Stabilized reverse transcriptase fusion proteins

Stabilized reverse transcriptase fusion proteins including a thermostable reverse transcriptase connected to a stabilizer protein are described. Attaching the stabilizer protein to the thermostable reverse transcriptase stabilizes the fusion protein and can aid in its purification, provide increased solubility, allow for longer storage, or allow the fusion protein to be used under more rigorous conditions such as higher temperature. The stabilized reverse transcriptase fusion protein can also include a linker between the stabilizer protein and the thermostable reverse temperature. The stabilized reverse transcriptase fusion proteins are suitable for use in nucleic acid amplification methods such as the reverse transcription polymerase chain reaction and other applications involving cDNA synthesis.Board of Regents, University of Texas Syste

Group II Intron Protein Localization and Insertion Sites Are Affected by Polyphosphate

Mobile group II introns consist of a catalytic intron RNA and an intron-encoded protein with reverse transcriptase activity, which act together in a ribonucleoprotein particle to promote DNA integration during intron mobility. Previously, we found that the Lactococcus lactis Ll.LtrB intron-encoded protein (LtrA) expressed alone or with the intron RNA to form ribonucleoprotein particles localizes to bacterial cellular poles, potentially accounting for the intron's preferential insertion in the oriC and ter regions of the Escherichia coli chromosome. Here, by using cell microarrays and automated fluorescence microscopy to screen a transposon-insertion library, we identified five E. coli genes (gppA, uhpT, wcaK, ynbC, and zntR) whose disruption results in both an increased proportion of cells with more diffuse LtrA localization and a more uniform genomic distribution of Ll.LtrB-insertion sites. Surprisingly, we find that a common factor affecting LtrA localization in these and other disruptants is the accumulation of intracellular polyphosphate, which appears to bind LtrA and other basic proteins and delocalize them away from the poles. Our findings show that the intracellular localization of a group II intron-encoded protein is a major determinant of insertion-site preference. More generally, our results suggest that polyphosphate accumulation may provide a means of localizing proteins to different sites of action during cellular stress or entry into stationary phase, with potentially wide physiological consequences.This work was supported by National Institutes of Health R01 grants GM037949 to AML and GM076536 to EMM, Welch Foundation grants F-1607 to AML and F-1515 to EMM, and a Packard Foundation fellowship to EMM.Cellular and Molecular Biolog

Mucosal protection by phosphatidylcholine

The colonic mucus serves a first barrier towards invasion of commensal bacteria in stools to prevent inflammation. One essential component of intestinal mucus is phosphatidylcholine (PC) which represents more than 90% of the phospholipids in mucus indicative for a selective transport of PC into this compartment. It is arranged in lamellar structures as surfactant-like particles which provide a hydrophobic surface on top of the hydrated mucus gel to prevent the invasion of bacteria from intestinal lumen. In ulcerative colitis (UC), the mucus PC content is reduced by 70%, irrespective of the state of inflammation. Thus, it could represent an intrinsic primary pathogenetic condition predisposing to bacterial invasion and the precipitation of inflammation. Since PC was shown to be mainly secreted by the ileal mucosa from where it is assumed to move distally to the colon, the PC content along the colonic wall towards the rectum gradually thins, with the least PC content in the rectum. This explains the start of the clinical manifestation of UC in the rectum and the expansion from there to the upper parts of the colon. In three clinical trials, when missing mucus PC in UC was supplemented by an oral, delayed release PC preparation, the inflammation improved and even resolved after a 3-month treatment course. The data indicate the essential role of the mucus PC content for protection against inflammation in colon. Copyright (C) 2012 S. Karger AG, Base

Improvements in Attitude Determination and Control of the Small Satellite Flying Laptop

