bluuebunny/crossref_metadata_2025_split huggingface dataset:
Triazine dyes are agonists of the NAADP receptor
Authors: Billington Richard A , Bak Judit , Martinez‐Coscolla Ana , Debidda Marcella , Genazzani Armando A 
Published: August 2004

NAADP has been shown to be a potent calcium‐releasing second messenger in a wide variety of cell types to date. However, research has been hampered by a lack of pharmacological agents, with which to investigate NAADP‐induced calcium release, and by the molecular identity of its cellular target protein being unknown.
In the present paper, the sea urchin egg model was used to investigate whether triazine dyes, which can act as nucleotide mimetics, can bind to the NAADP receptor, induce Ca2+ release and be used for affinity chromatography of the receptor.
Indeed, all the triazine dyes tested (Reactive Red 120 (RR120), Reactive Green 19 (RG19), Reactive Green 5 (RG5), Cibacron Blue 3GA and Reactive Yellow 86) displayed micromolar affinities, except for Reactive Orange 14. Furthermore, unlike NAADP, RR120, RG19 and RG5 did not bind in an irreversible manner.
The compound that displayed the highest affinity, RR120, was tested in a 45Ca2+ efflux assay. This compound released Ca2+ via the NAADP receptor, as shown by the ability of subthreshold NAADP concentrations to inhibit this release. Furthermore, heparin and ruthenium red were unable to block RR120‐induced Ca2+ release.
We have also shown that RG5 and RG19, immobilised on resins, retain the ability to bind to the receptor, and that this interaction can be disrupted by high salt concentrations. As a proof of principle, we have shown that this can be used to partially purify the NAADP receptor by at least 75‐fold.
In conclusion, triazine dyes interact with the NAADP receptor, and this could be exploited in future to create a new generation of pharmacological tools to investigate this messenger and, in combination with other techniques, to purify the receptor.

British Journal of Pharmacology (2004) 142, 1241–1246. doi:10.1038/sj.bjp.0705886
Increased wakefulness, motor activity and decreased theta activity after blockade of the 5‐HT2B receptor by the subtype‐selective antagonist SB‐215505
Authors: Kantor Sandor , Jakus Rita , Balogh Brigitta , Benko Anita , Bagdy Gyorgy 
Published: August 2004

Serotonin‐2 receptor antagonists, like ritanserin, greatly enhance deep slow wave sleep (SWS‐2) and low‐frequency EEG power in humans and rodents. 5‐HT2A and 5‐HT2C receptors may be involved in these effects, but the role of the 5‐HT2B receptor is still unclear.
To investigate the role of the 5‐HT2B receptor in regulation of the sleep–wake cycle, the subtype‐selective antagonist SB‐215505 (0.1, 0.3 and 1.0 mg kg−1 i.p.) was administered to Sprague–Dawley rats at light onset (beginning of passive phase). EEG, EMG and motor activity were recorded during the subsequent 8 h.
SB‐215505 dose‐dependently increased wakefulness (W) at the expense of the intermediate stage of sleep, paradoxical sleep (PS) and SWS‐2 in the first hour. Parallel to increased W, significantly increased motor activity was found. Spectral analysis of the EEG in W showed a dose‐dependent decrease in power density in the 3–8 Hz frequency range (maximum effect at 6 Hz). In light slow wave sleep and SWS‐2, the drug reduced low‐frequency (<8 Hz) EEG power, suggesting decreased sleep intensity after SB‐215505 treatment. In PS, the drug dose‐dependently decreased EEG power solely in the theta (6–9 Hz) band, primarily affecting the peak power value (7 Hz).
The well‐known SWS‐2 enhancing effect of 5‐HT2 receptor antagonists is mediated by 5‐HT2A and/or 5‐HT2C receptors. In contrast, blockade of 5‐HT2B receptors increases motor activity and W along with decreased theta activity during W and PS. Activation of 5‐HT2B receptors may contribute to initiation of sleep and to theta generation during W and PS under physiological conditions.

