component analysis (PCA) results validated the metabolomic analysis data, indicating that hydroxylation was more crucial than acylation for controlling the flux toward TAX biosynthesis. Furthermore, the PCA contribution comparison showed that two undefined genes of OHX1 and ACX3 might have good potential in TAX upregulation and MAT downregulation. To the best of our knowledge, this study provides the first experimental evidence on the contribution of total hydroxylation to taxane biosynthesis. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:269–280, 2014
Parallel‐optimizing SPH fluid simulation for realistic VR environments
Authors: Huang Chen , Zhu Jian , Sun Hanqiu , Wu Enhua
Published: December 2013

In virtual environments, real‐time simulation and rendering of dynamic fluids have always been the pursuit for virtual reality research. In this paper, we present a real‐time framework for realistic fluid simulation and rendering on graphics processing unit. Because of the high demand for interactive fluids with larger particle set, the computational need is becoming higher. The proposed framework can effectively reduce the computational burden through avoiding the computation in inactive areas, where many particles with similar properties and low local pressure cluster together. While in active areas, the computation is fully carried out; thus, the fluid dynamics are largely preserved. Here, a robust particle classification technique is introduced to classify particles into either active or inactive. The test results have shown that the technique improves the time performance of fluid simulation largely. We then incorporate parallel surface reconstruction technique using marching cubes to extract the surfaces of the fluid. The introduced histogram pyramid‐based marching cubes technique is fast and memory efficiency. As a result, we are able to produce plausible and interactive fluids with the proposed framework for large‐scale virtual environments. Copyright © 2013 John Wiley & Sons, Ltd.
Social normalisation: Using marketing to make green normal
Authors: Rettie Ruth , Burchell Kevin , Barnham Chris
Published: December 2013

This paper proposes a novel environmental marketing approach in which the adoption of greener consumer behaviours is encouraged by repositioning them as normal. The research was undertaken in order to help explain the disappointing performance of green marketing initiatives. The methodology was qualitative, with focus groups and a wide range of stimulus materials. The study illuminates the ways in which consumers conceptualise and adopt pro‐environmental behaviours and highlights the importance of consumer ideas about what is normal. The research indicates that consumers are more likely to adopt behaviour and products that they think are normal and that what is regarded as normal changes over time. New activities and products that are initially seen as different, and as outside normal behaviour, can eventually become mainstream and accepted as normal, in a process of ‘social normalisation’. As part of this process, other behaviours, which have been mainstream everyday ways of doing things, can become marginalised over time. The research suggests that companies and policy makers tend to position green marketing initiatives as targeting a green niche and that this inhibits social normalisation and mainstream adoption. Green marketing can potentially play an important role in the social normalisation of green practices and products by portraying these as normal and everyday instead of emphasizing their greenness. The study contributes to understanding of consumer behaviour and the adoption of more sustainable products and practices and identifies practical ways to improve green initiatives. Copyright © 2013 John Wiley & Sons, Ltd.
Consumer response to interruption features and need for cognitive closure
Authors: Niculescu Mihai , Payne Collin R. , Luna‐Nevarez Cuauhtémoc
Published: December 2013

Prior research on interruptions focuses entirely on the process being interrupted and assumes interruption homogeneity. Across two studies, we examine how heterogeneous features of interruptions (i.e., timing, frequency, and perceived pleasantness) and consumer individual differences (i.e., need for cognitive closure (NFCC)) impact consumer response toward a product. We find interruption features have opposing effects on consumer response for consumers high versus low in NFCC—depending upon the perceived valence of the interruption. Specifically, individuals with high NFCC respond better to a product when interruptions are perceived to be pleasant and occur late or infrequently. In contrast, those who have low NFCC respond better to a product when interruptions are perceived to be pleasant and occur early or perceived to be unpleasant and occur infrequently. The role of interruption pleasantness is discussed in terms of its predictive power for perceived pleasant but not perceived unpleasant interruptions. Finally, study findings are placed within marketing contexts that guide managerial implications. Copyright © 2013 John Wiley & Sons, Ltd.
The direct and interactive effects of store‐level promotions on impulse purchase: Moderating impact of category familiarity and normative influences
Authors: Shukla Paurav , Banerjee Madhumita
Published: December 2013

