Cannabinoids stimulate fibroblastic colony formation by bone marrow cells indirectly via CB2 receptors

Recently, the cannabinoid receptors CB1 and CB2 were shown to modulate bone formation and resorption in vivo, although little is known of the mechanisms underlying this. The effects of cannabinoids on mesenchymal stem cell (MSC) recruitment in whole bone marrow were investigated using either the fibroblastic colony-forming unit (CFU-f) assay or high-density cultures of whole bone marrow. Levels of the CB1 and CB2 receptors were assessed by flow cytometry. Treatment of CFU-f cultures with the endocannabinoid 2-arachidonylglycerol (2-AG) dose-dependently increased fibroblastic and differentiated colony formation along with colony size. The nonspecific agonists CP 55,940 and WIN 55,212 both increased colony numbers, as did the CB2 agonists BML190 and JWH015. The CB1-specific agonist ACEA had no effect, whereas the CB2 antagonist AM630 blocked the effect of the natural cannabinoid tetrahydrocannabivarin, confirming mediation via the CB2 receptor. Treatment of primary bone marrow cultures with 2-AG stimulated proliferation and collagen accumulation, whereas treatment of subcultures of MSC had no effect, suggesting that the target cell is not the MSC but an accessory cell present in bone marrow. Subcultures of MSCs were negative for CB1 and CB2 receptors as shown by flow cytometry, whereas whole bone marrow contained a small population of cells positive for both receptors. These data suggest that cannabinoids may stimulate the recruitment of MSCs from the bone marrow indirectly via an accessory cell and mediated via the CB2 receptor. This recruitment may be one mechanism responsible for the increased bone formation seen after cannabinoid treatment in vivo.

Both tumour-promoting and non-promoting phorbol esters inhibit 125I-EGF binding and stimulate the phosphorylation of a 80kd protein kinase C substrate protein in intact Swiss 3T3 cells

Sapintoxin A (SAP A) and 12-deoxyphorbol 13-phenylacetate (DOPP), are two biologically active but non-turnour-promoting phorbol esters that potently bind to and activate the phorbol ester receptor, protein kinase C (PKC). SAP A and DOPP cause a dose-dependent increase in the phosphorylation of an 80 kd (80K) substrate protein for PKC in Swiss 3T3 cells. A similar dose—response effect was seen with sapintoxin D (SAP D), the stage 2 promoting analogue of 12-O-tetradecanoylphorbol-13-acetate and the complete promoter phorbol 12,13-dibutyrate (PDB). The doses resulting in a half maximal phosphorylation of this protein (Ka were 20 nM (SAP A), 45 nM (DOPP), 23 nM (SAP D) and 37 nM (PDB). Both non-promoting and phorbol esters induced a dose-dependent inhibition of [125I]epidermal growth factor (EGF) binding to its receptor in Swiss 3T3 cells. The doses required for 50% inhibition of binding (Ki) were: 8 nM (SAP A), 16 nM (DOPP), 14 nM (SAP D) and 17 nM (PDB). The results clearly demonstrate that induction of phosphorylation of the Pu 80K phosphoprotein and inhibition of [125I]EGF binding in Swiss 3T3 cells following exposure to phorbol esters is independent of the tumour-promoting activity of these compounds. The fact that SAP A, DOPP, SAP D and PDB are mitogenic for a variety of cell types and that exposure to these compounds leads to 80K phosphorylation and inhibition of [125I]EGF binding, suggests that these early biological events may play a role in the mitogenic response induced by these compounds.

Does regional infrastructure investment stimulate extra housing and commercial building supply? Long-run evidence across the English regions

This paper uses long-term regional construction data to investigate whether increases infrastructure investment in the English regions leads to subsequent rises in housebuilding and new commercial property, using time series modeling. Both physical (roads and harbours) and social infrastructure (education and health) impacts are investigated across nine regions in England. Significant effects for physical infrastructure are found across most regions and, also, some evidence of a social infrastructure effect. The results are not consistent across regions, which may be due to geographical differences and to network and diversionary effects. However, the results do suggest that infrastructure does have some impact but follows differential lag structures. These results provide a test of the hypothesis of the economic benefits of infrastructure investment in an approach that has not been used before.