British Journal of Pharmacology (2004) 142, 1332–1342. doi:10.1038/sj.bjp.0705887
The NAADP receptor: commentary on Billington et al.
Authors: Galione A , Parrington J , Dowden J 
Published: August 2004

NAADP is a recently described calcium‐mobilizing messenger. First discovered as a potent calcium‐releasing molecule in sea urchin eggs, its actions have now been reported in several mammalian cell types. In the sea urchin egg, NAADP‐sensitive calcium release channels appear distinct from inositol trisphosphate or ryanodine receptors, and are mainly localized to acidic compartments. In this study, Billington et al. extend the pharmacology of the putative NAADP receptor utilizing molecules unrelated to NAADP itself. This work may provide an important step in developing selective NAADP receptor modulators that will help define the role of NAADP in cell signalling.British Journal of Pharmacology (2004) 142, 1203–1207. doi:10.1038/sj.bjp.0705888
Effects of cannabinoid receptor‐2 activation on accelerated gastrointestinal transit in lipopolysaccharide‐treated rats
Authors: Mathison Ronald , Ho Winnie , Pittman Quentin J , Davison Joseph S , Sharkey Keith A 
Published: August 2004

The biological effects of cannabinoids (CB) are mediated by CB1 and CB2 receptors. The role of CB2 receptors in the gastrointestinal tract is uncertain. In this study, we examined whether CB2 receptor activation is involved in the regulation of gastrointestinal transit in rats.
Basal and lipopolysaccharide (LPS)‐stimulated gastrointestinal transit was measured after instillation of an Evans blue‐gum Arabic suspension into the stomach, in the presence of specific CB1 and CB2 agonists and antagonists, or after treatment with inhibitors of mediators implicated in the transit process.
In control rats a CB1 (ACEA; 1 mg kg−1), but not a CB2 (JWH‐133; 1 mg kg−1), receptor agonist inhibited basal gastrointestinal transit. The effects of the CB1 agonist were reversed by the CB1 antagonist AM‐251, which alone increased basal transit. LPS treatment increased gastrointestinal transit. This increased transit was reduced to control values by the CB2, but not the CB1, agonist. This inhibition by the CB2 agonist was dose dependent and prevented by a selective CB2 antagonist (AM‐630; 1 mg kg−1).
By evaluating the inhibition of LPS‐enhanced gastrointestinal transit by different antagonists, the effects of the CB2 agonist (JWH‐133; 1 mg kg−1) were found to act via cyclooxygenase, and to act independently of inducible nitric oxide synthase (NOS) and platelet‐activating factor. Interleukin‐1β and constitutive NOS isoforms may be involved in the accelerated LPS transit.
The activation of CB2 receptors in response to LPS is a mechanism for the re‐establishment of normal gastrointestinal transit after an inflammatory stimulus.

British Journal of Pharmacology (2004) 142, 1247–1254. doi:10.1038/sj.bjp.0705889
Cannabinoids and intestinal motility: welcome to CB2 receptors
Authors: Izzo Angelo A 
Published: August 2004

Δ9‐Tetrahydrocannabinol (the active ingredient of marijuana), as well as endogenous and synthetic cannabinoids, exert many biological functions by activating two types of cannabinoid receptors, CB1 receptors (expressed by central and peripheral neurons) and CB2 receptors (that occur mainly in immune cells). Convincing evidence has accumulated in recent years that cannabinoids inhibit gastric and intestinal motility through activation of enteric CB1 receptors. However, a report in this issue of British Journal of Pharmacology has highlighted the possibility that CB2 receptors in the rat intestine could contribute to reducing the increase of intestinal motility induced by an endotoxic inflammation. By minimizing the adverse psychotropic effects associated with brain cannabinoid receptors, the CB2 receptor represents a new molecular target for the treatment of motility disorders associated with intestinal inflammation.British Journal of Pharmacology (2004) 142, 1201–1202. doi:10.1038/sj.bjp.0705890
Effects of phosphodiesterase inhibitors on interleukin‐4 and interleukin‐13 generation from human basophils
Authors: Eskandari Nahid , Wickramasinghe Thulani , Peachell Peter T 
Published: August 2004

