Ubiquitin-dependent proteolysis of trihydrophobin 1 (TH1) by the human papilloma virus E6-associated protein (E6-AP).

Human Papilloma virus E6-associated protein (E6-AP), which is known as an E3 ubiquitin ligase, mediates ubiquitination and subsequent degradation of a series of cellular proteins. In this paper, we identify here trihydrophobin 1 (TH1), an integral subunit of the human negative transcription elongation factor (NELF) complex, as a novel E6-AP interaction protein and a target of E6-AP-mediated degradation. Overexpression of E6-AP results in degradation of TH1 in a dose-dependent manner, whereas knock-down of endogenous E6-AP elevates the TH1 protein level. TH1 protein turnover is substantially faster, compared to controls, in cells that overexpressed E6-AP. Wild-type E6-AP promotes the ubiquitination of TH1, while a catalytically inactive point mutant of E6-AP abolishes its ubiquitination. Furthermore, in vitro ubiquitination assay also demonstrates that TH1 can be ubiquitinated by E6-AP. The degradation is blocked by treatment with proteasome inhibitor MG132. Herein, we provide strong evidence that TH1 is a specific substrate that is targeted for degradation through E6-AP-catalyzed polyubiquitination.

William Herschel Telescope observations of distant comets

We present results from broad-band V- and R-filter observations obtained at the 4.2-m William Herschel Telescope on La Palma on 2002 July 12-14. A total of six comets were imaged, and their heliocentric distances ranged from 2.8 to 6.1 au. The comets observed were 43P/Wolf-Harrington, 129P/Shoemaker-Levy 3, 133P/Elst-Pizarro, 143P/Kowal-Mrkos, P/1998 U4 (Spahr) and P/2001 H5 (NEAT). A detailed surface brightness profile analysis indicates that three of the targeted comets (43P/Wolf-Harrington, 129P/Shoemaker-Levy 3 and P/1998 U4) were visibly active, and the remaining three comets were stellar in appearance. Further analysis shows that for the three `stellar-like' comets the possible coma contribution to the observed flux does not exceed 12.2 per cent, and in the case of comet 143P/Kowal-Mrkos the coma contribution is expected to be as low as 1 per cent, and so the resulting photometry most likely represents that of the projected nucleus surface. Effective radii for the inactive comets range from 1.02 to 4.56 km, and the effective radius upper limits for the active comets range from 1.94 to 4.15 km. We assume an albedo and phase coefficient of 0.04 and 0.035 mag deg-1, respectively, with the exception of comets 133P/Elst-Pizarro and 143P/Kowal-Mrkos for which phase coefficients were previously measured. These values are compared with previous measurements, and for comet 43P/Wolf-Harrington we find that the nucleus axial ratio a/b could be as large as 2.44. For the active comets we measured dust production levels in terms of the Af? quantity. Spectral gradients were extracted for two of the inactive comets from their measured broad-band colour indices, and compared with the rest of the comet population for which (V-R) colour and spectral gradient values exist. We find a spectral gradient for 143P/Kowal-Mrkos of 9.9 +/- 8.1 per cent/100 nm, which is very typical of Jupiter-family comets, the majority of which have reflectivity gradients in the range 0-13 per cent (100 nm)-1. The spectral gradient for comet 133P/Elst-Pizarro is amongst the bluest yet measured. We measure a (V-R) colour index value of 0.14 +/- 0.11 for the nucleus of 133P/Elst-Pizarro which is considerably lower than previous measurements. A possible explanation for this difference is considered.

Optical observations of 23 distant Jupiter Family Comets, including 36P/Whipple at multiple phase angles

We present photometry on 23 Jupiter Family Comets (JFCs) observed at large heliocentric distance, primarily using the 2.5-m Isaac Newton Telescope (INT). Snapshot images were taken of 17 comets, of which five were not detected, three were active and nine were unresolved and apparently inactive. These include 103P/Hartley 2, the target of the NASA Deep Impact extended mission, EPOXI. For six comets we obtained time-series photometry and use this to constrain the shape and rotation period of these nuclei. The data are not of sufficient quantity or quality to measure precise rotation periods, but the time-series do allow us to measure accurate effective radii and surface colours. Of the comets observed over an extended period, 40P/Väisälä 1, 47P/Ashbrook-Jackson and P/2004 H2 (Larsen) showed faint activity which limited the study of the nucleus. Light curves for 94P/Russell 4 and 121P/Shoemaker-Holt 2 reveal rotation periods of around 33 and 10h, respectively, although in both cases these are not unique solutions. 94P was observed to have a large range in magnitudes implying that it is one of the most elongated nuclei known, with an axial ratio a/b >= 3. 36P/Whipple was observed at five different epochs, with the INT and ESO's 3.6-m NTT, primarily in an attempt to confirm the preliminary short rotation period apparent in the first data set. The combined data set shows that the rotation period is actually longer than 24h. A measurement of the phase function of 36P's nucleus gives a relatively steep ß = 0.060 +/- 0.019. Finally, we discuss the distribution of surface colours observed in JFC nuclei, and show that it is possible to trace the evolution of colours from the Kuiper Belt Object (KBO) population to the JFC population by applying a `dereddening' function to the KBO colour distribution.

