Independence from kinetoplast DNA maintenance and expression is associated with multi-drug resistance in Trypanosoma brucei in vitro

It is well known that several antitrypanosomatid drugs accumulate in the parasite's mitochondrion, where they often bind to the organellar DNA, the kinetoplast. To what extent this property relates to the mode of action of these compounds has remained largely unquantified. Here we show that single point mutations that remove the dependence of laboratory strains of the sleeping sickness parasite Trypanosoma brucei on a functional kinetoplast result in significant resistance to the diamidine and phenanthridine drug classes

The Pursuit of Virtue in Outdoor Sport and Recreation

This thesis presents for consideration six virtues that can be used to determine how adventure sports, outdoor recreation and expeditions can maintain their integrity when their methods are challenged by novel influences. Sports must adapt to social change, such as cultural, technological, or monetary influences. When they do so, mountaineering and adventure sports should be guided by ethical and aesthetic values that I argue can be maintained by adhering to the virtues of collaboration, education, athleticism, stewardship, sport empowerment and temperance. Some methods of pursuing these activities have alienated athletes from the natural component of outdoor sports and have moved toward aiming to control the environment rather than seeking to achieve human excellence in collaboration with nature. Further, using and accessing support and assistance strains the moral integrity of the activity and the unique value of accomplishments. In some cases, athletes have shifted their values toward prioritizing being first or completing an achievement by any means necessary. I examine how this has undermined the value of outdoor pursuits. The implication of a competitive ethos and the acceptance of high levels of assistance is that some past and modern achievements and expeditions have yielded unethical accomplishments that compromise the integrity of the sport

Unicast Ad-Hoc Routing in Vehicular City Scenarios

Within Vehicular Ad-Hoc Networking (VANET), i.e., networking between radio-equipped vehicles, unicast packet forwarding can be separated into the one-dimensional highway case and the two-dimensional city case. In this report, we survey the routing methods developed in the FleetNet and Network-on-Wheels projects plus a novel combination of two wellknown methods called PBR-DV or Position-Based Routing with Distance-Vector recovery. On the quest for a city-capable candidate routing algorithm as a possible standard, we discuss the usability and performance of the protocols in city scenarios. Finally, we conclude proposing PBR-DV as a candidate protocol for small-hop-count unicast VANET scenarios

Identification of novel components of Trypanosoma brucei editosomes

The editosome is a multiprotein complex that catalyzes the insertion and deletion of uridylates that occurs during RNA editing in trypanosomatids. We report the identification of nine novel editosome proteins in Trypanosoma brucei. They were identified by mass spectrometric analysis of functional editosomes that were purified by serial ion exchange/gel permeation chromatography, immunoaffinity chromatography specific to the TbMP63 editosome protein, or tandem affinity purification based on a tagged RNA editing ligase. The newly identified proteins have ribonuclease and/or RNA binding motifs suggesting nuclease function for at least some of these. Five of the proteins are interrelated, as are two others, and one is related to four previously identified editosome proteins. The implications of these findings are discussed

Editosome Accessory Factors KREPB9 and KREPB10 in Trypanosoma brucei

Multiprotein complexes, called editosomes, catalyze the uridine insertion and deletion RNA editing that forms translatable mitochondrial mRNAs in kinetoplastid parasites. We have identified here two new U1-like zinc finger proteins that associate with editosomes and have shown that they are related to KREPB6, KREPB7, and KREPB8, and thus we have named them Kinetoplastid RNA Editing Proteins, KREPB9 and KREPB10. They are conserved and syntenic in trypanosomatids although KREPB10 is absent in Trypanosoma vivax and both are absent in Leishmania. Tandem affinity purification (TAP)-tagged KREPB9 and KREPB10 incorporate into ∼20S editosomes and/or subcomplexes thereof and preferentially associate with deletion subcomplexes, as do KREPB6, KREPB7, and KREPB8. KREPB10 also associates with editosomes that are isolated via a chimeric endonuclease, KREN1 in KREPB8 RNA interference (RNAi) cells, or MEAT1. The purified complexes have precleaved editing activities and endonuclease cleavage activity that appears to leave a 5′ OH on the 3′ product. RNAi knockdowns did not affect growth but resulted in relative reductions of both edited and unedited mitochondrial mRNAs. The similarity of KREPB9 and KREPB10 to KREPB6, KREPB7, and KREPB8 suggests they may be accessory factors that affect editing endonuclease activity and as a consequence may affect mitochondrial mRNA stability. KREPB9 and KREPB10, along with KREPB6, KREPB7, and KREPB8, may enable the endonucleases to discriminate among and accurately cleave hundreds of different editing sites and may be involved in the control of differential editing during the life cycle of T. brucei