Precise attitude control is a key factor of many payloads with high ground resolutions, small fields of view or narrow beams such as an optical data downlink. The small satellite Flying Laptop (FLP), launched in July 2017, was developed by graduate and undergraduate students at the Institute of Space Systems of the University of Stuttgart with support by the space industry and research institutions. The satellite is three-axis stabilized with reaction wheels as main actuators. FLP is equipped with the OSIRIS optical data downlink which was built by the German Aerospace Center (DLR). As this instrument is body mounted on an optical bench, the attitude determination and control system (ACS) is required to point the whole satellite in the direction of the ground station with a high pointing accuracy of 150 arcseconds. At the time of launch the ACS did not reach this precision. This paper describes how the attitude determination and control were improved to achieve the required performance. The improvements can be divided into two parts. The first part includes the enhancement of on-board sensor processing and attitude control. In the second part, in-orbit data were utilized to increase the accuracy of parameters which are used to control the spacecraft. The first part includes the addition of a Kalman filter, an improved position propagation, and the introduction of adaptive gains to the on-board ACS. The FLP simulation test bed was used to verify the changes. The test bed was also used to find adequate initial values for the Kalman filter and to find inaccuracies in the sensor processing. In the second part, the adaptive gains and the Kalman initial values were validated in-orbit after the upload of the new sensor processing. Moreover, the on-board component orientation settings were corrected for the star trackers, the multi-spectral camera system, and the OSIRIS instrument on FLP. As a result, the satellite fulfills the pointing requirement of less than 150 arcsecond deviation from the target attitude for a sufficient period of time during a pass over the target. Successful links with the optical data downlink were demonstrated with the DLR ground station in Oberpfaffenhofen

Der Kampf um Köpfe: verbessern Betriebe durch Weiterbildung ihre Attraktivität als Arbeitgeber?

Ausgehend von Überlegungen der Signaling-Theorie und des Reputationsansatzes untersucht der Beitrag anhand der Daten einer Betriebsbefragung in Deutschland aus dem Jahr 2011, ob ein Zusammenhang zwischen dem betrieblichen Weiterbildungsengagement und dem Erfolg von Betrieben, qualifizierte Fachkräfte zu rekrutieren, nachgewiesen werden kann. Die Ergebnisse einer multivariaten Analyse zeigen, dass weiterbildungsaktive Betriebe einen höheren Rekrutierungserfolg als Betriebe ohne Weiterbildungsengagement haben. Damit bestätigt sich, dass Maßnahmen der betrieblichen Weiterbildung nicht nur ein Instrument der internen Personalentwicklung darstellen, sondern in besonderem Maße auch bei der externen Personalgewinnung eine Wirkung entfalten

Zugang und Legitimität in der EU: vorläufige Ergebnisse der Befragung deutscher Interessenverbände, politischer Parteien, Ministerien und politischer Stiftungen zur Außenhandelspolitik in der Europäischen Union

"Dieser Bericht stellt erste Ergebnisse der deutschen Teilstudie des Forschungsprojekts 'Access and legitimacy' vor. Ausgehend von der wachsenden Bedeutung europäischer Politik untersucht dieses Projekt, wie Interessengruppen und soziale Bewegungen auf die EU-Politik im Allgemeinen und die EU-Handelspolitik im Besonderen reagieren. Der Bericht präsentiert Daten von insgesamt 113 strukturierten face-to-face Interviews mit Vertreterinnen und Vertretern deutscher Interessenorganisationen (Wirtschaftsverbände, Gewerkschaften und NGOs) sowie Vertreterinnen und Vertreter politischer Parteien, Ministerien und politischer Stiftungen in Deutschland. Wir konzentrieren uns auf die Meinungen, Einstellungen und Präferenzen der Akteure in der Handelspolitik und untersuchen die unterschiedlichen Formen politischer Aktivität, die zur Einflussnahme auf politische Entscheidungen verwendet werden. Die Daten zeigen, dass Europa für die deutschen Akteure insgesamt eine sehr große Rolle spielt und dass diese einen beachtlichen Teil ihrer Ressourcen zur Einflussnahme auf der EU-Ebene einsetzen. Gleichwohl wird der größte Anteil der Ressourcen für die politische Interessenvertretung auf nationaler Ebene verwendet. Bei der Frage, mit welchen Strategien Interessenorganisationen vorgehen, um Einfluss zu nehmen, kann auf die recht verbreitete Nutzung von Netzwerkstrategien verwiesen werden. Bei der Nutzung von Öffentlichkeitsstrategien lassen sich dagegen größere Unterschiede zwischen den Akteursgruppen festhalten. Schließlich zeigt diese erste Auswertung, dass eine der Hauptkonfliktlinien im Bereich der Handelspolitik zwischen der Forderung nach stärkerer wirtschaftlicher Liberalisierung und der Forderung nach besserem Schutz des Sozialstaates verläuft." (Autorenreferat)"This paper presents preliminary results on the German data that is part of the larger research project 'Access and Legitimacy'. Given the increasing significance of European policy-making, this project investigates how interest groups and social movements respond and adapt to EU-policies in general and trade policy in particular. The paper presents data generated in 113 structured, face-to-face interviews with representatives of German interest groups (economic associations, labour unions, and NGOs) as well as representatives of political parties, ministries, and political foundations in Germany. Our focus is on the ideas, attitudes and preferences of specific actors regarding trade policy, and we investigate the different forms of political action that actors employ in order to exert influence on policy outcomes. The data indicates that Europe has great influence on German actors and that a majority of these actors have invested resources in order to influence European policies directly at the EU-level. Nevertheless, actors tend to spend the lion's share of their resources at the national level. In terms of which strategies actors employ, the data shows that most actors to a large extent utilize network strategies, while a considerable difference with regard to the use of public strategies across different types of actors can be detected. Finally, our initial research findings show that one main cleavage in the trade domain exists between economic liberalism and welfare state protectionism." (author's abstract

