160 research outputs found

    Anti-Saccharomyces cerevisiae antibodies in patients with inflammatory bowel disease and their first-degree relatives: Potential clinical value

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    Anti-Saccharomyces cerevisiae antibodies (ASCA) have been described as specific markers in Crohn's disease and their healthy first-degree relatives. 171 patients with Crohn's disease, their 105 first-degree relatives, 145 patients with ulcerative colitis and 101 first-degree relatives of patients with ulcerative colitis, 50 patients with infectious enterocolitis and 100 healthy controls were tested for ASCA employing the ELISA technique. When compared with the healthy controls (p < 0.0001) and patients with infectious enterocolitis (p < 0.0001) the prevalence of ASCA was significantly increased in patients with Crohn's disease and their first-degree relatives (p < 0.01). Further significant differences concerning the frequency of ASCA within the different groups of our study population were not observed. In particular, ASCA were not found in increased prevalence in infectious enterocolitis. These observations are compatible with a role of ASCA as a marker of genetic predisposition to Crohn's disease. Copyright (C) 2002 S. Karger AG, Basel

    Determination of −3858G→A and −164C→A genetic polymorphisms of CYP1A2 in blood and saliva by rapid allelic discrimination: large difference in the prevalence of the −3858G→A mutation between Caucasians and Asians

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    Introduction: Two mutations in CYP1A2, −164C→A (allele CYP1A2*F) and −3858G→A (allele CYP1A2*C), affecting the inducibility of the enzyme, have been published. The aim of this study was to develop a high throughput allelic discrimination assay for these mutations in both saliva and blood and to determine their frequency in Caucasians. Methods: An allelic discrimination assay, based on the fluorogenic 5â€Č-nuclease activity (TaqMan), was developed for the two mutations. Genomic DNA extracted from 17 saliva and 100 blood samples from Caucasians was analysed. Results and conclusions: For the −164C→A mutation, we found an allelic frequency of 68% in the Caucasian population, comparable with data published for Asians and Caucasians. For the −3858G→A mutation, the allele frequency was only 2% in Caucasians, a much lower value than the ~25% reported in Asians (P<0.001). The presented allelic discrimination allows fast and accurate detection of these two mutations. Genotype calls were 100% identical for DNA from saliva and blood. Saliva is easily accessible and represents an excellent alternative to the traditionally used venous blood for genotypin

    Modulation of transendothelial permeability and expression of ATP-binding cassette transporters in cultured brain capillary endothelial cells by astrocytic factors and cell-culture conditions

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    Confluent cell monolayers of brain capillary endothelial cells (BCEC) are used widely as an in vitro cell culture model of the blood-brain barrier. The present study describes the influence of cell-culture conditions on tight junctions, filamentous-actin cytoskeleton, and expression of ATP-binding cassette (ABC) transporters in primary cell cultures of porcine BCEC. Astrocyte as well as C6 glioma-conditioned cell culture medium was used in combination with retinoic acid, dexamethasone, cyclic adenosine monophosphate (cAMP) analogs, or 1,25-dihydroxyvitamin D3. It was shown that C6-conditioned medium led to a reorganization of filamentous actin and to an improved staining of zonula occludens-associated protein-1 (ZO-1). Further optimization of these culture conditions was achieved with cAMP analogs and dexamethasone. Retinoic acid, as well as 1,25-dihydroxyvitamin D3, did not improve cellular tight junctions as judged by filamentous actin, ZO-1 rearrangement, and transcellular electrical resistance (TER) measurements. However, these morphological changes did not influence the paracellular permeability of the extracellular marker sucrose. Expression of ABC transporters such as P-glycoprotein, multidrug resistance-associated protein-1( MRP1), and MRP2 were compared by measuring messenger RNA (mRNA) levels in whole-brain tissue, isolated brain capillaries, and cultured cells. In freshly isolated BCEC, mRNA levels of MRP2 and P-glycoprotein dropped by two- to sevenfold, respectively, whereas MRP1 mRNA levels were slightly increased. During cell culture, mRNA levels of MRP1 and MRP2 decreased by up to fivefold, while P-glycoprotein levels remained constant. These results were unaltered by different cell-culture conditions. In conclusion, the present study suggests that paracellular permeability, as well as mRNA expression of the studied ABC transporters in primary cultures, of porcine BCEC are insensitive toward changes in cell-culture condition