Marketing literature conceptually and empirically establishes the direct effects of different variables on impulse purchase. However, the simultaneous interactions between variables influencing impulse purchase are yet to be studied. This paper measures the direct effects of store‐level promotions, brand equity, and price consciousness and also examines the interactive effects of store‐level promotions and the moderating influence of category familiarity and normative influences. The results demonstrate the importance of simultaneous examination of interplay between different consumer and store‐level variables. Collectively, the results provide substantial segmentation opportunities for manufacturers of branded goods and retailers. Copyright © 2013 John Wiley & Sons, Ltd.
TiO2–B Nanosheets/Anatase Nanocrystals Co‐Anchored on Nanoporous Graphene: In Situ Reduction–Hydrolysis Synthesis and Their Superior Rate Performance as an Anode Material
Authors: Chen Chaoji , Hu Xianluo , Jiang Yan , Yang Ze , Hu Pei , Huang Yunhui
Published: December 2013

A unique hybrid, TiO2–B nanosheets/anatase nanocrystals co‐anchored on nanoporous graphene sheets, can be synthesized by a facile microwave‐induced in situ reduction–hydrolysis route. The as‐formed nanohybrid has a hierarchically porous structure, involving both mesopores of approximately 4 nm and meso‐/macropores of 30–60 nm in the graphene sheets, and a large surface area. Importantly, electrodes composed of the nanohybrid exhibit superior rate capability (160 mA h g−1 at ca. 36 C; 154 mA h g−1 at ca. 72 C) and excellent cyclability. The synergistic effects of conductive graphene with numerous nanopores and the pseudocapacitive effect of ultrafine TiO2–B nanosheets and anatase nanocrystals endow the hybrid a superior rate capability.
Chemical Analysis at Atomic Resolution of Isolated Extended Defects in an Oxygen‐Deficient, Complex Manganese Perovskite
Authors: Ruiz‐González M. Luisa , Cortés‐Gil Raquel , Torres‐Pardo Almudena , González‐Merchante Daniel , Alonso José M. , González‐Calbet José M.
Published: December 2013

A general approach to the structural and analytical characterization of complex bulk oxides that exploits the advantage of the atomic spatial resolution and the analytical capability of aberration‐corrected microscopy is described. The combined use of imaging and spectroscopic techniques becomes necessary to the complete characterization of the oxygen‐deficient colossal magnetoresistant La0.56Sr0.44MnO2.5‐related perovskite. In this compound, the formation of isolated (La/Sr)O and MnO rock‐salt‐type planar defects are identified from atomically resolved High Angle Annular Dark Field (HAADF) images. The location of the oxygen atomic columns from Annular Bright Field (ABF) images indicates edge‐sharing MnO6 octahedra in the MnO planes and the study performed by Electron Energy Loss Spectroscopy (EELS) reveals different Mn oxidation states derived from the corner‐ or edge‐sharing MnO6 octahedra environment.
Controlling Electron Rebound within Four‐Base π‐Stacks in Z‐DNA by Changing the Sugar Moiety from Deoxy‐ to Ribonucleotide
Authors: Sannohe Yuta , Kizaki Seiichiro , Kanesato Shuhei , Fujiwara Ayaka , Li Yue , Morinaga Hironobu , Tashiro Ryu , Sugiyama Hiroshi
Published: December 2013