The aim of the present study was to determine whether inhibition of cyclic nucleotide phosphodiesterase (PDE) modulates the stimulated generation of the cytokines, interleukin‐4 (IL‐4) and IL‐13, from human basophils. This was addressed by evaluating the effects of both nonselective and selective inhibitors of PDEs on the generation of cytokines from basophils.
The nonselective PDE inhibitors, isobutyl‐methylxanthine (IBMX) and theophylline, attenuated the IgE‐mediated generation of IL‐4 and IL‐13 and, also, the release of histamine from basophils.
The effects of the isoform‐selective inhibitors, 8‐methoxymethyl‐IBMX (PDE 1 inhibitor), siguazodan (PDE3 inhibitor), rolipram (PDE4 inhibitor), denbufylline (PDE4 inhibitor), Org 30029 (mixed PDE3 and 4 inhibitor) and zaprinast (PDE5 inhibitor), were studied. Of these selective compounds, only rolipram, denbufylline and Org 30029 inhibited the IgE‐dependent generation of IL‐4, IL‐13 and histamine from basophils to a statistically significant (P<0.05) degree.
The effects of isoform‐selective inhibitors on basophils activated by IL‐3 were evaluated. The IL‐3‐induced generation of IL‐4, IL‐13 and histamine was inhibited to a statistically significant (P<0.05) extent, only by compounds that act as inhibitors of PDE4.
These data suggest that inhibition of PDE4 can regulate the generation of cytokines from human basophils.

British Journal of Pharmacology (2004) 142, 1265–1272. doi:10.1038/sj.bjp.0705892
Dopamine D3 receptor‐mediated inhibition of Na+/H+ exchanger activity in normotensive and spontaneously hypertensive rat proximal tubular epithelial cells
Authors: Pedrosa Rui , Gomes Pedro , Zeng Chunyu , Hopfer Ulrich , Jose Pedro A , Soares‐da‐Silva Patrício 
Published: August 2004

This study evaluated the response of the Na+/H+ exchanger (NHE) to dopamine D1‐ and D2‐like receptor stimulation in immortalized renal proximal tubular epithelial cells and freshly isolated renal proximal tubules from the spontaneously hypertensive rat (SHR) and their normotensive controls (Wistar Kyoto rats; WKY).
Stimulation of D1‐like receptors with SKF 38393 attenuated NHE activity in WKY cells (IC50=151 nM), but not in SHR cells. Stimulation of D2‐like receptors with quinerolane (IC50=120 nM) attenuated NHE activity in SHR cells, but not in WKY cells. Forskolin was equipotent in SHR and WKY cells in inhibiting NHE activity. The effect of SKF 38393 was abolished by overnight treatment of WKY cells with cholera toxin (CTX, 500 ng ml−1), but not with pertussis toxin (PTX, 100 ng ml−1). The effect of quinerolane (1 μM) was abolished by overnight treatment of SHR cells with PTX, but not with CTX.
The D3 receptor agonist 7‐OH‐DPAT (IC50=0.8 μM) attenuated NHE activity in SHR cells only. This effect was abolished by S‐sulpiride and by overnight treatment with PTX. The D4 receptor agonist RBI 257 did not affect NHE activity.
The 7‐OH‐DPAT inhibited NHE activity in freshly isolated renal proximal tubules from 4‐ and 12‐week‐old SHR and 12‐week‐old WKY, but not in freshly isolated renal proximal tubules from 4‐week‐old WKY.
It is concluded that D3 receptors coupled to a Gi/o protein play a role in the handling of tubular Na+, namely through inhibition of the NHE activity, this being of particular relevance in the SHR, which fail to respond to D1‐like dopamine receptor stimulation.

