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J Roelofs

Publications and source records attributed to J Roelofs.

12 recordsLinked to original sources
PubMed2002

Dutch version of the Pain Vigilance and Awareness Questionnaire: validity and reliability in a pain-free population.

The current study examined validity and reliability of the Pain Vigilance and Awareness Questionnaire (PVAQ) in two samples of healthy college students. Exploratory and confirmatory factor analysis showed that a two-factor model of the PVAQ was most suitable in the present study. The first factor could be referred to as attention to pain whereas the second factor could be specified as attention to changes in pain. With regard to the convergent and divergent validity, the PVAQ was found to correlate highly with related constructs like catastrophising (PCS) and general body vigilance (BVQ). The correlation between PVAQ and pain-related fear (FPQ) was moderate, whereas correlations with unrelated constructs like trait anxiety (STAI-T) and fear of spiders (FSQ) were low. Furthermore, the PVAQ showed good internal consistency and fair test-retest reliability. Altogether, these findings suggest that the PVAQ is a valid and reliable measure of pain vigilance in healthy individuals. The results of this study can be regarded as a starting point for further validation of the PVAQ in clinical pain populations. Implications for future research and treatment interventions are discussed.

Adult↗
PubMed2001

GTPgammaS regulation of a 12-transmembrane guanylyl cyclase is retained after mutation to an adenylyl cyclase.

DdGCA is a Dictyostelium guanylyl cyclase with a topology typical for mammalian adenylyl cyclases containing 12 transmembrane-spanning regions and two cyclase domain. In Dictyostelium cells heterotrimeric G-proteins are essential for guanylyl cyclase activation by extracellular cAMP. In lysates, guanylyl cyclase activity is strongly stimulated by guanosine 5'-3-O-(thio) triphosphate (GTPgammaS), which is also a substrate of the enzyme. DdGCA was converted to an adenylyl cyclase by introducing three point mutations. Expression of the obtained DdGCA(kqd) in adenylyl cyclase-defective cells restored the phenotype of the mutant. GTPgammaS stimulated the adenylyl cyclase activity of DdGCA(kqd) with properties similar to those of the wild-type enzyme (decrease of K(m) and increase of V(max)), demonstrating that GTPgammaS stimulation is independent of substrate specificity. Furthermore, GTPgammaS activation of DdGCA(kqd) is retained in several null mutants of Galpha and Gbeta proteins, indicating that GTPgammaS activation is not mediated by a heterotrimeric G-protein but possibly by a monomeric G-protein.

Adenylyl Cyclases↗
PubMed2001

The Dictyostelium homologue of mammalian soluble adenylyl cyclase encodes a guanylyl cyclase.

A new Dictyostelium discoideum cyclase gene was identified that encodes a protein (sGC) with 35% similarity to mammalian soluble adenylyl cyclase (sAC). Gene disruption of sGC has no effect on adenylyl cyclase activity and results in a >10-fold reduction in guanylyl cyclase activity. The scg- null mutants show reduced chemotactic sensitivity and aggregate poorly under stringent conditions. With Mn(2+)/GTP as substrate, most of the sGC activity is soluble, but with the more physiological Mg(2+)/GTP the activity is detected in membranes and stimulated by GTPgammaS. Unexpectedly, orthologues of sGC and sAC are present in bacteria and vertebrates, but absent from Drosophila melanogaster, Caenorhabditis elegans, Arabidopsis thaliana and Saccharomyces cerevisiae.

Adenylyl Cyclases↗
PubMed2001

Guanylate cyclase in Dictyostelium discoideum with the topology of mammalian adenylate cyclase.