New ways to break an old bond: the bacterial carbon-phosphorus hydrolases and their role in biogeochemical phosphorus cycling

Phosphonates are organophosphorus molecules that contain the highly stable C-P bond, rather than the more common, and more labile, C-O-P phosphate ester bond. They have ancient origins but their biosynthesis is widespread among more primitive organisms and their importance in the contemporary biosphere is increasingly recognized; for example phosphonate-P is believed to play a particularly significant role in the productivity of the oceans. The microbial degradation of phosphonates was originally thought to occur only under conditions of phosphate limitation, mediated exclusively by the poorly characterized C-P lyase multienzyme system, under Pho regulon control. However, more recent studies have demonstrated the Pho-independent mineralization by environmental bacteria of three of the most widely distributed biogenic phosphonates: 2-aminoethylphosphonic acid (ciliatine), phosphonoacetic acid, and 2-amino-3-phosphonopropionic acid (phosphonoalanine). The three phosphonohydrolases responsible have unique specificities and are members of separate enzyme superfamilies; their expression is regulated by distinct members of the LysR family of bacterial transcriptional regulators, for each of which the phosphonate substrate of the respective degradative operon serves as coinducer. Previously no organophosphorus compound was known to induce the enzymes required for its own degradation. Whole-genome and metagenome sequence analysis indicates that the genes encoding these newly described C-P hydrolases are distributed widely among prokaryotes. As they are able to function under conditions in which C-P lyases are inactive, the three enzymes may play a hitherto-unrecognized role in phosphonate breakdown in the environment and hence make a significant contribution to global biogeochemical P-cycling.

Catalytic role of gold in gold-based catalysts: A density functional theory study on the CO oxidation on gold

Gold-based catalysts have been of intense interests in recent years, being regarded as a new generation of catalysts due to their unusually high catalytic performance. For example, CO oxidation on Au/TiO2 has been found to occur at a temperature as low as 200 K. Despite extensive studies in the field, the microscopic mechanism of CO oxidation on Au-based catalysts remains controversial. Aiming to provide insight into the catalytic roles of Au, we have performed extensive density functional theory calculations for the elementary steps in CO oxidation on Au surfaces. O atom adsorption, CO adsorption, O-2 dissociation, and CO oxidation on a series of Au surfaces, including flat surfaces, defects and small clusters, have been investigated in detail. Many transition states involved are located, and the lowest energy pathways are determined. We find the following: (i) the most stable site for O atom on Au is the bridge site of step edge, not a kink site; (ii) O-2 dissociation on Au (O-2-->20(ad)) is hindered by high barriers with the lowest barrier being 0.93 eV on a step edge; (iii) CO can react with atomic O with a substantially lower barrier, 0.25 eV, on Au steps where CO can adsorb; (iv) CO can react with molecular O-2 on Au steps with a low barrier of 0.46 eV, which features an unsymmetrical four-center intermediate state (O-O-CO); and (v) O-2 can adsorb on the interface of Au/TiO2 with a reasonable chemisorption energy. On the basis of our calculations, we suggest that (i) O-2 dissociation on Au surfaces including particles cannot occur at low temperatures; (ii) CO oxidation on Au/inactive-materials occurs on Au steps via a two-step mechanism: CO+O-2-->CO2+O, and CO+O-->CO2; and (iii) CO oxidation on Au/active-materials also follows the two-step mechanism with reactions occurring at the interface.

Disposable, stable media for reproducible surface-enhanced Raman spectroscopy

Large numbers of identical and stable SE(R)RS [surface-enhanced (resonance) Raman]-active media, which are convenient to handle and manipulate but sufficiently inexpensive that they can be used once and then discarded, have been prepared by isolating nanoparticles from Ag and Au sols in hydrophilic polymer gels. The preparation simply involves mixing a suitable polymer with the sol to give a viscous suspension that can be coated onto a substrate and dried to form a hard translucent film. The films remain inactive until they are treated with aqueous analyte solution, which causes the film to swell and brings the analyte into contact with the active metal particles. The swollen films give strong SERS spectra which are effectively identical to those obtained from simple sols. The advantage of this method is that the dried polymers can be stored indefinitely before use and that they give a high degree of spectral reproducibility.