Charge transfer through DNA is of great interest because of the potential of DNA to be a building block for nanoelectronic sensors and devices. The photochemical reaction of 5‐halouracil has been used for probing charge‐transfer processes along DNA. We previously reported on unique charge transfer following photochemical reaction of 5‐bromouracil within four‐base π‐stacks in Z‐DNA. In this study, we incorporated a guanosine instead of a deoxyguanosine into Z‐DNA, and found that electron transfer occurs in a different mechanism through four‐base π‐stacks.
Discovery of 5′′‐Chloro‐N‐[(5,6‐dimethoxypyridin‐2‐yl)methyl]‐2,2′:5′,3′′‐terpyridine‐3′‐carboxamide (MK‐1064): A Selective Orexin 2 Receptor Antagonist (2‐SORA) for the Treatment of Insomnia
Authors: Roecker Anthony J. , Mercer Swati P. , Schreier John D. , Cox Christopher D. , Fraley Mark E. , Steen Justin T. , Lemaire Wei , Bruno Joseph G. , Harrell C. Meacham , Garson Susan L. , Gotter Anthony L. , Fox Steven V. , Stevens Joanne , Tannenbaum Pamela L. , Prueksaritanont Thomayant , Cabalu Tamara D. , Cui Donghui , Stellabott Joyce , Hartman George D. , Young Steven D. , Winrow Christopher J. , Renger John J. , Coleman Paul J.
Published: December 2013

The field of small‐molecule orexin antagonist research has evolved rapidly in the last 15 years from the discovery of the orexin peptides to clinical proof‐of‐concept for the treatment of insomnia. Clinical programs have focused on the development of antagonists that reversibly block the action of endogenous peptides at both the orexin 1 and orexin 2 receptors (OX1R and OX2R), termed dual orexin receptor antagonists (DORAs), affording late‐stage development candidates including Merck’s suvorexant (new drug application filed 2012). Full characterization of the pharmacology associated with antagonism of either OX1R or OX2R alone has been hampered by the dearth of suitable subtype‐selective, orally bioavailable ligands. Herein, we report the development of a selective orexin 2 antagonist (2‐SORA) series to afford a potent, orally bioavailable 2‐SORA ligand. Several challenging medicinal chemistry issues were identified and overcome during the development of these 2,5‐disubstituted nicotinamides, including reversible CYP inhibition, physiochemical properties, P‐glycoprotein efflux and bioactivation. This article highlights structural modifications the team utilized to drive compound design, as well as in vivo characterization of our 2‐SORA clinical candidate, 5′′‐chloro‐N‐[(5,6‐dimethoxypyridin‐2‐yl)methyl]‐2,2′:5′,3′′‐terpyridine‐3′‐carboxamide (MK‐1064), in mouse, rat, dog, and rhesus sleep models.
Nitrate enrichment alters a Daphnia–microparasite interaction through multiple pathways
Authors: Dallas Tad , Drake John M.
Published: December 2013

Nutrient pollution has the potential to alter many ecological interactions, including host–parasite relationships. One of the largest sources of nutrient pollution comes from anthropogenic alteration of the nitrogen (N) cycle, specifically the increased rate of nitrate (NO3 ‐N) deposition to aquatic environments, potentially altering host–parasite relationships. This study aimed to assess the mechanisms through which nitrate may impact host–pathogen relationships using a fungal pathogen (Metschnikowia bicuspidata) parasitic to crustacean zooplankton (Daphnia dentifera) as a tractable model system. First, the influence of nitrate on host population dynamics was assessed along a gradient of nitrate concentrations. Nitrate decreased host population size and increased infection prevalence. Second, the influence of nitrate on host reproduction, mortality, and infection intensity was assessed at the individual host level by examining the relationship between pathogen dose and infection prevalence at ambient (0.4 mg NO3 ‐N*L−1) and intermediate (12 mg NO3 ‐N*L−1) levels of nitrate. Host fecundity and infection intensity both decreased with increasing pathogen dose, but increased nitrate levels corresponded to greater infection intensities. Nitrate had no effect on host growth rate, suggesting that hosts do not alter feeding behavior in nitrate‐treated media compared with ambient conditions. This study suggests that nutrient enrichment may enhance disease through increased transmission and infection intensity, but that high levels of nitrate may result in smaller epidemics through reduced transmission caused by smaller population sizes and increased pathogen mortality.
Morphological development and environmental degradation of superhydrophobic aspen and black locust leaf surfaces
Authors: Tranquada George Christopher , Erb Uwe
Published: December 2013

