Biosynthesis of acridone alkaloids formation of rutacridone by cell-free extracts of Ruta graveolens cell suspension cultures

AbstractMicrosomes prepared either by ultracentrifugation or MgCl2 precipitation from cultured Ruta graveolens cells catalyze the condensation of 1,3-dihydroxy-N-methylacridone and isopentenylpyrophosphate or dimethylallylpyrophosphate. In the presence of NADPH and oxygen rutacridone was identified as reaction product. By omission of NADPH glycocitrine-II is accumulated. The results suggest that at first a prenylated acridone is formed which in turn is cyclized giving the dihydrofuran part of rutacridone

Business and services models for electric vehicles

The paper presents the Green eMotion Project, the objectives of the business models analysis and preliminary results of such analysis.This paper introduces the approach for the business models analysis for electric vehicles, as followed in the FP7 EU-funded Green eMotion project. The main goal of Green eMotion is to enable a mass deployment of electric mobility in Europe. For that purpose, Green eMotion will connect ongoing regional and national electric mobility initiatives leveraging the results and comparing different technology approaches to ensure the best solutions prevail for the European market. A virtual marketplace will be created to enable the different actors to interact and to allow for new highvalue transportation services as well as electric vehicle (EV) user convenience in billing (EU Clearing House). In addition, the Green eMotion project will demonstrate the integration of electric mobility into electricity networks and contribute to the improvement and development of new and existing standards for electric mobility interfaces. In order to facilitate large-scale EVs roll-out in terms of social acceptance, commercial viability and system/environmental impact, the most suited business models should be identified and assessed according to a methodology taking into account all economic transactions between the different participating stakeholders.European Commission's FP

P and S wave velocity measurements of water-rich sediments from the Nankai Trough, Japan

Acoustic velocities were measured during triaxial deformation tests of silty clay and clayey silt core samples from the Nankai subduction zone (Integrated Ocean Drilling Program Expeditions 315, 316, and 333). We provide a new data set, continuously measured during pressure increase and subsequent axial deformation. A new data processing method was developed using seismic time series analysis. Compressional wave velocities (V-p) range between about 1450 and 2200 m/s, and shear wave velocities (V-s) range between about 150 and 800 m/s. V-p slightly increases with rising effective confining pressure and effective axial stress. Samples from the accretionary prism toe show the highest Vp, while fore-arc slope sediments show lower Vp. Samples from the incoming plate, slightly richer in clay minerals, have the lowest values for V-p. V-s increases with higher effective confining pressures and effective axial stress, irrespective of composition and tectonic setting. Shear and bulk moduli are between 0.2 and 1.3 GPa, and 3.85 and 8.41 GPa, respectively. Elastic moduli of samples from the accretionary prism toe and the footwall of the megasplay fault (1.50 and 3.98 GPa) are higher than those from the hanging wall and incoming plate (0.59 and 0.88 GPa). This allows differentiation between normal and overconsolidated sediments. The data show that in a tectonosedimentary environment of only subtle compositional differences, acoustic properties can be used to differentiate between stronger (accretionary prism toe) and weaker (fore-arc slope, incoming plate) sediments. Especially V-p/V-s ratios may be instrumental in detecting zones of low effective stress and thus high pore fluid pressur

Development of a test setup for the characterization of an optical microscope for high precision length metrology applications

A test setup to qualify the performance of optical microscopes has been designed and optimized using FEM calculations to exhibit a minimal susceptibility to thermal and mechanical influences of the ambient environment. The alignment is performed using an alignment autocollimator and alignment targets. The data acquisition of the camera and the position sensors of the stage is synchronized. The short-term repeatability (1s) of the line position and -width measurement obtained with the integrated UV microscope are 1 nm and 0.2 nm respectively. In long-term measurements the maximum lateral and focus drift rate observed were 30- and 20 nm / hour respectively. The measured point spread function contained only radial symmetric optical aberrations. Using the Zernike-Nijboer theory including only the defocus and spherical aberrations, fit residuals were obtained that contain systematic deviations in the order of the noise level

Identification of genes involved in Ca(2+ )ionophore A23187-mediated apoptosis and demonstration of a high susceptibility for transcriptional repression of cell cycle genes in B lymphoblasts from a patient with Scott syndrome