    Die Strandaster-Seidenbiene Colletes halophilus Verhoeff, 1944 (Hymenoptera: Colletidae) ist weit verbreitet an der OstseekĂŒste von DĂ€nemark und Deutschland

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    The Sea Aster mining bee Colletes halophilus Verhoeff, 1944 is known as an endemic of the western European coasts of the Atlantic and the North Sea. The species has specific habitat requirements and is restricted to coastal habitats with populations of the Sea Aster (Tripolium pannonicum), its preferred host plant. Due to the late summer activity of adults and habitat specialisation, this solitary bee species is easily overlooked. In 2019, C. halophilus was first found at the Baltic Sea coast of the island of Fyn (Denmark), 2020 on the island of RĂŒgen (Mecklenburg-Vorpommern, Germany) and in 2021 in Sehlendorf (Schleswig-Holstein, Germany), demonstrating that the species is far more widespread and also occurring along the more continental Western Baltic Sea coast and Kattegat. Targeted sampling in 2022 revealed that C. halophilus is now present in most of the potentially suitable coastal habitats in Schleswig-Holstein and at least in some parts of Mecklenburg-Vorpommern. We here present data on the currently known distribution of C. halophilus along the Baltic Sea coast of Denmark and Germany and discuss hypotheses of a potential recent range extension.Die Strandaster-Seidenbiene Colletes halophilus Verhoeff, 1944 ist ein Endemit der Atlantik- und NordseekĂŒste Westeuropas und aufgrund ihrer spezifischen HabitatansprĂŒche an KĂŒstenbiotope mit Vorkommen der Strandaster (Tripolium pannonicum) als bevorzugter Futterpflanze gebunden. Aufgrund der spĂ€ten Flugzeit und ihrer HabitatspezifitĂ€t kann die Art leicht ĂŒbersehen werden. Durch Funde der Art auf der Insel FĂŒnen (DK) 2019, RĂŒgen (MV) 2020 und Sehlendorf (SH) 2021 wurde deutlich, dass die Art deutlich weiter verbreitet ist und auch im Bereich der kontinentaleren OstseekĂŒste vorkommt. Durch gezielte Nachsuche im Jahr 2022 konnte die Art in den meisten potentiell geeignet erscheinenden KĂŒstenhabitaten in Schleswig-Holstein und einigen Gebieten in Mecklenburg-Vorpommern festgestellt werden. Der aktuelle Kenntnisstand zur Verbreitung von C. halophilus an der OstseekĂŒste wird vorgestellt und Hypothesen einer möglichen rezenten Aus- breitung der Art werden diskutiert

    TRANSPORT OF THE ␀-LACTAM ANTIBIOTIC BENZYLPENICILLIN AND THE DIPEPTIDE GLYCYLSARCOSINE BY BRAIN CAPILLARY ENDOTHELIAL CELLS IN VITRO