Non-D9-tetrahydrocannabinol phytocannabinoids stimulate feeding in rats

Cannabinoid type 1 receptor-mediated appetite stimulation by D9tetrahydrocannabinol (D9THC) is well understood. Recently, it has become apparent that non-D9THC phytocannabinoids could also alter feeding patterns. Here, we show definitively that non-D9THC phytocannabinoids stimulate feeding. Twelve male, Lister-Hooded rats were prefed to satiety prior to administration of a standardized cannabis extract or to either of two mixtures of pure phytocannabinoids (extract analogues) comprising the phytocannabinoids present in the same proportions as the standardized extract (one with and one without D9THC). Hourly intake and meal pattern data were recorded and analysed using two-way analysis of variance followed by one-way analysis of variance and Bonferroni post-hoc tests. Administration of both extract analogues significantly increased feeding behaviours over the period of the test. All three agents increased hour-one intake and meal-one size and decreased the latency to feed, although the zero-D9THC extract analogue did so to a lesser degree than the high-D9THC analogue. Furthermore, only the analogue containing D9THC significantly increased meal duration. The data confirm that at least one non-D9THC phytocannabinoid induces feeding pattern changes in rats, although further trials using individual phytocannabinoids are required to fully understand the observed effects.

Collagen, convulxin, and thrombin stimulate aggregation-independent tyrosine phosphorylation of CD31 in platelets. Evidence for the involvement of Src family kinases

Platelet endothelial cell adhesion molecule-1 (CD31) is a 130-kDa glycoprotein receptor present on the surface of platelets, neutrophils, monocytes, certain T-lymphocytes, and vascular endothelial cells. CD31 is involved in adhesion and signal transduction and is implicated in the regulation of a number of cellular processes. These include transendothelial migration of leukocytes, integrin regulation, and T-cell function, although its function in platelets remains unclear. In this study, we demonstrate the ability of the platelet agonists collagen, convulxin, and thrombin to induce tyrosine phosphorylation of CD31. Furthermore, we show that this event is independent of platelet aggregation and secretion and is accompanied by an increase in surface expression of CD31. A kinase capable of phosphorylating CD31 was detected in CD31 immunoprecipitates, and its activity was increased following activation of platelets. CD31 tyrosine phosphorylation was reduced or abolished by the Src family kinase inhibitor PP2, suggesting a role for these enzymes. In accordance with this, each of the Src family members expressed in platelets, namely Fyn, Lyn, Src, Yes, and Hck, was shown to co-immunoprecipitate with CD31. The involvement of Src family kinases in this process was confirmed through the study of mouse platelets deficient in Fyn.

Agonists of protease-activated receptors 1 and 2 stimulate electrolyte secretion from mouse gallbladder

Cholecystitis is one of the most common gastrointestinal diseases. Inflammation induces the activation of proteases that can signal to cells by cleaving protease-activated receptors (PARs) to induce hemostasis, inflammation, pain, and repair. However, the distribution of PARs in the gallbladder is unknown, and their effects on gallbladder function have not been fully investigated. We localized immunoreactive PAR(1) and PAR(2) to the epithelium, muscle, and serosa of mouse gallbladder. mRNA transcripts corresponding to PAR(1) and PAR(2), but not PAR(4), were detected by RT-PCR and sequencing. Addition of thrombin and a PAR(1)-selective activating peptide (TFLLRN-NH(2)) to the serosal surface of mouse gallbladder mounted in an Ussing chamber stimulated an increase in short-circuit current in wild-type but not PAR(1) knockout mice. Similarly, serosally applied trypsin and PAR(2) activating peptide (SLIGRL-NH(2)) increased short-circuit current in wild-type but not PAR(2) knockout mice. Proteases and activating peptides strongly inhibited electrogenic responses to subsequent stimulation with the same agonist, indicating homologous desensitization. Removal of HCO(3)(-) ions from the serosal buffer reduced responses to thrombin and trypsin by >80%. Agonists of PAR(1) and PAR(2) increase intracellular Ca(2+) concentration in isolated and cultured gallbladder epithelial cells. The COX-2 inhibitor meloxicam and an inhibitor of CFTR prevented the stimulatory effect of PAR(1) but not PAR(2). Thus PAR(1) and PAR(2) are expressed in the epithelium of the mouse gallbladder, and serosally applied proteases cause a HCO(3)(-) secretion. The effects of PAR(1) but not PAR(2) depend on generation of prostaglandins and activation of CFTR. These mechanisms may markedly influence fluid and electrolyte secretion of the inflamed gallbladder when multiple proteases are generated.