BACKGROUND: In contrast to other agents able to induce apoptosis of cultured cells, Ca(2+ )ionophore A23187 was shown to elicit direct activation of intracellular signal(s). The phenotype of the cells derived from patients having the hemorrhagic disease Scott syndrome, is associated with an abnormally high proportion of apoptotic cells, both in basal culture medium and upon addition of low ionophore concentrations in long-term cultures. These features are presumably related to the mutation also responsible for the defective procoagulant plasma membrane remodeling. We analyzed the specific transcriptional re-programming induced by A23187 to get insights into the effect of this agent on gene expression and a defective gene regulation in Scott cells. RESULTS: The changes in gene expression upon 48 hours treatment with 200 nM A23187 were measured in Scott B lymphoblasts compared to B lymphoblasts derived from the patient's daughter or unrelated individuals using Affymetrix microarrays. In a similar manner in all of the B cell lines, results showed up-regulation of 55 genes, out of 12,000 represented sequences, involved in various pathways of the cell metabolism. In contrast, a group of 54 down-regulated genes, coding for histones and proteins involved in the cell cycle progression, was more significantly repressed in Scott B lymphoblasts than in the other cell lines. These data correlated with the alterations of the cell cycle phases in treated cells and suggested that the potent effect of A23187 in Scott B lymphoblasts may be the consequence of the underlying molecular defect. CONCLUSION: The data illustrate that the ionophore A23187 exerts its pro-apoptotic effect by promoting a complex pattern of genetic changes. These results also suggest that a subset of genes participating in various steps of the cell cycle progress can be transcriptionally regulated in a coordinated fashion. Furthermore, this research brings a new insight into the defect in cultured Scott B lymphoblasts, leading to hypothesize that a mutated gene plays a role not only in membrane remodeling but also in signal transduction pathway(s) leading to altered transcriptional regulation of cell cycle genes

Autoantibodies in a Subgroup of Patients with Linear IgA Disease React with the NC16A Domain of BP1801

Linear IgA disease is an autoimmune subepidermal blistering disease characterized by IgA deposits at the cutaneous basement membrane zone. IgA antibodies from linear IgA disease sera react with antigens of 97 kDa (LABD97) and 120 kDa (LAD-1), both of which appear to be fragments of the extracellular domain of bullous pemphigoid 180 (type XVII collagen). The aim of this study was to determine whether linear IgA disease sera react with the immunodominant region of BP180 (NC16A domain), which is a major target of IgG autoanti-bodies produced by patients with bullous pemphigoid. Indeed, 11 of 50 linear IgA disease sera were found to contain IgA autoantibodies that recognized a recombinant form of NC16A by immunoblotting. The same sera also reacted with NC16A by enzyme-linked immunosorbent assay. An epitope mapping analysis uncovered four linear IgA disease-associated epitopes located within the 45 amino acid N-terminal stretch of NC16A, all of which were previously identified as antigenic sites targeted by bullous pemphigoid autoantibodies. Eight of the linear IgA disease sera that were reactive with NC16A also recognized LAD-1 secreted by the SCC-25 cell line, and five sera recognized BP180 extracted from keratinocytes. Linear IgA disease sera depleted of reactivity to NC16A by immunoadsorption continued to react with both the LAD-1 antigen and BP180 by immunoblotting and with the basement membrane zone by indirect immunofluorescence microscopy. Our results demonstrate that IgA autoantibodies from a subset of linear IgA disease patients react with the same sites on BP180 that are targeted by IgG autoantibodies in bullous pemphigoid

A randomized trial of a transglutaminase 2 inhibitor for celiac disease

BACKGROUND In celiac disease, small intestinal transglutaminase 2 causes deamidation of glutamine residues in gluten peptides, which enhances stimulation of T cells and leads to mucosal injury. Inhibition of transglutaminase 2 is a potential treatment for celiac disease. METHODS In a proof-of-concept trial, we assessed the efficacy and safety of a 6-week treatment with ZED1227, a selective oral transglutaminase 2 inhibitor, at three dose levels as compared with placebo, in adults with well-controlled celiac disease who underwent a daily gluten challenge. The primary end point was the attenuation of gluten-induced mucosal damage, as measured by the ratio of villus height to crypt depth. Secondary end points included intraepithelial lymphocyte density, the Celiac Symptom Index score, and the Celiac Disease Questionnaire score (for assessment of health-related quality of life). RESULTS Of the 41 patients assigned to the 10-mg ZED1227 group, the 41 assigned to the 50-mg group, the 41 assigned to the 100-mg group, and the 40 assigned to the placebo group, 35, 39, 38, and 30 patients, respectively, had adequate duodenal-biopsy samples for the assessment of the primary end point. Treatment with ZED1227 at all three dose levels attenuated gluten-induced duodenal mucosal injury. The estimated difference from placebo in the change in the mean ratio of villus height to crypt depth from baseline to week 6 was 0.44 (95% confidence interval [CI], 0.15 to 0.73) in the 10-mg group (P=0.001), 0.49 (95% CI, 0.20 to 0.77) in the 50-mg group (P<0.001), and 0.48 (95% CI, 0.20 to 0.77) in the 100-mg group (P<0.001). The estimated differences from placebo in the change in intraepithelial lymphocyte density were -2.7 cells per 100 epithelial cells (95% CI, -7.6 to 2.2) in the 10-mg group, -4.2 cells per 100 epithelial cells (95% CI, -8.9 to 0.6) in the 50-mg group, and -9.6 cells per 100 epithelial cells (95% CI, -14.4 to -4.8) in the 100-mg group. Use of the 100-mg dose may have improved symptom and quality-of-life scores. The most common adverse events, the incidences of which were similar across all groups, were headache, nausea, diarrhea, vomiting, and abdominal pain. Rash developed in 3 of 40 patients (8%) in the 100-mg group. CONCLUSIONS In this preliminary trial, treatment with ZED1227 attenuated gluten-induced duodenal mucosal damage in patients with celiac disease.publishedVersionPeer reviewe