The core of adenylate and guanylate cyclases is formed by an intramolecular or intermolecular dimer of two cyclase domains arranged in an antiparallel fashion. Metazoan membrane-bound adenylate cyclases are composed of 12 transmembrane spanning regions, and two cyclase domains which function as a heterodimer and are activated by G-proteins. In contrast, membrane-bound guanylate cyclases have only one transmembrane spanning region and one cyclase domain, and are activated by extracellular ligands to form a homodimer. In the cellular slime mould, Dictyostelium discoideum, membrane-bound guanylate cyclase activity is induced after cAMP stimulation; a G-protein-coupled cAMP receptor and G-proteins are essential for this activation. We have cloned a Dictyostelium gene, DdGCA, encoding a protein with 12 transmembrane spanning regions and two cyclase domains. Sequence alignment demonstrates that the two cyclase domains are transposed, relative to these domains in adenylate cyclases. DdGCA expressed in Dictyostelium exhibits high guanylate cyclase activity and no detectable adenylate cyclase activity. Deletion of the gene indicates that DdGCA is not essential for chemotaxis or osmo-regulation. The knock-out strain still exhibits substantial guanylate cyclase activity, demonstrating that Dictyostelium contains at least one other guanylate cyclase.

Adenylyl Cyclases↗
PubMed2001

Identification and characterization of DdPDE3, a cGMP-selective phosphodiesterase from Dictyostelium.

In Dictyostelium cAMP and cGMP have important functions as first and second messengers in chemotaxis and development. Two cyclic-nucleotide phosphodiesterases (DdPDE 1 and 2) have been identified previously, an extracellular dual-specificity enzyme and an intracellular cAMP-specific enzyme (encoded by the psdA and regA genes respectively). Biochemical data suggest the presence of at least one cGMP-specific phosphodiesterase (PDE) that is activated by cGMP. Using bioinformatics we identified a partial sequence in the Dictyostelium expressed sequence tag database that shows a high degree of amino acid sequence identity with mammalian PDE catalytic domains (DdPDE3). The deduced amino acid sequence of a full-length DdPDE3 cDNA isolated in this study predicts a 60 kDa protein with a 300-residue C-terminal PDE catalytic domain, which is preceded by approx. 200 residues rich in asparagine and glutamine residues. Expression of the DdPDE3 catalytic domain in Escherichia coli shows that the enzyme has Michaelis-Menten kinetics and a higher affinity for cGMP (K(m)=0.22 microM) than for cAMP (K(m)=145 microM); cGMP does not stimulate enzyme activity. The enzyme requires bivalent cations for activity; Mn(2+) is preferred to Mg(2+), whereas Ca(2+) yields no activity. DdPDE3 is inhibited by 3-isobutyl-1-methylxanthine with an IC(50) of approx. 60 microM. Overexpression of the DdPDE3 catalytic domain in Dictyostelium confirms these kinetic properties without indications of its activation by cGMP. The properties of DdPDE3 resemble those of mammalian PDE9, which also shows the highest sequence similarity within the catalytic domains. DdPDE3 is the first cGMP-selective PDE identified in lower eukaryotes.

1-Methyl-3-isobutylxanthine↗
PubMed2001

Reduced protein diffusion rate by cytoskeleton in vegetative and polarized dictyostelium cells.

Fluorescence recovery after photobleaching measurements with high spatial resolution are performed to elucidate the impact of the actin cytoskeleton on translational mobility of green fluorescent protein (GFP) in aqueous domains of Dictyostelium discoideum amoebae. In vegetative Dictyostelium cells, GFP molecules experience a 3.6-fold reduction of their translational mobility relative to dilute aqueous solutions. In disrupting the actin filamentous network using latrunculin-A, the intact actin cytoskeletal network is shown to contribute an effective viscosity of 1.36 cP, which accounts for 53% of the restrained molecular diffusion of GFP. The remaining 47% of hindered protein motions is ascribed to other mechanical barriers and the viscosity of the cell liquid. A direct correlation between the density of the actin network and its limiting action on protein diffusion is furthermore established from measurements under different osmotic conditions. In highly locomotive polarized cells, the obstructing effect of the actin filamentous network is seen to decline to 0.46 cP in the non-cortical regions of the cell. Our results indicate that the meshwork of actin filaments constitutes the primary mechanical barrier for protein diffusion and that any noticeable reorganization of the network is accompanied by altered intracellular protein mobility.

Actins↗
PubMed1998

Expression and temperature-dependent regulation of the beta2-microglobulin (Cyca-B2m) gene in a cold-blooded vertebrate, the common carp (Cyprinus carpio L.).