The complete adaxial surface development and degradation due to weathering of two superhydrophobic leaf species with (black locust, Robinia pseudoacacia and bigtooth aspen, Populus grandidentata) and one without (quaking aspen, Populus tremuloides) a protective downy layer of trichomes was followed over one entire growing season using scanning electron microscopy and wetting property measurements. Scanning electron microscopy micrographs illustrate the initial development period of a dual‐scale surface structure consisting of micrometre‐sized papillae and nanoscale wax crystals, which was followed by a period of environmental degradation as nanoscale wax features were subjected to environmental stress. It was discovered that robust micropapillae protect nanoscale wax morphologies in‐between them for quite some time allowing for a high tolerance to wear and erosion. Weather data was used to demonstrate the correlation between cumulative environmental stresses (temperature, relative humidity, precipitation and wind) and the loss of superhydrophobicity at the end of the growing season. Analysis of all binary combinations of the aforementioned environmental stress factors propose that cumulatively hot and humid weather have the most significant impact on degradation of nanoscale wax crystals and therefore result in the final loss of superhydrophobic behaviour. These findings were also tentatively linked to their importance in ecohydrology suggesting that canopy storage capacity, which has been previously correlated with leaf hydrophobicity, of superhydrophobic leaves may fluctuate considerably throughout an entire growing season. Copyright © 2013 John Wiley & Sons, Ltd.
Leveling the Playing Field: Fostering Collaborative Governance Towards On‐Going Reconciliation
Authors: Zurba Melanie
Published: December 2013

This paper looks at the potential for collaborative governance for land and resources to become a form of on‐going reconciliation in societies transitioning from past oppression and on‐going social injustice. Structural violence in the form of capacity disparities and policy configuration are explored in order to frame the discussion. Canada and South Africa are presented as two countries that have been going through different experiences in terms of transition, but have had similar experiences in regards to a need to foster new ways of working towards more equitable forms of governance. Here, much like reconciliation, governance is presented as a process, which under the right conditions can lead to relationship building and mutually desirable outcomes. This process creates a platform for establishing common ground, as well as the potential to negotiate solutions to structural violence. Copyright © 2013 John Wiley & Sons, Ltd and ERP Environment.
Legitimacy in Co‐Management: The Impact of Preexisting Structures, Social Networks and Governance Strategies
Authors: Sandström Annica , Crona Beatrice , Bodin Örjan
Published: December 2013

With the ambition to contribute to the endeavour of co‐management, this paper focuses on the critical aspect of legitimacy and sets out to explain stakeholder acceptance in natural resource governance. A comparative study of five coastal and marine areas in Sweden is conducted. The empirical results demonstrate, first, how the past and the present institutional landscape set the underlying conditions and affect stakeholders’ acceptance of new co‐management initiatives. Second, the results point to the critical function of network governance. Conscious choices regarding what composition of actors to involve, and in particular the inclusion and commitment of government actors, have significant bearing on stakeholder acceptance. Furthermore, deliberative efforts to reframe the process, adjusting the agenda to ongoing collaborative processes and key stakeholder goals, are seemingly as important. Thus, strivings towards legitimate co‐management require skilful manoeuvring of the present institutional landscape as well as deliberate strategies for the evolution of social networks. Copyright © 2013 John Wiley & Sons, Ltd and ERP Environment
Environmental Policy Can Happen: Shuttle Diplomacy and the Reality of Reg Neg Lite
Authors: Rinfret Sara , Cook Jeffrey
Published: December 2013

In an era of congressional gridlock, innovation and pathways for the future of environmental policymaking are needed. The US Environmental Protection Agency (EPA) has long been an innovator in environmental decisionmaking, reconciling stakeholder differences. Yet, with the decline of negotiated rulemaking (‘reg neg’), the purpose of our examination is to offer insights into how a contemporary US EPA creates rules. This exploratory study examines original interview data from actors involved during the creation of EPA rules to suggest that a present‐day EPA uses a new approach, shuttle diplomacy, to develop rules. The findings from our research suggest that this new rule development method can help agencies that might operate in an adversarial environment to reconcile stakeholder differences while writing the language of a proposed rule before publishing a Notice of Proposed Rulemaking. We argue that this new rule development model is important for understanding the next generation of environmental rulemaking, not only in the US, but internationally as well. Copyright © 2013 John Wiley & Sons, Ltd and ERP Environment.
Implantable device diagnostics on day of discharge identify heart failure patients at increased risk for early readmission for heart failure
Authors: Small Roy S. , Whellan David J. , Boyle Andrew , Sarkar Shantanu , Koehler Jodi , Warman Eduardo N. , Abraham William T.
Published: December 2013