Hypothyreose bei Frühgeborenen - Retrospektive Analyse der Thyreotropin- und freien Thyroxin-Werte einer 7 Jahreskohorte Frühgeborener < 32 SSW

Bei transienten Hypothyreosen im Rahmen von Frühgeburtlichkeit (THOP) ist weiterhin Diskussionsgegenstand, ob eine Hormonersatztherapie zur Verbesserung kognitiver Einschränkungen beitragen kann. Vorliegende Dissertation hatte es zum Ziel, zunächst Schilddrüsenreferenzwerte speziell für das Tübinger Patientenkollektiv zu generieren sowie im Sinne der Qualitätssicherung die Therapieergebnisse der Tübinger Schilddrüsenleitlinie aus dem Jahr 2005 auszuwerten. In der Folge sollte, unter Berücksichtigung resultierender Ergebnisse sowie der aktuellen Literatur, eine Überarbeitung der Tübinger Leitlinie und Formulierung neuer Therapierichtlinien erfolgen. In diese retrospektive, beobachtende Analyse wurden alle Frühgeborenen mit einem Gestationsalter unter 32 Schwangerschaftswochen eingeschlossen, welche während des Auswertezeitraums zwischen 01.01.2007 und 20.09.2014 im Perinatalzentrum Tübingen zur Welt kamen oder direkt nach einer externen Geburt dorthin verlegt wurden. Insgesamt wurden 822 Frühgeborene (408 weiblich) entsprechend o.g. Kriterien eingeschlossen. Aus vorliegenden Analysen ließen sich folgende Kernempfehlungen für die Generierung einer neuen Leitlinie der Tübinger Neonatologie ableiten: Kontrollen von TSH und FT4 im chronologischen Alter von 2 Wochen, im postmenstruellen Alter (PMA) von 32 und 36 Wochen sowie im Rahmen einer ambulanten Kontrolle im korrigierten Alter von 4 Monaten. Unter L-Thyroxin (LT4) Therapie zusätzliche Kontrollen 2 und 4 Wochen nach Therapiestart zur Dosisanpassung und dann in 4-6 Wochen-Intervallen. Eine prophylaktische Routinesupplementierung bei Frühgeborenen ist nicht empfohlen, die Indikationsstellung für eine Therapie erfolgt anhand des FT4-Werts bei 12 mU/l unter 36 Wochen PMA, > 8 mU/l zwischen 36 Wochen PMA und korrigiert 4 Monaten und > 6 mU/l für ein korrigiertes Alter von über 4 Monaten. Die LT4-Startdosis beträgt (4)-5-(8) µg/kg/d, eine probatorische Dosisreduktion und Schilddrüsen-Funktionstestung kann bei einer LT4-Dosis von < 3,3 µg/kg/d im chronologischen Alter von 12 Monaten oder bei Borderline-Befunden nach 3 Jahren erfolgen. Aufgrund des retrospektiv-beobachtenden Designs vorliegender Studie lassen sich keine kausalen Zusammenhänge ableiten, sondern lediglich Assoziationen beschreiben. Insbesondere hinsichtlich der Fragestellungen Einfluss von Störfaktoren, neurologisches Outcome bei THOP durch verschiedene Therapieformen, Unterscheidungsmerkmale zwischen kongenitaler und transienter Hypothyreose wäre die Durchführung großer, multizentrischer, prospektiver, ggf. interventioneller Studien notwendig