Strong sediments at the deformation front, and weak sediments at the rear of the Nankai accretionary prism, revealed by triaxial deformation experiments

Nineteen whole-round core samples from the Nankai accretionary prism (IODP Expeditions 315, 316, and 333) from a depth range of 28–128 m below sea floor were experimentally deformed in a triaxial cell under consolidated and undrained conditions at confining pressures of 400–1000 kPa, room temperature, axial displacement rates of 0.01–9.0 mm/min, and up to axially compressive strains of ∼64%. Despite great similarities in composition and grain size distribution of the silty clay samples, two distinct “rheological groups” are distinguished: The first group shows deviatoric peak stress after only a few percent of compressional strain (10%), or does not weaken at all. This is characteristic of structurally strong material. The strong samples tend to be overconsolidated and are all from the drillsites at the accretionary prism toe, while the weak and normally consolidated samples come from the immediate hanging wall of a megasplay fault further upslope. Sediments from the incoming plate are also structurally weak. The observed differences in mechanical behavior may hold a key for understanding strain localization and brittle faulting within the uniform silty and clayey sedimentary sequence of the Nankai accretionary prism

Proteomics based analysis of interactions between CEA cell adhesion molecule 1 (CEACAM1) and intracellular proteins in transfected tumor cells

CEA Cell Adhesion Molecule 1 (CEACAM1), a type 1 transmembrane protein belonging to the CEA gene family, is alternatively spliced to produce four major isoforms with a long (CEACAM1-L) or a short cytoplasmic domain (CEACAM1-S). Immunoprecipitations using the murine adenocarcinoma MC38 cell line transfected with human CEACAM1-L were performed to isolate cytoplasmic proteins binding to the unphosphorylated or tyrosine phosphorylated CEACAM1 long cytoplasmic domain. Mass spectrometric analysis of immunoprecipitates separated on two-dimensional gels revealed actin, tropomyosin, myosin, vimentin and cytokeratin as the major proteins interacting with CEACAM1-L after sodium pervanadate induced tyrosine phosphorylation of CEACAM1-L. Surface plasmon resonance studies were performed to characterize actin and tropomyosin binding to the membrane-proximate region of the CEACAM1-L cytoplasmic domain and to its short splicing variant CEACAM1-S. A CEACAM1-L derived peptide bound F-actin and tropomyosin with KDs of 1.3x10–5 M, and 1.8x10-5 M, respectively, while an equivalent CEACAM1-S peptide bound less than 10% of the long form to F-actin and tropomyosin with a KD <10-4 M. GST-long or short cytoplasmic domain fusion proteins bound actin and tropomyosin with KDs of 3.0x10-8 M and 4.1x10-7 M, respectively, for the long form, and KDs of 6.2x10-8 M and 2.6x10-7 M, respectively, for the short form. Calmodulin, a known CEACAM1 binding protein, or EDTA inhibited binding of the CEACAM1-L peptide and both GST-CEACAM1 fusion proteins to actin, while calmodulin and actin, but not EDTA, stimulated binding of GST-CEACAM1-L to tropomyosin. Confocal microscopy studies showed a sodium pervanadate dependent colocalization of F-actin and CEACAM1-L at cell-cell adhesion sites in CEACAM1-L transfected MC38 cells, confirming the initial immunoprecipitation results. The identification of an association of both CEACAM1 cytoplasmic domains with actin and tropomyosin and its potential regulation by calmodulin and Ca2+ could lead to a better understanding of the mechanisms and the functions of interactions between transmembrane cell adhesion proteins and the cytoskeleton