Aims

We hypothesized that diagnostic data in implantable devices evaluated on the day of discharge from a heart failure hospitalization (HFH) can identify patients at risk for HF readmission (HFR) within 30 days.

Methods and results

In this retrospective analysis of four studies enrolling patients with CRT devices, we identified patients with a HFH, device data on the day of discharge, and 30‐day post‐discharge clinical follow‐up. Four diagnostic criteria were evaluated on the discharge day: (i) intrathoracic impedance >8 Ω below reference impedance; (ii) AF burden >6 h; (iii) CRT pacing <90%; and (iv) night heart rate >80 b.p.m. Patients were considered to have higher risk for HFR if ≥2 criteria were met, average risk if 1 criterion was met, and lower risk if no criteria were met. A Cox proportional hazards model was used to compare the groups. The data cohort consisted of a total of 265 HFHs in 175 patients, of which 36 (14%) were followed by HFR. On the discharge day, ≥2 criteria were met in 43 (16% of 265 HFHs), only 1 criterion was met in 92 (35%), and none of the four criteria were met in 130 HFHs (49%); HFR rates were 28, 16, and 7%, respectively. HFH with ≥2 criteria met was five times more likely to have HFR compared with HFH with no criteria met (adjusted hazard ratio 5.0; 95% confidence interval 1.9–13.5, P = 0.001).

Conclusion

Device‐derived diagnostic criteria evaluated on the day of discharge identified patients at significantly higher risk of HFR.
The DNA damage response induces antigen presenting cell‐like functions in fibroblasts
Authors: Tang Melissa Li Fang , Khan Muznah Khatoo Nazar , Croxford John Ludovic , Tan Kar Wai , Angeli Veronique , Gasser Stephan
Published: December 2013

The DNA damage response (DDR) alerts the immune system to the danger posed by DNA damage through the induction of damage‐associated molecular pattern molecules, chemokines, and ligands for activating immune receptors such as lymphocyte function‐associated antigen 1 (LFA‐1), NKG2D, and DNAX accessory molecule 1 (DNAM‐1). Here we provide evidence that OVA257–264‐pulsed fibroblasts gain the ability to activate naïve OT‐I CD8+ T cells in response to DNA damage. The ability of fibroblasts to activate OT‐I CD8+ T cells depended on the upregulation of ICAM‐1 on fibroblasts and DNAM‐1 expression of CD8+ T cells. OVA257–264‐pulsed fibroblasts were able to induce a protective T‐cell response against B16‐OVA cells in a DDR‐dependent manner. Hence, the DDR may alert the immune system to the presence of potentially dangerous cells by upregulating the expression of ligands that can induce the activation of innate and adaptive immune cells.
Human regulatory T cells are selectively activated by low‐dose application of the CD28 superagonist TGN1412/TAB08
Authors: Tabares Paula , Berr Susanne , Römer Paula S. , Chuvpilo Sergej , Matskevich Alexey A. , Tyrsin Dmitry , Fedotov Yury , Einsele Hermann , Tony Hans‐Peter , Hünig Thomas
Published: December 2013

