bluuebunny/crossref_metadata_2025 huggingface dataset:
The ACh‐induced contraction in rat aortas is mediated by the Cys Lt1 receptor via intracellular calcium mobilization in smooth muscle cells
Authors: Mazzetti Luca , Franchi‐Micheli Sergio , Nistri Silvia , Quattrone Silvia , Simone Ruggero , Ciuffi Mario , Zilletti Lucilla , Failli Paola
Published: February 2003

Our previously published data indicate that an endogenously produced 5‐lipoxygenase metabolite can strongly contract isolated endothelium‐preserved rat aortic strips when cyclo‐oxygenase isoenzymes are inhibited. Therefore, we decided to investigate if cysteinyl‐containing leukotrienes (Cys Lts) are involved in this endothelium‐dependent contraction.

The isometric contraction of endothelium‐preserved rat aortic strips was recorded in preparations preincubated with 5 μM indomethacin and precontracted with phenylephrine, adjusting resting tension at 0.7 g. Acetylcholine (ACh) contracted control strips. Montelukast and MK‐571, selective type 1 Cys Lts receptor (Cys Lt1) antagonists and the Cys Lt1/Cys Lt2 (type 2 Cys Lts receptor) antagonist BAYu9773 dose‐dependently prevented ACh‐induced contraction, their IC50s being 2.2, 3.1 and 7.9 nM respectively. The leukotriene B4 receptor antagonist U75302 was far less potent (IC50 1.5 μM).

In rat aorta smooth muscle cells (RASMs), Western blot analysis showed the presence of Cys Lt1 and Cys Lt2 receptors, the Cys Lt1 receptor being predominantly expressed.

In fura‐2 loaded RASMs, LTD4 (0.01–100 nM) and LTC4 (200–800 nM) dose‐dependently increased intracellular calcium concentration ([Ca2+]i). Montelukast (1–100 nM) reduced LTD4‐induced [Ca2+]i increase, its IC50 being approximately 10 nM. BAY u9773 exhibited significantly low effectiveness.

LTD4 (10 nM) induced a redistribution of smooth muscle actin fibres throughout the cytoplasm as visualized by confocal microscopy.

In conclusion, Cys Lt1 activation by endogenously produced Cys Lts, can contract rat aortas, while Cys Lt2 only marginally influences aortic tone. Intracellularly, this effect is mediated by an increase in [Ca2+]i. Therefore, Cys Lts, by inducing vascular contraction, can contribute to systemic hypertension.

British Journal of Pharmacology (2003) 138, 707–715. doi:10.1038/sj.bjp.0705087
Comparison of human recombinant adenosine A2B receptor function assessed by Fluo‐3‐AM fluorometry and microphysiometry
Authors: Patel H , Porter R H P , Palmer A M , Croucher M J
Published: February 2003

The aim of this study was to establish the utility of a fluorometric imaging plate reader (FLIPR) assay to assess human adenosine A2B receptor function by characterizing its receptor pharmacology and comparing this profile to that obtained using a microphysiometer.

FLIPR was used, in conjunction with a Ca2+‐sensitive dye (Fluo‐3‐AM), to measure rapid rises in intracellular calcium in a Chinese Hamster Ovary (CHO‐K1) cell line stably transfected with both the human A2B receptor and a promiscuous Gα16 protein. Microphysiometry was used to measure rapid changes in the rate of extracellular acidification in a Human Embryonic Kidney (HEK‐293) cell line also stably transfected with human A2B receptor.

Activation of A2B receptors by various ligands caused a concentration‐dependent increase in both the intracellular calcium concentration and the extracellular acidification rate in the cells tested, with a similar rank order of potency for agonists: NECA>N6‐Benzyl NECA>adenosineR‐PIA>CPA>S‐PIA>CHA>CGS 21680. No comparable effects were observed in the non‐transfected control cell lines.

The rank order of potency of the agonists examined was the same in all studies, whereas absolute potency and efficacy varied. Thus, all compounds exhibited greater potency in FLIPR than the microphysiometer and the efficacies obtained with CHO‐K1+Gα16+A2B cell line and FLIPR were greater than those obtained with HEK‐293+A2B cell line in the microphysiometer.

ZM‐241385 was the most potent of a range of adenosine antagonists tested with a pA2 of 8.0 in both the FLIPR and microphysiometer assays.

In conclusion, the profile of the responses to both A2B receptor agonists and antagonists in FLIPR were similar to those obtained by the microphysiometer, although both potency and efficacy values were higher in the FLIPR assay. With this caveat in mind, this study shows that FLIPR coupled with a cell line transfected with both the human A2B receptor and a promiscuous Gα16 protein provides a useful, high throughput method for the assessment of A2B receptor function.

British Journal of Pharmacology (2003) 138, 671–677. doi:10.1038/sj.bjp.0705091
Expression of prostanoid receptors in human ductus arteriosus
Authors: Leonhardt Andreas , Glaser Alexander , Wegmann Markus , Schranz Dietmar , Seyberth Hannsjörg , Nüsing Rolf
Published: February 2003

Prostaglandins play a major role in maintaining ductal patency in utero. Ductal tone is regulated by both locally released and circulating vasodilatory prostaglandins. In infants with ductus arteriosus‐dependent congenital heart disease, ductal patency is maintained by intravenous administration of prostaglandin (PG) E1. Little information is available regarding the expression of prostaglandin receptors in man.

By means of RT–PCR and immunohistochemistry we studied the expression of the PGI2 receptor (IP), the four different PGE2 receptors (EP1, EP2, EP3 and EP4), and the receptors for thromboxane (Tx) A2 (TP), PGD2 (DP) and PGF2α (FP) in the ductus arteriosus of three newborn infants with ductus arteriosus‐dependent congenital heart disease and intravenous infusion of PGE1 and of one 8 month old child with a patent ductus arteriosus.