Does infrastructure investment stimulate building supply? The case of the English regions

Does infrastructure investment stimulate building supply? The case of the English regions, Regional Studies. Policies to improve infrastructure to stimulate regional growth remain common. This paper investigates whether increases in infrastructure investment in the English regions lead to subsequent rises in new commercial and residential property, using time-series modelling. Both physical (roads and harbours) and social infrastructure (education and health) impacts are investigated. Hardly any infrastructure effects with respect to commercial property investment were found, which raises doubts about whether extra infrastructure creates employment, though some impact was related to residential building. Overall, these results raise doubts about the supposed direct effects of infrastructure policies on regional jobs and growth.

Endothelin-1 and fibroblast growth factors stimulate the mitogen-activated protein kinase signaling cascade in cardiac myocytes. The potential role of the cascade in the integration of two signaling pathways leading to myocyte hypertrophy.

Maximally effective concentrations of endothelin-1 (ET-1), acidic FGF (aFGF), or 12-O-tetradecanoylphorbol-13-acetate (TPA) activated mitogen-activated protein kinase (MAPK) by 3-4-fold in crude extracts of myocytes cultured from neonatal rat heart ventricles. Maximal activation was achieved after 5 min. Thereafter, MAPK activity stimulated by ET-1 or aFGF declined to control values within 1-2 h, whereas activation by TPA was more sustained. Two peaks of MAPK activity (a 42- and a 44-kDa MAPK) were resolved in cells exposed to ET-1 or aFGF by fast protein liquid chromatography on a Mono Q column. One major and one minor peak of MAPK kinase (MAPKK) was stimulated by ET-1 or aFGF. Cardiac myocytes expressed protein kinase C (PKC)-alpha, -delta, -epsilon and -zeta as shown immunoblotting. Exposure to 1 microM TPA for 24 h down-regulated PKC-alpha, -delta, and -epsilon, but not PKC-zeta. This maneuver wholly abolished the activation of MAPK on re-exposure to TPA but did not affect the response to aFGF. The effect of ET-1 was partially down-regulated. ET-1 stimulated phospho[3H]inositide hydrolysis 18-fold, whereas aFGF stimulated by only 30%. Agonists which initially utilize dissimilar signaling pathways may therefore converge at the level of MAPKK/MAPK and this may be relevant to the hypertrophic response of the heart.

Pro-inflammatory cytokines stimulate mitogen-activated protein kinase subfamilies, increase phosphorylation of c-Jun and ATF2 and upregulate c-Jun protein in neonatal rat ventricular myocytes.

Pro-inflammatory cytokines may be important in the pathophysiological responses of the heart. We investigated the activation of the three mitogen-activated protein kinase (MAPK) subfamilies ¿c-Jun N-terminal kinases (JNKs), p38-MAPKs and extracellularly-responsive kinases (ERKs) by interleukin-1 beta (IL-1 beta) or tumour necrosis factor alpha (TNF alpha) in primary cultures of myocytes isolated from neonatal rat ventricles. Both cytokines stimulated a rapid (maximal within 10 min) increase in JNK activity. Although activation of JNKs by IL-1 beta was transient returning to control values within 1 h, the response to TNF alpha was sustained. IL-1 beta and TNF alpha also stimulated p38-MAPK phosphorylation, but the response to IL-1 beta was consistently greater than TNF alpha. Both cytokines activated ERKs, but to a lesser degree than that induced by phorbol esters. The transcription factors, c-Jun and ATF2, are phosphorylated by the MAPKs and are implicated in the upregulation of c-Jun. IL-1 beta and TNF alpha stimulated the phosphorylation of c-Jun and ATF2. However, IL-1 beta induced a greater increase in c-Jun protein. Inhibitors of protein kinase C (PKC) (Ro318220, GF109203X) and the ERK cascade (PD98059) attenuated the increase in c-Jun induced by IL-1 beta, but LY294002 (an inhibitor of phosphatidylinositol 3' kinase) and SB203580 (an inhibitor of p38-MAPK, which also inhibits certain JNK isoforms) had no effect. These data illustrate that some of the pathological effects of IL-1 beta and TNF alpha may be mediated through the MAPK cascades, and that the ERK cascade, rather than JNKs or p38-MAPKs, are implicated in the upregulation of c-Jun by IL-1 beta.

