The HLA-E Gene encodes two differentially regulated Transcripts and a Cell Surface Protein

An HLA-E-specific oligonucleotide probe was used to study the expressioonf HLA-E. This probed etects two HLA-E transcripts, 1.8 and 2.7 kb in size, which are present in varying ratios in allt issues and cell lines investigated. We demonstrate that alternative poly(A) site usage accounts for the differential regulation of the two HLA-E mRNA species. Sequence analysis of three cDNA clones, representing the two transcripts of HLA-E, and of anH LA-E gene encoded by cosmid cd3.14, revealed identity of gene and cDNA in the 3’ untranslated region. S1 nuclease protection assays confirmed that the two HLA-E transcripts are not alternative splicing products. Introduction of cd3.14, together with human ,&m into the murine myeloma cell line P3X63-Ag8.653, resulted in a cell surface expresosf ioan HLA-class I heavy chain detectablbey indirect immunofluorescence whereas transfection into the humBaznr n expressing mouse L cell line, 527 was negative with regard to cell surface expressionC. ell surface labeling of transfectants and immunoprecipitation with a monomorphic HLA class I-specific antibodyo r an antibody against human &m confirmed the presence of an HLA-E H chain on the cell surface. These results indicate that the HLA-E gene codes for a class I H chain that can be expressed on the cell surface

Measurement of Gluten in Food Products: Proficiency‐Testing Rounds as a Measure of Precision and Applicability

In 2008, Codex Alimentarius endorsed the R5 Enzyme‐Linked Immunosorbent Assay (ELISA) method as Method Type 1 for gluten measurement in gluten‐free foods. The most recognized R5 ELISA test kit is the RIDASCREEEN® Gliadin (R7001; manufacturer R‐Biopharm). Beside collaborative tests that led to several international approved methods of this test kit, proficiency‐testing (PT) rounds are regularly performed in Europe by different PT providers. Results from these rounds were analyzed regarding the number of participating labs with acceptable results for the RIDASCREEN® Gliadin. All PT rounds document the excellent consistency and comparability of results. The data show that the RIDASCREEN® Gliadin R5 ELISA is also applicable to cake mix, oat‐based foodstuff, infant soya formula, cookies, canned boiled sausage, gravy thickener, pasta, and potato dumpling. These rounds also included the analysis of blank matrices. It was found that more than 95% of all participating laboratories correctly detected these samples as negative. Other gluten test kit manufacturers were analyzed as well, but due to the low number of participants using these test kits results were often only analyzed in a qualitative manner questioning the comparability of these kits to the RIDASCREEN® Gliadin R5 ELISA

Eine «‹sektorielle› lückenhafte Heimpolitik»?: rechtliche Steuerung des Heimwesens seit den 1940er Jahren in föderaler Perspektive

Using the cantons of Geneva, St. Gallen and Uri as examples, this article describes the development of legal instruments within the residential care sector for children and adolescents since 1940, with a strong emphasis on the means of the state to secure the quality of care. The goal of this article is to show that the quality standards resulting from the legislation processes were a product of political debates in which child protection was weighed against organizational and financial considerations. Furthermore, this article draws on a comparative perspective to illustrate the significant differences between cantons when quality standards were implemented. Central to this was not only federalism, but also the concept of subsidiarity as a mode of statehood – meaning that the lowest possible level should be activated for a task – which resulted in a complex division of responsibility regarding control and financing between the different governmental and private actors

Is it really always only the others who are to blame? GP’s view on medical overuse. A questionnaire study