The EP3, EP4, FP, IP and TP receptor were markedly expressed at the mRNA and protein level, whereas the EP2 receptor was weakly expressed and the EP1 receptor was detected in two out of four tissue specimens only. The DP receptor was not detected in any of the samples. The most pronounced expression, which was located in the media of the ductus arteriosus, was observed for the EP4 and TP receptors followed by IP and FP receptor protein.

These data indicate that ductal patency during the infusion of PGE1 in infants with ductus arteriosus‐dependent congenital heart disease might be mediated by the EP4 and IP receptor. The data further suggest that a heterogeneous population of prostanoid receptors may contribute to the regulation of ductus arteriosus tone in humans.

British Journal of Pharmacology (2003) 138, 655–659. doi:10.1038/sj.bjp.0705092
Activation of c‐fos expression in the heart after morphine but not U‐50,488H withdrawal
Authors: González‐Cuello Ana , Milanés M Victoria , Castells M Teresa , Laorden M Luisa
Published: February 2003

In the present work we have studied in the heart the expression of Fos, the protein product of the c‐fos proto‐oncogene and the adaptive changes in noradrenergic neurons after naloxone or nor‐binaltorphimine (nor‐BNI) administration to morphine or U‐50,488H pretreated rats.

Male rats were implanted with placebo (naïve) or morphine (tolerant/dependent) pellets for 7 days. On day 8 rats received saline s.c., naloxone (5 mg kg−1 s.c.) or nor‐BNI (5 mg kg−1 i.p.). Other groups of rats were rendered tolerant/dependent on U‐50,488H by injecting the drug twice daily (15 mg kg−1 i.p.) for 4 days. Control animals received saline. On day 5 the animals were injected with vehicle i.p. or nor‐BNI (5 mg kg−1 i.p.).

Using immunohistochemical staining of Fos, present results indicate that morphine withdrawal induced marked Fos immunoreactivity (Fos‐IR) within the cardiomyocyte nuclei. Moreover, Western blots analysis revealed a peak expression of c‐fos in right and left ventricle after naloxone induced withdrawal in parallel with an increase in noradrenaline (NA) turnover.

However, after nor‐BNI administration to rats chronically treated with U‐50,488H, we found a decrease in the NA turnover. In addition, the administration of nor‐BNI to rats chronically treated with U‐50,488H or morphine did not induce modifications in the Fos‐IR, in the heart.

These results demonstrated that morphine withdrawal induces the expression of Fos protein, as well as an enhancement of noradrenergic activity in the heart. In contrast to morphine U‐50,488 withdrawal produces no changes in Fos‐IR in parallel with a decrease in NA turnover, indicating that the kappa–opioid receptors are not involved in the molecular adaptive mechanisms responsible for the development of opioid dependence in the heart.

British Journal of Pharmacology (2003) 138, 626–633. doi:10.1038/sj.bjp.0705093
CC chemokines induce P‐selectin‐dependent neutrophil rolling and recruitment in vivo: intermediary role of mast cells
Authors: Wan Ming Xiu , Wang Yusheng , Liu Qing , Schramm Rene , Thorlacius Henrik
Published: February 2003

Based on in vitro chemotaxis experiments, it is widely held that CC chemokines, such as macrophage inflammatory protein‐1α (MIP‐1α) and macrophage chemotactic protein‐1 (MCP‐1) mainly support lymphocyte trafficking.

The objective of the present study was to examine the role of MIP‐1α and MCP‐1 in neutrophil recruitment in vivo by use of intravital microscopy of the mouse cremaster microcirculation.

MIP‐1α and MCP‐1 caused a dose‐dependent increase in leukocyte rolling, adhesion and recruitment. Indeed, neutrophils comprised more than 85% of the leukocyte response to MIP‐1α and MCP‐1. An anti‐P‐selectin antibody reduced MIP‐1α and MCP‐1‐provoked leukocyte rolling by more than 94%. Concomitantly, firm adhesion and extravasation of neutrophils in response to MIP‐1α and MCP‐1 challenge were significantly decreased by more than 78 and 84%, respectively. In contrast, an anti‐E‐selectin antibody had no influence on CC chemokine‐induced neutrophil recruitment.

Flow cytometric analysis revealed that MIP‐1α and MCP‐1 had no effect on P‐selectin expression on endothelial cells, suggesting that neutrophil recruitment elicited by CC chemokines in vivo is not mediated via a direct effect on the endothelium but rather via an indirect effect involving activation of an intermediary tissue cell. Indeed, it was found that MIP‐1α‐induced neutrophil accumulation was significantly decreased by 58% in mast cell‐deficient mice.

These findings demonstrate that CC chemokines trigger P‐selectin‐dependent rolling and tissue recruitment of neutrophils via tissue mast cells in vivo and suggest that CC chemokines may also be important targets in neutrophil‐mediated tissue damage in multicellular organs.

British Journal of Pharmacology (2003) 138, 698–706. doi:10.1038/sj.bjp.0705094
Thrombin‐induced conversion of fibrinogen to fibrin results in rapid platelet trapping which is not dependent on platelet activation or GPIb
Authors: Jarvis Gavin E , Atkinson Ben T , Frampton Jon , Watson Steve P
Published: February 2003

Activation of human platelets by thrombin is mediated by the proteolytic cleavage of two G‐protein coupled protease‐activated receptors, PAR‐1 and PAR‐4. However, thrombin also binds specifically to the platelet surface glycoprotein GPIb. It has been claimed that thrombin can induce aggregation of platelets via a novel GPIb‐mediated pathway, which is independent of PAR activation and fibrinogen binding to αIIbβ3 integrin, but dependent upon polymerizing fibrin and the generation of intracellular signals.