British Journal of Pharmacology (2004) 142, 1343–1353. doi:10.1038/sj.bjp.0705893
Cellular actions of somatostatin on rat periaqueductal grey neurons in vitro
Authors: Connor Mark , Bagley Elena E , Mitchell Vanessa A , Ingram Susan L , Christie MacDonald J , Humphrey Patrick P A , Vaughan Christopher W 
Published: August 2004

Functional studies indicate that the midbrain periaqueductal grey (PAG) is involved in the analgesic actions of somatostatin; however, the cellular actions of somatostatin in this brain region are unknown. In the present study, whole‐cell patch clamp recordings were made from rat PAG neurons in vitro. In 93% of acutely isolated neurons, somatostatin inhibited Ca2+‐channel currents. This effect was mimicked by the sst‐2 selective agonist BIM‐23027, but not by the sst‐1 and sst‐5 selective agonists CH‐275 and L‐362855. In brain slices, 81% of neurons responded to somatostatin (300 nM) with an increase in K+ conductance that reversed polarity at −114 mV. A greater proportion of somatostatin‐sensitive neurons (93%) than somatostatin‐insensitive neurons (53%) responded to the opioid agonist met‐enkephalin (10 μM). Somatostatin also reduced the amplitude of evoked GABAA‐mediated inhibitory postsynaptic currents (IPSCs). The actions of somatostatin in brain slices were mimicked by BIM‐23027, but not by CH‐275. Somatostatin had a variable effect on the rate of spontaneous miniature IPSCs in normal external potassium solutions. In high external potassium solutions, somatostatin reduced the rate of miniature IPSCs in all neurons, and this inhibition was abolished by addition of Cd2+ (30 μM). Somatostatin had no effect on the amplitude of miniature IPSCs. These results indicate that somatostatin acts via sst‐2 receptors to directly inhibit a subpopulation of PAG neurons by activating a potassium conductance and inhibits GABA release within PAG via a presynaptic Ca2+‐dependent mechanism. Thus, like opioids, somatostatin has the potential to exert pre‐ and postsynaptic disinhibitory effects within the PAG.British Journal of Pharmacology (2004) 142, 1273–1280. doi:10.1038/sj.bjp.0705894
Intestinal Na+–K+–ATPase activity and molecular events downstream of interferon‐γ receptor stimulation
Authors: Magro Fernando , Fraga Sónia , Ribeiro Tomé , Soares‐da‐Silva Patrício 
Published: August 2004

This study evaluated the effects of human interferon‐γ (IFN‐γ) on Na+–K+–ATPase activity and the intracellular signaling pathways involved in human intestinal epithelial Caco‐2 cells.
Na+–K+–ATPase activity was determined as the difference between total and ouabain‐sensitive ATPase. p38 MAP kinase activity was analyzed by Western blotting using the p38 MAP kinase assay kit. Total and phosphorylated STAT1 protein levels were detected using the PhosphoPlus® Stat1.
IFN‐γ decreased Na+–K+–ATPase activity in a time‐ and concentration‐dependent manner. The IFN‐γ‐induced decrease in Na+–K+–ATPase activity was accompanied by no changes in the abundance of α1 subunit Na+–K+–ATPase. Downregulation of protein kinase C (PKC) with phorbol‐12,13‐dibutyrate (PDBu) prevented the inhibitory effect of IFN‐γ on Na+–K+–ATPase activity. Inhibition of Raf‐1, mitogen‐activated protein kinase kinase (MAPKK/MEK), p38 MAPK and STAT1 with, respectively, GW 5074, PD 98059, SB 203580 and epigallocatechin gallate prevented inhibition of Na+–K+–ATPase activity by IFN‐γ.
Treatment with IFN‐γ markedly increased the expression of total and phospho‐STAT1, this being accompanied by activation of p38 MAPK. Activation of phospho‐STAT1 by IFN‐γ was almost abolished by epigallocatechin gallate and markedly reduced by SB 203580, but insensitive to downregulation of PKC.
The increase in short circuit current (Isc) by 1.0 and 2.5 μg ml−1 amphotericin B was markedly attenuated in IFN‐γ‐treated cells. However, the inhibitory effect of PDBu on the amphotericin B‐induced increase in Isc was of similar magnitude in vehicle‐ and IFN‐γ‐treated cells.
It is concluded that IFN‐γ markedly attenuates Na+–K+–ATPase activity. The transduction mechanisms set into motion by IFN‐γ involve the activation of PKC downstream STAT1 phosphorylation and Raf‐1, MEK, ERK2 and p38 MAPK pathways, in a complex sequence of events.