CD28 superagonists (CD28SAs) are potent T‐cell‐activating monoclonal antibodies (mAbs). In contrast to their benign behavior and marked therapeutic efficacy as activators of regulatory T (Treg) cells in preclinical rodent models, a phase I trial of the human CD28SA TGN1412 (now called TAB08) in 2006 resulted in a life‐threatening cytokine release syndrome (CRS). We studied TAB08‐mediated Treg‐cell activation in a recently developed in vitro system of human PBMCs, which also reproduces the CRS experienced by the healthy volunteers. We show that just as in rodents, CD28SAs are potent activators and expanders of Treg cells from healthy donors and rheumatoid arthritis patients, even under effective blockade of pro‐inflammatory cytokine release by a corticosteroid. Moreover, CD28SA titration identifies a dose range where pro‐inflammatory cytokine secretion from conventional T cells is absent while appreciable Treg‐cell activation is maintained. Finally, we report that low‐dose application of TAB08 to healthy volunteers results in dose‐dependent systemic release of the Treg‐cell signature cytokine IL‐10 in the absence of the pro‐inflammatory factors associated with the CRS of the 2006 TGN1412 study. These results demonstrate the potential of appropriately dosed CD28SA and corticosteroid comedication to mobilize human Treg cells for the treatment of autoimmune and inflammatory conditions.
Accounting for Word‐of‐Mouth Effects in Preference‐Based Market Forecasts
Authors: Pescher Christian , Spann Martin
Published: December 2013

Consumers differ in their involvement in new product purchase decisions. Opinion leaders usually show a higher involvement in their purchase decisions than other consumers. This leads to a higher stability in their answers when being asked about their preferences. An important question that previous research has not analyzed yet is whether and how to capture this finding in preference‐based market forecasts. The authors study these aspects for a representative sample of 364 consumers in the mobile phone market of a large European country. They find that assigning higher weights to the preferences of opinion leaders in aggregate market forecasts results in estimates that are more consistent with observed market shares than forecasts in which all consumers are given equal weights. The authors further test different measures of opinion leadership and find that sociometric indicators outperform psychographic constructs to account for the outcome of opinion leadership in preference‐based market forecasts. Copyright © 2013 John Wiley & Sons, Ltd.
A tiered area‐of‐review framework for geologic carbon sequestration
Authors: Birkholzer Jens , Cihan Abdullah , Bandilla Karl
Published: December 2013

This paper discusses the current guidance given by the United States Environmental Protection Agency (EPA) on delineating the so‐called Area of Review (AoR) for the permitting of geologic carbon sequestration (GCS) projects. According to the EPA's regulatory framework for GCS, the AoR refers to the region surrounding the CO2 injection well(s) wherein leakage of CO2 and/or the migration of formation fluids could possibly endanger overlying groundwater resources. Our evaluation of the current framework for delineating the size of this area finds unnecessary conservatism in the definition of the critical pressure, which could lead to a heavy burden on permit applicants that seek to get regulatory compliance, in particular for very large GCS projects. We propose a risk‐based re‐interpretation of this framework, separating the total Area of Review into different sub‐areas with different regulatory requirements depending on whether the concern is about free‐phase CO2 or pressure‐driven brine migration. This leads to a tiered AoR definition in which the projected region of CO2 plume extent would have the highest regulatory standards regarding site characterization, monitoring, and corrective action. The requirements in the AoR outside this central region would be less burdensome because of a narrower focus on major pathways for brine leakage such as unplugged wellbores and large faults. We expect that this revised framework would allow for a reduction in the cost of regulatory compliance for projects with very large injection volumes, while ensuring that the objective of protecting valuable groundwater resources is preserved.
LIQUIDITY SHOCKS AND THE SUPPLY OF CREDIT AFTER THE 2007–2008 CRISIS
Authors: Kapan Tümer , Minoiu Camelia
Published: December 2013

We examine the impact of heightened liquidity risk in bank funding markets during the 2007–2008 subprime crisis on the supply of credit to firms internationally. Our sample comprises almost 800 banks lending in the syndicated loan market to borrowers from 48 advanced and emerging market economies. To control for simultaneous changes in demand, we relate within‐borrower changes in bank credit from multiple banks to banks' exposure to liquidity risk. To capture banks' vulnerability to the liquidity shocks that occurred during 2007–2008, we use measures of pre‐crisis reliance on wholesale funding. Our results suggest that banks that were more exposed to liquidity risk during the crisis reduced syndicated lending more than other banks. Our results are robust to alternate measures of vulnerability to liquidity risk, choice of sample period and simultaneous movements in exchange rates. Copyright © 2013 John Wiley & Sons, Ltd.
MANAGING THE FRAGILITY OF THE EUROZONE BY PAUL De GRAUWE
Authors: Borgy Vladimir , Bouthevillain Carine , Dufrénot Gilles
Published: December 2013