In the presence of both fibrinogen and the αIIbβ3 receptor antagonist lotrafiban, thrombin induced a biphasic platelet aggregation response. The initial primary response was small but consistent and associated with the release of platelet granules. The delayed secondary response was more substantial and was abolished by the fibrin polymerization blocking peptide GPRP.

Cleavage of the extracellular portion of GPIb by mocarhagin partially inhibited thrombin‐induced αIIbβ3‐dependent aggregation and release, but had no effect on the secondary fibrin‐dependent response.

Fixing of the platelets abolished αIIbβ3‐dependent aggregation and release of adenine nucleotides, whereas the fibrin‐dependent response remained, indicating that platelet activation and intracellular signalling are not necessary for this secondary ‘aggregation’.

In conclusion, the secondary fibrin‐dependent ‘aggregation’ response observed in the presence of fibrinogen and lotrafiban is a platelet trapping phenomenon dependent primarily on the conversion of soluble fibrinogen to polymerizing fibrin by thrombin.

British Journal of Pharmacology (2003) 138, 574–583. doi:10.1038/sj.bjp.0705095 doi:0705095a/sj.bjp.0704832
A possible mechanism for improvement by a cognition‐enhancer nefiracetam of spatial memory function and cAMP‐mediated signal transduction system in sustained cerebral ischaemia in rats
Authors: Takeo Satoshi , Niimura Makiko , Miyake‐Takagi Keiko , Nagakura Akira , Fukatsu Tomoko , Ando Tsuyoshi , Takagi Norio , Tanonaka Kouichi , Hara Junko
Published: February 2003

Accumulated evidence indicates that the adenylyl cyclase (AC)/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP‐responsive element binding protein (CREB) signal transduction system may be linked to learning and memory function.

The effects of nefiracetam, which has been developed as a cognition enhancer, on spatial memory function and the AC/cAMP/PKA/CREB signal transduction system in rats with sustained cerebral ischaemia were examined.

Microsphere embolism (ME)‐induced sustained cerebral ischaemia was produced by injection of 700 microspheres (48 μm in diameter) into the right hemisphere of rats. Daily oral administration of nefiracetam (10 mg kg−1 day−1) was started from 15 h after the operation.

The delayed treatment with nefiracetam attenuated the ME‐induced prolongation of the escape latency in the water maze task that was examined on day 7 to 9 after ME, but it did not reduce the infarct size.

ME decreased Ca2+/calmodulin (CaM)‐stimulated AC (AC‐I) activity, cAMP content, cytosolic PKA Cβ level, nuclear PKA Cα and Cβ levels, and reduced the phosphorylation and DNA‐binding activity of CREB in the nucleus in the right parietal cortex and hippocampus on day 3 after ME. The ME‐induced changes in these variables did not occur by the delayed treatment with nefiracetam.

These results suggest that nefiracetam preserved cognitive function, or prevented cognitive dysfunction, after sustained cerebral ischaemia and that the effect is, in part, attributable to the prevention of the ischaemia‐induced impairment of the AC/cAMP/PKA/CREB signal transduction pathway.

British Journal of Pharmacology (2003) 138, 642–654. doi:10.1038/sj.bjp.0705096
Group III metabotropic glutamate receptors as autoreceptors in the cerebellar cortex
Authors: Lorez Matthias , Humbel Urs , Pflimlin Marie‐Claire , Kew James N C
Published: February 2003

Group III metabotropic glutamate receptors (mGluRs) of the subtype 4a are localized within presynaptic active zones of cerebellar parallel fibre (PF)‐Purkinje cell (PC) synapses. In order to investigate the conditions necessary for group III mGluR autoreceptor‐activation by synaptically released glutamate, we characterized the effects of selective agonists and antagonists on excitatory postsynaptic currents (EPSCs) evoked by several distinct PF stimulation patterns.

The group III mGluR‐selective agonist L‐AP4 depressed evoked EPSCs at PF‐PC synapses in rat brain slices with an EC50 of 2.4 μM and maximum inhibition of 80%. This L‐AP4‐induced depression was antagonized by the group III mGluR‐selective antagonist MSOP with an estimated equilibrium dissaciation constant of 12.5 μM.

Paired‐pulse or four‐pulse PF stimulations did not activate presynaptic group III mGluRs as revealed by the lack of effect of 1 mM MSOP on relative test EPSC amplitudes with latencies of 250–500 ms. The potentiation of a test EPSC evoked 200–500 ms after a short tetanic burst (100 Hz for 60 ms), was also unchanged in the presence of MSOP.

Endogenous autoreceptor‐activation was revealed only during prolonged stimulation trains (10 Hz for 4.4 s), where, in the presence of 1 mM MSOP, the EPSC amplitudes were enhanced by 15%.

These observations support an autoreceptor function of group III mGluRs and a role in short‐term synaptic plasticity at PF synapses. However, the low to moderate activation levels observed, despite the close spatial relation with glutamate release sites, suggests that additional mechanisms regulate receptor activation.

British Journal of Pharmacology (2003) 138, 614–625. doi:10.1038/sj.bjp.0705099
The CB1 receptor antagonist SR141716A selectively increases monoaminergic neurotransmission in the medial prefrontal cortex: implications for therapeutic actions
Authors: Tzavara Eleni T , Davis Richard J , Perry Kenneth W , Li Xia , Salhoff Craig , Bymaster Frank P , Witkin Jeffrey M , Nomikos George G
Published: February 2003

In order to explore potential therapeutic implications of cannabinoid antagonists, the effects of the prototypical cannabinoid antagonist SR141716A on monoamine efflux from the medial prefrontal cortex and the nucleus accumbens of the rat were investigated by in vivo microdialysis.