British Journal of Pharmacology (2004) 142, 1281–1292. doi:10.1038/sj.bjp.0705895
Anandamide acts as a vasodilator of dural blood vessels in vivo by activating TRPV1 receptors
Authors: Akerman S , Kaube H , Goadsby P J 
Published: August 2004

Migraine pathophysiology is believed to involve the release of neuropeptides via the activation of trigeminal afferents that innervate the cranial vasculature.
Anandamide, the endogenous ligand to the cannabinoid receptor, is able to inhibit neurogenic dural vasodilatation, calcitonin gene‐related peptide (CGRP)‐induced and nitric oxide‐induced dural vessel dilation in the intravital microscopy model. In an in vitro setting anandamide is also able to activate the vanilloid type 1 (TRPV1) receptor and cause vasodilation, via the release of CGRP.
In this study we used intravital microscopy to study whether anandamide behaves as a TRPV1 receptor agonist in the trigeminovascular system. We examined if anandamide‐induced dural vasodilation involves CGRP release that can be reversed by the CGRP receptor antagonist, CGRP8–37, and whether like capsaicin the anandamide effect could be reversed by the TRPV1 receptor antagonist, capsazepine.
Anandamide 1 (19±9%, n=12), 3 (29±5%, n=37), 5 (74±7%, n=13) and 10 mg kg−1 (89±18%, n=6) was able to cause a dose‐dependent increase in dural vessel diameter. Capsazepine (3 mg kg−1, t5=6.2, P<0.05) and CGRP8–37 (300 μg kg−1, t6=11.1, P<0.05) attenuated the anandamide‐induced dural vessel dilation when compared to control (Student's paired t‐test). AM251 (3 mg kg−1), a cannabinoid type 1 (CB1) receptor antagonist, was unable to reverse this anandamide‐induced dilation.
The study demonstrates that anandamide acts as a TRPV1 receptor agonist in the trigeminovascular system, activating TRPV1 receptors that promote CGRP release and cause vasodilation independent of any action at the CB1 receptor. Anandamide has been shown previously to inhibit trigeminovascular neurons and prevent vasodilation, through an action at CB1 receptors.

British Journal of Pharmacology (2004) 142, 1354–1360. doi:10.1038/sj.bjp.0705896
The cannabinomimetic arachidonyl‐2‐chloroethylamide (ACEA) acts on capsaicin‐sensitive TRPV1 receptors but not cannabinoid receptors in rat joints
Authors: Baker Chris L , McDougall Jason J 
Published: August 2004

The vasoactive effects of the synthetic cannabinoid (CB) arachidonyl‐2‐chloroethylamide (ACEA) was tested in the knee joints of urethane‐anaesthetised rats. Experiments were also performed to determine whether these vasomotor responses could be blocked by the selective CB1 receptor antagonists AM251 (N‐(piperidin‐1‐yl)‐5‐(4‐iodophenyl)‐1‐(2,4‐dichlorophenyl)‐4‐methyl‐1H‐pyrazole‐3‐carboxamide) (10−9 mol) and AM281 (1‐(2,4‐dichlorophenyl)‐5‐(4‐iodophenyl)‐4‐methyl‐N‐4‐morpholinyl‐1H‐pyrazole‐3‐carboxamide) (10−8 mol), as well as the selective CB2 receptor antagonist AM630 (6‐iodo‐2‐methyl‐1‐[2‐4(morpholinyl)ethyl]‐[1H‐indol‐3‐yl](4‐methoxyphenyl)methanone) (10−8 mol).
Peripheral application of ACEA (10−14–10−9 mol) onto the exposed surface of the knee joint capsule caused a dose‐dependent increase in synovial blood flow. The dilator action of the CB occurred within 1 min after drug administration and rapidly returned to control levels shortly thereafter. The maximal vasodilator effect of ACEA corresponded to a 30% increase in articular perfusion compared to control levels.
The hyperaemic action of ACEA was not significantly altered by coadministration of AM251, AM281 or AM630 (P>0.05; two‐way ANOVA).
The transient receptor potential channel vanilloid receptor 1 (TRPV1) antagonist capsazepine (10−6 mol) significantly reduced the vasodilator effect of ACEA on joint blood vessels (P=0.002). Furthermore, destruction of unmyelinated and thinly myelinated joint sensory nerves by capsaicin (8‐methyl‐N‐vanillyl‐6‐nonenamide) treatment also attenuated ACEA responses (P<0.0005).
These data clearly demonstrate a vasodilator effect of the cannabinomimetic ACEA on knee joint perfusion. Rather than a classic CB receptor pathway, ACEA exerts its vasomotor influence by acting via TRPV1 receptors located on the terminal branches of capsaicin‐sensitive afferent nerves innervating the joint.