Background Medical overuse is a common problem in health care. Preventing unnecessary medicine is one of the main tasks of General Practice, so called quaternary prevention. We aimed to capture the current opinion of German General Practitioners (GPs) to medical overuse. Methods A quantitative online study was conducted. The questionnaire was developed based on a qualitative study and literature search. GPs were asked to estimate prevalence of medical overuse as well as to evaluate drivers and solutions of medical overuse. GPs in Bavaria were recruited via email (750 addresses). A descriptive data analysis was performed. Additionally the association between doctors’ attitudes and (1) demographic variables and (2) interest in campaigns against medical overuse was assessed. Results Response rate was 18%. The mean age was 54 years, 79% were male and 68% have worked as GP longer than 15 years. Around 38% of medical services were considered as medical overuse and nearly half of the GPs (47%) judged medical overuse to be the more important problem than medical underuse. Main drivers were seen in “patients´ expectations” (76%), “lack of a primary care system” (61%) and “defensive medicine” (53%), whereas “disregard of evidence/guidelines” (15%) and “economic pressure on the side of the doctor” (13%) were not weighted as important causes. Demographic variables did not have an important impact on GPs´ response pattern. GPs interested in campaigns like “Choosing Wisely” showed a higher awareness for medical overuse, although these campaigns were only known by 50% of the respondents. Discussion Medical overuse is an important issue for GPs. Main drivers were searched and found outside their own sphere of responsibility. Campaigns as “Choosing Wisely” seem to have a positive effect on GPs attitude, but knowledge is still limited

Boundary Zonal Flow in Rotating Turbulent Rayleigh-Bénard Convection

For rapidly rotating turbulent Rayleigh–Bénard convection in a slender cylindrical cell, experiments and direct numerical simulations reveal a boundary zonal flow (BZF) that replaces the classical large-scale circulation. The BZF is located near the vertical side wall and enables enhanced heat transport there. Although the azimuthal velocity of the BZF is cyclonic (in the rotating frame), the temperature is an anticyclonic traveling wave of mode one, whose signature is a bimodal temperature distribution near the radial boundary. The BZF width is found to scale like Ra1/4Ek2/3 where the Ekman number Ek decreases with increasing rotation rate

Regulation of the arachidonic acid mobilization in macrophages by combustion-derived particles

<p>Abstract</p> <p>Background</p> <p>Acute exposure to elevated levels of environmental particulate matter (PM) is associated with increasing morbidity and mortality rates. These adverse health effects, e.g. culminating in respiratory and cardiovascular diseases, have been demonstrated by a multitude of epidemiological studies. However, the underlying mechanisms relevant for toxicity are not completely understood. Especially the role of particle-induced reactive oxygen species (ROS), oxidative stress and inflammatory responses is of particular interest.</p> <p>In this <it>in vitro </it>study we examined the influence of particle-generated ROS on signalling pathways leading to activation of the arachidonic acid (AA) cascade. Incinerator fly ash particles (MAF02) were used as a model for real-life combustion-derived particulate matter. As macrophages, besides epithelial cells, are the major targets of particle actions in the lung murine RAW264.7 macrophages and primary human macrophages were investigated.</p> <p>Results</p> <p>The interaction of fly ash particles with macrophages induced both the generation of ROS and as part of the cellular inflammatory responses a dose- and time-dependent increase of free AA, prostaglandin E₂/thromboxane B₂(PGE₂/TXB₂), and 8-isoprostane, a non-enzymatically formed oxidation product of AA. Additionally, increased phosphorylation of the mitogen-activated protein kinases (MAPK) JNK1/2, p38 and ERK1/2 was observed, the latter of which was shown to be involved in MAF02-generated AA mobilization and phosphorylation of the cytosolic phospolipase A₂. Using specific inhibitors for the different phospolipase A₂isoforms the MAF02-induced AA liberation was shown to be dependent on the cytosolic phospholipase A₂, but not on the secretory and calcium-independent phospholipase A₂. The initiation of the AA pathway due to MAF02 particle exposure was demonstrated to depend on the formation of ROS since the presence of the antioxidant N-acetyl-cysteine (NAC) prevented the MAF02-mediated enhancement of free AA, the subsequent conversion to PGE₂/TXB₂via the induction of COX-2 and the ERK1/2 and JNK1/2 phosphorylation. Finally we showed that the particle-induced formation of ROS, liberation of AA and PGE₂/TXB₂together with the phosphorylation of ERK1/2 and JNK1/2 proteins was decreased after pre-treatment of macrophages with the metal chelator deferoxamine mesylate (DFO).</p> <p>Conclusions</p> <p>These results indicate that one of the primary mechanism initiating inflammatory processes by incinerator fly ash particles seems to be the metal-mediated generation of ROS, which triggers via the MAPK cascade the activation of AA signalling pathway.</p