Expression of beta2-microglobulin (beta2m) in the common carp was studied using a polyclonal antibody raised against a recombinant protein obtained from eukaryotic expression of the Cyca-B2m gene. Beta2m is expressed on peripheral blood Ig+ and Ig lymphocytes, but not on erythrocytes and thrombocytes. In spleen and pronephros, dull- and bright-positive populations could be identified correlating with the presence of erythrocytes, thrombocytes, and mature leucocytes or immature and mature cells from the lympho-myeloid lineage, respectively. Thymocytes were shown to be comprised of a single bright-positive population. The Cyca-B2m polyclonal antiserum was used in conjunction with a similarly produced polyclonal antiserum to an MHC class I (Cyca-UA) alpha chain to investigate the expression of class I molecules on peripheral blood leucocytes (PBL) at different permissive temperatures. At 12 degrees C, a temporary downregulation of class I molecules was demonstrated, which recovered to normal levels within 3 days. However, at 6 degrees C, a lasting absence of class I cell-surface expression was observed, which could be restored slowly by transfer to 12 degrees C. The expression of immunoglobulin molecules on B cells was unaffected by temperature changes. The absence of the class I cell-surface expression was shown to be the result of a lack of sufficient Cyca-B2m gene transcription, although Cyca-UA mRNA was present at comparable levels at all temperatures. This suggests that class I expression is regulated by a temperature-sensitive transcription of the Cyca-B2m gene.

Animals↗
PubMed1995

Evaluation of antral motility in humans using manometry and scintigraphy.

Recent studies suggest that scintigraphy can be used to evaluate non-invasively antral motility in humans, although scintigraphic techniques have not yet been compared with more conventional measurements of intraluminal pressures by manometry. Simultaneous scintigraphic and manometric measurements of antral motility were performed in nine healthy volunteers. After intubation with a sleeve/sidehole catheter which incorporated five pressure sideholes located at 1.5 cm intervals spanning the antrum, each subject ingested 100 g minced beef labelled with 100 MBq 99mTc-chicken liver and 150 ml water. Between 40-43, 60-63, 80-83, and 100-103 minutes after meal ingestion, radioisotopic data were acquired in two second frames. Time-activity curves showing antral 'contractions' resulting from wall motion were derived by drawing small regions of interest over the antrum to coincide with the position of the antral manometric sideholes. Scintigraphic contraction rates approximated 3/minute, whereas antral pressure waves that occluded the lumen were less frequent (p < 0.01 for all), particularly in the proximal antrum. The amplitude of wall motion, evaluated scintigraphically, and the amplitude of pressure waves were both inversely related to the distance from the pylorus (r > -0.32, p < 0.05) and antral volume r > -0.29 (p < 0.05). There were significant relationships between the amplitude of contractions assessed scintigraphically and the number of lumen-occlusive antral pressure waves in the distal antrum (r -0.48, p < 0.05) but not in the more proximal antral regions. It is concluded that scintigraphy can detect antral wall motion with greater sensitivity than manometry, particularly in the proximal antrum. As manometry gives information on the amplitude as well as the temporal and spatial organisation of those contractions which result in lumen occlusion, the combination of scintigraphic and manometric techniques in the evaluation of antral motility shows considerable promise.

Adult↗
PubMed1994

[Social isolation in the psychiatric department].

In this article account is given of a validation study, according to the nursing diagnosis 'social isolation'. The label, definition and cluster of defining characteristics according to cluster of defining characteristics according to Townsend (1990) are the starting point, the related factors are left out of consideration. The validation mentioned above was carried out in accordance with the Delphi-method, that consisted of three Delphi rounds. A panel of experts was asked particularly to compile defining characteristics that are relevant for 'social isolation' and to indicate the critical defining characteristics. This procedure generated a cluster of seven defining characteristics for 'social isolation' of which the content validity turned out to be considerably increased with regard to the cluster formulated by Townsend. The distinction between critical defining characteristics and supporting defining characteristics was not clarified by this study.

Delphi Technique↗