SR141716A moderately increased serotonin efflux and concentrations of its metabolite 5‐HIAA, both in the medial prefrontal cortex and the nucleus accumbens, and increased norepinephrine, dopamine and their metabolites in the medial prefrontal cortex. In contrast, it had no effect on norepinephrine, dopamine and their metabolites in the nucleus accumbens.

At the same doses, SR141716A increased acetylcholine efflux in the medial prefrontal cortex, in agreement with previous studies; contrary to the effects in cortex, SR141716A had no effect on acetylcholine efflux in the nucleus accumbens.

The efficacy of SR141716A in the psychostimulant‐induced hyperlocomotion and the forced swimming paradigms was also explored in mice. SR141716A attenuated phenylcyclidine‐ and d‐amphetamine‐induced hyperlocomotion, without affecting locomotor activity when administered alone, and decreased immobility in the forced swimming test.

These results suggest that the cortical selectivity in the release of catecholamines, dopamine in particular, induced by the cannabinoid antagonist SR141716A, its procholinergic properties, together with its mild stimulatory effects on serotonin and norepinephrine efflux make similar compounds unique candidates for the treatment of psychosis, affective and cognitive disorders.

British Journal of Pharmacology (2003) 138, 544–553. doi:10.1038/sj.bjp.0705100
G protein‐coupled receptor kinase 6 (GRK6) selectively regulates endogenous secretin receptor responsiveness in NG108‐15 cells
Authors: Ghadessy Roxana S , Willets Jonathon M , Kelly Eamonn
Published: February 2003

To determine the role of G protein‐coupled receptor kinases (GRKs) in the regulation of endogenous secretin receptor responsiveness, we have transiently overexpressed both wild‐type (WT) and dominant negative mutant (DNM) GRKs in NG108‐15 mouse neuroblastoma×rat glioma hybrid cells and investigated the effects of this on agonist‐stimulated adenylyl cyclase activity.

Overexpression of WT GRK6 selectively inhibited secretin‐stimulated cyclic AMP accumulation (fold stimulation of cyclic AMP above basal following 15 min incubation with 100 nMsecretin was 12.1±2.0 and 6.2± 0.8 in control and WT GRK overexpressing cells, respectively) without affecting cyclic AMP responses mediated by the adenosine A2receptor agonist 5′‐(N‐ethylcarboxamido) adenosine (NECA) or the prostanoid‐IP receptor agonist iloprost, or the direct activator of adenylyl cyclase, forskolin. On the other hand DNM GRK6 (Lys215Arg) overexpression produced the opposite effect – a selective increase in the secretin‐stimulated cyclic AMP response was observed in cells overexpressing DNM GRK6 compared to plasmid‐transfected cells (fold stimulation of cyclic AMP above basal following 15 min incubation with 100 nMsecretin was 12.6±2.7 and 29.6±5.6 for control and DNM GRK6‐overexpressing cells, respectively).

Overexpression of WT GRK5 likewise inhibited the secretin‐stimulated cyclic AMP response, however, this effect was not as selective as with GRK6, since adenosine A2receptor responsiveness was also suppressed by GRK5 overexpression. Unlike DNM GRK6, overexpression of DNM GRK5 failed to modulate secretin or A2adenosine receptor signalling suggesting that endogenous GRK5 is unlikely to regulate desensitization of these receptors in NG108‐15 cells.

Overexpression of WT GRK2 did not affect secretin‐stimulated cyclic AMP accumulation. Instead, GRK2 overexpression selectively inhibited A2adenosine receptor responsiveness, confirming our previous findings.

Together these results suggest a selective role of endogenous GRK6 in regulating secretin receptor responsiveness in NG108‐15 cells. In addition, these data indicate that GRKs exert a surprising degree of selectivity in the regulation of natively expressed GPCR responses.

British Journal of Pharmacology(2003)138, 660–670. doi:10.1038/sj.bjp.0705101
Spinal amino acid release and repeated withdrawal in spinal morphine tolerant rats
Authors: Ibuki Takae , Marsala Martin , Masuyama Takashi , Yaksh Tony L
Published: February 2003

We used spinal microdialysis in awake rats to investigate whether the repeated withdrawal with naloxone during continuous spinal infusion of morphine would lead to a progressively greater spinal glutamate release and a more pronounced intrathecal tolerance.

Rats received lumbar intrathecal (IT) infusion of morphine (IT‐M: 20 nmol μl−1 h−1) or saline (IT‐S: 1 μl h−1) continuously for 3 days. Both groups were further subdivided to receive intraperitoneal (i.p.) injection of naloxone (IP‐N: 0.6 mg kg−1) or saline (IP‐S: 3 ml kg−1) every 24 h after the beginning of IT infusion. Daily thermal escape latencies, withdrawal signs, the resting basal release of spinal amino acids before IP injection and the release immediately after the injection (evoked) were measured.

Rats receiving IT morphine showed a maximum increase in thermal escape latency on day 1, after which this value declined, with the fastest decline observed in IT morphine+IP naloxone group. On day 1, no significant difference was observed among groups in the resting basal release of amino acids. Rats in IT morphine+i.p. naloxone group displayed a progressive increase in this value. The release was not significantly altered in other groups.

For the IT‐M+IP‐N group, basal resting dialysate concentrations of Glu, Asp and Tau rose steadily over the 3‐day infusion interval. No change in basal resting release was noted for any other treatment.