This paper discusses sources of self‐fulfilling equilibria in the Eurozone when some governments are highly susceptible to movements of distrust by investors who fear some payment difficulty. Self‐fulfilling prophecies occur when countries become insolvent only because investors fear insolvency. They induce multiple equilibria, some of which correspond to bad equilibria and others to good equilibria. An important issue then is to solve this problem, notably to eliminate the bad equilibria. In the short‐run, the role of the central bank as a lender of last resort is key. But this raises issues about the risk inherent to its intervention (inflation, solvency). In the medium run, macroeconomic policies in the euro are central (structural reforms and the reduction of external imbalances). In the long run, it may be worth proceeding to the consolidation of national budgets and debts, which would protect the countries of being forced with default by the financial markets. Copyright © 2013 John Wiley & Sons, Ltd.
GLOBAL IMBALANCES AND FINANCIAL SECTOR INSTABILITIES: INTRODUCTION
Authors: Borgy Vladimir , Bouthevillain Carine , Dufrénot Gilles
Published: December 2013

This special issue provides several views about the sources of the current crisis and policy solutions to cope with it. It brings together papers from academic institutions, international organizations and central banks. The first three papers argue that the crisis was triggered by the lack of confidence of the investors in the markets. This was reflected, for instance, in the pricing of the public debt (with an increase in the sovereign debt spreads) and in the reduced syndicated lending in wholesale lending markets. The other three papers focus on policy aspects by analyzing indicators that could serve as early warning signals of increasing stress and vulnerability. The authors propose three set of indicators: policy‐based indicators, some variables used in the macro‐prudential literature and financial indexes. The papers are a selection of papers presented at a Conference on Macroeconomic and financial vulnerability indicators in advanced economies co‐organizes by the Banque de France and the University of Strasbourg on 13–14 September 2013. Copyright © 2013 John Wiley & Sons, Ltd.
PRICING SOVEREIGN BOND RISK IN THE EUROPEAN MONETARY UNION AREA: AN EMPIRICAL INVESTIGATION
Authors: Afonso António , Arghyrou Michael G. , Kontonikas Alexandros
Published: December 2013

We use a panel of 10 euro area countries to assess the determinants of long‐term sovereign bond yield spreads over the period 1999.01–2010.11. We find that government bond yield spreads are well explained by fiscal fundamentals over the crisis period. We also find that the menu of risk factors priced by markets has been significantly enriched since March 2009, including international risk, liquidity risk and the risk of the crisis' transmission among European Monetary Union member states. Finally, we find that transmission risk has increased considerably since spring 2009 because of rapidly increasing risk of investing in periphery bonds relative to core ones. Copyright © 2013 John Wiley & Sons, Ltd.
Osteoblasts Protect AML Cells From SDF‐1‐Induced Apoptosis
Authors: Kremer Kimberly N. , Dudakovic Amel , McGee‐Lawrence Meghan E. , Philips Rachael L. , Hess Allan D. , Smith B. Douglas , van Wijnen Andre J. , Karp Judith E. , Kaufmann Scott H. , Westendorf Jennifer J. , Hedin Karen E.
Published: December 2013