Immersive Projection for biochemical pattern matching

In all biological processes, protein molecules and other small molecules interact to function and form transient macromolecular complexes. This interaction of two or more molecules can be described by a docking event. Docking is an important phase for structure-based drug design strategies, as it can be used as a method to simulate protein-ligand interactions. Various docking programs exist that allow automated docking, but most of them have limited visualization and user interaction. It would be advantageous if scientists could visualize the molecules participating in the docking process, manipulate their structures and manually dock them before submitting the new conformations to an automated docking process in an immersive environment, which can help stimulate the design/docking process. This also could greatly reduce docking time and resources. To achieve this, we propose a new virtual modelling/docking program, whereby the advantages of virtual modelling programs and the efficiency of the algorithms in existing docking programs will be merged.

Pesticides and other micro-organic contaminants in freshwater sedimentary environments - a review

A wide range of issues relating to the presence and fate of pesticides and other micro-organic contaminants (MOCs) in surface freshwater sedimentary environments is reviewed. These issues include the sources, transport and occurrence of MOCs in freshwater environments; their ecological effects; their interaction with sedimentary material; and a range of processes related to their fate, including degradation, diffusion in bed sediments, bioturbation and slow contaminant release. An emphasis is placed on those processes-chemical, physical or biological-in which sediments play a role in determining the fate of micro-organics in freshwater environments. The issues of occurrence, source and transport, and the ecological effects of micro-organics are introduced more briefly, the focus where these aspects are concerned being largely on pesticides. In the concluding section, key points and issues relating to the study of micro-organics in freshwater environments are summarised and areas where initial or further research would be welcome are highlighted. It is hoped that this paper will both form a useful reference for workers in the field of micro-organic contaminants, and also stimulate new work in the freshwater environment and beyond. (C) 2003 Elsevier Science Ltd. All rights reserved.

Integrating methods for developing sustainability indicators that can facilitate learning and action

Bossel's (2001) systems-based approach for deriving comprehensive indicator sets provides one of the most holistic frameworks for developing sustainability indicators. It ensures that indicators cover all important aspects of system viability, performance, and sustainability, and recognizes that a system cannot be assessed in isolation from the systems upon which it depends and which in turn depend upon it. In this reply, we show how Bossel's approach is part of a wider convergence toward integrating participatory and reductionist approaches to measure progress toward sustainable development. However, we also show that further integration of these approaches may be able to improve the accuracy and reliability of indicators to better stimulate community learning and action. Only through active community involvement can indicators facilitate progress toward sustainable development goals. To engage communities effectively in the application of indicators, these communities must be actively involved in developing, and even in proposing, indicators. The accuracy, reliability, and sensitivity of the indicators derived from local communities can be ensured through an iterative process of empirical and community evaluation. Communities are unlikely to invest in measuring sustainability indicators unless monitoring provides immediate and clear benefits. However, in the context of goals, targets, and/or baselines, sustainability indicators can more effectively contribute to a process of development that matches local priorities and engages the interests of local people.