Evoked release (after i.p. naloxone) in IT‐M animals displayed a progressive increase over the three repeated exposures. Evoked release did not change significantly in other treatment groups.

The degree of precipitated withdrawal significantly correlated with the increase in glutamate acutely evoked by i.p. injection.

The present results show that periodic transient withdrawal of spinal opiate agonist activity leads to a progressive increase in glutamate outflow and withdrawal signs, in a manner consistent with an enhanced development of spinal tolerance.

British Journal of Pharmacology (2003) 138, 689–697. doi:10.1038/sj.bjp.0705102
Renal targeting of captopril selectively enhances the intrarenal over the systemic effects of ACE inhibition in rats
Authors: Haverdings R G F , Haas M , Navis G J , Van Loenen‐Weemaes A M A , Meijer D K F , De Zeeuw D , Moolenaar F
Published: February 2003

British Journal of Pharmacology (2003) 138, 531. doi:10.1038/sj.bjp.0705183
P2Ys go neuronal: modulation of Ca2+ and K+ channels by recombinant receptors
Authors: Boehm Stefan
Published: February 2003

British Journal of Pharmacology (2003) 138, 716. doi:10.1038/sj.bjp.0705201
Growth‐rate dependent RNA polyadenylation in Escherichia coli
Authors: Jasiecki Jacek , Wçgrzyn Grzegorz
Published: February 2003

RNA polyadenylation occurs not only in eukaryotes but also in bacteria. In prokaryotes, polyadenylated RNA molecules are usually degraded more efficiently than non‐modified transcripts. Here we demonstrate that two transcripts, which were shown previously to be substrates for poly(A) polymerase I (PAP I), Escherichia coli lpp messenger RNA and bacteriophage λ oop RNA, are polyadenylated more efficiently in slowly growing bacteria than in rapidly growing bacteria. Intracellular levels of PAP I varied in inverse proportion to bacterial growth rate. Moreover, transcription from a promoter for the pcnB gene (encoding PAP I) was shown to be more efficient under conditions of low bacterial growth rates. We conclude that efficiency of RNA polyadenylation in E. coli is higher in slowly growing bacteria because of more efficient expression of the pcnB gene. This may allow regulation of the stability of certain transcripts (those subjected to PAP I‐dependent polyadenylation) in response to various growth conditions.
A novel, inducible, eukaryotic gene expression system based on the quorum‐sensing transcription factor TraR
Authors: Neddermann Petra , Gargioli Cesare , Muraglia Ester , Sambucini Sonia , Bonelli Fabio , De Francesco Raffaele , Cortese Riccardo
Published: February 2003

Bacteria adapt their pattern of gene expression in response to a variety of external cues, including fluctuations in population density. This type of bacterial cell‐to‐cell communication is referred to as quorum‐sensing. Quorum‐sensing systems are present in many bacterial species and constitute a large collection of ligands and cognate receptors. The availability of such diversity offers interesting opportunities for biotechnological exploitation. We describe here the transformation of the quorum‐sensing system of Agrobacterium tumefaciens into a transcription regulatory system that works in mammalian cells. The A. tumefaciens TraR protein was fused to the eukaryotic activation domain of NF‐κB p65, generating a novel chimaeric transcriptional activator that stimulates gene transcription in different human cell lines from a minimal promoter containing the TraR DNA recognition sequence in the presence of the Agrobacterium quorum‐sensing signal molecule N‐(3‐oxo‐octanoyl)homoserine lactone (3‐oxo‐C8‐HSL). The basal level of transcription was low in the absence of 3‐oxo‐C8‐HSL, and gene expression was stimulated up to 1,000‐fold at a saturating concentration of 3‐oxo‐C8‐HSL.
Branching Out: A Molecular Fingerprint of Endothelial Differentiation into Tube‐Like Structures Generated by Affymetrix Oligonucleotide Arrays
Authors: GERRITSEN MARY E. , SORIANO ROBERT , YANG SUYA , ZLOT CONSTANCE , INGLE GLADYS , TOY KAREN , WILLIAMS P. MICKEY
Published: February 2003

The process of endothelial differentiation into a network of tube‐like structures with patent lumens requires an integrated program of gene expression. To identify genes upregulated in endothelial cells during the process of tube formation, RNA was prepared from several different time points (0, 4, 8, 24, 40, and 48 hours) and from three different experimental models of human endothelial tube formation: in collagen gels and fibrin gels driven by the combination of PMA (80), bFGF (40 ng/ml) and bFGF (40 ng/ml) or in collagen gels driven by the combination of HGF (40 ng/ml) and VEGF (40 ng/ml). Gene expression was evaluated using Affymetrix® Gene Chip® oligonucleotide arrays. Over 1000 common genes were upregulated greater than twofold over baseline at one or more time points in the three different models. In the present study, we discuss the identified genes that could be assigned to major functional classes: apoptosis, cytoskeleton, proteases, matrix, and matrix turnover, pumps and transporters, membrane lipid turnover, and junctional molecules or adhesion proteins.
The effect of infant colic on maternal self‐perceptions and mother‐‐infant attachment
Authors: Stifter C A , Bono M A
Published: February 2003

Mothers of infants who developed colic were compared with mothers of non‐colic infants on their perceptions of parenting self‐efficacy, and separation anxiety as well as their attachment relationship with their infants. Colic was identified prospectively through telephone contacts with mothers. Questionnaires on self‐efficacy and separation anxiety were completed when infants were 5 months of age. At 18 months of age mothers and infants (colic and non‐colic) participated in a laboratory situation to measure attachment. Results revealed that mothers of colic infants reported feeling less competent as mothers. In addition, while mothers of colicky infants tended to have more separation anxiety than mothers of non‐colic infants, they felt that these separations did not have a negative effect on their child. Finally, no differences were revealed for attachment classifications between colic and non‐colic infants at 18 months.
Links between maternal care and persistent infant crying in the early months
Authors: St James‐Roberts I , Conroy S , Wilsher K
Published: February 2003