Credit and community: working class debt in the UK since 1880

This book examines credit in working class communities since 1880, focusing on forms of borrowing that were dependent on personal relationships and social networks. It provides an extended historical discussion of credit unions, legal and illegal moneylenders (loan sharks), and looks at the concept of ‘financial exclusion’. Initially, the book focuses on the history of tallymen, check traders, and their eventual movement into moneylending following the loss of their more affluent customers, due to increased spending power and an increasingly liberalized credit market. They also faced growing competition from mail order companies operating through networks of female agents, whose success owed much to the reciprocal cultural and economic conventions that lay at the heart of traditional working class credit relationships. Discussion of these forms of credit is related to theoretical debates about cultural aspects of credit exchange that ensured the continuing success of such forms of lending, despite persistent controversies about their use. The book contrasts commercial forms of credit with formal and informal co-operative alternatives, such as the mutuality clubs operated by co-operative retailers and credit unions. It charts the impact of post-war immigration upon credit patterns, particularly in relation to the migrant (Irish and Caribbean) origins of many credit unions and explains the relative lack of success of the credit union movement. The book contributes to anti-debt debates by exploring the historical difficulties of developing legislation in relation to the millions of borrowers who have patronized what has come to be termed the sub-prime sector.

An empirical study of green supply chain management practices amongst UK manufacturers

Purpose: The purpose of this paper is to examine the extent and nature of greening the supply chain (SC) in the UK manufacturing sector; and the factors that influence the breadth and depth of this activity.

Design/methodology/approach: Based on the findings from a sample of manufacturing organisations drawn from the membership of The Chartered Institute for Purchasing and Supply. Data are collected using a questionnaire, piloted and pre-tested before distribution with responses from 60 manufacturing companies.

Findings: On average manufacturers perceive the greatest pressure to improve environmental performance through legislation and internal drivers (IDs). The least influential pressures are related to societal drivers and SC pressures from individual customers. Green supply chain management (GSCM) practices amongst this “average” group of UK manufacturing organisations are focusing on internal, higher risk, descriptive activities, rather than proactive, external engagement processes. Environmental attitude (EA) is a key predictor of GSCM activity and those organisations that have a progressive attitude are also operationally very active. EA shows some relationship to legislative drivers but other factors are also influential. Operational activity may also be moderated by organisational contingencies such as risk, size, and nationality.

Research limitations/implications: The main limitation to this paper is the relatively small manufacturing sample.

Practical implications: This paper presents a series of constructs that identify GSCM operational activities companies to benchmark themselves against. It suggests which factors are driving these operational changes and how industry contingencies may be influential.

Originality/value: This paper explores what is driving environmental behaviour amongst an “average” sample of manufacturers, what specific management practices take place and the relationships between them.

Keywords: Manufacturing industries, Environmental management, Supply chain management, Sustainable development, United Kingdom
Paper type: Research paper

An allosteric potentiator of M4 mAChR modulates hippocampal synaptic transmission.

Muscarinic acetylcholine receptors (mAChRs) provide viable targets for the treatment of multiple central nervous system disorders. We have used cheminformatics and medicinal chemistry to develop new, highly selective M₄ allosteric potentiators. VU10010, the lead compound, potentiates the M₄ response to acetylcholine 47-fold while having no activity at other mAChR subtypes. This compound binds to an allosteric site on the receptor and increases affinity for acetylcholine and coupling to G proteins. Whole-cell patch clamp recordings revealed that selective potentiation of M₄ with VU10010 increases carbachol-induced depression of transmission at excitatory but not inhibitory synapses in the hippocampus. The effect was not mimicked by an inactive analog of VU10010 and was absent in M₄ knockout mice. Selective regulation of excitatory transmission by M₄ suggests that targeting of individual mAChR subtypes could be used to differentially regulate specific aspects of mAChR modulation of function in this important forebrain structure.

Understanding online customer repurchasing intention and the mediating role of trust - an empirical investigation in two developed countries

Although e-commerce adoption and customers initial purchasing behavior have been well studied in the literature, repeat purchase intention and its antecedents remain understudied. This study proposes a model to understand the extent to which trust mediates the effects of vendor-specific factors on customers intention to repurchase from an online vendor. The model was tested and validated in two different country settings. We found that trust fully mediates the relationships between perceived reputation, perceived capability of order fulfillment, and repurchasing intention, and partially mediates the relationship between perceived website quality and repurchasing intention in both countries. Moreover, multi-group analysis reveals no significant between-country differences of the model with regards to the antecedents and outcomes of trust, except the effect of reputation on trust. Academic and practical implications and future research are discussed. © 2009 Operational Research Society Ltd.