British Journal of Pharmacology (2004) 142, 1361–1367. doi:10.1038/sj.bjp.0705902
SMEs in the New Economy — Evidence from Selected Italian Districts
Authors: Piscitello Lucia , Sgobbi Francesca 
Published: August 2004

The paper investigates SMEs' responses to the challenges posed by the new economy, i.e. the joint action of the increasing globalisation and the diffusion of ICTs. Indeed, while on the one hand the new economy may weaken the traditional obstacles to the transformation of SMEs into global players, on the other, it may also confront SMEs with new obstacles and threats. The present paper provides empirical evidence on these issues by analysing the behaviour of 277 SMEs located in two of the largest Italian industrial districts. The empirical findings (i) show the existence of differentiated behavioural patterns in approaching ICTs and e-business applications; (ii) suggest that, at the firm level, the attitude towards ICTs and e-business reflects the company strategy in facing global competition.
BlastAlign: a program that usesblastto align problematic nucleotide sequences
Authors: Belshaw Robert , Katzourakis Aris 
Published: August 2004

AbstractSummary: BlastAlign uses NCBI blastn to build a multiple nucleotide alignment and is intended for use with sequences that have large indels or are otherwise difficult to align globally. The program builds a matrix representing regions of homology along the sequences, from which it selects the ‘most representative’ sequence and then extracts the blastn query-anchored multiple alignment for this sequence. The matrix is printed and allows subgroups to be identified visually and an option allows other sequences to be used as the ‘most representative’. The program contains elements of both Perl and Python and will run on UNIX (including Mac OSX) and DOS. An additional Perl program BlastAlignP uses tblastn to align nucleotide sequences to a single amino acid sequence, thus allowing an open reading frame to be maintained in the resulting multiple alignment.Availability: It is freely available at http://www.bio.ic.ac.uk/research/belshaw/BlastAlign.tar and at http://evolve.zoo.ox.ac.uk/software/blastalignContact: r.belshaw@imperial.ac.uk
Economic and Legal Issues in Reducing the Eurosystem's Excess of International Reserves
Authors: Badinger Harald , Dutzler Barbara 
Published: August 2004

AbstractEconomic studies suggest that the Eurosystem's international reserves ($370 billion) could be reduced by up to half of its existing level. The article discusses the likely size and distribution of excess reserves and proposals for their uses. Small economic gains can be expected from a reserve reduction, as well as an elimination of incompatibilities and conflicts of interest between the conduct of monetary and investment policy. A careful and co‐ordinated reserve reduction would pose no threat to financial stability, making it also admissible from a legal perspective against the background of Art. 31 of the ESCB (European System of Central Banks) Statute. Finally, transferring reserves as an extraordinary gain to the government does not constitute monetary financing as prohibited by Art. 101 EC Treaty.
The Logic of Access to the European Parliament: Business Lobbying in the Committee on Economic and Monetary Affairs
Authors: Bouwen Pieter 
Published: August 2004

AbstractThis article is an attempt to test empirically a theory of access that investigates the logic behind the lobbying behaviour of business interests in the European Parliament. The theoretical framework tries to explain the degree of access of different organizational forms of business interest representation (companies, associations and consultants) to the supranational assembly in terms of a theory of the supply and demand of ‘access goods’. On the basis of 14 exploratory and 27 semi‐structured interviews, the hypotheses are checked in the Committee on Economic and Monetary Affairs (ECON) of the European Parliament. Surprisingly, European and national associations enjoy a similar degree of access to the Parliament. Individual companies and consultants have a much lower degree of access than the two collective forms of interest representation. In the conclusion, these results are analysed in the light of the existing literature on party cohesion and coalition formation in the European Parliament.