A community sample was screened to select three groups of infants and their mothers according to how much the babies cried at 6 weeks of age, the peak age for infant crying. The three groups — of moderate (n = 55), evening (n = 38) and persistent criers (n = 67) and their mothers— were assessed by diary, observation and questionnaire measures of mother and infant characteristics and interactions at 6 weeks and 5 months of infant age. At 6 weeks, mothers of persistent criers spent more time interacting with and physically stimulating their babies. Below‐optimum maternal sensitivity/affection was linked to moderately increased crying in the infants overall. However, most mothers of persistent criers showed optimum sensitivity and affection, while no significant links between maternal sensitivity/affection and infant crying were found in the persistent crying group. By 5 months, when infant crying declined, the range and size of differences between mothers of persistent criers and other mothers declined. Home observations and a standard play measure failed to show group differences in maternal sensitivity, affection and intrusiveness at this age. The findings show that persistent infant crying in the early months often occurs in spite of high quality maternal care, so that in most cases the crying is probably not due to inadequate parenting. The need to distinguish general community cases from those at social or medical risk is emphasized and the findings’ implications for professionals are discussed.
Relationships between infant crying, birth complications, and maternal variables
Authors: Keller H , Lohaus A , VÎlker S , Cappenberg M , Chasiotis A
Published: February 2003

The aim of the study is to identify factors influencing infants’ crying behaviour (total crying duration, length of crying bouts, and frequency of crying). The searching for variables explaining inter‐individual differences requires a sufficient stability of the cry parameters at least over short time intervals. Thus, a second aim is to assess the intra‐individual stability of the crying behaviour in an age range from 3 to 12 months. Sixty‐two mother–child dyads participated in the study. The results show substantial stabilities of the crying behaviour of infants between 3 and 12 months of age. This is related to the amount of crying as well as to the pattern of crying shown by the children over a 24 h interval. The typical cry pattern is characterized by peaks at 12.00 h and early in the evening, although there are large individual differences between the children with regard to cry durations at each hour of the day. Regarding the cry durations at 3 months of age, birth complications and the perceived emotional atmosphere in the mothers’ family of origin are the major contributing factors.
Basal ganglia metabolite concentrations in idiopathic Parkinson's disease and multiple system atrophy measured by proton magnetic resonance spectroscopy
Authors: Clarke C. E. , Lowry M.
Published: February 2003

Six patients with idiopathic Parkinson's disease (IPD), six with clinically probable multiple system atrophy and six control subjects underwent quantitative proton magnetic resonance spectroscopy (MRS). The concentrations of the three major metabolites, N‐acetylaspartate (NAA), creatine and choline, were quantified in the lentiform nucleus using tissue water content as an internal concentration reference. Glutamate was assessed as the (glutamate + glutamine)/creatine peak area ratio (Glx/Cre). In the control subjects the mean (± SD) concentrations of the three metabolites were 15.2 ± 2.9 µmol/g wet weight for NAA, 12.0 ± 1.4 for creatine and 2.4 ± 0.3 for choline. The Glx/Cre ratio was 1.28 ± 0.32. The only significant difference in any metabolite concentration was in the lentiform nucleus of patients with IPD compared with controls, with an increase in choline which lead to a significant reduction in the NAA/choline ratio. The relevance of this finding is uncertain. The results of the present pilot study, combined with the conflicting findings from previous work, suggest that further, much larger, studies are required to evaluate the diagnostic capability of proton MRS.
Effects of the non‐selective phosphodiesterase inhibitor pentoxifylline on regional cerebral blood flow and large arteries in healthy subjects
Authors: Kruuse C. , Jacobsen T. B. , Thomsen L. L. , Hasselbalch S. G. , Frandsen E. K. , Dige‐Petersen H. , Olesen J.
Published: February 2003

The vasodilating properties of the non‐selective phosphodiesterase (PDE) inhibitor pentoxifylline were evaluated. Pentoxifylline has been reported to increase cerebral blood flow (CBF) and improve recovery rate of stroke patients. Whether these results are due to a dilating effect on arteries or to other mechanisms is not clear. In the present double‐blind crossover study, 10 healthy subjects received pentoxifylline 300 mg or placebo intravenously on separate days. Blood flow velocity in the middle cerebral artery (Vmca) was recorded by transcranial Doppler and rCBF was measured using 133Xenon‐inhalation SPECT. High‐frequency ultrasound was used for measurements of temporal and radial artery diameter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) concentrations were assessed in plasma. Except for increased heart rate (P < 0.05), systolic blood pressure (P < 0.05) and plasma cAMP (P < 0.001), no significant differences in CBF, rCBFmca or plasma cGMP were seen between placebo and pentoxifylline infusion. During pentoxifylline infusion, Vmca decreased 7.2% (SD 12.0; P < 0.05) and temporal artery diameter increased 9.0% (SD 7.0; P < 0.001), suggesting minor dilatation of the large arteries. However, this change was not significantly different from placebo. In conclusion, pentoxifylline 300 mg had no effect on rCBF. A possible minor dilatation of the middle cerebral artery and the temporal artery cannot be excluded. Any potential clinical effect of pentoxifylline is most likely mediated through non‐vascular mechanisms.
Identification of Gal(β1–3)GalNAc bearing glycoproteins in cerebrospinal fluid of amyotrophic lateral sclerosis (ALS) patients
Authors: Niebroj‐Dobosz I. , Mickielewicz A. , Rowiñska‐Marciñska K. , Kwieciñski H.
Published: February 2003