Pesticides and other micro-organic contaminants in freshwater sedimentary environments - a review

A wide range of issues relating to the presence and fate of pesticides and other micro-organic contaminants (MOCs) in surface freshwater sedimentary environments is reviewed. These issues include the sources, transport and occurrence of MOCs in freshwater environments; their ecological effects; their interaction with sedimentary material; and a range of processes related to their fate, including degradation, diffusion in bed sediments, bioturbation and slow contaminant release. An emphasis is placed on those processes-chemical, physical or biological-in which sediments play a role in determining the fate of micro-organics in freshwater environments. The issues of occurrence, source and transport, and the ecological effects of micro-organics are introduced more briefly, the focus where these aspects are concerned being largely on pesticides. In the concluding section, key points and issues relating to the study of micro-organics in freshwater environments are summarised and areas where initial or further research would be welcome are highlighted. It is hoped that this paper will both form a useful reference for workers in the field of micro-organic contaminants, and also stimulate new work in the freshwater environment and beyond. (C) 2003 Elsevier Science Ltd. All rights reserved.

The expert teacher of English

The Expert Teacher of English is for all passionate teachers – both novice and expert - who aspire to become outstanding professionals. It considers what we mean by ‘expert’ and ‘expertise’, explores concepts that are vital to understanding what expertise in teaching is ‘for’, and discusses the characteristics of excellent teaching. As increasing attention is being paid to the concept of the professional who can model excellent teaching and mentor and develop others, it provides a critical analysis of The Advanced Skills Teacher and the Excellent Teacher, as well as the Chartered Teacher in Scotland and the ‘highly accomplished teacher’ in the US. Drawing on the views, ideas and experiences of a group of skilful teachers, The Expert Teacher of English aims to stimulate personal and professional development, help you reflect on the concept of expertise, and support you as you develop as a highly accomplished teacher.

Proceedings of the West Africa Built Environment Research (WABER) Conference 2010

FOREWORD Welcome to this West Africa Built Environment Research (WABER) conference taking place here in Ghana. Thank you for coming and welcome to Accra. The main aims of the WABER conference are: to help young researchers and early-career scholars in West Africa to develop their research work and skills through constructive face-to-face interaction with experienced academics; to provide a platform for networking and collaborative work among senior built environment academics in West Africa; and to serve as a vehicle for developing the field of construction management and economics in Africa. Waber 2009 The WABER event in 2009 was held at the British Council in Accra, Ghana on 2-3 June. The event was a resounding success. It attracted participation from 32 researchers, from 12 different institutions, who presented their work to an audience of approximately 100 people. Each presenter received immediate and constructive feedback from an international panel. The event was opened by Professor K.K. Adarkwa, Vice Chancellor of KNUST, Kumasi, Ghana, with several senior academics and researchers from universities, polytechnics, and other institutions in Ghana and Nigeria in attendance. There was also a significant level of attendance by senior construction practitioners in Ghana. Thank you to the School of Construction Management and Engineering, University of Reading, UK for funding the inaugural event in 2009. We are also grateful to all of you who helped to make the event a success and to those of you who have joined us here today to build upon the success and legacy of WABER 2009. Waber 2010 This year, we have 60+ peer-reviewed papers and presentations on topics relating to Building services and maintenance, Construction costs, Construction design and technology, Construction education, Construction finance, Construction procurement, Contract administration, Contract management, Contractor development, Decision support systems, Dispute resolution, Economic development, Energy efficiency, Environment and sustainability, Health and safety, Human resources, Information technology, Marketing, Materials science, Organisation strategy and business performance, Productivity, Project management, Quantity surveying, Real estate and planning, Solar energy systems, Supply chain management and Urban development. We hope that these papers will generate interest among delagates and stimulate discussion here and beyond the conference into the wider community of academia and industry. The delegates at this conference come from 10 different countries. This provides a rich international and multicultural blend and a perfect platform for networking and developing collaborations. This year we are blessed to have three high profile keynote speakers in the persons of Professor George Ofori (National University of Singapore), Dr Roine Leiringer (University of Reading, UK) and Professor Will Hughes (University of Reading, UK). We are also thankful to Dr Chris Harty (University of Reading, UK) who is facilitating the Research Skills Workshop on ‘Writing a scientific article’. Thank you to Dr Sena Agyepong of our conference organising team for her capable management of local organising arrangements. And above all, thank you to all of you for coming to this conference. Enjoy and have a safe journey back home. Dr Samuel Laryea School of Construction Management and Engineering University of Reading, July 2010