Homogeneous habitat can meet the discrete and varied resource requirements of hares but may set an ecological trap

In common with other farmland species, hares (Lepus spp.) are in widespread decline in agricultural landscapes due to agricultural intensification and habitat loss. We examined the importance of habitat heterogeneity to the Irish hare (Lepus timidus hibernicus) in a pastoral landscape. We used radio-tracking during nocturnal active and diurnal inactive periods throughout one year. In autumn, winter and spring, hares occupied a heterogeneous combination of improved grassland, providing food, and Juncus-dominated rough pasture, providing refuge. In summer, hares significantly increased their use of improved grassland. This homogeneous habitat can fulfil the discrete and varied resource requirements of hares for feeding and shelter at certain times of year. However, improved grassland may be a risky habitat for hares as silage harvesting occurs during their peak birthing period of late spring and early summer. We therefore posit the existence of a putative ecological trap inherent to a homogeneous habitat of perceived high value that satisfies the hares' habitat requirements but which presents risks at a critical time of year. To test this hypothesis in relation to hare populations, work is required to provide data on differential leveret mortality between habitat types.

Relationship between rooting behaviour and foraging in growing pigs

The objective of the present study was to investigate the relationship between rooting behaviour and foraging in growing pigs. In study 1, forty-eight 11-week-old pigs were housed in eight groups of six with access to a rooting substrate in the form of spent mushroom compost. In half of the groups the rooting substrate contained food rewards, and in the other half of the groups it did not. All pigs had ad libitum access to feed. In study 2, one hundred and ninety-two 11-week-old pigs were housed in thirty-two groups of six, all with access to spent mushroom compost, and eight groups were each fed to 70, 80, 90 or 100% appetite. Treatments were applied over a two-week period in both studies. The number of pigs involved in active rooting (rooting in substrate while standing), inactive rooting (rooting in substrate while sitting or lying) or non-rooting activity (standing in substrate area and involved in any activity except rooting) was recorded by scan sampling. These behaviours tended to reach a peak in the morning and again in the afternoon. Inactive rooting was not significantly affected by treatments in study I or study 2. Food rewards in the rooting substrate led to a significant reduction in active rooting behaviour and in non-rooting activity during peak periods of the day (P

Influence of environmental enrichment on welfare-related behavioural and physiological parameters in growing pigs

Three hundred and twenty pigs were reared from birth to slaughter at 21 weeks in either barren or enriched environments. The barren environments were defined as intensive housing (slatted floors and minimum recommended space allowances) and the enriched environments incorporated extra space including an area which contained peat and straw in a rack. Behavioural observations showed that environmental enrichment reduced time spent inactive and rime spent involved in harmful social and aggressive behaviour (P

Influence of environmental enrichment on the behaviour, performance and meat quality of domestic pigs

Three hundred and twenty pigs were reared from birth to slaughter at 21 weeks in either barren or enriched environments. The barren environments were defined as intensive housing (slatted floors and minimum recommended space allowances) and the enriched environments incorporated extra space, an area which contained peat and straw in a rack. Behavioural observations showed that environmental enrichment reduced time spent inactive and time spent involved in harmful social and aggressive behaviour while increasing the time spent in exploratory behaviour. During the finishing period (15-21 weeks) mean daily food intakes were higher and food conversion ratios were lower for pigs in enriched environments compared with their counterparts in barren environments (P

Mechanism of Activation of a G Protein-coupled Receptor, the Human Cholecystokinin-2 Receptor

G protein-coupled receptors (GPCRs) represent a major focus in functional genomics programs and drug development research, but their important potential as drug targets contrasts with the still limited data available concerning their activation mechanism. Here, we investigated the activation mechanism of the cholecystokinin-2 receptor (CCK2R). The three-dimensional structure of inactive CCK2R was homology-modeled on the basis of crystal coordinates of inactive rhodopsin. Starting from the inactive CCK2R modeled structure, active CCK2R (namely cholecystokinin-occupied CCK2R) was modeled by means of steered molecular dynamics in a lipid bilayer and by using available data from other GPCRs, including rhodopsin. By comparing the modeled structures of the inactive and active CCK2R, we identified changes in the relative position of helices and networks of interacting residues, which were expected to stabilize either the active or inactive states of CCK2R. Using targeted molecular dynamics simulations capable of converting CCK2R from the inactive to the active state, we delineated structural changes at the atomic level. The activation mechanism involved significant movements of helices VI and V, a slight movement of helices IV and VII, and changes in the position of critical residues within or near the binding site. The mutation of key amino acids yielded inactive or constitutively active CCK2R mutants, supporting this proposed mechanism. Such progress in the refinement of the CCK2R binding site structure and in knowledge of CCK2R activation mechanisms will enable target-based optimization of nonpeptide ligands.