Glycoproteins in cerebrospinal fluid of 55 patients with amyotrophic lateral sclerosis (ALS), six disease controls (multifocal motor neuropathy, sensorimotor neuropathy, Guillain–Barré syndrome, spinal muscular atrophy type II, motor neuropathy with monoclonal gammopathy) and 20 healthy controls were separated by PAGE electrophoresis and then detected immunochemically with peanut agglutinin (PNA). In 36 amyotrophic lateral sclerosis patients the 262 kDa glycoprotein was significantly increased (over the normal mean ± SD × 2), which was associated with a decrease in the 114 kDa fraction. In the remaining patients, both fractions were either equal in concentration or the 114 kDa glycoprotein predominated. In normal cerebrospinal fluid, the 114 kDa glycoprotein predominated over the other glycoproteins. The total amount of separated glycoproteins was increased in 15 amyotrophic lateral sclerosis patients. In 12 of them it was followed by an increase in the percentage of the 262 kDa glycoprotein. There was no correlation between the content of the peanut agglutinin‐labelled glycoproteins and the patients' age, duration and severity of the disease. There was a correlation between the 262 kDa glycoprotein being increased in cerebrospinal fluid and the electrophysiological pattern of denervation seen in electromyographic study. The glycoproteins change, similar to that occurring in amyotrophic lateral sclerosis patients, was also observed in one case of multifocal motor neuropathy (MMN). We suggest that in amyotrophic lateral sclerosis and multifocal motor neuropathy, the peanut agglutinin‐labelled glycoproteins are released in excess from the nervous tissues into the cerebrospinal fluid as a result of neuronal degeneration. The question to be answered is, whether the released glycoproteins are becoming targets for auto‐antibodies.
Eosinophilic pleocytosis and myelitis related to Toxocara canis infection
Authors: Goffette S. , Jeanjean A. P. , Duprez T. P. J. , Bigaignon G. , Sindic C. J. M.
Published: February 2003

Toxocara canis causes the visceral larva migrans syndrome in which central nervous involvement is rare. We report the case of a 40‐year‐old woman presenting with a subacute weakness of the right leg and dysaesthesiae in the right Th8–Th10 dermatomas. Spinal magnetic resonance imaging examination showed abnormal hyperintensity within the spinal cord. Cerebrospinal fluid analysis revealed eosinophilic pleocytosis. Antibody titres to Toxocara canis were higher in the cerebrospinal fluid than in the serum. Treatment using mebendazole led to a complete clinical recovery, normalization of cerebrospinal fluid parameters and improvement in spinal magnetic resonance imaging abnormalities.
Heart rate variability in patients with untreated Parkinson’s disease
Authors: Kallio M. , Haapaniemi T. , Turkka J. , Suominen K. , Tolonen U. , Sotaniemi K. , Heikkilä V.‐P. , Myllylä V.
Published: February 2003

The aim of this study was to evaluate cardiovascular responses as a marker of autonomic nervous system (ANS) disturbances in patients with untreated Parkinson’s disease (PD) and to assess the relationship between them and the clinical characteristics of PD. The ANS functions were investigated in 50 patients with PD and 55 healthy subjects by measuring standard cardiovascular autonomic reflexes and heart rate variability (HRV) at rest using spectral analysis (the autoregressive model and the fast Fourier transformation), the percentage of the counts of beat‐to‐beat variation greater than 50 ms and the fractal dimension. Significantly attenuated HRV and deficient blood pressure reaction to tilting were found in the PD patient group. The patients with hypokinesia/rigidity as the initial symptom of PD had a more pronounced HRV deficit than those with tremor onset. Untreated PD patients suffer significant failure in cardiovascular nervous system regulation, and in patients with hypokinesia/rigidity as their initial disease manifestation the risk of this ANS dysfunction is high. However, in the early stages of PD these changes did not reach significance at individual level.
Clinical epidemiology of Guillain–Barré syndrome in adults in Sweden 1996–97: a prospective study
Authors: Cheng Q. , Jiang G.‐X. , Press R. , Andersson M. , Ekstedt B. , Vrethem M. , Liedholm L. J. , Lindsten H. , Brattström L. , Fredrikson S. , Link H. , de Pedro‐Cuesta J.
Published: February 2003

We described clinical manifestations, outcomes, prognostic indicators and clinico‐epidemiological subgroups for 53 adult patients with Guillain–Barré syndrome (GBS) in Sweden during the period 1996–97. These patients were identified from a population of 2.8 million inhabitants and prospectively followed up for one year by a network of neurologists. An additional 10 cases, of whom five were adults who had not been prospectively followed up, were not included in the analyses. At 6 months after onset 80% of the patients could walk without aid, while at 1 year 46% were fully recovered, 42% had mild residual signs or symptoms, 4% had moderate and 6% severe disabilities, and 2% had died. Intravenous human immunoglobulin or plasmapheresis were used in 72% of the patients. The sum of the Medical Research Council (MRC) score at nadir was found as the only significant predictor for residual signs at 1 year in a multivariate model. Three subgroups, with different clinico‐epidemiological characteristics, were identified by using cluster analysis. In conclusion, GBS in Sweden is frequently preceded by a respiratory infection, is often treated with immunomodulatory therapies, and exhibits a high recovery rate and a low fatality rate.
The high frequency of coeliac disease among children with neurological disorders
Authors: Salur L. , Uibo O. , Talvik I. , Justus I. , Metsküla K. , Talvik T. , Uibo R.
Published: February 2003