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    This paper is available online at http://www.dmd.org ABSTRACT: Peripherally administered ␀-lactam antibiotics, which are structural analogs of tripeptides, may cause neurotoxic reactions or induce seizures. Previous in vivo studies provided evidence for brain uptake of these antibiotics. In the present work, we studied the extent and mechanism of the uptake of benzylpenicillin and glycylsarcosine by brain microvessel endothelial cells in vitro, using freshly isolated and cultured porcine brain capillary endothelial cells. Characterization of the cell culture model demonstrated the functional expression of the system transporting the neutral amino acids leucine and phenylalanine. The initial rate of uptake of benzylpenicillin was &gt;3-fold greater than the rate of uptake of the extracellular marker sucrose (ratio, 3.29 ێ 0.37), whereas uptake of glycylsarcosine did not differ from that of sucrose. The differences in cellular uptake correlated with the octanol/buffer partition coefficients for glycylsarcosine and benzylpenicillin (1.16 ۋ 10 ۊ3 for glycylsarcosine and 6.83 ۋ 10 ۊ2 for benzylpenicillin). The concentration-dependent uptake of benzylpenicillin (1-2000 M) was not saturable and was not sensitive to shifts in pH or temperature. The permeability-surface area product for the uptake of benzylpenicillin at pH 7.4 was determined from these experiments and was found to be 8.1 ۋ 10 ۊ5 ml/sec/g of brain. This value was very close to the value determined in in vivo studies. Uptake of benzylpenicillin and glycylsarcosine was not reduced in the presence of 1 mM ceftibuten or 100 M probenecid. The findings with cultured cell monolayers were confirmed using freshly isolated endothelial cells. These in vitro data are compatible with benzylpenicillin, but not glycylsarcosine, being able to penetrate endothelial cells. Uptake of benzylpenicillin by brain capillary endothelial cells occurs by a slow nonsaturable process, with no evidence for carriermediated transport

    Identification of reference markers for characterizing honey bee (Apis mellifera) hemocyte classes

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    Cell mediated immunity of the honey bee (Apis mellifera) involves the activity of several hemocyte populations, currently defined by morphological features and lectin binding characteristics. The objective of the present study was to identify molecular markers capable of characterizing subsets of honey bee hemocytes. We developed and employed monoclonal antibodies with restricted reactions to functionally distinct hemocyte subpopulations. Melanizing cells, known as oenocytoids, were defined by an antibody to prophenoloxidase, aggregating cells were identified by the expression of Hemolectin, and phagocytic cells were identified by a marker expressed on granulocytes. We anticipate that this combination of antibodies not only allows for the detection of functionally distinct hemocyte subtypes, but will help to further the exploration of hematopoietic compartments, as well as reveal details of the honey bee cellular immune defense against parasites and microbes

    Ultrafast glutamate sensors resolve high-frequency release at Schaffer collateral synapses

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    Glutamatergic synapses display a rich repertoire of plasticity mechanisms on many different time scales, involving dynamic changes in the efficacy of transmitter release as well as changes in the number and function of postsynaptic glutamate receptors. The genetically encoded glutamate sensor iGluSnFR enables visualization of glutamate release from presynaptic terminals at frequencies up to ∌10 Hz. However, to resolve glutamate dynamics during high-frequency bursts, faster indicators are required. Here, we report the development of fast (iGluf) and ultrafast (iGluu) variants with comparable brightness but increased Kd for glutamate (137 ÎŒM and 600 ÎŒM, respectively). Compared with iGluSnFR, iGluu has a sixfold faster dissociation rate in vitro and fivefold faster kinetics in synapses. Fitting a three-state model to kinetic data, we identify the large conformational change after glutamate binding as the rate-limiting step. In rat hippocampal slice culture stimulated at 100 Hz, we find that iGluu is sufficiently fast to resolve individual glutamate release events, revealing that glutamate is rapidly cleared from the synaptic cleft. Depression of iGluu responses during 100-Hz trains correlates with depression of postsynaptic EPSPs, indicating that depression during high-frequency stimulation is purely presynaptic in origin. At individual boutons, the recovery from depression could be predicted from the amount of glutamate released on the second pulse (paired pulse facilitation/depression), demonstrating differential frequency-dependent filtering of spike trains at Schaffer collateral boutons

    Resonant second harmonic generation in random dielectric structures

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    International audience; We show that resonant second harmonic generation can be obtained in random dielectric structures. The scheme is based on internal resonances due to the optical counterpart of Anderson localization. By making use of different localization lengths at the fundamental and at the second harmonic frequencies, we predict a conversion efficiency that is four orders of magnitude higher than a bulk material and even one order of magnitude higher than an ideal phase matched slab of the same size. The method is highly insensitive to fabrication tolerances, and provides excellent angle tunability. [DOI:10.2971/jeos.2006.06021
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