The bone marrow provides a protective environment for acute myeloid leukemia (AML) cells that often allows leukemic stem cells to survive standard chemotherapeutic regimens. Targeting these leukemic stem cells within the bone marrow is critical for preventing relapse. We recently demonstrated that SDF‐1, a chemokine abundant in the bone marrow, induces apoptosis in AML cell lines and in patient samples expressing high levels of its receptor, CXCR4. Here we show that a subset of osteoblast lineage cells within the bone marrow can protect AML cells from undergoing apoptosis in response to the SDF‐1 naturally present in that location. In co‐culture systems, osteoblasts at various stages of differentiation protected AML cell lines and patient isolates from SDF‐1‐induced apoptosis. The differentiation of the osteoblast cell lines, MC3T3 and W‐20‐17, mediated this protection via a cell contact‐independent mechanism. In contrast, bone marrow‐derived mesenchymal cells, the precursors of osteoblasts, induced apoptosis in AML cells via a CXCR4‐dependent mechanism and failed to protect AML cells from exogenously added SDF‐1. These results indicate that osteoblasts in the process of differentiation potently inhibit the SDF‐1‐driven apoptotic pathway of CXCR4‐expressing AML cells residing in the bone marrow. Drugs targeting this protective mechanism could potentially provide a new approach to treating AML by enhancing the SDF‐1‐induced apoptosis of AML cells residing within the bone marrow microenvironment. J. Cell. Biochem. 115: 1128–1137, 2014. © 2013 Wiley Periodicals, Inc.
Expression of P2X7 ATP Receptor Mediating the IL8 and CCL20 Release in Human Periodontal Ligament Stem Cells
Authors: Trubiani Oriana , Horenstein Alberto L. , Caciagli Francesco , Caputi Sergio , Malavasi Fabio , Ballerini Patrizia
Published: December 2013

ATP is released by human periodontal ligament cells (hPDLCs) and has been shown to regulate PDL regeneration and responses to mechanical stress through activation of P2Y receptors. This nucleotide, however, has also been reported to trigger the pro‐inflammatory cascade by inducing the maturation and/or release of chemokines/cytokines from various cell types mainly via P2X7 receptors. Much less is known on the possible role of ATP in stem cells deriving from PDL (hPDLSCs) which are considered to be a promising tool for cell‐based therapy to restore lesions. Given the role played by P2X7 in pathophysiological conditions, in this study we investigated the expression of P2X7 ATP receptors in hPDLSCs. The results obtained showed that hPDLSCs express P2X7 receptors evaluated by means of cytofluorimetric, immunohistochemistry, reverse transcriptase‐PCR, and Western blot analyses. P2X7 ligation by 2′,3′‐(benzoyl‐4‐benzoyl)‐ATP (BzATP), a specific receptor agonist, was followed by an increase in intracellular Ca2+ and in the uptake of ethidium bromide. These effects were dramatically reduced by oxidized ATP (oATP), the P2X7 irreversible inhibitor, suggesting that the P2X7 is the functional receptor involved. At 24 h treatment of hPDLSCs with BzATP it enhanced the release of the pro‐inflammatory agents IL8 and CCL20, without influencing cell viability. These effects were counteracted by pre‐treating the cells with oATP or with A‐740003, a selective and potent P2X7 competitive antagonist. Collectively, these results indicated that extracellular ATP mediate a pro‐inflammatory response via P2X7 receptors in hPDLSCs opening a further approach to control hPDLSCs behavior in their possible application as therapeutic tool. J. Cell. Biochem. 115: 1138–1146, 2014. © 2013 Wiley Periodicals, Inc.
Issues in Identifying Germ Tube Positive Yeasts by Conventional Methods
Authors: Yazdanpanah Atta , Khaithir Tzar Mohd Nizam
Published: December 2013

Candida speciation is vital for epidemiology and management of candidiasis. Nonmolecular conventional methods often fail to identify closely related germ tube positive yeasts from clinical specimens. The present study was conducted to identify these yeasts and to highlight issues in conventional versus molecular methods of identification. A total of 98 germ tube positive yeasts from high vaginal swabs were studied over a 12‐month period. Isolates were examined with various methods including growth at 42°C and 45°C on Sabouraud dextrose agar (SDA), color development on CHROMagar Candida medium, chlamydospore production on corn meal agar at 25°C, carbohydrate assimilation using ID 32C system, and polymerase chain reaction using a single pair of primers targeting the hyphal wall protein 1 (Hwp1) gene. Of all the isolates studied, 97 were molecularly confirmed as C. albicans and one isolate was identified as C. dubliniensis. No C. africana was detected in this study. The molecular method used in our study was an accurate and useful tool for discriminating C. albicans, C. dubliniensis, and C. africana. The conventional methods, however, were less accurate and riddled with many issues that will be discussed in further details.