In the present study we have explored antigliadin (AGA) and antireticulin (ARA) antibody tests for the serological screening for coeliac disease (CD) of 206 children with neurological disorders. IgA‐ or/and IgG‐type AGA was discovered in 17 (8.3%) patients and IgA‐type ARA in one (0.5%) patient. A small intestinal biopsy was performed in all 18 antibody‐positive patients, and villous atrophy compatible with CD was revealed in three cases (patients with either epilepsy, retardation of psychomotor development or Down's syndrome). The CD prevalence rate of 14.6 per 1000 (95% CI 7.0–22.2) found in the present study was higher than could have been anticipated on the basis of the results of our previous population studies, which indicate that CD occurs more frequently among children with neurological disorders (OR = 37.6; 95% CI 9.7–146.9). Whether this finding reflects certain immunopathogenic links between CD and particular neurological diseases needs to be studied further. In this study, we were unable, using indirect immunofluorescence testing, to demonstrate the presence of autoantibodies against brain tissue in CD and AGA‐positive patients.
A two‐fold difference in the age‐adjusted prevalences of Parkinson’s disease between the island of Als and the Faroe Islands
Authors: Wermuth L. , von Weitzel‐Mudersbach P. , Jeune B.
Published: February 2003

With the aim of comparing the previously found high prevalence of idiopathic Parkinson’s disease (PD) in the Faroe Islands with the prevalence of PD in an area of Denmark, we used the same case‐finding methods for case ascertainment and the same strict criteria to diagnose PD on the island of Als. During the last year before the prevalence date (1 January 1998), we found in various registries from pharmacies, hospital, private neurologist and general practioners 121 patients with suspected Parkinsonism out of 56 839 inhabitants on the island of Als. After exclusion of those who had other diseases, a total of 79 patients were left for further examinations. Among these we found 58 with PD. The overall prevalence of PD was estimated to be 102.0 and the age‐adjusted prevalence to be 98.3 per 100 000 persons compared with 187.6 and 209.0 in the Faroe Islands. Compared with the previous results from the Faroe Islands (prevalence date 1 July 1995) we found an even lower mean age at onset of PD symptoms and at onset of treatment, a lower proportion of definite PD and a lower average dose of levodopa. We therefore conclude that the two‐fold higher prevalence in the Faroe Islands than on the island of Als was not due to an early diagnosis and a higher ascertainment of cases with mild PD, which was suggested as being one possible explanation for our previous finding of a high prevalence of PD in the Faroe Islands.
Excitatory effect of thiamin on CA1 pyramidal neurones in rat hippocampal slices in vitro
Authors: Tallaksen C. M. E. , Taubøll E.
Published: February 2003

Clinically, acute thiamin deficiency may lead to Wernicke encephalopathy and fulminant cardial beriberi. Both diseases respond to high parenteral doses of thiamin. The cofactor role of intracellular thiamin diphosphate has been thoroughly investigated, but an additional acute effect of unphosphorylated extracellular thiamin has been postulated but not elucidated.

In order to investigate the role of thiamin at the membrane level in the central nervous system, a study using a well‐established in vitro rat hippocampal slice model was designed. Hippocampal slices were perfused with 0.5, 0.75 and 1 mM thiamin solutions for 30 min and the pre‐synaptic volley, field excitatory post‐synaptic potential and population spike amplitudes were recorded continuously. The results showed an acute, excitatory effect of high‐dose thiamin on hippocampal neurones by significantly increasing the number of repetitive afterdischarges. Additional experiments with low concentrations of the potassium channel blocker 4‐aminopyridine showed similar findings. The results support previous evidence of thiamin affecting membrane ion channel activity, probably involving potassium channels, although the precise mechanisms of action are still unclear.
Interactive videoconsultation is a feasible method for neurological in‐patient assessment
Authors: Craig J. , Patterson V. , Russell C. , Wootton R.
Published: February 2003

To evaluate the feasibility of interactive videoconsultation (IATV) as a means by which neurologists might assess patients admitted with neurological symptoms to hospitals distant from a neurological centre, we studied 25 unselected patients using interactive videoconsultation (IATV) and then validated the IATV diagnoses and management plans at a later face‐to‐face consultation. IATV consultation led to an eventual diagnosis in 23 out of 25 patients, with one diagnosis being changed and one remaining uncertain. The IATV management plans were felt to be appropriate for all patients in study. Twelve patients were able to be discharged from hospital on the same day as IATV on the advice of the neurologist. It is therefore practical to assess patients admitted with neurological symptoms to distant hospitals using IATV and this may result in more efficient use of in‐patient resources.
European Stroke Initiative (EUSI) Recommendations for  Stroke Management The European Stroke Initiative Writing Committee
Authors: Hacke W. , Kaste M. , Skyhoj Olsen T. , Orgogozo J.‐M. , Bogousslavsky J.
Published: February 2003

The European Stroke Initiative (EUSI) is the common body of stroke‐related activities within the European Federation of Neurological Societies (EFNS), the European Neurological Society (ENS) and the European Stroke Council (ESC). The Executive committee of the EUSI has authorized the writing committee of the EUSI to create recommendations for stroke management covering all areas of stroke treatment. The recommendations are listed according to levels of evidence pre‐specified and modified according to several proposals in the literature. The recommendations have been approved by the executive committees of the EUSI, the ESC, the EFNS and the ENS. They are called recommendations rather than guidelines in order to underline the large amount of individual decision making due to the fact that for many important questions, no data of high evidence level is available. The EUSI plans to review and update the recommendations on a regular basis.