Gesellschaftlicher Wandel und personale Identität in der Spätmoderne: von den Schwierigkeiten Veränderung und Persistenz gleichermaßen zu erfassen

Dieser Artikel zeichnet kritisch nach, wie soziologische Identitätstheorien in Anlehnung an Becks Individualisierungsthese die Folgen des gesellschaftlichen Wandels für die Konstruktionsbedingungen personaler Identität beschreiben. Wir argumentieren jedoch dafür, den Blick auf Ambivalenzen im gesellschaftlichen Wandel zu richten und plädieren für eine systematische Einbeziehung soziale Ungleichheitskategorien in aktuelle Konzeptionen personaler Identität.This paper highlights how current sociological theories of identity discuss social change of individualization and its relevance for the conditions constituting individual identity. We would, however, propose an approach that goes beyond the focus on individualization but includes ambivalent and contradictory tendencies and a systematic comprehension of social inequalities

Protocol for the saMS trial (supportive adjustment for multiple sclerosis): a randomized controlled trial comparing cognitive behavioral therapy to supportive listening for adjustment to multiple sclerosis

BackgroundMultiple Sclerosis (MS) is an incurable, chronic, potentially progressive and unpredictable disease of the central nervous system. The disease produces a range of unpleasant and debilitating symptoms, which can have a profound impact including disrupting activities of daily living, employment, income, relationships, social and leisure activities, and life goals. Adjusting to the illness is therefore particularly challenging. This trial tests the effectiveness of a cognitive behavioural intervention compared to supportive listening to assist adjustment in the early stages of MS.MethodsThis is a two arm randomized multi-centre parallel group controlled trial. 122 consenting participants who meet eligibility criteria will be randomly allocated to receive either Cognitive Behavioral Therapy or Supportive Listening. Eight one hour sessions of therapy (delivered over a period of 10 weeks) will be delivered by general nurses trained in both treatments. Self-report questionnaire data will be collected at baseline (0 weeks), mid-therapy (week 5 of therapy), post-therapy (15 weeks) and at six months (26 weeks) and twelve months (52 weeks) follow-up. Primary outcomes are distress and MS-related social and role impairment at twelve month follow-up. Analysis will also consider predictors and mechanisms of change during therapy. In-depth interviews to examine participants’ experiences of the interventions will be conducted with a purposively sampled sub-set of the trial participants. An economic analysis will also take place. DiscussionThis trial is distinctive in its aims in that it aids adjustment to MS in a broad sense. It is not a treatment specifically for depression. Use of nurses as therapists makes the interventions potentially viable in terms of being rolled out in the NHS. The trial benefits from incorporating patient input in the development and evaluation stages. The trial will provide important information about the efficacy, cost-effectiveness and acceptability of the interventions as well as mechanisms of psychosocial adjustment.Trial registrationCurrent Controlled Trials ISRCTN91377356<br/